Avaliação do estresse oxidativo induzido por superfícies de titânio

Commercially pure Titanium (cp Ti) is a material largely used in orthopedic and dental implants due to its biocompatibility properties. Changes in the surface of cp Ti can determine the functional response of the cells such as facilitating implant fixation and stabilization, and increased roughness of the surface has been shown to improve adhesion and cellular proliferation. Various surface modification methods have been developed to increase roughness, such as mechanical, chemical, electrochemical and plasma treatment. An argon plasma treatment generates a surface that has good mechanical proprieties without chemical composition modification. Besides the topography, biological responses to the implant contribute significantly to its success. Oxidative stress induced by the biomaterials is considered one of the major causes of implant failure. For this reason the oxidative potential of titanium surfaces subjected to plasma treatment was evaluated on this work. CHO-k1 cells were cultivated on smooth or roughed Ti disks, and after three days, the redox balance was investigated measuring reactive oxygen species (ROS) generation, total antioxidant capacity and biomarkers of ROS attack. The results showed cells grown on titanium surfaces are subjected to intracellular oxidative stress due to hydrogen peroxide generation. Titanium discs subjected to the plasma treatment induced less oxidative stress than the untreated ones, which resulted in improved cellular ability. Our data suggest that plasma treated titanium may be a more biocompatible biomaterial.

Identificação de genes em Chromobacterium violaceum relacionados à resposta ao estresse

The sequencing of the genome of Chromobacterium violaceum identified one single circular chromosome of 4.8 Mb, in which approximately 40% of the founded ORFs are classified as hypothetical conserved or hypothetical. Some genic regions of biotechnological and biological interest had been characterized, e. g., environmental detoxification and DNA repair genes, respectively. Given this fact, the aim of this work was to identify genes of C. violaceum related to stress response, as the ones involved with mechanisms of DNA repair and/or genomic integrity maintenance. For this, a genomic library of C. violaceum was built in Escherichia coli strain DH10B (RecA-), in which clones were tested to UVC resistance, resulting in five candidates clones. In the PLH6A clone were identified four ORFs (CV_3721 to 3724). Two ORFs, CV_3722 and CV_3724, were subcloned and a synergic complementation activity was observed. The occurrence of an operon was confirmed using cDNA from C. violaceum in a RT-PCR assay. Further, it was observed the induction of the operon after the treatment with UVC. Thus, this operon was related to the stress response in C. violaceum. The mutagenesis assay with rifampicin after the treatment with UVC light showed high frequency of mutagenicity for the ORF CV_3722 (Pol III δ subunit). In this way, we propose that the C. violaceum δ subunit can act in DH10B in the translesion synthesis using Pol IV in a RecA independent-manner pathway. In growth curve assays other four clones (PLE1G, PLE7B, PLE10B and PLE12H) were able to complement the function at the dose 5 J/m2 and in mutagenicity assays PLE7B, PLE10B and PLE12H showed frequencies of mutation with significant differences upon the control (DH10B), demonstrating that in some way they are involved with the stress response in C. violaceum. These clones appear to be interrelated, probably regulated by a messenger molecule (eg., nucleotide c-di-GMP) and/or global regulatory molecule (eg., σS subunit of RNA polymerase).The results obtained contribute for a better genetic knowledge of this specie and its response mechanisms to environmental stress.

Proteômica diferencial da Chromobacterium violaceum em resposta ao estresse oxidativo induzido por peróxido de hidrogênio

