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Produção recombinante, caracterização enzimática e estudos sobre a ocorrência de pectinases no "bicudo da cana-de-açúcar" (Sphenophorus levis, Curculionidae)
(Universidade Federal de São Carlos, 2012-04-04) Evangelista, Danilo Elton; Silva, Flávio Henrique da; https://lattes.cnpq.br/1757309852446263; https://lattes.cnpq.br/6691516072529137
Plant cell wall confers to the cell plant, structural support as well as protection against pathogens and phytophagous. Among the cell wall polysaccharides includes pectic substances, which are composed of partially methyl-esterified galacturonic acid residues linked by α-1,4 glycosidic bonds. These enzymes are often synthesized by phytophatogenic micro-organisms for invasion of host plant or by own plant for modeling plant cell wall. The pectic substances are the major component of middle lamella and are natural degraded by pectinases action. Pectin methylesterase (PME) catalysis removes methyl-ester groups, and the Endo-polygalacturonase (Endo-PG) promoves the randomly hydrolysis reaction of α-1,4 bonds. One of the most importante agricultural pest species of the family Curculionidae (Coleoptera: Curculionidae) is Sphenophorus levis, the sugarcane weevil. The larvae of this insect penetrate into the rhizome and build galleries in the stem, decreasing productivity and causing the death of the plant. Large damages to the crop like that are significant in the costs of products derived from sugarcane. Considering the impact of this pest in the sugarcane crop and the absence of efficient method for control, new strategies for controlling are still necessary. The analysis of the cDNA library of S. levis larvaes shows the presence of one PME and one Endo-PG genes that we called Sl-PME and Sl-EndoPG respectively. Considering the importance of studies of insect pests and the extensively use of theses pectinases in different industry fields, we performed the characterization of genomic sequences coding for S. levis pectinases (Sl-Pectinases). It was also carried out the production and characterization of a Sl-PME and a Sl-EndoPG recombinant, expressed in heterologous system. We also accomplished analysis of gene expression by qRTPCR in different stages of development as well as different tissues, and phylogenetic studies between Sl-Pectinases and other pectinases from different kingdoms. The Sl- Pectinases sequences identified, show more similar to homologous insect genes deposited in the GenBank, especially with Sitophilus oryzae. The phylogenetic analysis indicates that the insect group is more correlated with bacteria group and fungi group respectively to PMEs and EndoPGs sequences. Pectinases genomic sequences revealed two introns for Sl-EndoPG gene with 53 and 166 bp, but no one for Sl-PME gene. Both of Sl-Pectinases Recombinant showed catalytic activity. The recombinant Sl-EndoPG shows optimal activity at pH 5,06 ± 0,27 and 49,74 ± 2,49 oC, but extremely low thermostability. For the polygalacturonic acid no-methylated as substract, the enzyme revealed Km = 3,88 mg.mL-1, Vmax = 21.96 μM.s-1 e Kcat = 3.137 s-1; for the citrus pectin partially methylated as substract, the enzyme presented Km = 4,98 mg.mL-1, Vmax = 17,19 μM.s-1 e Kcat = 2.456 s-1. Results in expression analysis suggest that S. levis pectinases have a digestive enzymes role, actting on the midgut. The present work represents the first pectinases of insect produced in Pichia pastoris heterologous system and characterized as optimal conditions of activity, thermostability and kinetic parameters.
Análise discursiva de documentos oficiais sobre o sujeito surdo
(Universidade Federal de São Carlos, 2012-04-04) Carvalho, Lílian Pereira de; Baronas, Roberto Leiser; https://lattes.cnpq.br/4613001301744682; https://lattes.cnpq.br/7492175179598682
According to the census that was made in Brazil in 2000, around 5.800.000 people have some degree of hearing loss or deafness. Among them, the numbers of deaf people who don t hear anything, classified as incapable of hearing and that use the sign language to communicate was 176.067 people. Our research aims to analyze the images discursively that the official documents compose about this important part of society that can not hear. To do this, we will consider the lexical choices made, through words or statements, when those document refer to the deaf. It was mobilized the notion of Imaginary Formation, which will be used to analyze the build images from the self image and the deaf people, subject of the discourse. We still propose to analyze these documents as genre of discourse, as it was such proposed by Mikail Bakhtin (1997), because we believe that if we ignore the peculiarities of the genre that we will be led to formalism and abstraction, to disregard the historicity of the study and the relation between language and life. From this notion, we will also discuss the scenographies permeating these documents, in order to understand the tone of the discourse. To compose the corpus, some laws or decrees were chosen dealing with several issues about deaf cause such as: the sign language, their accessibility to media, theaters, public institutions, education and health, among others. These documents produced by government departments have a normative nature that aims to regulate specific actions for these subjects, especially about the guarantee of their rights. From this perspective, we will take as theoretical and methodological focus the Discourse Analysis of French orientation and the Dialogical Analysis of Discourse, through Michel Pêcheux, Dominique Maingueneau and Mikail Bakhtin studies.
O Programa "Minha Casa, Minha Vida" na política habitacional brasileira: continuidades, inovações e retrocessos
(Universidade Federal de São Carlos, 2012-04-04) Romagnoli, Alexandre José; Vargas, Marcelo Coutinho; https://lattes.cnpq.br/3816397515831545
This research aims to analyze the National Housing Policy of the Lula government throught the program Minha Casa, Minha Vida (PMCMV). The analizes concentrates on the continuities, innovations or any setbacks presents in PMCMV in comparison to Housing Policy historically built in Brazil. We assume that the housing issue is taken up in the country with the beginning of the Lula government, which shall enable a process of change in the area, such as the creation of a new Ministry (Ministry of Cities), changes in regulations sector, and programs with larger goals to those already found in the country. Implemented in this context that the PMCMV, although placed initially as a tool "anti-cyclical" government facing the international economic crisis, also stands outfor its social proposal. With the goal of building 1 million houses, being part of those destined for low-income population (historically one of the main bottlenecks Housing Policy) and considerable investment of resources of their own Federal Budget, the program becomes relevant and representative for the policy housing as a whole. From the neoinstitutionalist literature and public policy, this dissertation shows, in general, the PMCMV innovates with respect to funding issues and exploitation of the resources. The main result is the break with some logical housing production in the country, as the financing of housing policy from resources requiring immediate return, seeing a new trend in this direction. However, timidly intervenes in matters already well discussed in the country, such as urban planning and land tenure.