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.

Estudo do papel da proteína multifuncional APE1/Ref-1 sobre a resposta inflamatória na meningite bacteriana

Despite advances in antibiotic therapy, bacterial meningitis (BM) remains with high mortality and morbidity rates in worldwide. One important mechanism associated to sequels during disease is the intense inflammatory response which promotes an oxidative burst and release of reactive oxygen species, consequently leading to cell death. Activation of DNA repair enzymes during oxidative stress has been demonstrated in several neurological disorders. APE1/Ref-1 is a multifunctional protein involved in DNA repair and plays a redox function on transcription factors such as NFkB and AP-1.The aim of this study was assess the role of APE1/Ref-1 on inflammatory response and the possibility of its modulation to reduce the sequels of the disease. Firstly it was performed an assay to measure cytokine in cerebrospinal fluid of patients with BM due to Streptococcus pneumoniae and Neisseriae meningitides. Further, a cellular model of inflammation was used to observe the effect of the inhibition of the endonuclease and redox activity of APE1/Ref-1 on cytokine levels. Additionally, APE1/Ref-1 expression in cortex and hippocampus of rat with MB after vitamin B6 treatment was evaluated. Altogether, results showed a similar profile of cytokines in the cerebrospinal fluid of patients from both pathogens, although IFNy showed higher expression in patients with BM caused by S. pneumoniae. On the other hand, inhibitors of APE1/Ref-1 reduced cytokine levels, mainly TNF-α. Reduction of oxidative stress markers was also observed after introduction of inhibitors in the LPS-stimulated cell. In the animal model, BM increased the expression of the protein APE1/Ref-1, while vitamin B6 promoted reduction. Thereby, this data rise important factors to be considered in pathogenesis of BM, e.g., IFNy can be used as prognostic factor during corticosteroid therapy, APE1/Ref-1 can be an important target to modulate the level of inflammation and VIII oxidative stress, and vitamin B6 seems modulates several proteins related to cell death. So, this study highlights a new understanding on the role of APE1/Ref-1 on the inflammation and the oxidative stress during inflammation condition

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

A fé e a religiosidade na cova de Pedro Ribeirão do largo -Bahia

At this research we attempted to understand what feeds and supports the faith and popular religiosity at Pedro s Cave in Ribeirao do Largo, located in Bahia State. For this we studied the visitation ritual at Pedro s Cave on June 29th and tried to identify the importance of this ritual in the construction and reconstruction of believer s faith. Moreover, we endeavored to understand which factors led Pedro Afonso Nascimento, a cowhand whose lift was taken in a violent manner, to the roll of popular saints and how it was built around his tomb a sanctuary of constant visitation

Ideal Mariano e docência: a identidade feminina da Proposta Educativa Marista

In the teaching history course it s found the female teacher entering, reflecting significantly in the continuous process of her identity redefinitions and reconstruction. This entering brings, even in a silent and not explicit way, the confluence of gender identity to professional identity, in which are present the relations of the proper to female. In the mastership case, these relations are found imbricated in the teaching identity construction and in the translation of this profession as female, based in Virgin Mary, through Mariano ideal diffused by the catholic church. Therefore there is a lanck in educational historiography towards the register of female teachers presence, mainly in male religious institute, the reason of this study, in which we seek to investigate: how to explain that a female identity can give support to a highly male educational project? What relations can we establish between this female identity and the entering of female teachers in Maristas Province Schools of North Brazil? We performed a bibliographic research in Maristas Schools Libraries, of Marista historial, in Recife, of Pontificia Universidade Católica do Paraná (Catholic University), in Curitiba-PR. We searched for support in studies about pedagogy history, of Marista Institute, Catholic Church, Brasilian Education and of Women; Mariologic studies, the referring to Marista Educational Proposal, to feminization and to teaching identity; and feminist theology productions. The empirical research included 10 schools from referred province and Marista Schools from Lisbon Portugal. Through semi-estructured interviews, we interviewed pioneer female teachers, Marista former Brothers and Brothers from those schools, due to their presence and action within the study period and for the concern to save their memories about the theme. The study reveals the particularities of Mariano ideal as female model and teaching, and her presence in Marista Educational Proposal. This study, in a historical approach, aims to contribute to give visibility to the woman in history and teaching construction in Brazil, mainly in religious institutions from male origin

