Caracterização funcional de dois cDNAs homólogos à ciclofilina e à proteína reprimida por auxina (ARP) identificados em bibliotecas subtrativas a para floração em tomateiro

Flowering is a fundamental process in the life cycle for plant. This process is marked by vegetative to reproductive apical meristem conversion, due to interactions between several factors, both internal and external to plant. Therefore, eight subtractive libraries were constructed using apical meristem induced or not induced for two contrasting species: Solanum lycopersicum cv. Micro-Tom and Solanum pimpinellifolium. Several cDNAs were identified and among these, were selected two cDNAs: one homologous cDNA to cyclophilin (LeCYP1) and the other to Auxin repressed protein (ARP). It has observed that LeCYP1 and ARP genes are important in the developmental process to plants. In silico analysis, were used several databases with the exclusion criterion E-value <1.0x10-15. As a result, conservation was observed for proteins analyzed by means of multiple alignments and the presence of functional domains. Then, overexpression cassettes were constructed for the ARP cDNA in sense and antisense orientations. For this step, it was used the CaMV35S promoter. The cDNA orientation (sense or antisense) in relation to the promoter was determined by restriction enzymes and sequencing. Then, this cassette was transferred to binary vector pZP211 and these cassettes were transferred into Agrobacterium tumefaciens LBA4404. S. lycopersicum cv. Micro-Tom (MT) and MT-Rg1 plants were transformed. In addition, seedlings were subjected to hormone treatments using a synthetic auxin (- naphthalene acetic acid) and cyclosporin A (cyclophilin inhibitor) treatments and it was found that the hormone treatment there were changes in development of lateral roots pattern, probably related to decreases in auxin signaling caused by reduction of LeCYP1 in MT-dgt plants while cyclosporin A treatments, there was a slight delay in flowering in cv. MT plants. Furthermore, assay with real-time PCR (RT-qPCR) were done for expression level analysis from LeCYP1 and ARP in order to functionally characterize these sequences in tomato plants.

Prospecção de genes associados ao processo de floração em tomateiro.

Flowering is a process marked by switch of shoot apical meristem to floral meristem, and it involves a complex regulation by endogenous and environmental factors. Analyses of key flowering genes have been carried out primarily in Arabidopsis thaliana and have provided a foundation for understanding the underlying molecular genetic mechanisms controlling different aspects of floral development. Several homologous have been found in other species, but for crops species such as tomatoes this process is not well known. The aim of this work was to use the genetic natural variation associated to the flowering process and use molecular tools such as subtractive libraries and real time PCR in order to identify and analyze the expression from genes that may be associated to flowering in these two species: L. esculentum cv Micro-Tom and L. pimpinellifolium. Our results showed there were identified many genes related to vegetative and possibly to the flowering process. There were also identified many sequences that were unknown. We ve chosen three genes to analyze the expression by real time PCR. The histone H2A gene gave an expression higher in L. pimpinellifolium, due to this the expression of this gene may be associated to flowering in this specie. It was also analyzed the expression of an unknown gene that might be a key factor of the transition to flowering, also in L. pimpinellifolium. For the elongation factor 1-α expression, the expression results were not informative, so this gene may have a constitutive expression in vegetative and flowering state. The results observed allowed us to identify possible genes that may be related to the flowering process. For further results it will be necessary a better characterization of them.

Caracterização funcional de dois cDNAs homólogos à ciclofilina e à proteína reprimida por auxina (ARP) identificados em bibliotecas subtrativas a para floração em tomateiro

