A NEW METHOD TO OBTAIN GOOD ANATOMICAL SLIDES OF HETEROGENEOUS PLANT PARTS

A new method is presented to prepare anatomical slides of plant materials including a combination of soft and hard tissues, such as stems with cambial variants, arboreal monocotyledons, and tree bark The method integrates previous techniques aimed at softening the samples and making them thereby more homogeneous, with the use of anti-tearing polystyrene foam solution In addition, we suggest two other alternatives to protect the sections from tearing adhesive tape and/or Mayer`s albumin adhesive, both combined with the polystyrene foam solution This solution is cheap and easy to make by dissolving any packaging polystyrene m butyl acetate It is applied before each section is cut on a sliding microtome and ensures that all the tissues in the section will hold together This novel microtechnical procedure will facilitate the study of heterogeneous plant portions, as shown in some illustrated examples

Long-Term Culture of Mouse Embryonic Stem Cell-Derived Adherent Neurospheres and Functional Neurons

Innumerous protocols, using the mouse embryonic stem (ES) cells as model for in vitro study of neurons functional properties and features, have been developed. Most of these protocols are short lasting, which, therefore, does not allow a careful analysis of the neurons maturation, aging, and death processes. We describe here a novel and efficient long-lasting protocol for in vitro ES cells differentiation into neuronal cells. It consists of obtaining embryoid bodies, followed by induction of neuronal differentiation with retinoic acid of nonadherent embryoid bodies (three-dimensional model), which further allows their adherence and formation of adherent neurospheres (AN, bi-dimensional model). The AN can be maintained for at least 12 weeks in culture under repetitive mechanical splitting, providing a constant microenvironment (in vitro niche) for the neuronal progenitor cells avoiding mechanical dissociation of AN. The expression of neuron-specific proteins, such as nestin, sox1, beta III-tubulin, microtubule-associated protein 2, neurofilament medium protein, Tau, neuronal nuclei marker, gamma-aminobutyric acid, and 5-hydroxytryptamine, were confirmed in these cells maintained during 3 months under several splitting. Additionally, expression pattern of microtubule-associated proteins, such as lissencephaly (Lis1) and nuclear distribution element-like (Ndel1), which were shown to be essential for differentiation and migration of neurons during embryogenesis, was also studied. As expected, both proteins were expressed in undifferentiated ES cells, AN, and nonrosette neurons, although presenting different spatial distribution in AN. In contrast to previous studies, using cultured neuronal cells derived from embryonic and adult tissues, only Ndel1 expression was observed in the centrosome region of early neuroblasts from AN. Mature neurons, obtained from ES cells in this work, display ionic channels and oscillations of membrane electrical potential typical of electrically excitable cells, which is a characteristic feature of the functional central nervous system (CNS) neurons. Taken together, our study demonstrated that AN are a long-term culture of neuronal cells that can be used to analyze the process of neuronal differentiation dynamics. Thus, the protocol described here provides a new experimental model for studying neurological diseases associated with neuronal differentiation during early development, as well as it represents a novel source of functional cells that can be used as tools for testing the effects of toxins and/or drugs on neuronal cells.

p53 Mutant Human Glioma Cells Are Sensitive to UV-C-Induced Apoptosis Due to Impaired Cyclobutane Pyrimidine Dimer Removal

The p53 protein is a key regulator of cell responses to DNA damage, and it has been shown that It sensitizes glioma cells to the alkylating agent temozolomide by up-regulating the extrinsic apoptotic pathway, whereas it increases the resistance to chloroethylating agents, such as ACNU and BCNU, probably by enhancing the efficiency of DNA repair. However, because these agents induce a wide variety of distinct DNA lesions, the direct Importance of DNA repair is hard to access. Here, it is shown that the Induction of photoproducts by UV light (UV-C) significantly Induces apoptosis In a p53-mutated glioma background. This Is caused by a reduced level of photoproduct repair, resulting In the persistence of DNA lesions in p53-mutated glioma cells. UV-C-Induced apoptosis in p53 mutant glioma cells Is preceded by strong transcription and replication inhibition due to blockage by unrepaired photolesions. Moreover, the results Indicate that UV-C-induced apoptosis of p53 mutant glioma cells Is executed through the intrinsic apoptotic pathway, with Bcl-2 degradation and sustained Bax and Bak up-regulation. Collectively, the data Indicate that unrepaired DNA lesions Induce apoptosis In p53 mutant gliomas despite the resistance of these gliomas to temozolomide, suggesting that efficiency of treatment of p53 mutant gliomas might be higher with agents that Induce the formation of DNA lesions whose global genomic repair is dependent on p53. (Mol Cancer Res 2009;7(2):237-46)