O ensino da história e o pensamento reflexivo-crítico da professora no 3º ano do Ensino Fundamental

This collaborative research is a qualitative approach, from historical and cultural perspective, made about a teacher of third grade of basic education in the area of history, in the municipality of Caicó / RN. It has as objective to investigate, in a collaborative action, if the design process of teaching and learning the discipline of history based on the relationship before / after allows the development of critical reflective thinking of the teacher in school practices. The theoretical and methodological approaches are supported in the postulates of Vygotsky (1998), Rubinstein (1973) and Linblinskaia (1979), among others, whose understanding has led us to reflect if the teacher develops the reflective critical-thinking in history discipline classes. The complexity of the study led us to an analysis exercise, using different methodological procedures, such as: bibliographic review of the literature, considering also the literature of the history area, interviews, observation in the classroom, video recordings and reflective sessions, enabling clarify the construction and reconstruction of thought that the teacher had been developing during the process of teaching and learning. The test results point to a dichotomy between theory and practice, and also to a certain fragility in the position of professor in teaching and learning the discipline of history, at the third grade of basic education. The teacher recognizes that she requires a theoretical deepening with more intensity, as a process of continuous training to improve the practice of teaching in history school, though, stating in her speech that her teaching practice is based on critical reflection. However, she presents serious limitations in classroom practices. We conclude that, although she has shown willing to work on a critical perspective of reality, showed also poor change at school practices, starting to reflect about her own actions, pointing her limits and the changes needed that didn t become reality yet. It s necessary a formative process for her. The study therefore showed that even with the sessions and reflective of deepening theoretical studies, the teacher does not change its profile, while maintaining its traditional vision in any pedagogical action. This research recommends the formation of school groups for further studies and discussion on the practices of education in history area from a reflective-critical thought perspective as a mean of personal and professional development

Formação de professores, saberes, reflexividade e apropriação da cultura digital no Projeto Um Computador por Aluno (UCA)

This study investigates teacher training and cognitive practice of teachers in a Basic Education school that adopted the Project One Computer per Student (OCS) in their school routine. Its relevance consists in provide directions for the continuation of training activities on the Project and guide the teachers with their pedagogical practices using the laptop model one to one. The thesis defended is that the educator formation for social using of digital media (specially the laptops from the Project UCA) gives space to establish new sociotechnical relationships, of new social and professionals practices, new identitary components and a process of reflexivity and knowledge reconstruction to teach. We reaffirm the importance of reflexivity and appropriation of digital culture for the better development of teaching practice using the Information and Communication Technologies (ICTs), giving focus to the aspects of social and professional use of the technology. The study is part of the qualitative aspect and is a procedural tracking based on principles of ethnographic research. As procedures and methodological tools, were used: intensive observation of school environments, documental analysis, focal group, semi-structured questionnaires and semi-structured individual interviews. The research was held in a public school in the city of Parnamirim - RN. The subject sample relates to 17 teachers, coming from the elementary school I and II, Youth and Adult Education and High School, who went through the process of training UCA and having entered the laptops in their teaching. The research corpus is structured based on the messages built into the process of data collection and is analyzed based on principles of Content Analysis, specified by Laurence Bardin (2011). Was taken as theoretical reference studies by Tardif (2000; 2011), Pimenta (2009), Gorz (2004, 2005), Giddens (1991), Dewey, J. (1916), Boudieu (1994; 1999), Freire (1996; 2005), among others. The analysis indicates a process of reconstruction / revision of knowledge to teach and work in digital culture, being these knowledges guided by the experience of the subjects investigated. The reconstructed knowledges will be revealed from a categorization process. The following groups of knowledges: "technical knowledges", "didactic-methodological knowledges and knowledges of professionalization" were built on the assumption of ownership of digital culture in the educational context. The analysis confirms the appearance of new ways of sociability when acquiring other forms of acting and thinking ICTs, despite the environment adverse to the reflexivity shared among the teachers. Also reveals, based on the ownership concept present on the data analysis, the construction of meanings of belonging and transformation of individuals into social routes from the interweaving of the teaching practice with the digital culture. Emphasizes, finally, the importance of a training for use of ICTs that exceeds the instrumentation, in other words, what we call "technical knowledges", but taking on its structural basis the shared reflection, the opening for the ressignificance (new meaning) and reconstruction of new knowledges and practices and that really allows, to the teacher, the living of an experience capable of providing socio-technical transformations of their relationships

