Avaliação funcional e estrutural de um novo peptídeo antimicrobiano do escorpião Tityus stigmurus

In Brazil, there is a high incidence of venomous animals. Among them, scorpions are highlighted by their medical importance, and for being their venom a source of several molecules with biological and pharmacological activity not yet fully understood, including several bioactive peptides. Antimicrobial peptides (AMPs) are components of the immune system in prokaryotes and eukaryotes, used in the first line of defense against microorganisms. In the present study, we characterized the first PAM previously identified through transcriptome of the venom gland of the scorpion Tityus stigmurus, named Stigmurin. The characteristics of Stigmurin were investigated by computational modeling and construction of dendrogram. In vitro tests investigated the antibacterial, antifungal, haemolytic and cytotoxic effects of crude venom and Stigmurin. In addition, the structural characteristics of Stigmurin were investigated by circular dochroism in water, 2, 2 , 2- trifluoethanol (TFE) and sodium dodecyl sulfate (SDS) and the models were refined by molecular dynamics simulations. The results showed that the selected sequence encodes a mature protein of 17 amino acid residues and the dendrogram reveals a case of convergent evolution. The crude venom showed no antimicrobial activity, however, Stigmurin exhibited a broad spectrum of antibacterial activity, with minimal inhibitory concentrations (MIC) ranging from 31.25 and 250 µg/mL for different strains, while the hemolytic activity at these concentrations was low. In cytotoxicity studies, the crude venom was unable to reduce cell viability in VERO E6 cells; in contrast, its activity in SiHa cells was significantly higher, corresponding to IC50 of 3.6 µg/mL. For Stigmurin the concentration sable to decrease cell viability of Vero E6 and SiHa cells in 50% were 275.67 µg/mL and 212.54 µg/mL, respectively. The dichroism spectra revealed the conformational flexibility, with predominating extended and β–sheet structures, as well as a remark able renaturation ability. The results suggest that Stigmurin could be considered as a potential antiinfective drug

Caracterização funcional de dois cDNAs de cana-de- açúcar : PKCI e SHAGGY

Flowering is controlled by several environmental and endogenous factors, usually associated with a complex network of metabolic mechanisms. The gene characterization in Arabidopsis model has provided much information about the genetic and molecular mechanisms that control flowering process. Some of these genes had been found in rice and maize. However, in sugarcane this processe is not well known. It is known that early flowering may reduce its production up to 60% at northeast conditions. Considering the impact of early flowering in sugarcane production, the aim of this work was to make the gene characterization of two cDNAs previously identified in subtractive cDNA libraries: scPKCI and scSHAGGY. The in silico analysis showed that these two cDNAs presented both their sequence and functional catalytic domains conserved. The results of transgenic plants containing the overexpression of the gene cassette scPKCI in sense orientation showed that this construction had a negative influence on the plant development as it was observed a decrease in plant height and leaf size. For the scPKCI overexpression in antisense orientation it was observed change in the number of branches from T1 transgenic plants, whereas transgenic T2 plants showed slow development during germination and initial stages of development. The other cDNA analyzed had homology to SHAGGY protein. The overexpression construct in sense orientation did not shown any effect on development. The only difference observed it was an increase in stigma structure. These results allowed us to propose a model how these two genes may be interact and affect floweringdevelopment.

Metagenômica: busca de novos genes envolvidos com a biodegradação de hidrocarbonetos

Industrial activities, oil spills and its derivatives, as well as the incomplete combustion of fossil fuels have caused a great accumulation of hydrocarbons in the environment. The number of microorganisms on the planet is estimated at 1030 and prokaryotes the most abundant. They colonized diverse environments for thousands of years, including those considered extreme and represent an untapped source of metabolic and genetic diversity with a large biotechnological potential. It is also known that certain microorganisms have the enzymatic capacity to degrade petroleum hydrocarbons and, in many ecosystems, there is an indigenous community capable of performing this function. The metagenomic has revolutionized the microbiology allowing access uncultured microbial communities, being a powerful tool for elucidation of their ecological functions and metabolic profiles, as well as for identification of new biomolecules. Thus, this study applied metagenomic approaches not only for functional selection of genes involved in biodegradation and emulsification processes of the petroleum-derived hydrocarbons, but also to describe the taxonomic and metabolic composition of two metagenomes from aquatic microbiome. We analyzed 123.116 (365 ± 118 bp) and 127.563 sequences (352 ± 120 bp) of marine and estuarine metagenomes, respectively. Eight clones were found, four involved in the petroleum biodegradation and four were able to emulsify kerosene indicating their abilities in biosurfactants synthesis. Therefore, the metagenomic analyses performed were efficient not only in the search of bioproducts of biotechnological interest and in the analysis of the functional and taxonomic profile of the metagenomes studied as well