Flowering is a fundamental process in the life cycle for plant. This process is marked by vegetative to reproductive apical meristem conversion, due to interactions between several factors, both internal and external to plant. Therefore, eight subtractive libraries were constructed using apical meristem induced or not induced for two contrasting species: Solanum lycopersicum cv. Micro-Tom and Solanum pimpinellifolium. Several cDNAs were identified and among these, were selected two cDNAs: one homologous cDNA to cyclophilin (LeCYP1) and the other to Auxin repressed protein (ARP). It has observed that LeCYP1 and ARP genes are important in the developmental process to plants. In silico analysis, were used several databases with the exclusion criterion E-value <1.0x10-15. As a result, conservation was observed for proteins analyzed by means of multiple alignments and the presence of functional domains. Then, overexpression cassettes were constructed for the ARP cDNA in sense and antisense orientations. For this step, it was used the CaMV35S promoter. The cDNA orientation (sense or antisense) in relation to the promoter was determined by restriction enzymes and sequencing. Then, this cassette was transferred to binary vector pZP211 and these cassettes were transferred into Agrobacterium tumefaciens LBA4404. S. lycopersicum cv. Micro-Tom (MT) and MT-Rg1 plants were transformed. In addition, seedlings were subjected to hormone treatments using a synthetic auxin (- naphthalene acetic acid) and cyclosporin A (cyclophilin inhibitor) treatments and it was found that the hormone treatment there were changes in development of lateral roots pattern, probably related to decreases in auxin signaling caused by reduction of LeCYP1 in MT-dgt plants while cyclosporin A treatments, there was a slight delay in flowering in cv. MT plants. Furthermore, assay with real-time PCR (RT-qPCR) were done for expression level analysis from LeCYP1 and ARP in order to functionally characterize these sequences in tomato plants.

Henrique Novaes: técnica, território e cidade em uma trajetória profissional: Brasil, primeira metade do século XX

The career of civil engineer Henrique de Novaes, a remarkable figure in the academic, technical and political fields, demonstrates its extensive and relevant work across Brazil in the first half of the twentieth century. It covered from the design of water supply and sewage works, road and rail transport networks, works against drought in the Northeast, hydroelectric and harbor facilities to the urbanization planning and architectural projects, which displays a systematic and multiple production. City and territory conformed to his fields of verification, practical and above all, transformation. The search for understanding of the inclusion of a social actor in this process thus contributes to the specific analysis of the doings of Henrique de Novaes, who graduated at Polytechnic School of Rio de Janeiro in 1906. From a polymorphic activity in different scales, one tries to figure how, through academic education or professional practice, urban history in Brazil can be told or built. The introduction of technological innovations matched the purposes of planning and urban sprawl, as well as met the specifications of regulation and institutionalization of public infrastructure services at the time. The overall plans proposed by the engineer thought of the city as a whole, interconnected to the structural networks. At the same time, the knowledge of a larger physical scale the territory bounces back in the urban in a relationship of reciprocity and completeness. The objective research, therefore, tries to understand the roles played by Henrique de Novaes s works and academic education in the accomplishment of systematic modernization of Brazilian urban space and territory, recovering a little known historical figure by current historiography. It is proposed, as methodological axis, that the study of this professional career configures itself as an essential element for understanding the idea of progress embodied in the technical studies and proposals for improvements and sanitation nationwide in the first half of the twentieth century . The primary sources for the construction of this analysis were technical articles in journals of the period ( Clube de Engenharia , Viação e Revista Brasileira de Engenharia ), and technical reports, government messages, newspaper articles published at the time, autobiographical reports and the engineer s verbal communications with relatives. The work is structured in three chapters: "Biographical traces, academic education and 'technical and political' activities" illustrates the initiation into the technical, public and political environment; Chapter 2, "Technique and territory" outlines his network understanding through sanitation and transport services; Chapter 3 "Technique and City" describes the influence of polytechnics knowledge on the propositions of modernization of cities; Finally, "Final Thoughts: An Evaluation," presents an overview of the affiliations and practice of an engineer in the different scales, and its contribution to the modernization of Brazilian urban and territorial space

Avaliação da ação bioinseticida de SBTI e vicilina de Erythrina velutina em enzimas digestivas e membrana peritrófica de larvas de Plodia interpunctella (Lepidoptera: Pyralidae)