The β-proteobacterium Chromobacterium violaceum is a Gram-negative, free-living, saprophytic and opportunistic pathogen that inhabits tropical and subtropical ecosystems among them, in soil and water of the Amazon. It has great biotechnological potential, and because of this potential, its genome was completely sequenced in 2003. Genome analysis showed that this bacterium has several genes with functions related to the ability to survive under different kinds of environmental stresses. In order to understand the physiological response of C. violaceum under oxidative stress, we applied the tool of shotgun proteomics. Thus, colonies of C. violaceum ATCC 12472 were grown in the presence and absence of 8 mM H2O2 for two hours, total proteins were extracted from bacteria, subjected to SDS-PAGE, stained and hydrolysed. The tryptic peptides generated were subjected to a linear-liquid chromatography (LC) followed by mass spectrometer (LTQ-XL-Orbitrap) to obtain quantitative and qualitative data. A shotgun proteomics allows to compare directly in complex samples, differential expression of proteins and found that in C. Violaceum, 131 proteins are expressed exclusively in the control condition, 177 proteins began to be expressed under oxidative stress and 1175 proteins have expression in both conditions. The results showed that, under the condition of oxidative stress, this bacterium changes its metabolism by increasing the expression of proteins capable of combating oxidative stress and decreasing the expression of proteins related processes bacterial growth and catabolism (transcription, translation, carbon metabolism and fatty acids). A tool with of proteomics as an approach of integrative biology provided an overview of the metabolic pathways involved in the response of C. violaceum to oxidative stress, as well as significantly amplified understanding physiological response to environmental stress. Biochemical and "in silico" assays with the hypothetical ORF CV_0868 found that this is part of an operon. Phylogenetic analysis of superoxide dismutase, protein belonging to the operon also showed that the gene is duplicated in genome of C. violaceum and the second copy was acquired through a horizontal transfer event. Possibly, not only the SOD gene but also all genes comprising this operon were obtained in the same manner. It was concluded that C. violaceum has complex, efficient and versatile mechanisms in oxidative stress response

Análise da Expressão de APE1 após Estresse Oxidativo Induzido em Células Proficientes e Deficientes na Via de Reparo por Excisão de Nucleotídeos

Riboflavin is a vitamin very important in aerobic organisms, as a precursor of many coenzymes involved in the electron transporter chain. However, after photosensitization of riboflavin with UV or visible light, it generates reactive oxygen species (ROS), which can oxidize the DNA. The repair of oxidative lesions on DNA occurs through the base excision repair pathway (BER), where APE1 endonuclease plays a central role. On the other hand, the nucleotide excision repair pathway (NER) repairs helix-distorting lesions. Recently, it was described the participation of NERproteins in the repair of oxidative damage and in stimulation of repair function fromAPE1. The aim of this research was to evaluate the cytotoxic effects of photosensitized riboflavin (RF*) in cells proficient and deficient in NER, correlating with APE1 expression. For this propose, the cells were treated with RF* and it was performed the cell viability assay, extraction of whole proteins, cells fractionation, immunoblotting, indirect immunofluorescence and analysis of polymorphisms of BER gens. The results evidenced that cells deficient in XPA and CSB proteins were more sensitive to RF*. However, XPC-deficient cells presented similar resistance to MRC5- SV cells, which is proficient in NER. These results indicate that XPA and CSB proteins have an important role on repair of oxidative lesions induced by RF*. Additionally, it was evidenced that single nucleotide polymorphisms (SNPs) in BER enzymes may influence in sensitivity of NER-deficient cell lines. Concerning the APE1 expression, the results showed that expression of this protein after treatment with RF* only changed in XPC-deficient cells. Though, it was observed that APE1 is recruited and is bound to chromatin in MRC5-SV and XPA cells after treatment with RF*. The results also showed the induction of DNA damage after treatment with RF*, through the analysis of-H2AX, since the treatment promoted an increase of endogenous levels of this phosphorylated protein, which acts signaling double strand-break on DNA. On the other hand, in XPC-deficient cells, regardless of resistance of RF*, the endogenous levels of APE1 are extremely reduced when compared with other cell lines and APE1 is not bound to chromatin after treatment with RF*. These results conclude that RF* was able to induce cell death in NERdeficient cells, where XPA and CSB cells were more sensitive when compared with MRC5-SV and XPC-deficient cells. This last result is potentially very interesting, since XPC-deficient cell line presents low levels of APE1. Additionally, the results evidenced that APE1 protein can be involved in the repair of oxidative damage induced by RF*, because APE1 is recruited and bound strongly to chromatin after treatment.

