Endophytic bacteria in long-term in vitro cultivated axenic pineapple microplants revealed by PCR-DGGE

In vitro propagated plants are believed to be free of microbes. However, after 5 years of in vitro culture of pineapple plants, without evidence of microbial contamination, the use of culture-independent molecular approach [classifying heterogeneous nucleic acids amplified via universal and specific 16S rRNA gene by polymerase chain reaction (PCR)], and further analysis by denaturing gradient gel electrophoresis (DGGE) revealed endophytic bacteria in roots, young and mature leaves of such plants. The amplification of 16S rRNA gene (Bacteria domain) with the exclusion of the plant chloroplast DNA interference, confirmed the presence of bacterial DNA, from endophytic microorganisms within microplant tissues. PCR-DGGE analysis revealed clear differences on bacterial communities depending on plant organ. Group-specific DGGE analyses also indicated differences in the structures of Actinobacteria, Alphaproteobacteria and Betaproteobacteria communities in each part of plants. The results suggest the occurrence of a succession of bacterial communities colonizing actively the microplants organs. This study is the first report that brings together evidences that pineapple microplants, previously considered axenic, harbor an endophytic bacterial community encompassing members of Actinobacteria, Alphaproteobacteria and Betaproteobacteria group which is responsive to differences in organs due to plant development.

Antimicrobial compounds produced by Lactobacillus sakei subsp sakei 2a, a bacteriocinogenic strain isolated from a Brazilian meat product

Bacteriocins produced by lactic acid bacteria are gaining increased importance due to their activity against undesirable microorganisms in foods. In this study, a concentrated acid extract of a culture of Lactobacillus sakei subsp. sakei 2a, a bacteriocinogenic strain isolated from a Brazilian pork product, was purified by cation exchange and reversed-phase chromatographic methods. The amino acid sequences of the active antimicrobial compounds determined by Edman degradation were compared to known protein sequences using the BLAST-P software. Three different antimicrobial compounds were obtained, P1, P2 and P3, and mass spectrometry indicated molecular masses of 4.4, 6.8 and 9.5 kDa, respectively. P1 corresponds to classical sakacin P, P2 is identical to the 30S ribosomal protein S21 of L. sakei subsp. sakei 23 K, and P3 is identical to a histone-like DNA-binding protein HV produced by L. sakei subsp. sakei 23 K. Total genomic DNA was extracted and used as target DNA for PCR amplification of the genes sak, lis and his involved in the synthesis of P1, P2 and P3. The fragments were cloned in pET28b expression vector and the resulting plasmids transformed in E. coli KRX competent cells. The transformants were active against Listeria monocytogenes, indicating that the activity of the classical sakacin P produced by L. sakei 2a can be complemented by other antimicrobial proteins.

Innovation as a Meta-Attribute for Graduate Engineers

This paper reviews the attitudes, skills and knowledge that engineering innovators should possess. It critically analyses and compares sets of graduate attributes from the USA, Australia and Malaysia in terms of which of these relate to the ability to innovate. Innovation can be described as an integrative, meta attribute that overarches most of the other graduate attributes. Due to the “graduate attribute paradox”, it is shown how meeting the stated attributes of graduates by industry does not necessarily satisfy the requirements of industry. It is argued that the culture of the engineering school is an important influence on fostering innovation in engineers.

B-1 cells temper endotoxemic inflammatory responses

Sepsis syndrome is caused by inappropriate immune activation due to bacteria and bacterial components released during infection. This syndrome is the leading cause of death in intensive care units. Specialized B-lymphocytes located in the peritoneal and pleural cavities are known as B-1 cells. These cells produce IgM and IL-10, both of which are potent regulators of cell-mediated immunity. It has been suggested that B-1 cells modulate the systemic inflammatory response in sepsis. In this study, we conducted in vitro and in vivo experiments in order to investigate a putative role of B-1 cells in a murine model of LPS-induced sepsis. Macrophages and B-1 cells were studied in monocultures and in co-cultures. The B-1 cells produced the anti-inflammatory cytokine IL-10 in response to LPS. In the B-1 cell-macrophage co-cultures, production of proinflammatory mediators (TNF-alpha, IL-6 and nitrite) was lower than in the macrophage monocultures, whereas that of IL-10 was higher in the co-cultures. Co-culture of B-1 IL-10(-/-) cells and macrophages did not reduce the production of the proinflammatory mediators (TNF-alpha, IL-6 and nitrite). After LPS injection, the mortality rate was higher among Balb/Xid mice, which are B-1 cell deficient, than among wild-type mice (65.0% vs. 0.0%). The Balb/Xid mice also presented a proinflammatory profile of TNF-alpha, IL-6 and nitrite, as well as lower levels of IL-10. In the early phase of LPS stimulation, B-1 cells modulate the macrophage inflammatory response, and the main molecular pathway of that modulation is based on IL-10-mediated intracellular signaling. (C) 2010 Elsevier GmbH. All rights reserved.

