Residues located inside the Escherichia coli FepE protein oligomer are essential for lipopolysaccharide O-antigen modal chain length regulation

The Escherichia coli O157 : H7 FepE protein regulates lipopolysaccharide (LPS) O-antigen (Oag) chain length to confer a very long modal chain length of >80 Oag repeat units (RUs). The mechanism by which FepE regulates Oag modal chain length and the regions within it that are important for its function remain unclear. Studies on the structure of FepE show that the protein oligomerizes. However, the exact size of the oligomer is in dispute, further hampering our understanding of its mechanism. Guided by information previously obtained for regions known to be important for Oag modal chain length determination in the homologous Shigella flexneri WzzBSF protein, a set of FepE mutant constructs with single amino acid substitutions was created. Analysis of the resulting LPS conferred by these mutant His6-FepE proteins showed that amino acid substitutions of leucine 168 (L168) and aspartic acid 268 (D268) resulted in LPS with consistently shortened Oag chain lengths of <80 Oag RUs. Substitution of FepE’s transmembrane cysteine residues did not affect function. Chemical cross-linking experiments on mutant FepE proteins showed no consistent correlation between oligomer size and functional activity, and MS analysis of FepE oligomers indicated that the in vivo size of FepE is consistent with a maximum size of a hexamer. Our findings suggest that different FepE residues, mainly located within the internal cavity of the oligomer, contribute to Oag modal chain length determination but not the oligomeric state of the protein.Elizabeth Ngoc Hoa Tran and Renato Moron

Identification of Shigella flexneri IcsA residues affecting interaction with N-WASP, and evidence for IcsA-IcsA co-operative interaction

The Shigella flexneri IcsA (VirG) protein is a polarly distributed outer membrane protein that is a fundamental virulence factor which interacts with neural Wiskott-Aldrich syndrome protein (N-WASP). The activated N-WASP then activates the Arp2/3 complex which initiates de novo actin nucleation and polymerisation to form F-actin comet tails and allows bacterial cell-to-cell spreading. In a previous study, IcsA was found to have three N-WASP interacting regions (IRs): IR I (aa 185–312), IR II (aa 330–382) and IR III (aa 508–730). The aim of this study was to more clearly define N-WASP interacting regions II and III by site-directed mutagenesis of specific amino acids. Mutant IcsA proteins were expressed in both smooth lipopolysaccharide (S-LPS) and rough LPS (R-LPS) S. flexneri strains and characterised for IcsA production level, N-WASP recruitment and F-actin comet tail formation. We have successfully identified new amino acids involved in N-WASP recruitment within different N-WASP interacting regions, and report for the first time using co-expression of mutant IcsA proteins, that N-WASP activation involves interactions with different regions on different IcsA molecules as shown by Arp3 recruitment. In addition, our findings suggest that autochaperone (AC) mutant protein production was not rescued by another AC region provided in trans, differing to that reported for two other autotransporters, PrtS and BrkA autotransporters

The role of bacterial protein tyrosine phosphatases in the regulation of the biosynthesis of secreted polysaccharides

SIGNIFICANCE: Tyrosine phosphorylation and associated protein tyrosine phosphatases are gaining prominence as critical mechanisms in the regulation of fundamental processes in a wide variety of bacteria. In particular, these phosphatases have been associated with the control of the biosynthesis of capsular polysaccharides and extracellular polysaccharides, critically important virulence factors for bacteria.RECENT ADVANCES: Deletion and over-expression of the phosphatases result in altered polysaccharide biosynthesis in a range of bacteria. The recent structures of associated auto-phosphorylating tyrosine kinases has suggested that the phosphatases may be critical for the cycling of the kinases between monomers and higher order oligomers. CRITICAL ISSUES: Additional substrates of the phosphatases apart from cognate kinases are currently being identified. These are likely to be critical to our understanding of the mechanism by which polysaccharide biosynthesis is regulated. FUTURE DIRECTIONS: Ultimately, these protein tyrosine phosphatases are an attractive target for the development of novel anti-microbials. This is particularly the case for the polymerase and histidinol phosphatase family, which are predominantly found in bacteria. Furthermore, the determination of bacterial tyrosine phosphoproteomes will likely help to uncover the fundamental roles, mechanism and critical importance of these phosphatases in a wide range of bacteria.Alistair James Standish and Renato Moron

IcsA autotransporter passenger promotes increased fusion protein expression on the cell surface