Proteômica comparativa de linhagens celulares humanas expostas a estresse oxidativo induzido por riboflavina fotossensibilizada

Reactive oxygen species (ROS) are continuously generated and can be derived from cellular metabolism or induced by exogenous factors, in addition, have the capacity to damage molecules like DNA and proteins. BER is considered the main route of DNA damage oxidative repair, however, several studies have demonstrated the importance of the proteins participation of other ways to correct these injuries. NER enzymes deficiency, such as CSB and XPC, acting in the damage recognition step in the two subways this system influences the effectiveness of oxidative damage repair. However, the mechanisms by which cells deficient in these enzymes respond to oxidative stress and its consequences still need to be better understood. Thus, the aim of this study was to perform a proteomic analysis of cell lines proficient and deficient in NER, exposed to oxidative stress, in order to identify proteins involved, directly or not, in response to oxidative stress and DNA repair. For this, three strains of human fibroblasts, MRC5-SV, CS1AN (CSBdeficient) and XP4PA (XPC-deficient) were treated with photosensitized riboflavin and then carried out the differentially expressed proteins identification by mass spectrometry. From the results, it was observed in MRC5-SV increase expression in most of the proteins involved in cellular defense, an expected response to a normal cell line subjected to stress. CS1AN showed a response disjointed, it is not possible to establish many interactions between the proteins identified, may be one explanation for their sensitivity to treatment with riboflavin and other oxidants and increased cell death probably by induction of pro-apoptotic pathways. Already XP4PA showed higher expression of apoptosis-blocking proteins, as there was inhibition or reduced expression of others involved with the activation of this process, suggesting the activation of an anti-apoptotic mechanism in this lineage, which may help explain the high susceptibility to develop cancers in XPC individuals. These results also contribute to elucidate action mechanisms of NER in oxidative damage and the understanding of important routes in the oxidative stress correlation, repair and malignant tumors formation

Avaliação de parâmetros de estresse oxidativo em plantas de cana-de-açúcar tratadas com peróxico de hidrogênio

The genus Saccharum belongs to Poaceae family. Sugarcane has become important monocultures in Brazil due to their products: ethanol and sugar. The production may change between different regions from Brazil. This difference is related to soil, climatic conditions and temperature that promotes oxidative stress that may induce an early flowering. The aim of this work was to identify the effects of oxidative stress. In order to analyse this, sugarcane plants were submitted to oxidative stress using hydrogen peroxide. After this treatment, the oxidative stress were analyzed Then, the plant responses were analyzed under different approaches, using morphophysiological, biochemical and molecular tools. Thus, sugarcane plants were grown under controlled conditions and until two months they were subjected first to a hydroponics condition for 24 hours in order to acclimation. After this period, these plants were submitted to oxidative stresse using 0 mM, 10 mM, 20 mM and 30 mM hydrogen peroxide during 8 hours. The histomorphometric analysis allowed us to verify that both root and leaf tissues had a structural changes as it was observed by the increased in cell volume, lignin accumulation in cell walls. Besides, this observation suggested that there was a change in redox balance. Also, it was analyzed the activity of the SOD, CAT and APX enzymes. It was observed an increase in the SOD activity in roots and it was also observed a lipid peroxidation in leaves and roots. Then, in order to identify proteins that were differently expressed in this conditions it was used the proteomic tool either by bidimensional gel or by direct sequencing using the Q-TOF EZI. The results obtained with this approach identified more than 3.000 proteins with the score ranging from 100-5000 ions. Some of the proteins identified were: light Harvesting; oxygenevolving; Thioredoxin; Ftsh-like protein Pftf precusor; Luminal-binding protein; 2 cys peroxiredoxin e Lipoxygenase. All these proteins are involved in oxidative stress response, photsynthetic pathways, and some were classified hypothetical proteins and/or unknown (30% of total). Thus, our data allows us to propose that this treatment induced an oxidative stress and the plant in response changed its physiological process, it made changes in tissue, changed the redox response in order to survival to this new condition

Relação entre estresse oxidativo e oesteopenia diabética em ratas pré-menopausadas