O currículo e as práticas pedagógicas na EJA: concepções e crenças dos professores da escola municipal Francisca Leonísia Cruz

The present investigation deals with the Curriculum and Pedagogical Practices of Young and Adults, EJA, and articulates with the conceptions and beliefs of teachers sitting. The empirical scope of this investigation consisted in the Municipal School Francisca Leonísia, located in the Agrarian Reform Settlement Serra Nova, city of Florânia/RN. Seeks to analyze the relationship between the Curriculum and Educational Practices of Young and Adult EJA - conceptions beliefs of these teachers. This work is based on qualitative research, interpretative character reflective and makes use of documentary analysis, structured interviews and reflective sessions as methodological procedures which ensure the achievement of our goals in research. These procedures allowed us to enter the practice setting and curriculum for teachers to understand how they think, prepare and practice of adult education in school curriculum research field. The documentary analysis provided the rethinking of the curriculum selected references, Political Pedagogical Project and Proposed Course of EJA, from a critical reading of concepts and conceptions given in these references, with a view to construction and reconstruction of concepts that reclaim the identity of young people and adult field, inserted in the Rural Workers Movement landless/MST. As reflective sessions were constituted in spaces of collective training and allowed the group to selfreflection e collective reflection about the ideas and beliefs that permeate the curriculum and guide their practices education in adult education. In these areas of training are also discussed current problems of adult education, the construction of an educational project of the adult education field to, warranties of learning and cultural identity of the adult education field and placed in the context of the MST. Under this view, it is concluded that the conceptions and beliefs of teachers seated directly relate to the curriculum designed, developed and practiced in adult education, as well as the educational practices that permeate the curriculum. This relation is in the midst of adversities of Rural Education, inserted in the Movement of Landless Rural Workers-MST, and requires the commitment of teachers to the necessary changes to an education are critical and emancipatory

Modelo de gerenciamento de estoques de peças de reposição para empresas de transporte urbano de passageiros

This work searches to offer a model to improve spare parts stock management for companies of urban passenger transport by bus, with the consequent progress in their maintenance management. Also known as MRO items (Maintenance, Repair and Operations), these spare parts, according their consumption and demand features, cost, criticity to operation, lead-time, quantity of suppliers, among other parameters, shouldn´t have managed their inventory like normal production items (work in process e final products), that because their features, are managed by more predictable models based, for example, in economic order quantity. In the case specifically of companies of urban passenger transport by bus, items MRO have significant importance in their assets and a bad management of these inventories can cause serious losses to company, leading it even bankrupticy business, in more severe situations which missing spare part provokes vehicles shutdown indefinitely. Given slight attention to the issue, which translates in little literature available about it when compared to that literature about normal items stocks, and due the fact that MRO items be critical to bus urban transport of passengers companies´, it is necessary, so, deepen in this theme searching to give technical and scientific subsidies to companies that work, in many times, empirically, with these so decisive inputs to their business. As a typical portfolio problem, in which there are n items, separated into critical and noncritical, while competing for the same resource, it was developed a new algorithm to aid in a better inventory management of spare parts used only in corrective maintenance (whose failures are unpredictable and random), by analyzing the cost-benefit ratio, which compares the level of service versus cost of each item. The model was tested in a company of urban passenger transport by bus from the city of Natal, who anonymously provided their real data to application in this work