Cutaneous Mycobacterium haemophilum infection in a kidney transplant recipient after acupuncture treatment

P>Mycobacterium haemophilum is a slow-growing nontuberculous mycobacterium that can cause disease in both immunocompetent and immunocompromised patients. The most common clinical presentations of infection are the appearance of suppurative and ulcerated skin nodules. For the diagnosis, samples collected from suspected cases must be processed under the appropriate conditions, because M. haemophilum requires lower incubation temperatures and iron supplementation in order to grow in culture. In this case report, we describe the occurrence of skin lesions in a kidney transplant recipient, caused by M. haemophilum, associated with acupuncture treatment. The diagnosis was established by direct smear and culture of material aspirated from cutaneous lesions. Species identification was achieved by characterization of the growth requirements and by partial sequencing of the hsp65 gene. The patient was successfully treated with clarithromycin and ciprofloxacin for 12 months. Considering that the number of patients receiving acupuncture treatment is widely increasing, the implications of this potential complication should be recognized, particularly in immunosuppressed patients.

Anandamide prior to sensitization increases cell-mediated immunity in mice

The endocannabinoid system has become a topic of great interest in pharmacology due to its remarkable distribution in mammal organisms and capacity to play a modulatory role on several physiological systems, including modulation of immunity. Many studies have shown that administration of cannabinoids causes inhibitory effects on immune cells, including decreased proliferation and antigen-presenting cell (APC) costimulatory activity. In contrast, other groups have shown that some cannabinoids might present stimulatory actions on macrophage activity and T cell activation. Therefore, we aimed to investigate whether a treatment in vivo with a low dose of anandamide (0.1 mg/kg) immediately prior to sensitization would have an immunosuppressive or immunostimulatory effect on cell-mediated immunity (Th1 response) in mice. We report here that anandamide, prior to sensitization, was able to increase the Th1 response to ovalbumin in vivo and ex vivo. Anandamide increased delayed type hypersensitivity (DTH), splenocyte proliferation, and IFN-gamma production in a co-culture of adherent and non-adherent splenocytes. Moreover, anandamide prior to sensitization increased both the expression of DC co-stimulatory molecules (CD80/CD86) and IL-12/IL23 (p40) production ex vivo. We have also assessed direct effects of anandamide in the IFN-gamma/IL-4 balance of ConA-stimulated splenocytes in vitro. Anandamide at nanomolar concentrations increased the production of IFN-gamma, while such production decreased at micromolar range. Thus, anandamide induced both the increment of DC activation and IFN-gamma production, which are likely the mechanisms involved in the increase of Th1 response reported here. (C) 2010 Elsevier B.V. All rights reserved.

Cytotoxic effects and pulpal response caused by a mineral trioxide aggregate formulation and calcium hydroxide

Purpose: To evaluate: the in vivo pulpal response after pulpotomy with different capping agents. In addition, the in vitro cytotoxic effects of both materials were assessed by applying them on culture of pulp cells. Methods: For the in vivo test, the coronal pulp of 28 teeth of dogs was mechanically removed and the root pulps were capped with the following dental materials: Group 1: Pro-Root NITA (PRMTA); and Group 2 (control): calcium hydroxide saline paste (CH). After 60 days, the animals were sacrificed and the teeth processed for histological analysis. In the in vitro test, experimental extracts obtained from both capping agents were applied on the cultured MDPC-23 odontoblast-like cells. Results: In the root pulps capped with PRMTA or CH, coagulation necrosis partially replaced by dystrophic calcification as well as tubular dentin matrix laid down by elongated pulp cells was observed. None or mild inflammatory response occurred beneath the capped pulpal wound. Regarding the pulpal response, PRMTA and CH presented no statistical difference. However, the teeth capped CH presented greater healthy pulp loss which resulted in convex shape of the hard barrier than PRMTA. When applied on the cultured cells, it was demonstrated that PRMTA and CH solutions decreased the cell metabolic activity by 9.9% and 29.4%, respectively. CH caused higher cytotoxic effects to the MDPC-23 cells as well as deeper healthy pulp tissue loss than PRMTA. However, similar sequence of healing occurred after pulpotomy with both dental materials.