INDEX OF AN IMPLICIT DIFFERENTIAL EQUATION

In this paper we introduce the concept of the index of an implicit differential equation F(x,y,p) = 0, where F is a smooth function, p = dy/dx, F(p) = 0 and F(pp) = 0 at an isolated singular point. We also apply the results to study the geometry of surfaces in R(5).

SINGULARITY THEORY AND FORCED SYMMETRY BREAKING IN EQUATIONS

A theory of bifurcation equivalence for forced symmetry breaking bifurcation problems is developed. We classify (O(2), 1) problems of corank 2 of low codimension and discuss examples of bifurcation problems leading to such symmetry breaking.

A Hopping Mechanism for Cargo Transport by Molecular Motors on Crowded Microtubules

Most models designed to study the bidirectional movement of cargos as they are driven by molecular motors rely on the idea that motors of different polarities can be coordinated by external agents if arranged into a motor-cargo complex to perform the necessary work Gross, Hither and yon: a review of bidirectional microtubule-based transport (Gross in Phys. Biol. 1:R1-R11, 2004). Although these models have provided us with important insights into these phenomena, there are still many unanswered questions regarding the mechanisms through which the movement of the complex takes place on crowded microtubules. For example (i) how does cargo-binding affect motor motility? and in connection with that-(ii) how does the presence of other motors (and also other cargos) on the microtubule affect the motility of the motor-cargo complex? We discuss these questions from a different perspective. The movement of a cargo is conceived here as a hopping process resulting from the transference of cargo between neighboring motors. In the light of this, we examine the conditions under which cargo might display bidirectional movement even if directed by motors of a single polarity. The global properties of the model in the long-time regime are obtained by mapping the dynamics of the collection of interacting motors and cargos into an asymmetric simple exclusion process (ASEP) which can be resolved using the matrix ansatz introduced by Derrida (Derrida and Evans in Nonequilibrium Statistical Mechanics in One Dimension, pp. 277-304, 1997; Derrida et al. in J. Phys. A 26: 1493-1517, 1993).

O curso GQT e a implantação da qualidade total na administração pública indireta no RGS : o caso PROSAC

O Programa Gaúcho da Qualidade e Produtividade – PGQP, criado pelo Governo do Estado do Rio Grande do Sul (1991-1994), reconhece o Curso Gestão da Qualidade Total (GQT) como conteúdo básico para os programas de desenvolvimento de recursos humanos que visem à implantação da qualidade total. Neste contexto, o presente estudo objetiva analisar as contribuições do Curso GQT para a implantação da qualidade total no âmbito da Administração Pública Indireta, mais especificamente, nas empresas integrantes do PROSAC – Programa de Satisfação e Atenção ao Cliente, desenvolvido pela Secretaria de Energia, Minas e Comunicações do Estado do RS, no período de 1993-1994. A metodologia adotada inclui pesquisa documental e de campo. Primeiramente, sob o enfoque qualitativo, analisa-se o conteúdo curricular do Curso GQT frente à realidade organizacional dos participantes. Numa segunda etapa, sessenta funcionários, de empresa integrante do PROSAC, avaliam o processo ensino-aprendizagem no contexto organizacional. A análise dos dados envolveu estatísticas descritivas, análise fatorial, mapeamento conceitual e análise de conteúdo. Os resultados demonstram que os pesquisados reconhecem a utilidade dos métodos e técnicas da qualidade para o controle e melhoria dos processos de trabalho, e a importância das informações apresentadas para o exercício de uma reflexão crítica sobre a sua realidade profissional. Por outro lado, indicam que foi enfatizada a aquisição de conhecimentos técnicos em detrimento do desenvolvimento das habilidades conceituais e da competência interpessoal, o que contraria os princípios da qualidade total que salientam igualmente a importância destes aspectos. Os resultados permitem, ainda, inferir uma ausência de associação conceitual - condição necessária para a aprendizagem significativa - entre o conteúdo curricular do Curso GQT, estruturado a partir da realidade do setor industrial, e as peculiaridades do serviço público.