Purificação e caracterização estrutural de homogalactanas sulfatadas anticoagulantes da alga verde Codium isthmocladum

Conselho Nacional de Desenvolvimento Científico e Tecnológico

A correlação locução adjetiva/adjetivo: uma análise funcional

This research deals with the relationship locução adjetiva / adjective, focusing the reasons for using one form or the other one, in situations where the speaker has a choice between an adjective or a locução adjetiva virtually correspondent and vice versa. In general, the objective is to investigate the possibility of correlation between locução adjetiva and adjective, and to identify motivations for the use of one form or the other one. The theoretical support of this paper is the Linguística Funcional Centrada no Uso (LFCU), which groups together theoretical and methodological assumptions of North American Functional Linguistics, represented by authors as Givón (1984), Bybee (2010), Traugott (2011), and Cognitive Linguistics, as developed by Lakoff (1987), Langacker (1987) and Lakoff and Johnson (2002). As regards the methodological procedures, this research is of qualitative and quantitative character: at first, we did the survey data in the texts that made up the corpus, followed by analysis of each them, especially the locução adjetiva, by considering the semantic, morphosyntactic, cognitive and pragmatic aspects. The corpus is composed of 36 texts, taken from 12 issues of the Veja magazine, published between January 2013 and March 2013. As for the results, in the case of locução adjetiva with related adjectives, this research has shown that the choice of former is due to semantic change, the use of lexicalized forms, to priming cases or to stylistic motivations. It means that the choices of one of these two forms go beyond lexical issues, since there are other motivations, such as syntactic, semantic and pragmatic ones.

Ajuste estrutural e educação superior no Brasil: princípios negados

SILVA, Hiran Francisco Oliveira Lopes da. A juste estruturale educação superior no Brasil: princípios negados. 206 f., 2007. Tese (Doutorado em Educação) - Universidade Federal da Paraiba, João Pessoa, 2007.

Atuação do estado como empregador de última instância: uma proposta para eliminar o desemprego estrutural do Brasil

GOMES, Z. B. ; LOURENÇO, André Luís Cabral de . Atuação do estado como empregador de última instância: uma proposta para eliminar o desemprego estrutural do Brasil. In: Encontro Nacional de Economia Política,13., 2008, João Pessoa/PB. Anais... João Pessoa, 2008.

Desempenho internacional de empresas exportadoras: um modelo estrutural baseado em capacidades organizacionais e ambiente

The internationalization as an organizational phenomenon fundamentally strategic had as theoretical contributions some Schools that throughout the decades 60, 70, and 80 developed behavioral and economic approaches in order to explain the process. The behavioral approach deals with the perception of phenomenon as a gradual process from the perspective of the executives behavior (JOHANSON and VAHLNE, 1977; HALLÉN and WIEDERSHEIM - PAUL, 1979; CZINKOTA, 1985). This phenomenon in permanent theoretical and managerial evolution made an opportunity to build this investigation, whose goal is to analyse the impact comes from organizational capabilities and the external environment on the international performance of exporting firms. For both, were used as theoretical basis two types of analysis for the comprehension of international performance: Strategic Management - Industrial Organization and Resource-Based View and International Businesses - Current Economic and Behavioral. It was made a cross-sectional survey-based explanatory research, including 150 exporting companies with operations in the Northeast of Brazil. A conceptual model was made with eight constructs and eight research hypotheses, representative of the effects of external factors on international performance. The data were processed using the Exploratory Factor Analysis and Structural Equation Modeling. The structural equations model was reespecified and estimated through the use of the maximum-likelihood method up to achieve adequated values of indexes of adjustment. As the main theoretical contribution, were identified organizational and physical resources which shows the importance of the management skills development, of the learning capability and capability to establish strategic alliances abroad. That because the knowledge, as the operational point of view as in its strategic application, offers to organization conditions of market positioning which can create opportunities sustainable competitive advantages and which impact the performance of international companies