Nitric oxide-enhanced resistance to oral candidiasis

A marine model of oral candidiasis was used to show that nitric oxide (NO) is involved in host resistance to infection with Candida albicans in infection-'resistant' BALB/c and infection-'prone' DBA/2 mice. Following infection, increased NO production was detected in saliva. Postinfection samples of saliva inhibited the growth of yeast in vitro. Treatment with N-G-monomethyl-L-arginine (MMLA), an inhibitor of NO synthesis, led to reduced NO production, which correlated with an increase in C. albicans growth. Reduction in NO production following MMLA treatment correlated with an abrogation of interleukin-4 (IL-4), but not interferon-gamma (IFN-gamma), mRNA gene expression in regional lymph node cells. Down-regulation of IL-4 production was accompanied with an increase in IFN-gamma production in infection-'prone' DBA/2 mice. There was a functional relationship between IL-4 and NO production in that mice treated with anti-IL-4 monoclonal antibody showed a marked inhibition of NO production in saliva and in culture of cervical lymph node cells stimulated with C albicans antigen. The results Support previous conclusions that IL-4 is associated with resistance to oral candidiasis and suggest that NO is involved in controlling colonization of the oral mucosal surface with C albicans.

Interspecific hybrids between the homothallic Phytophthora sojae and Phytophthora vignae

An interspecific cross was attempted between two homothallic species of Phytophthora, P. sojae and P. vignae. From 1640 single-oospore cultures isolated, DNA was extracted from 800, and two interspecific F-1 hybrids (F(1)1121 and F(1)1426) were putatively identified using RAPD markers. The true hybrid nature of these F-1 hybrids was confirmed using additional AFLP analysis. Single- zoospore cultures were generated for each F-1 hybrid and one single-zoospore culture of each was used in pathogenicity and virulence tests. Both F-1 hybrids were pathogenic to soybean and cowpea, causing symptoms including lesions, wilting and death of susceptible soybean and cowpea cultivars. However, the aggressiveness of the F-1 hybrids was reduced and was substantially more variable when compared with that of the parental isolates on their respective hosts. The F-1 hybrids were reisolated from infected seedlings and their hybrid nature confirmed using RAPD and AFLP analysis. These results provide a basis for further research aimed at obtaining an increased understanding of the genetics of host specificity in the Oomycetes.

Teachers and productive pedagogies: Contextualising, conceptualising, utilising

Improving students' outcomes from schooling requires schools to be learning organisations, where both students and teachers are engaged in learning. As such, knowledge and talk about pedagogy need to be at the core of the professional culture of schools. This article argues that this will require the valuing of teachers' work, that is, their pedagogical practices, to be a central focus of educational policy. Dangers are associated with this argument in terms of understating the impacts of poverty, lack of funding to disadvantaged schools and other social factors such as the pressures of globalisation upon students' educational opportunities. Hence, while acknowledging the importance of pedagogy to students' outcomes, the article contextualises the argument through a recognition of the policy and structural conditions that work against the valuing of teachers and their work. It then conceptualises how, within this context, a focus on pedagogies can make a difference to students' academic and social outcomes from schooling. This conceptualisation utilises the productive pedagogies model of classroom practice, developed in a large Australian study of school reform, as an example of the forms of pedagogical practices that support students' achievement of academic and social outcomes. It is argued that such pedagogical practices ought to be a concern of teachers, school administrators, education systems and local communities interested in schools as learning organisations.

A educação ambiental nos espaços de convivência : tecendo a cultura da sustentabilidade nas redes cotidianas de uma escola municipal de Vila Velha/ES

