Introdução: O exercício resistido em nível atlético pode promover estresse oxidativo crônico, fato que implica em uma resposta imuno-inflamatória exacerbada com consequente redução de desempenho e efeitos à saúde. Ao mesmo tempo, exercícios de caráter intenso elevam o consumo de glutamina por células e tecidos, reduzindo assim a disponibilidade deste aminoácido ao organismo, Todavia, estudos avaliando o metabolismo da glutamina em exercício do tipo resistido ainda são escassos. A síntese de antioxidantes, tais como a glutationa (GSH) e proteínas citoprotetoras como proteínas de choque térmico (HSPs) podem ser influenciadas pela disponibilidade de glutamina. Objetivo: Avaliar o efeito da suplementação oral crônica com L-glutamina e L-alanina, ambas na forma livre ou como Dipeptídeo sobre o estresse oxidativo e citoproteção mediado pela HSP 27 em ratos submetidos a exercício resistido. Métodos: Cinquenta ratos Wistar machos adultos (n = 10 por grupo) foram distribuídos em 5 grupos experimentais: Sedentário (SED), Treinado (CTRL) e suplementados com DIP, solução com L-glutamina e L-alanina livres (GLN+ALA) e somente L-Alanina (ALA), e foram submetidos ao protocolo de subida em escada durante 6 semanas, suplementados na água de beber em uma solução à 4%, nos últimos 21 dias do experimento. Foram analisados: teste de carga máxima, lactato sanguíneo, glutamina e glutamato (plasma, fígado e músculo -Tibial e EDL), creatina kinase e mioglobina (plasma) transaminases (plasma), glutationa oxidada (GSSG) e reduzida (GSH) (papa de hemácias, fígado e músculo - Tibial e EDL), TBARS (fígado e músculo- Tibial e EDL) e, expressão de HSP-27 e Glutamina Sintetase (músculo Tibial). Resultados: Os resultados demonstraram que o protocolo de exercício resistido reduziu a concentração de glutamina no músculo (p<0,05), aumentou a razão [GSSG/GSH] no fígado, papa de hemácia e músculo (p<0,05), e consequentemente houve aumento de TBARS nos tecidos. Já as suplementações com L-glutamina e L-alanina livres e como dipeptídeo aumentaram as concentrações de glutamina no plasma e tecidos (p<005), melhoraram a razão de [GSSG/GSH] no fígado, papa de hemácias e músculo (p<0,05). Também foi encontrado aumento da expressão de HSP 27 no Tibial, redução de TBARS nos tecidos, e creatina kinase no plasma (p<0,05). Conclusão: As suplementações com L- glutamina e L- alanina livres ou como dipeptídeo aumentam a síntese de GSH e a expressão de HSP 27, atenuando assim o estresse oxidativo causado pelo exercício resistido. / Introduction: Athletic Resistance exercise way promotes chronic oxidative stress, which implies in exacerbated immune inflammatory response with consequent reduction in performance and health effects. At the same time, intense exercise improves consumption of glutamine for cells and tissues, thereby reducing the availability of this amino acid to the body. However, studies evaluating the glutamine metabolism in resistance exercise are still scarce. The synthesis of antioxidants such as glutathione (GSH) and cytoprotective proteins such as heat shock proteins (HSPs) can be influenced by the availability of glutamine. Objective: To evaluate the effect of chronic oral supplementation with L-glutamine and L-alanine, both in its free form or as dipeptide on oxidative stress and the cytoprotection mediated by HSP 27 in rats subjected to resistance exercise. Methods: Fifty (n = 10 per group) Wistar adult rats were divided into 5 groups: Sedentary (SED), Trained (CTRL) and supplemented with DIP, solution with L-glutamine and free L-alanine (GLN + ALA) and L-alanine (ALA). The trained groups were underwent to climb stairs protocol for six weeks. Supplementations were offered in 4% solution in drinking water in the last 21 days of the experiment. Were analyzed: maximum load test, blood lactate, glutamine and glutamate (in plasma, liver and muscle Tibialis and EDL), creatine kinase and myoglobin (plasma), transaminase (plasma), oxidized (GSSG) and reduced (GSH) glutathione (erythrocytes, liver and muscle- Tibialis and EDL), TBARS (liver and muscle Tibialis- and EDL), HSP-27 and Glutamine Synthetase expression (Tibialis muscle). Results: The results showed that resistance exercise protocol reduced glutamine concentration in muscle (p <0.05) increased the ratio [GSSG / GSH] in the liver, erythrocytes and muscle (p <0.05), and TBARS increase the tissue. The Supplementation with L-glutamine and L-alanine and free dipeptide and increased glutamine concentrations in the plasma and tissues (p <0.05), improved the ratio of [GSSG / GSH] in liver, erythrocytes and muscle (p <0.05). HSP 27 expression was also increased in Tibialis Muscle. There was reduction of TBARS in tissues and creatine kinase in plasma (p <0.05). Conclusion: Supplementations with L-glutamine and L-alanine in its free form or as dipeptide increase the synthesis of GSH and Expression HSP 27, thus reducing oxidative stress caused by resistance exercise.