Plodia interpunctella (Indian meal moth) is a cosmopolitan pest that attacks not only a wide range of stored grain as well other food products. Due to its economic importance several researches have focused in a method with ability to control this pest with few or no damage to the environment. The study of digestive enzymes inhibitors, lectins and chitin-binding proteins, has often been proposed as an alternative to reduce insect damage. In this study we report the major classes of digestive enzymes during larval growth in P. Interpunctella, being those proteinases actives at pH 9.5 and optimum temperature of 50 oC to both larvae of the 3rd instar and pre-pupal stage of development. In vitro and zymogram assays presented the effects of several inhibitors, such as SBTI, TLCK and PMSF to intestinal homogenate of 3rd instar larvae of 62%, 92% and 87% of inhibition and In pre-pupal stage of 87%, 62 % and 55% of inhibition, respectively. Zymograms showed inhibition of two low molecular masses protein bands by TLCK and that in presence of SBTI were retarded. These results are indicative of predominance of digestive serine proteinases in gut homogenate from Plodia interpunctella larvae. This serine proteinase was then used as a target to evaluate the effect of SBTI on larvae in in vivo assay. Effect of SBTI on mortality and larval mass was not observed at until 4% of concentration (w/w) in diets. Chitin, another target to insecticidal proteins, was observed by chemical method. Moreover, optic microscopy confirmed the presence of a peritrophic membrane. Established this target, in vivo effect of EvV, a chitin binding vicilin, evaluated during the larval development of P. interpunctella and was obtained a LD50 of 0,23% and WD50 of 0,27% to this protein. Mechanism of action was proposed through of the in vivo digestibility of EvV methodology. During the passage through the larval digestive tract was observed that EvV was susceptible to digestive enzymes and a reactive fragment, visualized by Western blotting, produced by digestion was recovered after dissociation of the peritrophic membrane. The bound of EvV to peritrophic membrane was confirmed by immunohystochemical assays that showed strong immunofluorescent signal of EvV-FITC binding and peritrophic membrane. These results are a indicative that vicilins could be utilized as potential insecticide to Plodia interpunctella and a control methods using EvV as bioinsecticide should be studied to reduce lost caused by storage insect pests

O controle social do judiciário: a experiência do Observatório da Justiça e Cidadania no Estado do Rio Grande do Norte

This dissertation deals with the possibility of build an effective social control of the judiciary Brazilians. The theme was bounded by a cut geographic and secular: the experience of the Center of Justice and Citizenship in the state of Rio Grande do Norte (OJC/RN), which begins formally in august 2003. The research approach and leave of experience in judicial practice and policy specific substrates to theorize about the subject. We collected documents about cases, the judicial diagnoses, reports, news material, in addition to lifting bibliographic. Therefore, it is working with about notions of a democratic state of right in the light of the Brazilian Constitution of 1988, in order to contextualize the insertion of the judicial system, by the prospect of legitimacy, which is considered by a look formal and material. It is a brief analysis of the system of official control of the judiciary (internal and external), is emphasizing its shortcomings functional and its corporate character, which suffers from poor conformation democratic. Then there is a discussion about the need to establish the social control of the judiciary, through the prism of relations of power that are locked in the judiciary, the lack of formal criteria for the guarantee of obtaining the correct judgment (laws, precedents and conscience of the judge), the problems of impunity and justice class, and from the examination of some cases, as the body of search. From this conjuncture, prepares to be an outline of shapes and the limits of social control, consonant the proposal erected in certain sectors of organized civil society, represented by the movement s social OJC. In the end, considerations are made on the legitimacy and constitutionality of OJC

Instituições, agricultura familiar e crédito rural: avaliação de resultados do Pronaf B nos territórios rurais do Rio Grande do Norte