A maturação biológica e a seleção de atletas no minibasquetebol

Este trabalho tem como objeto principal investigar o nível da maturação biológica e o processo de seleção dos atletas de minibasquetebol. A amostra é constituída por 43 minibasquetebolistas do sexo masculino, com idades entre 11 e 12 anos, pertencentes às quatro equipes finalistas do Campeonato Gaúcho de Minibasquetebol de 1999. Para avaliar o nível de maturação biológica dos atletas, utilizamos o Teste de Tanner que determina o grau de desenvolvimento da característica sexual secundária - pêlos pubianos. O método utilizado é o da auto-avaliação, conforme descrito em MATSUDO & MATSUDO (1991). Para identificar a presença do aspecto maturação biológica, nos planejamentos de treinos e participação em competições, elaboramos um instrumento em forma de questionário, respondido pelos técnicos das equipes participantes da amostra. Para definir quais os critérios utilizados para a seleção dos atletas, preparamos uma entrevista semi-estruturada feita com os técnicos. Verificamos que, na determinação do nível de maturação biológica dos atletas, a concentração de freqüência é mais elevada no nível P3 que, junto com o nível P4 representam 74,4% da amostra, indicando como característica importante do grupo um estágio de maturação continuada. Os técnicos referem no questionário que não levam em consideração o aspecto maturacional biológico dos atletas na elaboração dos trabalhos juntos aos mesmos. Através da entrevista com os profissionais, constatamos que o desempenho técnico é o principal critério por eles utilizado para a seleção dos atletas. A principal conclusão deste estudo sugere que os atletas selecionados alcançam tal posição por apresentarem condição de maturados ou pós-maturados.

Optimisation of solid-phase microextraction combined with gas chromatography–mass spectrometry based methodology to establish the global volatile signature in pulp and skin of Vitis vinifera L. grape varieties

The volatiles (VOCs) and semi-volatile organic compounds (SVOCs) responsible for aroma are mainly present in skin of grape varieties. Thus, the present investigation is directed towards the optimisation of a solvent free methodology based on headspace-solid-phase microextraction (HS-SPME) combined with gas chromatography–quadrupole mass spectrometry (GC–qMS) in order to establish the global volatile composition in pulp and skin of Bual and Bastardo Vitis vinifera L. varieties. A deep study on the extraction-influencing parameters was performed, and the best results, expressed as GC peak area, number of identified compounds and reproducibility, were obtained using 4 g of sample homogenised in 5 mL of ultra-pure Milli-Q water in a 20 mL glass vial with addition of 2 g of sodium chloride (NaCl). A divinylbenzene/carboxen/polydimethylsiloxane fibre was selected for extraction at 60 °C for 45 min under continuous stirring at 800 rpm. More than 100 VOCs and SVOCs, including 27 monoterpenoids, 27 sesquiterpenoids, 21 carbonyl compounds, 17 alcohols (from which 2 aromatics), 10 C13 norisoprenoids and 5 acids were identified. The results showed that, for both grape varieties, the levels and number of volatiles in skin were considerably higher than those observed in pulp. According to the data obtained by principal component analysis (PCA), the establishment of the global volatile signature of grape and the relationship between different part of grapes—pulp and skin, may be an useful tool to winemaker decision to define the vinification procedures that improves the organoleptic characteristics of the corresponding wines and consequently contributed to an economic valorization and consumer acceptance.