As glutationas transferases compõem uma superfamíla de proteínas que atuam na fase II do sistema de desintoxicação das células. Participam principalmente através do processo de conjugação da glutationa com moléculas hidrofóbicas e eletrofílicas, como por exemplo os herbicidas. No entanto, outras funções foram descritas como a tolerância ao estresse oxidativo, inseticidas, antibióticos microbianos, transporte de produtos secundários tóxicos, sinalização da célula durante as respostas ao estresse e fenômenos de resistência envolvendo agentes de quimioterapia contra o câncer. Nesta tese procurou-se estabelecer uma relação entre a seqüência, estrutura, função e afinidade das GSTs. A estrutura, de modo geral, determina a função da enzima, mas por si só, não dita sua especificidade. Esta última informação é fundamental para o desenvolvimento de novos agroquímicos ou para o desenho racional de novas proteínas. A relação entre a seqüência, estrutura, função e afinidade mostra que o paradigma estrutura-função deveria ser ampliado para incluir a seqüência de aminoácidos e a afinidade da enzima. Apesar da grande diversidade de substratos e seqüências encontradas nas GSTs há pelo menos um caso de convergência funcional em duas classes distintas desta superfamília. Uma encontrada apenas no reino Animalia (classe Pi) e outra exclusiva do reino Plantae (classe Phi). Ferramentas da bioinformática estrutural, como docking molecular e minimização de energia foram utilizadas para analisar as interações entre a enzima e o substrato. Estas ajudam a explicar como duas proteínas com aproximadamente 22% de identidade de seqüência apresentam afinidades semelhantes. Finalmente, foram propostos mutantes da GST de Saccharum officinarum utilizando a informação estrutural da enzima, visando uma alteração na afinidade da mesma. / Glutathione Transferases comprehend a superfamily of proteins that plays the phase II of the detoxification system of the cells. Their major catalysis is the conjugation of glutathione with hydrophobic and eletrophilic molecules, for example herbicides. However, other functions were described like oxidative stress, insecticides, microbial antibiotics, transport of secondary products, cells signalization during response to stress and resistance of chemotherapy drugs against cancer. This work aimed to establish a relation between sequence, structure, function and affinity of GSTs. The structure, in general, determines the function, but alone, can not determine the enzyme specificity. This last information is essential to the development of new agrochemicals or for the rational design of proteins. The relation between sequence, structure, function and affinity shows that the paradigm of structure-function should be enlarged in order to include the information of amino acid sequences and the enzyme affinities. Despite the wide variety of substrates and sequences found in the GSTs, there is at least one case of functional convergence between two distinct classes in this superfamily. One is found in the Animalia kingdom (class Pi) and the other is exclusively found in Plantae (class Phi). The structural bioinformatic tools, such as molecular docking and energy minimization were used to analyze the interactions between the enzyme and the substrate. These help to understand how two enzymes with approximately 22% of sequence identity can show the same affinities. Finally, GST mutants of Saccharum officinarum were proposed, using the enzyme structural information in order to modify the enzyme affinities.