Este trabalho tem como objetivo problematizar a ecoformação de professoras e alunos em espaços de convivência, potencializados com as experiências da IV Conferência Nacional Infanto-juvenil de Meio Ambiente (CNIJMA), visitas monitoradas, aulas de campo e saídas. Ancorada na Política Estruturante de Educação Ambiental, entendo que esses espaços de convivência expressam um processo educacional, permanente, continnum e transformador. Tendo como mediadora a COM-Vida (Comissão de Meio Ambiente e Qualidade de Vida) e o Tratado de Educação Ambiental para as Sociedades Sustentáveis e Responsabilidade Global, a Educação Ambiental visa fortalecer a cidadania ambiental em movimento escola-comunidade-escola. Impregnada de sentidos e significados, busquei pesquisar a ecoformação dessa coletividade pelo viés do Paradigma da Complexidade, que me apresentou as incertezas como um processo potencializador da criatividade, da amizade e da solidariedade que tecem a rede de saberes e fazeres que envolvem esses sujeitos investigados, além de evidenciar o cuidado de si com o outro e com o mundo. Ao trilhar os caminhos desta pesquisa, optei por abordagens qualitativas inspiradas na fenomenologia-existencial proposta por Martin Heidegger, Michèle Sato e Paulo Freire e, dessa forma, pude investigar os saberes ambientais que atravessam as redes cotidianas da escola pesquisada, potencializando a cultura da sustentabilidade. Valendo-me da observação participante das práticas pedagógicas, encontrei nas narrativas das professoras e alunos a expressividade de um processo ecoformativo que instiga outras racionalidades comprometidas com a ética, a coletividade, a afetividade, a solidariedade, as transformações sociais, a diversidade e a outridade.

A narratividade do Hip Hop e suas interfaces com o contexto educacional

A presente tese de doutoramento em Educação aborda a cultura do Hip Hop e sua relação com o contexto educativo. Insere-se nas discussões da linha de pesquisa em Educação e Linguagens e problematiza o espaço escolar, enquanto espaço de reprodução da ideologia hegemônica, analisando o projeto “Escola de Rimas”, desenvolvido na Grande Vitória, como movimento de resistência e ressignificação cultural na escola. Parte da pergunta: Como as práticas discursivas do Hip Hop podem ressignificar o contexto escolar? A presente tese aborda a cultura do Hip Hop como campo discursivo singular de uma experiência narrativa (BENJAMIN, 1986; BONDÍA, 2001) e de uma subjetividade eticamente responsável (BAKHTIN, 1992a; 1992b; 2010), fundada no princípio da alteridade (PONZIO, 2009), e o analisa a partir de uma perspectiva crítica e dialógica (FREIRE, 1981; 1994; 1995; GIROUX, 1986; 1987; BRANDÃO, 1986; BAKHTIN, 1992a; 1992b; 2010). Desenvolve a pesquisa em um contexto limiar entre a escola e a cultura hip hop e dimensiona o debate das culturas marginais nos contextos educativos, voltando-se para os sujeitos e suas experiências narrativas, avaliando a interação de algumas de suas práticas discursivas com o processo de ensino-aprendizagem. Para isso, analisa o projeto cultural “Escola de Rimas”, criado pelos próprios ativistas do movimento hip hop da Grande Vitória e desenvolvido em uma escola da rede pública estadual de ensino do Espírito Santo, com o objetivo de discutir o seu papel em um processo de ressignificação educacional. Como hipótese de trabalho defende-se que o espaço escolar, como espaço de disputas, é ressignificado com a introdução de outras práticas discursivas e culturais, entre elas o hip hop, que aponta para a necessidade de ouvir responsiva e responsavelmente as narrativas dos educandos, contribuindo, assim, para a formação crítica desses sujeitos e enfrentando, ao mesmo tempo, práticas de exclusão historicamente instituídas.

Práticas e saberes da professora pomerana : um estudo sobre interculturalidade

Esta pesquisa analisa práticas e saberes da professora pomerana em uma escola do campo de ensino fundamental em Santa Maria de Jetibá/Estado do Espírito Santo/Brasil, para problematizar questões sobre cultura e interculturalidade. Aprofundam-se reflexões sobre quais têm sido as contribuições dessas práticas para uma educação escolar intercultural. Beneficia-se do estudo de caso para análise de documentos, observações de aulas, observações participantes em planejamentos, realização de entrevista semiestruturada com a professora e, anotações no diário de campo. Situam-se brevemente questões da história dos descendentes de pomeranos, bem como sobre as principais produções acadêmicas referentes à educação desse povo tradicional. Discussões teórico-metodológicas contribuem para compreensão e embasamento de alguns conceitos-chave, como: cultura(s) (THOMPSON, 1995; GEERTZ, 2012), interculturalismo (CANDAU, 2005; FORNET-BETANCOURT, 2004), cultura escolar e cultura da escola (FORQUIN, 1993), práticas docentes (FREIRE, 1996), saberes docentes (TARDIF, 2012), parceria (FOERSTE, 2005) etc. Aponta-se que as práticas da professora pomerana faziam parte do contexto escolar antes da implementação do Programa de Educação Escolar Pomerana (PROEPO) no que se refere ao uso oral da língua pomerana na sala de aula. As práticas da professora se ressignificam na interação com os saberes culturais, experienciais e coletivos produzidos na dinâmica escolar e comunitária. Essas práticas possibilitam o diálogo da cultura escolar com a cultura da escola, promovendo a vivência das experiências locais e o acesso ao saber universal. Pode-se afirmar que no contexto da escola investigada existem algumas condições que favorecem a dimensão da interculturalidade nos processos pedagógicos. Cabe aprofundar estudos sobre as contribuições da educação escolar para o ensino bilíngue e seus impactos à valorização da cultura pomerana e a consequente manutenção da língua deste povo tradicional. Tensões e conflitos foram identificados quanto ao enfoque do PROEPO ao desenvolvimento das habilidades de leitura e escrita em língua pomerana.