Background: Autotransporters are attractive cell surface display vehicles as they lack complex adaptor proteins necessary for protein export. Recent reports have suggested that the native effector domain (α domain) and translocation domain (β domain) interact with each other to drive translocation of the effector domain to the outer membrane. In this report we compared the expression, surface localisation and folding of TEM-1 β-lactamase (Bla) and maltose binding protein (MalE or MBP) fused to either full length Shigella flexneri IcsA (IcsA) autotransporter or to the β domain alone (IcsAβ) to determine the contribution of the native IcsA α domain in presenting the fusion proteins on the surface of E. coli K-12 UT5600 (ΔompT). Results: Expression of IcsA-Bla was greater than IcsAβ-Bla. High levels of IcsA-MalE were detected but IcsAβ-MalE was not expressed. All fusion proteins other than IcsAβ-MalE were localised to the outer membrane and were detected on the surface of UT5600 via immunofluorescence microscopy. All bacteria expressing IcsA-MalE were labelled with both α-IcsA and α-MBP. UT5600 expressing IcsAβ-MalE was not labelled with α-MBP. A third of UT5600 expressing IcsA-Bla were detectable with α-Bla but only 5% of UT5600 (IcsAβ-Bla) were labelled with α-Bla. The correct folding of the Bla moiety when fused to IcsA and IcsAβ was also retained as UT5600 expressing either fusion protein exhibited a decreased zone of inhibition in the presence of ampicillin. UT5600 expressing IcsA-Bla was more resistant compared to UT5600 expressing IcsAβ-Bla. Conclusions: The export mechanism of autotransporters is not well understood but accumulating evidence suggest a critical role for the native effector or α domain in facilitating its own export via interactions with the translocation or β domain. This is the first report directly comparing expression of heterologous proteins fused to the full length IcsA autotransporter and fusion to the β domain alone. Protein expression and surface presentation of the fusion proteins were dramatically improved when fused to IcsA rather than IcsAβ. Future studies involved in designing autotransporters as cell surface display vehicles would benefit from including the native α domain. This work also provides further evidence for a key interaction between the autotransporter α and β domains.Mabel Lum and Renato Moron

Prevalence of Schistosoma mansoni and soil transmitted helminths and factors associated with uptake of preventive chemotherapy among school children in Sengerema District in north-western Tanzania

Background: The control of intestinal schistosomiasis and soil transmitted helminths (STH) in Tanzania focuses on reducing morbidities through the treatment of infected and at risk populations, especially schoolchildren with praziquantel (PZQ) and albendazole (ALB). However, in some areas, the uptake of interventions is low. The objective of this study was to determine factors associated with the uptake of preventive chemotherapy and, secondarily, the prevalence of Schistosoma mansoni and soil-transmitted helminths' infections in Sengerema District of north western Tanzania.Methods: This cross-sectional study was conducted among 625 randomly selected school children aged 8-18 years from Sengerema district. A questionnaire was used to collect information on the reported uptake of PZQ/ALB. Single stool samples were collected, processed and examined for the presence of eggs pf S. mansoni and soil-transmitted helminths using Kato Katz technique.Results: The self-reported uptake of preventive chemotherapy was 95.6% (95%CI; 92.78-98.49). Provision of food (AOR= 25.25, 95%CI: 5.28-120.49, p&lt;0.001) and information about the anti-helminthic drug prior to taking it (AOR =14.24, 95%CI: 3.23-62.72, p&lt;0.001) were associated with a high reported uptake of preventive chemotherapy. The overall prevalence of S. mansoni and geometrical mean of eggs per gram (EPG) of faeces were 36.64% (95%CI: 21.55 -62.29) and 229.47 EPG (202.73-259.86). The prevalence of STH was 10.88% (95%CI; 7.52-15.75).Conclusion: The high reported uptake of preventive chemotherapy was associated with provision of food and information about the drugs prior to their administration. However, S. mansoni and soil-transmitted infections are still a public health concern in the study area. Integrating health education in mass drug administration campaigns will allow provision of other complementary public preventive measures to reduce the burden of these infections

O papel da economia solidária no Brasil : frente à crise econômica atual

Orientador: Prof. Dr. Wilson LoureiroMonografia (especialização) - Universidade Federal do Paraná, Setor de Ciências Agrárias, Curso de Especialização em Economia e Meio AmbienteInclui referênciasEste trabalho visa analisar a importância da econômica solidária na geração de emprego no Brasil frente a crise econômica, a partir dos conceitos fundamentais de economia solidária, seus projetos e programas governamentais e movimentações sociais e fatores de influencia do desemprego nacional, assim como de geração de renda, abrangendo as atividades rurais, cooperativas e associações, utilizando referência bibliográfica de reportagens, estudos sobre a economia nacional e da economia solidária. Cabendo como ressalva a importância em manter a abordagem apartidária e tendo como resultado a importância econômica e a geração de emprego e renda para grupos familiares, cooperativas e sociedades organizadas, neste período de crise

Prepatterning and patterning of the thalamus along embryonic development of Xenopus laevis