Broadly anchored on Institutionalist literature this dissertation seeks to answer: Why rural credit granted to farming families in Rio Grande do Norte, Pronaf group B, has shown positive results in some territories while in others the same results are insatisfactory? Considering formal rules are the same, then why such results are so distinguished? This research supports the fact that these results are a direct consequence of the quality of institutions defined as longstanding social rules established and built in social interactions. For institutions one understands; organizational structures, formal and non-formal rules, social trust, thinking behaviors and its evolution as well as routines. Using Pronaf Rural Credit as a background this study measures and compares results obtained in the rural areas of Seridó, Sertão do Apodi, Mato Grande, Assú-Mossoró, Trairi, Potengi and Alto Oeste. The dimensions of measurement considered were: economics, social and guarantee of transparency. The study considers a sample of 402 Group B farming families. It also elaborates a socio-economic profile of these families in the considered areas and underlines the main causes for defaulted loans, difficulties to improve family income and results of access to Pronaf B credit line, as well as it proposes solutions to surpass obstacles to improve the efficiency of this credit line

Jornalismo cultural em texto, imagem e cor

O tema deste trabalho é o Jornalismo Cultural. Uma apresentação e discussão de temas pertinentes ao jornalismo especializado em cultura nas páginas do jornal diário. A partir da leitura de alguns jornais, se traça considerações gerais acerca dos caminhos do jornalismo e dos estudos sobre jornalismo com o objetivo de apresentar o jornal que hoje se produz no Brasil, em texto, imagem e cor. O resultado é a descoberta do jornal como forma, desde o texto, escolhas lexicais e sintáticas, passando pelas imagens, composição, escolha e determinação da pauta, ao arranjo destes elementos na página, em respeito ao projeto editorial de diagramação. A conclusão a que se chega que há vários "eus" que governam o jornal diário, e que fazem o jornalismo cultural inovador, porta de entrada às mudanças no jornalismo.

O poema e a canção em as coisas, de Arnaldo Antunes: imagens da primeiridade

Ce travail a pour but d analyser un corpus de six textes hybrides, que nous classons comme des poèmes-chansons/ poèmes-chantés à cause de leur double présence dans deux systèmes sémiotiques différents. Le premier, la littérature, ou plus spécifiquement la poésie, a comme support le livre As Coisas [Les Choses], d Arnaldo Antunes et l autre, la chanson, est enregistré dans les disques du même auteur. Notre travail lance un regard sur ce corpus, en essayant de vérifier un aspect recourrent dans l oeuvre d Arnaldo Antunes qui est la présence da priméité, catégorie theórique développée par Charles Sanders Peirce. Au-delà de l observation de cet aspect sémiothique, nous ferons une discussion sur la chanson populaire, et ses rapports avec la poésie et par conséquent avec la Litterature. La théorie sémiothique s appuyera sur deux piliers : En ce qui concerne l étude de la priméité, nous travaillerons avec les théories de Peirce, mais en nous servant aussi des ouvrages de Lúcia Santaella, Winfried Nöth, Júlio Plaza et Décio Pignatari ; Dans l autre voie, pour ce qui concerne l analyse des chansons, nous utiliserons la théorie de Luiz Tatit, fondée sur la sémiothique de Algirdas Julien Greimas. Tatit trace une méthode d analyse, où il est possible d analyser une chanson en exploitant et le texte et la mélodie, ce qui permet une meilleure compréhension de l étude des poèmes-chansons et ses variations. Comme support pour la discussion sur la musique, nous nous servirons des théories de José Miguel Wisnik, Claude Lévi-Strauss, Roland Barthes et Jean Fisette

Henrique Novaes: técnica, território e cidade em uma trajetória profissional: Brasil, primeira metade do século XX