Prospecção de genes de interesse biotecnológico : uma abordagem metagenômica

The total number of prokaryotic cells on Earth has been estimated at 4 to 6x1030 and only about 1% of microorganisms present in the environment can be cultivated by standard techniques of cultivation and plating. Therefore, it is a huge biological and genetic pool that can be exploited, for the identification and characterization of genes with biotechnological potential. Within this perspective, the metagenomics approach was applied in this work. Functional screening methods were performed aiming to identify new genes related to DNA repair and / or oxidative stress resistance, hydrocarbon degradation and hydrolytic activities (lipase, amylase and protease). Metagenomic libraries were built utilizing DNA extracted from soil samples collected in João Câmara RN. The libraries were analyzed functionally using specific substrate containing solid medium (hydrolytic activity), supplemented with H2O2 (DNA repair and / or resistance to oxidative stress) and liquid medium supplemented with light Arabian oil (activity, degradation of hydrocarbons). After confirmation of activity and exclusion of false-positive results, 49 clones were obtained, being 2 positive for amylase activity, 22 resistant to oxidative stress generated by H2O2 and 25 clones active for hydrocarbons degradation. Analysis of the sequences showed hypothetical proteins, dienelactona hydrolase, DNA polymerase, acetyltransferase, phosphotransferase, methyltransferase, endonucleases, among other proteins. The sequence data obtained matched with the functions tested, highlighting the success of metagenomics approaches combined with functional screening methods, leading to very promising results

Metagenoma de comunidades microbianas expostas à radiação natural e a metais

Many species have specialized to live in the most varied existing environments showing the remarkable adaptability of the microbial world the most diverse physicochemical conditions. Environments exposed to natural radiation and metals are scarce around the world, presenting a microbiota still unknown. With a total number estimated between 4 and 6 x 1030 microrganisms on earth, they constitute an enormous biological and genetic pool to be explored. Metagenomic approach independent of cultivation, provides a new form to access to the potential genomic environmental samples becoming a powerful tool for the elucidation of ecological functions, metabolic profiles, as well as to identify new biomolecules. In this context, the genetic material of environmental soil and water samples from Açude Boqueirao Parelhas-RN, under the influence of natural radiation and the presence of metals, was extracted, pirosequencing and the generated sequences were analyzed by bioinformatics programs (MG-RAST and STAMP). Taxonomic comparative profiles of both samples showed high abundance of Domain Bacteria, followed by a small portion attributable to Eucaryota Domains, Archaea and Viruses. Proteobacteria, Actinobacteria and Bacterioidetes phyla showed the greater dominance in both samples. Important genera and species associated with resistance to various stressors found in region were observed. Sequences related to oxidative and heat stress, DNA replication and repair, and resistance to toxic compounds were observed, suggesting a significant relationship between the microbiota and their metabolic profile, influenced by regional environmental variables. The results of this study add valuable and unpublished data on the composition of microbial communities in these regions

Representações discursivas de Câmara Cascudo por Mário de Andrade

This piece of research aims to address the issue on the construction and reconstruction of Câmara Cascudo´s discursive representations in Mário de Andrade´s discourse. In order to describe, analyze and interpret these representations, we recur to some semantic categories from the Discourse Textual Analysis (DTA) by articulating them with other categories, notably, Grize´s Logic (1996,1997), Text Linguistics and Semantics. Therefore, the purpose is to analyze how these representations are constructed discursively, in written letters, by means of semantic categories such as referentiation, predication, modification, connection and spatial and temporal locations. In the theoretical foundation, proposals of Textual Analysis of Discourses, conceived by the linguist Jean-Michel Adam (1990, 2008a, 2011a), Text Linguistics, Semantics and Logic, focusing, especially, on the phenomenon of discursive representations, are articulated. The research approach is of qualitative nature supported by some quantitative data (OLIVEIRA, 2012); option which makes the analysis richer and comprehensive. As a hypothesis, it is presented the fact that these categories used by Mário de Andrade, in his discourse, do not only enable the (re)construction of the interlocutor´s images, discursively constructed, but they also provide a multiplicity of information and viewpoints about the RN writer´s personality. The study corpus is constituted of 20 texts written by Mário de Andrade and sent to Câmara Cascudo between 1924 and 1944, of which 35 fragments were selected and analyzed. However, it can be verified that, in the analyzed corpus, a set of discursive representations is constructed for Câmara Cascudo, from semantic categories proposed for analysis and used in Mário de Andrade´s discourse. These categories enable to construct and reconstruct the representations that emerge in the texts. Therefore, the analysis points out the construction of a set of different representations, highlighting the representation of the writer, the intellectual and the friend.