Previous developmental studies of the thalamus (alar part of the diencephalic prosomere p2) have defined the molecular basis for the acquisition of the thalamic competence (preparttening), the subsequent formation of the secondary organizer in the zona limitans intrathalamica, and the early specification of two anteroposterior domains (rostral and caudal progenitor domains) in response to inducing activities and that are shared in birds and mammals. In the present study we have analyzed the embryonic development of the thalamus in the anuran Xenopus laevis to determine conserved or specific features in the amphibian diencephalon. From early embryonic stages to the beginning of the larval period, the expression patterns of 22 markers were analyzed by means of combined In situ hybridization (ISH) and immunohistochemical techniques. The early genoarchitecture observed in the diencephalon allowed us to discern the boundaries of the thalamus with the prethalamus, pretectum, and epithalamus. Common molecular features were observed in the thalamic prepatterning among vertebrates in which Wnt3a, Fez, Pax6 and Xiro1 expression were of particular importance in Xenopus. The formation of the zona limitans intrathalamica was observed, as in other vertebrates, by the progressive expression of Shh. The largely conserved expressions of Nkx2.2 in the rostral thalamic domain vs. Gbx2 and Ngn2 (among others) in the caudal domain strongly suggest the role of Shh as morphogen in the amphibian thalamus. All these data showed that the molecular characteristics observed during preparttening and patterning in the thalamus of the anuran Xenopus (anamniote) share many features with those described during thalamic development in amniotes (common patterns in tetrapods) but also with zebrafish, strengthening the idea of a basic organization of this diencephalic region across vertebrates

Estudo da viabilidade do uso de blendas de poliestireno reciclado e polipropileno

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro Tecnológico. Programa de Pós-graduação em Ciência e Engenharia de MateriaisO desenvolvimento e o crescimento tecnológico, sem dúvida, trouxeram grandes benefícios à sociedade. Mas, aliado a este crescimento encontra-se um comportamento de consumo desenfreado, que provoca vários efeitos adversos. Entre eles, destaca-se a poluição do meio ambiente. A interferência no meio ambiente gera resíduo e dentre estes se destaca o sólido, uma vez que seu grau de dispersão é bem menor que os gasosos e líquidos. Os resíduos sólidos plásticos demoram centenas de anos para se decompor. O consumo mundial projetado para 2004 somente de termoplásticos é da ordem de 183 milhões de toneladas. Para o Brasil o dado da transformação de poliestireno, material plástico inserido neste trabalho e usado na produção de descartáveis, é bastante significativo, apresentando consumo aparente de aproximadamente 320 000 ton/ano. Baseado nestes argumentos, este trabalho tem como objetivo principal verificar as propriedades mecânicas da blenda de poliestireno reciclado e polipropileno, utilizando como matérias-primas copos plásticos descartáveis pós-consumo e polipropileno virgem. O poliestireno de alto impacto pós-consumo utilizados no trabalho são copos descartáveis coletados nas cantinas e bebedouros da escola SATC/SENAI, através de um coletor seletivo. Para a caracterização foram misturados manualmente diferentes teores de poliestireno em polipropileno virgem e corpos de prova, normalizados, foram preparados por injeção sendo analisados quanto ao comportamento de suas propriedades mecânicas. As propriedades mecânicas analisadas foram: a resistência à tração, flexão, dureza, resistência ao impacto e densidade relativa, seguindo os métodos das normas ASTM. Embora a blenda de HIPS e PP seja uma mistura considerada imiscível, os resultados obtidos foram satisfatórios para algumas propriedades mecânicas e ótimas para outras, como é o caso da tensão na ruptura, que é semelhante às tensões do PP virgem. Já para o alongamento e resistência ao impacto o que se observou foi uma queda significativa em função do acréscimo de HIPS na blenda. O que se pode concluir é que as formulações são perfeitamente possíveis para determinados produtos, onde o esforço mecânico não é tão exigido. The development and the growth, without a doubt, had brought great benefits to the society. But, ally to this growth come across a behavior of wild consumption that provokes some adverse effect. Among them, it is distinguished pollution of the environment. The interference in the environment generates residue and amongst these if it detaches the solid residue, a time that its degree of dispersion is well lesser that gaseous and the liquid ones. In the solid residues we have plastics that they delay hundreds of years to putrefy itself. The world consumption projected for 2004 of thermoplastic is of the order of 183 million tons. For Brazil the data of the polystyrene transformation, inserted plastic material in this used work and in the production of discarded, are sufficiently significant, presenting apparent consumption of approximately 320 000 tons/years. Based in these arguments, this work has as objective main to verify the mechanical properties of the blend of recycled polystyrene and polypropylene, being used as raw materials discarded plastic cups after-I consume and virgin polypropylene. The polystyrene of high impact after-I consume used in the work are discarded cups collected in the canteens and #water throughs# of SATC/SENAI, through a selective collector. For the characterization different polystyrene texts had been mixed manually in virgin polypropylene and test bodies, normalized, had been prepared by injection being analyzed how much to the behavior of its mechanical properties. The analyzed mechanical properties had been: the tensile strenght, flexible, hardness, and resistance to the impact and relative density, following the methods of norms ASTM. Although the blend of HIPS and PP is a imiscíble mixture, the gotten results had been satisfactory for some mechanical and excellent properties for others, as it is the case of the tension in the rupture, that is similar to the tensions of the virgin PP. Already for the elongation and resistance to the impact what it was observed it was a significant fall in function of the addition of HIPS in the blend. What can be concluded is that the formularizations perfectly possible for are determined products, where the mechanical effort so is not demanded