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

Abstract Glutaredoxins (Grx) are small thiol disulphide oxidoreductases with a conserved active site sequence -CXXC/S- and a glutathione (GSH) binding site. They catalyze the reduction of protein disulphides, preferring protein-GSH mixed disulphides as substrates. The accumulation of protein-GSH mixed disulphides has been observed during oxidative stress, where they may serve both a regulatory and an antioxidant function by protecting the enzymes from irreversible oxidation. Once oxidative stress has been removed the GSH-protein mixed disulphides are reduced by GSH or, more efficiently, by Grx. The present study showed for the first time that Grx1 and Grx2 can be detected in healthy human lung. Highly specific expression of Grx1 was observed in alveolar macrophages, but it could also be detected from sputum supernatant. Grx1 levels in alveolar macrophages were lower in selected inflammatory diseases than in control lung samples. Grx1 was also mainly negative in the fibrotic areas in usual interstitial pneumonia, an aggressive fibrotic lung disease. Overall, the present study suggests that Grx1 is a potential redox modulatory protein regulating the intracellular as well as extracellular homeostasis of glutathionylated proteins and GSH not only in healthy lung, but also in inflammatory and fibrotic lung diseases. In order to study the mechanism of action of glutaredoxins in vitro, a new real-time fluorescence-based method for measuring the deglutathionylation activity of glutaredoxins using a glutathionylated peptide as a substrate was developed. The first reaction intermediate in the deglutathionylation reaction was shown to be exclusively Grx-GSH mixed disulphide and this specificity was solely dependent on the unusual γ-linkage present in glutathione. The study also demonstrated the role of conserved residues in the proximity of proposed GSH binding site to the GSH binding specificity of E. coli Grx1. Opening the binding groove and removing charged residues enabled Grx to form more readily mixed disulfides with other molecules besides GSH. Different members of the PDI family showed considerably lower activity levels compared to glutaredoxins and, in contrast to the glutaredoxin-GSH mixed disulphide, the only intermediate in the PDI catalysed reaction was PDI-peptide mixed disulphide.

Malignant melanoma is one of the most rapidly increasing cancers and accounts for about three-quarter of all skin cancer deaths worldwide. Despite compelling evidence that ultraviolet (UV) irradiation causes melanoma the knowledge how various wavelength spectra affect the balance between proliferation and apoptosis controlling the homeostasis of the melanocyte population is still limited. The aim of this thesis was to elucidate the regulation of UVA/B induced apoptotic signaling in human epidermal melanocytes in vitro in relation to redox alterations and antioxidant photoprotection. UVA irradiation induced changes in plasma membrane stability, decreased cell proliferation and increased apoptosis. In comparison, melanocyte plasma membrane was markedly resistant to UVB irradiation although apoptosis was triggered. Thus, UVA irradiation should not be overlooked as an etiologic factor in melanoma development. Further, after irradiation with UVA/B we found alterations in redox state manifested by a reduction of intracellular GSH levels, translocation of nuclear factor-κB from the cytosol to the nucleus, an increase of γ-glutamylcysteine synthetase, the rate-limiting enzyme in GSH synthesis, and an increased apoptosis frequency. α-Tocopherol provided photoprotection through several modes of action affecting redox alterations and signaling, stabilizing the plasma membrane, and decreased proliferation and apoptosis rate, while β-carotene did not show the same protective capacity. Altogether, α-tocopherol might be a useful substance in protecting melanocytes from UV induced damage. We demonstrate UVA/B irradiation to activate the intrinsic pathway of apoptosis in melanocytes where translocation of Bcl-2 family proteins to the mitochondria modulates the apoptosis signal. Interestingly, the anti-apoptotic Bcl-2 family proteins generally thought to be attached to membranes, were localized in the cytosol before UV irradiation and translocated to the mitochondria in the surviving population, which might be a critical event in preventing apoptotic cell death. Lysosomal cathepsins were released to the cytosol acting as pro-apoptotic mediators upstream of activation and translocation of Bax to the mitochondria. When melanocytes were exposed to UVA, p53 participated in apoptosis regulation through interaction with Bcl-2 family proteins, while UVB induced p53-transcriptional activity and apoptosis involving lysosomal membrane permeabilization. Thus, depending on the UV wavelength p53 mediated apoptosis in melanocytes by transcriptional dependent or independent activity. These results emphasize p53 as an important pro-apoptotic component in the regulation of apoptosis. This thesis gives new insight in the harmful and various effects of different wavelengths within the UV spectrum on human melanocytes in vitro. Improved knowledge of the apoptosis regulatory systems in melanocytes might lead to a better understanding of the formation of pigment nevi and malignant melanoma and, in the future, provide better strategies to prevent and eliminate tumor development and progression.