Retrievals of a size parameter for phytoplankton and spectral light absorption by colored detrital matter from water-leaving radiances at SeaWiFS channels in a continental shelf region off Brazil

Many efforts are currently oriented toward extracting more information from ocean color than the chlorophyll a concentration. Among biological parameters potentially accessible from space, estimates of phytoplankton cell size and light absorption by colored detrital matter (CDM) would lead to an indirect assessment of major components of the organic carbon pool in the ocean, which would benefit oceanic carbon budget models. We present here 2 procedures to retrieve simultaneously from ocean color measurements in a limited number of bands, magnitudes, and spectral shapes for both light absorption by CDM and phytoplankton, along with a size parameter for phytoplankton. The performance of the 2 procedures was evaluated using different data sets that correspond to increasing uncertainties: ( 1) measured absorption coefficients of phytoplankton, particulate detritus, and colored dissolved organic matter ( CDOM) and measured chlorophyll a concentrations and ( 2) SeaWiFS upwelling radiance measurements and chlorophyll a concentrations estimated from global algorithms. In situ data were acquired during 3 cruises, differing by their relative proportions in CDM and phytoplankton, over a continental shelf off Brazil. No local information was introduced in either procedure, to make them more generally applicable. Over the study area, the absorption coefficient of CDM at 443 nm was retrieved from SeaWiFS radiances with a relative root mean square error (RMSE) of 33%, and phytoplankton light absorption coefficients in SeaWiFS bands ( from 412 to 510 nm) were retrieved with RMSEs between 28% and 33%. These results are comparable to or better than those obtained by 3 published models. In addition, a size parameter of phytoplankton and the spectral slope of CDM absorption were retrieved with RMSEs of 17% and 22%, respectively. If these methods are applied at a regional scale, the performances could be substantially improved by locally tuning some empirical relationships.

Fecundidade em Macrobrachium acanthurus (Wiegmann, 1836) do Rio Ribeira de Iguape (Crustacea, Decapoda, Palaemonidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Spatial variability of aggregate stability in latosols under sugarcane

Solos submetidos ao mesmo sistema de manejo manifestam variabilidade espacial diferenciada de seus atributos. A variabilidade espacial da estabilidade de agregados foi caracterizada em um Latossolo Vermelho distrófico e Latossolo Vermelho eutroférrico, sob cultivo de cana-de-açúcar. Foram realizadas amostragens de terra nos pontos de interseção de uma malha de 10 x 10 linhas, espaçadas de 10 m, totalizando 100 pontos de amostragem por área, coletadas nas camadas de 0,0-0,2 e 0,2-0,4 m de profundidade, para determinação de diâmetro médio geométrico (DMG), diâmetro médio ponderado (DMP), agregados na classe >2,0 mm e teor de matéria orgânica do solo. O Latossolo Vermelho eutroférrico apresenta maior estabilidade de agregados quando comparado com o Latossolo Vermelho distrófico, condizente com o maior teor de argila, matéria orgânica e mineralogia gibbsítica. A evolução diferenciada dos Latossolos estudados explica os maiores alcances, o menor coeficiente de variação (CV) e a menor variabilidade, observados no Latossolo Vermelho eutroférrico para todos os atributos estudados.

Identificação e validação de marcadores microssatélites ligados ao gene Rpp5 de resistência à ferrugem-asiática-da-soja

O objetivo deste trabalho foi identificar novos marcadores microssatélites, ligados ao gene Rpp5 de resistência à ferrugem-da-soja, e validar os marcadores previamente mapeados, para que possam ser utilizados em programas de seleção assistida por marcadores moleculares (SAM). Para tanto, uma população F2 com 100 indivíduos, derivada do cruzamento entre a PI 200526 e a cultivar Coodetec 208, suscetível à ferrugem, foi artificialmente infectada e avaliada quanto à sua reação de resistência à ferrugem. Marcadores microssatélites foram testados nos genitores e em dois bulks contrastantes, para a identificação de marcadores ligados. Dois novos marcadores, potencialmente associados à resistência, foram testados em plantas individuais, e se constatou que eles estão ligados ao gene Rpp5 e estão presentes no grupo de ligação N da soja. A eficiência de seleção foi determinada em relação a todos os marcadores ligados ao gene Rpp5, e a combinação entre os marcadores Sat_275+Sat_280 foi de 100%.

MUC1 gene polymorphism in three Nelore lines selected for growth and its association with growth and carcass traits

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)