Perfil do estilo de vida e composição corporal em meninas do ensino médio e fundamental de uma escola de Criciúma - SC

Artigo apresentado para obtenção do grau de Bacharel, no Curso de Educação Física, da Universidade do Extremo Sul Catarinense, UNESC.Objetivo: Verificar o estilo de vida e a composição corporal de meninas do ensino médio e fundamental de uma escola particular da cidade de Criciúma, SC. Metodologia: Foram avaliadas 64 adolescentes, do sexo feminino com idade de 12 a 17 anos estudantes do ensino médio e fundamental. Os instrumentos utilizados para a coleta de dados foram: avaliação antropométrica: massa corporal (Kg), estatura (cm) e dobras cutâneas segundo padronização de Petroski. Para avaliar o Perfil de Estilo de Vida foi utilizado o questionário do “Pentáculo do Bem Estar” adaptado para adolescentes, o mesmo engloba 15 questões fechadas, sobre cinco aspectos do estilo de vida: nutrição, controle do stress, comportamento preventivo, relacionamento social e atividade física. Para análise estatística foi utilizado o pacote estatístico SPSS (20.1). Resultados e discussões: Os resultados das variáveis antropométricas demonstram que foram encontradas diferenças significativas em p<0,05 entre as séries para as variáveis: idade, peso, altura, e IMC. No entanto, não verificou diferença estatisticamente significativa na soma das dobras cutâneas e percentual de gordura.A média de IMC das adolescentes do EM foi de 21,39 (± 2,85) e das adolescentes do EF foi de 20,03 (± 2,52), a qual apresentou diferença estatisticamente significativa entre as médias (p=0,03). Ao avaliar o IMC das adolescentes observou-se uma alta frequência de IMC adequado (94% EM e 84% EF). Em relação à comparação do estilo de vida entre as adolescentes do EM e EF, as adolescentes encontram-se com um estilo de vida, de um modo geral positivos, deixando em alguns casos a desejar quanto aos componentes nutrição, atividade física e controle de stress. Conclusão: Através deste estudo evidencia-se a necessidade de intervenção nos hábitos que priorizem a promoção de saúde e qualidade de vida para os adolescentes. Por se tratar de fatores passíveis de intervenção, recomenda-se que estratégias de caráter educativo sejam adotadas, tanto em nível coletivo quanto individual, como forma de incentivar o consumo de alimentos saudáveis e estimular a prática de atividade física regular

Estudo da motivação dos funcionários dentro de uma organização para o alcance do sucesso nos resultados

Monografia apresentada à Diretoria de Pós-graduação da Universidade do Extremo Sul Catarinense- UNESC, para a obtenção do título de especialista em MBA Gestão de Desenvolvimento Humano e Organizacional.A motivação se faz presente nos ambientes organizacionais, onde o sucesso das organizações depende muito da satisfação de seus funcionários para melhor desempenho das tarefas. Ainda se faz presente o estudo da comunicação interna, no qual é de suma importância a organização ter essa ferramenta bem aperfeiçoada para melhores resultados, ainda destaca-se o clima organizacional, onde a empresa pode buscar melhores recursos para tornar seus resultados e satisfação ainda mais satisfatórios. Por isso esse estudo tem como objetivo estudo da motivação interna para que se tenha sucesso nos resultados, buscando assim maneiras de motivar pessoas para o alcance desse resultado. Quanto à metodologia foi utilizado um questionário qualitativo – quantitativo, que busca descrever os resultados obtidos, sendo assim foi classificado em descritiva.Levando em consideração que o presente trabalho apresentou uma pesquisa a respeito de satisfação, fatores motivacionais e benefícios, concluiu-se que: as pessoas estão satisfeitas com suas empresas, são motivadas para trabalhar, seu relacionamento é bom, a valorização e reconhecimento é bem vista pelos seus subordinados e chefes, o clima organizacional é satisfatório, pois os funcionários gostam do que fazem e são valorizados, porém os benefícios oferecidos é que precisa destaque, pois a empresa não disponibiliza benefícios, somente vale compras para 4 pessoas