The relationship between lipid peroxidation, antioxidant defense and diabetic osteopenia remains unclear. This study evaluated the relationship between lipid peroxidation index, antioxidant defense parameter and bone metabolism in a premenopausal diabetic model by measurements such as thiobarbituric acid-reactive substances concentration (TBARS) and reduced glutathione (GSH) content in brain homogenates, histomorphometric analysis, biomechanical testing and bone mineral density (BMD). Female Wistar rats with regular estrous cycle were divided into two groups: Group 1: control rats (n = 15) and Group 2: diabetic rats (n = 15). Diabetes mellitus was induced by alloxan and confirmed by glycemia 250 mg/dL. The experimental period understood 1 and 5 after days induction and 45, 75 and 120 days after the installation of diabetes mellitus.The lipid peroxidation index, measured by TBARS concentration, showed a significant increase (p<0.05) in diabetic animals in comparison to animals control. However, the antioxidant parameter, measured by GSH content, was significantly decrease (p<0.05) in diabetic animals. Histomorphometric analysis showed a significant increase (p<0.05) in femoral trabecular separation together with a significant decrease (p<0.05) in trabecular thickness and reduced trabecular bone volume in diabetic rats. Moreover, biomechanical testing and BMD values were significant decrease (p<0.05) in diabetic group. Thus, our results demonstrated that increased lipid peroxidation and altered antioxidant defense could be related to the development of oxidative stress and diabetic osteopenia in premenopausal rats

Representações sociais de enfermeiros brasileiros e portugueses sobre o estresse em serviço de urgência

This study aimed to analyze the social representations of Brazilian and Portuguese nurses on stress in the emergency service. A semi-structured interview and the free word association test, with "stress" as the inductive stimulus, were used as research instruments. Data were collected from 120 nurses, being 60 from an emergency hospital in the city of Natal, Brazil and 60 from an urgency hospital in the city of Aveiro, Portugal. Data from the word association test were analyzed with the EVOC 2002 program, after thematic categorical content analysis, enabling construction of a data bank. Data gathered from the interview were analyzed by ALCESTE 4.8 software. Nurse represent the stress in the urgency department as a generation of physical and mental detrition where adaptation is unsatisfactorily, resulting in the onset of fatigne, irritability, lack of concentration, lack of motivation, pessimism, impaired interpersonal relationship and low productivity. The solution is part of a complex whole, which demands an integrated way of acting that has demanded increasingly professional attitudes based on multidisciplinarity

Desenpenho funcional, carga alostática e biomarcadores do estresse em idosos residentes na comunidade

Conselho Nacional de Desenvolvimento Científico e Tecnológico

Aspectos socioeconômicos e demográficos relacionados ao estresse e a síndrome de Burnout em fisioterapeutas no Brasil

Dos problemas de saúde existentes no ambiente de trabalho, 18% afetam o aparelho psíquico, a exemplo do estresse, atingindo cerca de 30% dos trabalhadores em geral. A persistência e intensidade do estresse, associada às sucessivas tentativas de lidar com os mesmos, tornam os indivíduos vulneráveis ao surgimento da Síndrome deBurnout. O objetivo deste trabalho foi identificar a relação dos aspectos socioeconômicos e demográficos com o estresse e a Síndrome de Burnout em fisioterapeutas do Brasil. Para isso, este estudo teve uma abordagem do tipo transversal, com 1040 fisioterapeutas do Brasil, através de uma amostragem do tipo snowbolle não probabilística. Utilizou-se um questionário socioeconômico, demográfico e profissional, a Escala de Estresse no Trabalho (EET) e a adaptação do Cuestionario para la Evaluación del Síndrome de Quemarse por el Trabajo (CESQT). Na análise dos dados, foram utilizadas a estatística descritiva e inferencial. Dentre os principais resultados obtidos, percebeu-se uma maior representatividade da região Nordeste (48,7%), com idade média de 31anos, sexo feminino (75,7%), carga horária semanal de 35,4 horas, com 3-5 anos de atuação profissional. Observou-se que 37,0% apresentavam estresse relacionado estatisticamente com a idade (p=0,008),atividade física (p=0,039) e satisfação com a saúde (r=-0,322; p<0,001). Não foi observado nenhum caso de Burnout, porém houve uma média elevada nas dimensões, desgaste psíquico, indolência e culpa, totalizando 49,0% comtendência ao desenvolvimento da síndrome. Portanto, as variáveis, idade, prática da atividade física e satisfação com a saúde obtiveramrelação com o estresse. Para o Burnout, destacaram-se a região de moradia (centro-oeste), satisfação com a saúde, local de trabalho (clínicas e hospitais), além do maior número de locais de trabalho. Diante desse contexto, os estudos sobre o estresse e a Síndrome de Burnout se apresentam como elementos derelevância dentro do contexto da prevenção dos riscos laborais e da análise das condições de trabalho