Atuação e desempenho do Ministério Público do Estado do Rio Grande do Norte no contexto da gestão pública: perspectiva estratégica

The present dissertation analyzes the performance and acting of the Rio Grande do Norte Public Ministery, in the strategic perspective, while responsible Institution for the defense of the collective rights. The comprehension of this theme inserts in a context of modernization of the public administration, in which is inserted, as well as their functional and structural changes, in a reality of innovations there is passing the organizations, looking to rescue the legitimacy of the government organizations, aligning them to the democratic values of the society. It detaches the strategic administration and the public administration and it exposes the strategic performance about the central point of the study, focusing in the development of the organization in the last four years, and other relative subjects to the acting. It is a unic case study, framed in the characteristics of the qualitative approaches - descriptive and exploratory. The analysis showed, through the method of content analysis, by the criterion of thematic categorization, that MP / RN come developing in an intense way,and it detaches that the changes already happened reflect a good strategic acting of the Institution, especially in the structural and functional areas, showing the strategic conscience of this, although that is not still enough to consider it as a strategic organization, and it concludes that there is still a lot for doing, and that the occurrence of an administration typically strategic in the extent of MP/RN is possible, with the conscience and participation of all, members and servants

A concepção estrutural como expressão plástica: um olhar tectônico sobre arquitetura em madeira

This work regards to the structural conception as a formal element of design in wood architecture. On this approach, the group of projects studied is formed by some realized works of two important Brazilian architects: Severiano Porto and Marcos Acayaba. The time interval comprises the period from 1971 to 1997, which correspond respectively to the years of the creation of first and the last of the analyzed constructions. The research perspective concerns to the relationship between the parts, the whole and the building techniques adopted in each project. Moreover, the analysis focuses on the structure as the link among the different projectual ideas. Thus, the research method firstly includes a survey of specific bibliographies and documents which refer to the structural conception in wood architecture. Secondly,the buildings are analyzed according to the methods of architectural composition discussed through this work.

A contribuição de técnicas modernas de reabilitação estrutural para autenticidade e integridade de edificações patrimoniais

This work discusses the study of the application of structural reinforcement and rehabilitation using modern techniques for interventions in repair works of historical buildings as well as the verification of the behavior of works that have already undergone different structural intervention techniques, with the objective of obtaining data to allow a comparative analysis of the advantages/disadvantages of techniques adapted in each case, regarding the interferences related the authenticity and integrity of the buildings belong to the constructed patrimony so as to obtain a basis for the theoretical foundation for choosing the best solution to be adopted in the case study which consists in an intervention of structural rehabilitation of the old historic building in the primary school "Augusto Severo"

Potencial terapêutico de um heparinóide isolado do invertebrado marinho Litopenaeus vannamei

The occurrence of bioactive compounds in marine organisms comes awaking the interest of the pharmaceutical industry. Heparin, a sulfated polysaccharide which presence was already identified in several marine invertebrates, is very attractive due its remarkable functional versatility. Besides to intervene in blood coagulation, this molecule has a great anti-inflammatory potential. However, its strong anticoagulant activity difficult the clinical exploitation of its anti-inflammatory properties. Thus, the aims of this work were to evaluate the effect of a heparin-like compound (heparinoid), isolated from the cephalotorax of the Litopenaeus vannamei shrimp, on the inflammatory response, hemostasia and synthesis of antithrombotic heparan sulfate by endothelial cells, besides studying some aspects concerning its structure. The purified heparinoid was structurally characterized following an analytical boarding, involving electrophoresis and chromatography. The structural analysis have shown that this compound possess a high content of glucuronic acid residues and disulfated disaccharide units. In contrast to mammalian heparin, the heparinoid was incapable to stimulate the synthesis of heparan sulfate by endothelial cells in the tested concentrations, beyond to show reduced anticoagulant activity and hemorrhagic effect. In a model of acute inflammation, the compound isolated from the shrimp reduced more than 50% of the cellular infiltration. Besides reduce the activity of MMP-9 and proMMP-2 of the peritoneal lavage of inflamed animals, the heparinoid also reduced the activity of MMP-9 secreted by activated human leukocytes. These results demonstrate the potential of heparinoid from L. vannamei to intervene in the inflammatory response. For possessing reduced anticoagulant activity and hemorrhagic effect, this compound can serve as a structural model to direct the development of more specific therapeutical agents to the treatment of inflammatory diseases

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.

