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

A saúde psíquica de quem faz saúde: uma análise crítica sobre a humanização direcionada ao profissional de saúde

It is noticeable that pressure, tension and overwork are frequent in health professionals routine. The work related to the ward area demands deep attention and surveillance. Because of that, it is essential to have a specific look at the humanization directed at health professionals, considering that taking care of other human beings is the essence of their job. This study has analyzed the psychic health levels, as well as the stress health professionals are submitted to, providing a debate about the humanization in 06 public hospitals (03 of them awarded by actions of humanization, and 03 not awarded) in Rio Grande do Norte state, Brazil. A study with 126 active health professionals (doctors, nurses, psychologists, nutritionists and social workers) in ward areas in their respective institutions was carried out. The thesis presented, with multi-disciplinary characteristic, counted on the support of statisticians (to calculate samples and data analysis), psychologists, social workers and administrators (linked to the human resources sector in each hospital). A cross-sectional study was performed, taking into consideration both quantitative and qualitative factors. The tools used for that were a semistructured questionnaire with socio-demographic characteristics, work and humanization; Lipp's Stress Symptoms Inventory for Adults (ISSL), and the Goldberg s General Health Questionnaire (QSG). The workers are predominantly women (84,9%), married (54,8%), between 46 and 55 years old (40,5%), working in the same institution for more than 20 years (22,2%), and between 16 and 20 years (20,6%), respectively. They work 40 hours a week (71,4%) and have multiple jobs (61,9%). Although most of these individuals global psychic health is in a good level, there are a significant number of people that is gradually getting worse concerning psychic stress (F1) showed by QSG (54,7%), and stress showed by ISSL(42,1%). Observing the categories, nurses (41,5%). Nutritionists (20,8%), doctors and social workers (18,9%), were among the most affected. About general health (F6), 63% of the awarded hospitals and 70% of the not awarded ones, presented good health levels (ranging from 5 to 50%). It was also noticed that, in the groups mentioned above, 25 and 20% respectively, were inserted in scores between 55 to 90%, what means that they are in worsening phase. The fact that the hospital is awarded or well recognized doesn t interfere in health professionals stress level and in their psychic health. Through what was heard from these individuals, it was possible to verify that they know little about humanization, once few of them identify or know that the service they offer is in an adoption process by Ministerial Policies. It was also detected the necessity of developing actions aimed at worker s health. Such results showed the importance of have more investments in programs that are directed to workers well-being, because they deal with other people s health and it is known that it is difficult for them to offer high-quality assistance if there are not suitable physical, psychological and material conditions to help them develop their jobs. As a warning, it is fair to say that investments in actions that provide humanized care to health professionals, mainly concerning preventive care for their health and life quality in their work

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

Caracterização sócio-demográfica, ocupacional e de formação profissional de manipuladores de alimentos e bebidas em uma capital da região nordeste do Brasil

Universidade Federal do Rio Grande do Norte

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

Conselho Nacional de Desenvolvimento Científico e Tecnológico

Relações trabalhistas e estilo de vida do atleta profissional de futebol do Rio Grande do Norte

Brazilian Law n° 9.615 of 24 March, 1998 established new paradigms in the employment relations existent between the soccer athlete and the sport association, both of whom are accustomed to the former legislation. They do not seem to have organized themselves in the sense of understanding the practical effects of the legal precepts currently in force, raising doubts that hinder the professional practice of the former and the performance of the latter in relation to managing these human resources. The purpose of this study is to analyze the degree of knowledge displayed by the professional soccer athlete about the legal recourses at his disposal in relation to the employment relations established in his work contract, as well as investigating the stress habits and stress situations that he most often experiences. This descriptive study consisted of a sample of 105 players under contract with clubs participating in the final phase of the state soccer championship. A questionnaire was used as the data collection instrument, whose analysis allows us to verify the lack of knowledge, on the part of the subjects studied, about the laws that regulate their profession, as well as identifying the stress habits and stress situations that are most reflected in their lifestyle. The lack of knowedge of regulate norms of their profession influences, in a negative way, in the lifestyle of soccer professional athlete from Rio Grande do Norte, Brasil

A compreensão do corpo na formação profissional do fisioterapeuta

Physical therapy has suffered of a mechanistic influence, with the superspecialization and fragmentation of learning, which interfers directly in the professional s understanding of the body, besides affecting his therapeutic performance. Worried about this reality, this research analyzed perceptions of Physical therapy students from Universidade Federal do Rio Grande do Norte and Universidade Potiguar about the human body. This is a descriptive study where 167 students were evaluated through objective and subjective responses to questionnaires designited QUEB Questionnare of understanding body evaluation. Searching the complement of the datas the QUEB open-ended was created, which gives freedom to students to answer freely. This questionnnaire was applicated on a subgroup composed by 21 students of the Health Sociology subject of the Physical therapy course. The validation process of these questionnaires included strategies of a panel of experts and face validity. The theorical reference analyzed based on the studies which favor the rejoining of knowledge represented by transdisciplinary support. The analysis of the results were performed quantitativavely and qualitatively through categorization of the responses selectioning key-words and the most expressive discourses , besides using descritive statistics interpretation. According to the responses, the body thought only thought biologically, ignoring the understanding of man as a cultural and social construction, confirming the presence of the reductionist model, with overvalorization of early specialization, technical training and purely biological and mechanical considerations of the body and its mobility. Probably, the dialogic knowledge of the body human inside Physical Therapist s learning can promote a growth of health s concept and a true activeness of Physical therapy on it, being the body a link with social environment. So, through understanding of the body as a complex form, the physical therapy will be able to attend your patients considerating their biological characteristics, but so the religious, political, social and ethics. Finally, this reflection suggests a search for a less technical vision, which allows the professional to discern more than segments of the body and which will contribute to a wider understanding of the patient and his social context, leading to greater humanization of the body , improvement in services and consequently, in the quality of life of these patients

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.