>Magister Scientiae - MSc / Sugargraze is a moderately drought tolerant sweet sorghum hybrid which is ideal for grazing, winter stand over and pit silage. A major advantage that Sugargraze has over other forages is its very high sugar content which improves feed quality thus increasing palatability and results in significantly reduced feed wastage. This study explored the influence of hyperosmotic stress on plant development, ROS accumulation, antioxidant capacity and the extent of cell death. Heat shock protein (Hsp70) expression immunoblotting assays were used to demonstrate whether the various treatment conditions induced stress within natural physiological parameters for the experimental material. This was coupled with the separation, visualization and identification of abundant proteins in Sugargraze leaves in response to hyperosmotic stress using two-dimensional gel electrophoresis (2-DE) in combination with mass spectrometry (MS). The results showed that hyperosmotic stress significantly influences plant development by reducing plant biomass and increasing the levels of ROS accumulation, proline content and subsequently reducing total chlorophyll content. An over accumulation of ROS in the form of hydrogen peroxide and lipid peroxidation was observed in the stressed plants which was supported by the extent of cell death. Although an increase in antioxidant enzyme activity (in the form of total enzymatic activity or individual isoform activity) in response to hyperosmotic stress was observed, this increase was not sufficient to counter the deleterious effects caused by the stress conditions hence the decrease in plant biomass and increase in cell death. Western blotting analysis of Sugargraze leaf tissues using Hsp70 antibodies showed that hyperosmotic stress induced Hsp70 expression to levels significantly higher than observed for the control plants. A total of thirteen CBB stained spots were selected for mass spectrometric identification, owing to their good resolution and abundance levels, and of these, nine were positively identified. Identified proteins were divided into functional categories including both known and novel/putative stress responsive proteins. Molecular and physiological functions of some of the proteins of interest identified will be subjected to further investigation via bioinformatic and molecular biology approaches.

The Non-expressor of pathogenesis-related1 gene (NPR1) mediates the induction of pathogenesis-related (PR) gene products, vital for resistance in plants. In this study, the role of two previously isolated Cavendish banana NPR1-like genes (MNPR1A and MNPR1B) has been characterized in protection against Xanthomonas campestris, Hylaperonospora arabidopsidis, Botrytis cinerea and Pseudomonas syringae pathogens. The specific aim was to investigate if sequence differences in both genes are responsible for differential activity against pathogens because in a previous expression study, MNPR1A and not MNPR1B had been more responsive to the banana necrotrophic pathogen Fusarium oxysporum. By challenging Fusarium-tolerant GCTCV-218 and susceptible Grand Naine Cavendish banana plants (which had been used in a previous characterization study) with the hemi-biotrophic Xanthomonas pathogen (a very important economical pathogen of banana), the two MNPR1, PR-1 and PR-3 genes were found to be sequentially expressed. Expression of these genes was more pronounced in the tolerant GCTCV-218 banana cultivar than in the sensitive Grand Naine cultivar. Comparative sequence analysis further showed that these two banana NPR1-like coding sequences had dissimilarities even within conserved functional domains; they grouped closely with other defense-related NPR1-like sequences and harboured defense cis-regulatory elements. Transformation of the coding sequences of both genes under the control of the 35S CaMV promoter/terminator sequences into npr1-2 Arabidopsis mutant complimented the phenotype of this mutant following infection with distinct classes of pathogens (biotrophic Hyaloperonospora, necrotrophic Botrytis and hemi-biotrophic Pseudomonas pathogens). These Infected-MNPR1-expressing plants had higher PR-1 transcript amounts with more reduced pathogen growth compared to non-transgenic npr1-2 Arabidopsis mutant plants. However, the difference in the two banana coding sequences did not translate into a differential pattern of response against the three different classes of pathogens used in this study. Further detailed studies are suggested to investigate the role of the MNPR1 promoter-coding sequences in the differential response to pathogens using a bananapathogen system. This study also addressed the question of whether cystosolic glutathione (GSH) is necessary for NPR1 transcription during systemic acquired resistance. Using Arabidopsis mutants (clt1clt2clt3) defective in cytosolic GSH biosynthesis and following infection with either Pseudomonas or Botrytis, NPR1 and PR-1 transcription was much reduced rendering the mutants more sensitive to pathogens compared to infected-wild-type i>Arabidopsis plants. Results from this study therefore implicate cytosolic glutathione as an essential antioxidant for the establishment of an effective defense response cascade. / Thesis (PhD)--University of Pretoria, 2011. / Plant Science / unrestricted