Análise filogenética e funcional de dois genes de reparo homólogos a AP endonuclease em cana-de-açúcar: ScARP1 e ScARP3

The genome of all organisms constantly suffers the influence of mutagenic factors from endogenous and/or exogenous origin, which may result in damage for the genome. In order to keep the genome integrity there are different DNA repair pathway to detect and correct these lesions. In relation to the plants as being sessile organisms, they are exposed to this damage frequently. The Base Excision DNA Repair (BER) is responsible to detect and repair oxidative lesions. Previous work in sugarcane identified two sequences that were homologous to Arabidopsis thaliana: ScARP1 ScARP3. These two sequences were homologous to AP endonuclease from BER pathway. Then, the aim of this work was to characterize these two sequence using different approaches: phylogenetic analysis, in silico protein organelle localization and by Nicotiana tabacum transgenic plants with overexpression cassette. The in silico data obtained showed a duplication of this sequence in sugarcane and Poaceae probably by a WGD event. Furthermore, in silico analysis showed a new localization in nuclei for ScARP1 protein. The data obtained with transgenic plants showed a change in development and morphology. Transgenic plants had slow development when compared to plants not transformed. Then, these results allowed us to understand better the potential role of this sequence in sugarcane and in plants in general. More work is important to be done in order to confirm the protein localization and protein characterization for ScARP1 and ScARP3

Caracterização estrutural e avaliação das atividades farmacológicas da fucana B extraída da alga Dictyota menstrualis

Seaweeds are a major source of biologically active compounds . In the extracellular matrix of these organisms are sulfated polysaccharides that functions as structural components preventing it against dehydration. The fraction 0.9 (FucB) rich in sulfated fucans obtained from brown seaweed Dictyota menstrualis was chemical characterized and evaluated for pharmacological activity by testing anticoagulant activity, stimulatory action on the synthesis of an antithrombotic heparan sulfate, antioxidant activity and its effects in cell proliferation. The main components were FucB carbohydrates (49.80 ± 0.10 %) and sulfate (42.30 ± 0.015 %), with phenolic compounds ( 3.86 ± 0.016 %) and low protein contamination ( 0.58 ± 0.001 % ) . FucB showed polydisperse profile and analysis of signals in the infrared at 1262, 1074 and 930 cm -1 and 840 assigned to S = O bonds sulfate esters , CO bond presence of 3,6- anhydrogalactose , β -D- galactose non- sulfated sulfate and the axial position of fucose C4 , respectively. FucB exhibited moderate anticoagulant activity , the polysaccharides prolonged time (aPTT ) 200 ug ( > 90s ) partial thromboplastin FucB no effect on prothrombin time (PT), which corresponds to the extrinsic pathway of coagulation was observed. This stimulation promoted fraction of about 3.6 times the synthesis of heparan sulfate (HS) by endothelial cells of the rabbit aorta ( RAEC ) in culture compared with cells not treated with FucB . This has also been shown to compete for the binding site with heparin. The rich fraction sulfated fucans exhibited strong antioxidant activity assays on total antioxidant (109.7 and 89.5 % compared with BHT and ascorbic acid standards ) , reducing power ( 71 % compared to ascorbic acid ) and ferric chelation ( 71 , comparing with 5 % ascorbic acid). The fraction of algae showed cytostatic activity on the RAEC cells revealed that the increase of the synthesis of heparan sulfate is not related to proliferation. FucB showed antiproliferative action on cell lines modified as Hela and Hep G2 by MTT assay . These results suggest that FucB Dictyota menstrualis have anticoagulant , antithrombotic , antioxidant potential as well as a possible antitumor action, promoting the stimulation of the synthesis of antithrombotic HS by endothelial cells and is useful in the prevention of thrombosis, also due to its inhibitory action on species reactive oxygen ( ROS ) in some in vitro systems , being involved in promoting a hypercoagulable state