Cultura Organizacional e Relações de Poder : Uma análise de caso do Centro de Ciências da Saúde da Universidade Federal do Espírito Santo Vitória 2014

Esta dissertação analisa, por meio de um estudo de caso, a Cultura Organizacional do Centro de Ciências da Saúde - CCS - da Universidade Federal do Espírito Santo - UFES - e as relações de Poder dentro da universidade e do próprio CCS, no período de agosto de 2013 a janeiro de 2014. Para tanto, foi utilizado o método de Análise de Discurso Crítica, em entrevistas e documentos, visando compreender a influência interna da cultura organizacional, bem como sua influência junto à administração central. A partir da análise, percebe-se que o CCS possui uma dinâmica própria de funcionamento, que apesar de discordante dos princípios burocráticos buscados pela administração pública, basta para o funcionamento interno e permite influenciar, mesmo que tangencialmente, a estrutura de poder central da instituição. Além disso, percebe-se a força da influência cultural do curso de medicina dentro da própria organização do centro.

Iogurte simbiótico de açaí (Euterpe edulis Mart.): caracterização físico-química e viabilidade de bactérias láticas e probióticas

Os iogurtes simbióticos, que combinam microrganismos probióticos e substâncias prebióticas, adicionados de polpa de frutas são uma tendência crescente no mercado. O fruto açaí (Euterpe edulis) se destaca pela presença de compostos bioativos, como as antocianinas. Neste contexto, o presente trabalho teve como objetivo caracterizar e avaliar os parâmetros físico-químicos e a viabilidade microbiológica de iogurte simbiótico de açaí enriquecido com inulina e adicionado de cultura probiótica de Bifidobacterium animallis subsp. lactis BB-12. As formulações de iogurte atenderam aos requisitos físico-químicos exigidos pela IN no 46/2007 do MAPA apresentando teor de cinzas de 0,86 % ± 0,10, extrato seco total de 23,18 % ± 2,59, teor de gordura de 4,16 % ± 0,31, acidez de 0,70 % ± 0,05 e pH de 4,45 ± 0,10. Entre as formulações o conteúdo fenólico total variou de 18,17 a 117,84 mg de AGE/100 g, teor de antocianinas de 1,92 a 47,88 mg/100 g e atividade antioxidante de 0,71 a 6,95 μmol Trolox/g, observando-se um aumento de acordo com o aumento do teor de polpa de açaí adicionada. Ao final de 28 dias de armazenamento a 5 °C, observou-se uma redução no teor de antocianinas e da atividade antioxidante. Verificou-se a contribuição positiva da polpa de açaí na viabilidade das bactérias láticas totais, cujas contagens variaram de 4,56 a 7,04 log UFC.g-1 e de B. lactis BB-12 que variou de 3,17 a 6,34 log UFC.g-1, favorecendo a multiplicação dessas bactérias nos iogurtes. Nas formulações com 20 e 25 % de polpa de açaí as contagens das bactérias láticas totais e probiótica mantiveram-se viáveis de acordo com a IN no 46/2007 do MAPA e a Lista de Alegação de Propriedade Funcional (Anvisa), durante os 28 dias de armazenamento a 5 oC. Concluiu-se que a adição de polpa de açaí E. edulis, inulina e B. lactis BB-12 foi tecnologicamente viável na elaboração de iogurte simbiótico de açaí, sendo uma excelente alternativa de diversificação do produto no mercado.