Drug induced liver injury is a leading cause of human illness and a major cause of drug withdrawals from the market. A systems biology approach has the potential to aid toxicology research since toxicological responses are a consequence of multiple non-linear and interdependent biological responses. Here such an approach is developed.The glutathione pathway is a key hepatic defence mechanism and deactivates reactive metabolites before they have the chance to damage cellular proteins. However, glutathione availability is limited and can vary between individuals. As hepatic glutathione levels cannot be measured directly, two serum-based biomarkers, i.e. 5-oxoproline and ophthalmic acid, have been proposed in literature as a means of tracking glutathione depletion. This thesis aims to test the reliability of the correlation between biomarker concentration and decreasing glutathione concentration.In this study a spiral between experiments, model predictions and falsifications, model improvement, and experimental design is described. Using this approach a kinetic model of the hepatic glutathione pathway and biomarker metabolism was constructed and subsequently expanded by adding physiologically based pharmacokinetic (PBPK) models of paracetamol and the proposed biomarkers. These models have increased the understanding of the glutathione pathway. For example, the model predicted that Glutamyl-Cysteine Synthetase induction should be a highly effective way to increase the robustness of the liver to a paracetamol challenge.In addition, it was possible to qualify with increasing precision, the correlation between biomarkers and hepatic glutathione depletion. 5-Oxoproline and ophthalmic acid provide different information about the status of the glutathione pathway. 5-Oxoproline is correlated with paracetamol-glutathione conjugate formation, but not with extreme toxicity. Ophthalmic acid is a biomarker of a more advanced stage of toxicity, where the cell is unable to protect against glutathione depletion. However, care must be taken when inferring hepatic glutathione concentration. Both models demonstrate that the sensitivity of biomarkers to exposure of paracetamol, depends on the dynamics of exposure as well as on the concentrations of intracellular metabolites, such as methionine.I discuss how the methodology of biomarker assessment could be personalised with regards to individual patients and how systems toxicology could be further developed towards reliable tools for the pharmaceutical industry.

Previous studies on the medical use of ozone therapies show a very diverse array of results, from ozone reducing the amount of HIV virus in the blood, to no effect, to causing the death of several patients due to pulmonary embolism and infections. However, ozone therapies are widely used in Europe and considered medically safe. In the U.S., doctors in 28 states use ozone therapies. The objectives of this study were to investigate the effects of medical grade ozone at varying concentrations used in ozone therapies. These were achieved by evaluating the C-reactive protein, erythrocyte sedimentation rate, total reduced and oxidized glutathione content of erythrocytes which were all markers used to determine ozone injury/inflammation. Despite the fact that ozone is a very strong oxidant, previous research indicates that depending on the dose and the health status of the biological system, sometimes ozone can act as an antioxidant. The medical exposure range for ozone is between 20-80 mg/ml with an average of 50 mg/ml. The concentrations used in this study were 20, 40, 80 and 160 mg/ml. Ozone was generated in the "Breath Lab" at USF from medical grade oxygen obtained through electrical corona arc discharge using an OL80C ozone generator. De-identified blood samples of 10 ml blood/sample containing EDTA as anticoagulant were obtained from the James A. Haley VA Hospital patients. Equal volumes of blood and ozone gas mixture were allowed to mix in ozone-resistant syringes prior to dividing each sample into three parts, one for each corresponding parameter to be studied. The C-reactive protein was analyzed through ELISA using the colorimetric method available from Helica Biosystems; erythrocyte sedimentation rate was measured in graduated sedimentation tubes; the total reduced glutathione (GSH) and oxidized glutathione (GSSG) content of erythrocytes was determined according to the colorimetric method developed by the Oxford Biomedical Research. Overall, the concentrations of ozone used did not have a statistically significant effect on the parameters investigated. However, a small percentage of the blood samples showed an improvement in the parameters studied, especially at the highest ozone concentration.

Proteins with thiol groups interact with metal ions in the human body. They maintain their homeostasis, participate in cell signaling, protect the cell against the effects of toxic metals and detoxify them. This work is focused on proteins with thiol groups glutathione and metallothionein and their effects on selenium. The method of study is electrochemical.