A educação popular para todos de uma cidade educadora (Natal, Rio Grande do Norte, 1957-1964)

This work has as a research subject of popular education policies of the city of Natal, Rio Grande do Norte, in the years 1957 to 1964. It aims to identify and analyze popular education policies developed and implemented by the Municipality of Natal in these years. To get the historical data, we establish as a guiding reserch question the following: Which elaborated educational policies were implemented by the Municipality of Natal in the years 1957-1964? and took over as the method Evidential Paradigm as proposition in Pinheiro (2009). This is anchored in documentary sources of Educational Legislation at National, State and Municipal levels as well as in the newspapers Folha de Tarde and Jornal de Natal; in existing documents from the archives of the Historical and Geographical Institute of Rio Grande do Norte (IHGRN), the Municipal Public Archives of Natal; iconographic sources; interviews and academic publications. In addition to these sources, we were inspired by the works of Aristotles (2011), Hobbes (2009), Freire (2011), Góes (1980), Germano (1989), Cortez (2005) and Galvão (2004). This research allowed us to understand that policies of popular education of Natal (RN) were based on a democratic educational practice, supported on three pillars, namely: participation and involvement of Natal population; construction and reconstruction of teaching practices in prioritizing their action programs to mass literacy and the training of lay teachers; and the democratization of culture. This historical process made Natal on educating city.

Imunoexpressão das proteínas APE-1 e XRCC-1 em carcinoma epidermoide de língua oral

DNA repair systems play a critical role in protecting the human genome from damage caused by carcinogens present in the environment. Mutations in DNA repair genes may be responsible for tumor development and resistance of malignant cells to chemotherapeutic agents. The major pathway for oxidative DNA damage repair is the base excision repair pathway. The objective of this study was to investigate the immunoexpression of APE-1 and XRCC-1, which are proteins involved in DNA base excision repair and its association with clinical and histopathological parameters in oral tongue squamous cell carcinoma (OTSCC), in order to investigate a possible prognostic value for those proteins. The expression of APE-1 and XRCC-1 was evaluated semi-quantitatively by immunohistochemistry in 50 OTSCC cases. Clinical data was collected from patients’ medical charts and histopathological grading was performed for each case. Statistical analysis (Chi-square and Fisher’s exact tests; significance of 5%) was performed to determine the association between protein expressions and clinico-pathological characteristics. APE-1 was highly expressed in nucleus and cytoplasm in 56% of cases. XRCC-1 showed overexpression only in nucleus in 60% of cases. High expression of XRCC-1 was significantly associated to clinical stages I and II (P=0.02). Both proteins were not associated to other clinical parameters or histopathological grading. Our findings demonstrate that DNA base excision repair proteins APE-1 and XRCC-1 are upregulated in OTSCC, however, they are not related to clinical and histologic parameters, except for XRCC-1 association to better clinical staging. Our results indicate that the immunohistochemical expression of these proteins has no association with prognostic parameters in this tumor.

Imunoexpressão das proteínas APE-1 e XRCC-1 em carcinoma epidermoide de língua oral

DNA repair systems play a critical role in protecting the human genome from damage caused by carcinogens present in the environment. Mutations in DNA repair genes may be responsible for tumor development and resistance of malignant cells to chemotherapeutic agents. The major pathway for oxidative DNA damage repair is the base excision repair pathway. The objective of this study was to investigate the immunoexpression of APE-1 and XRCC-1, which are proteins involved in DNA base excision repair and its association with clinical and histopathological parameters in oral tongue squamous cell carcinoma (OTSCC), in order to investigate a possible prognostic value for those proteins. The expression of APE-1 and XRCC-1 was evaluated semi-quantitatively by immunohistochemistry in 50 OTSCC cases. Clinical data was collected from patients’ medical charts and histopathological grading was performed for each case. Statistical analysis (Chi-square and Fisher’s exact tests; significance of 5%) was performed to determine the association between protein expressions and clinico-pathological characteristics. APE-1 was highly expressed in nucleus and cytoplasm in 56% of cases. XRCC-1 showed overexpression only in nucleus in 60% of cases. High expression of XRCC-1 was significantly associated to clinical stages I and II (P=0.02). Both proteins were not associated to other clinical parameters or histopathological grading. Our findings demonstrate that DNA base excision repair proteins APE-1 and XRCC-1 are upregulated in OTSCC, however, they are not related to clinical and histologic parameters, except for XRCC-1 association to better clinical staging. Our results indicate that the immunohistochemical expression of these proteins has no association with prognostic parameters in this tumor.