Avaliação dos efeitos da atividade física na prevenção do estresse oxidativo e do câncer de mama

Conselho Nacional de Desenvolvimento Científico e Tecnológico

Avaliação dos parâmetros de estresse oxidativo no lúpus eritematoso

OBJECTIVE: The aim of this work was to analyse some oxidative stress parameters in patients of Systemic Lúpus Erythematosus. PATIENTS AND METHODS: Determinations of reduced glutathione content in whole blood were carried out. The activity of superoxide dismutase, gluthatione peroxidase and catalase in erythrocytes and the concentration of reactive substances of acid thiobarbituric in plasma of patients female (n =19) with SLE no activity of disease (Mex-SLEDAI < 2), with average ages of 32 ± 11 years, through the spectrophotometrical methods and from healthy individuals (n =30). Statistical data were analyzed by student t-test, p<0,05. RESULTS: Our data indicated a significant decrease on the activity of catalase and significant increase on the concentration of reactive substances of acid thiobarbituric in patients with SLE comparing with healthy individuals. There was no significant difference in other parameters. CONCLUSION: The results showed that oxidative stress has a role in the pathogenesis of the disease in SLE, even in patients without active disease.

Efeito do estresse hídrico sobre a eficiência do halosulfuron no controle de tiririca

Foi conduzido um trabalho em casa de vegetação com o objetivo de estudar o efeito de 10 dias de estresse hídrico (Yw de ³2,6; -2,2; -0,35 e -0,2 MPa) sobre a eficiência do halosulfuron (150g p.c./ha+Aterbane-0,25% v/v) no controle de tiririca em pós-emergência . A aplicação do halosulfuron em plantas de tiririca sob estresse de até -2,6 MPa controlou satisfatoriamente a tiririca desde que irrigadas até 24 horas após a aplicação, mas quando a irrigação ocorreu 48 horas após, houve comprometimento na translocação do herbicida nas plantas. Quando o estresse foi menor que -2,0 MPa, o herbicida manteve-se eficaz no controle de tiririca, principalmente para aquelas com Yw de -0,35 MPa.

Crescimento inicial e morfologia foliar em plantas de Enterolobium contortisiliquum (Vell.) Morong. E Erythrina velutina Mart. ex Benth, sob estresse hídrico

The Caatinga is the predominant vegetation type in semi-arid region of Brazil, where many inhabitants depend on hunting and gathering for survival, obtaining resources for: food and feed, folk medicine, timber production, etc. It‟s the dry ecosystem with highest population density in the world. The early stages of development are the most critical during the life cycle of a flowering plant and they‟re primordial to its establishment in environments exposed to water stress. Information about adjustments to the growth of the species, correlated with their studies of distribution in Seridó oriental potiguar, are an important ecological and economic standpoint, because they provide subsidies for the development of cultivation techniques, to programs of sustainable use and recovery of degraded areas. This thesis aimed to study the initial growth and foliar morphology in plants like Enterolobium contortisiliquum (Vell.) Morong. (tamboril) and Erythrina velutina Mart. ex Benth (mulungu), species of occurrence in the Caatinga, under water stress. After sowing and emergency, the seedlings were exposed to three water regimes: 450 (control), 225 (moderate stress) and 112.5 (severe stress) mm of water slide for 40 days. Seeding occurred in bags of 5 kg and after the establishment of seedlings thinning was carried out leaving a plantlet per bag. At the beginning the waterings occurred daily with distilled water, passing to be on alternate days after thinning. Twenty and forty days after the thinning seedlings collections were held to be done analysis of growth and biomass partition. When compared to the control group, the treatments with water stress showed reduction in the growth of the aerial part, growth of the greater root, number of leaves and leaflets, dry leaf area and total phytomass in both species, but in general, this effect was most marked for E. velutina. Regarding the partition of biomass, there were few changes throughout the experiment. Morphological changes in the leaves as a function of stress were not significant, however, there was a trend, in both species, to produce narrower leaves, that facilitate heat loss to the environment. It has not been possible to establish a positive relationship between inhibition of growth and distribution of species, whereas E. velutina is a species of most common occurrence in Seridó oriental potiguar. In this way, other aspects should be taken into account when studying the adaptation of species the dry environments, such as salinity, presence of heavy metals, wind speed, etc