The career of civil engineer Henrique de Novaes, a remarkable figure in the academic, technical and political fields, demonstrates its extensive and relevant work across Brazil in the first half of the twentieth century. It covered from the design of water supply and sewage works, road and rail transport networks, works against drought in the Northeast, hydroelectric and harbor facilities to the urbanization planning and architectural projects, which displays a systematic and multiple production. City and territory conformed to his fields of verification, practical and above all, transformation. The search for understanding of the inclusion of a social actor in this process thus contributes to the specific analysis of the doings of Henrique de Novaes, who graduated at Polytechnic School of Rio de Janeiro in 1906. From a polymorphic activity in different scales, one tries to figure how, through academic education or professional practice, urban history in Brazil can be told or built. The introduction of technological innovations matched the purposes of planning and urban sprawl, as well as met the specifications of regulation and institutionalization of public infrastructure services at the time. The overall plans proposed by the engineer thought of the city as a whole, interconnected to the structural networks. At the same time, the knowledge of a larger physical scale the territory bounces back in the urban in a relationship of reciprocity and completeness. The objective research, therefore, tries to understand the roles played by Henrique de Novaes s works and academic education in the accomplishment of systematic modernization of Brazilian urban space and territory, recovering a little known historical figure by current historiography. It is proposed, as methodological axis, that the study of this professional career configures itself as an essential element for understanding the idea of progress embodied in the technical studies and proposals for improvements and sanitation nationwide in the first half of the twentieth century . The primary sources for the construction of this analysis were technical articles in journals of the period ( Clube de Engenharia , Viação e Revista Brasileira de Engenharia ), and technical reports, government messages, newspaper articles published at the time, autobiographical reports and the engineer s verbal communications with relatives. The work is structured in three chapters: "Biographical traces, academic education and 'technical and political' activities" illustrates the initiation into the technical, public and political environment; Chapter 2, "Technique and territory" outlines his network understanding through sanitation and transport services; Chapter 3 "Technique and City" describes the influence of polytechnics knowledge on the propositions of modernization of cities; Finally, "Final Thoughts: An Evaluation," presents an overview of the affiliations and practice of an engineer in the different scales, and its contribution to the modernization of Brazilian urban and territorial space

Avaliação da ação bioinseticida de SBTI e vicilina de Erythrina velutina em enzimas digestivas e membrana peritrófica de larvas de Plodia interpunctella (Lepidoptera: Pyralidae)

Plodia interpunctella (Indian meal moth) is a cosmopolitan pest that attacks not only a wide range of stored grain as well other food products. Due to its economic importance several researches have focused in a method with ability to control this pest with few or no damage to the environment. The study of digestive enzymes inhibitors, lectins and chitin-binding proteins, has often been proposed as an alternative to reduce insect damage. In this study we report the major classes of digestive enzymes during larval growth in P. Interpunctella, being those proteinases actives at pH 9.5 and optimum temperature of 50 oC to both larvae of the 3rd instar and pre-pupal stage of development. In vitro and zymogram assays presented the effects of several inhibitors, such as SBTI, TLCK and PMSF to intestinal homogenate of 3rd instar larvae of 62%, 92% and 87% of inhibition and In pre-pupal stage of 87%, 62 % and 55% of inhibition, respectively. Zymograms showed inhibition of two low molecular masses protein bands by TLCK and that in presence of SBTI were retarded. These results are indicative of predominance of digestive serine proteinases in gut homogenate from Plodia interpunctella larvae. This serine proteinase was then used as a target to evaluate the effect of SBTI on larvae in in vivo assay. Effect of SBTI on mortality and larval mass was not observed at until 4% of concentration (w/w) in diets. Chitin, another target to insecticidal proteins, was observed by chemical method. Moreover, optic microscopy confirmed the presence of a peritrophic membrane. Established this target, in vivo effect of EvV, a chitin binding vicilin, evaluated during the larval development of P. interpunctella and was obtained a LD50 of 0,23% and WD50 of 0,27% to this protein. Mechanism of action was proposed through of the in vivo digestibility of EvV methodology. During the passage through the larval digestive tract was observed that EvV was susceptible to digestive enzymes and a reactive fragment, visualized by Western blotting, produced by digestion was recovered after dissociation of the peritrophic membrane. The bound of EvV to peritrophic membrane was confirmed by immunohystochemical assays that showed strong immunofluorescent signal of EvV-FITC binding and peritrophic membrane. These results are a indicative that vicilins could be utilized as potential insecticide to Plodia interpunctella and a control methods using EvV as bioinsecticide should be studied to reduce lost caused by storage insect pests