Insect chemistry and chirality

Examination of the chemistry of a number of Australian insect species provided examples of unusual structures and encouraged determinations of their absolute stereochemistry by stereocontrolled syntheses and chromatographic comparisons. Inter alia, studies with the fruit-spotting bug (Amblypelta nitida), certain parasitic wasps (Biosteres sp.), the aposematic shield bug (Cantao parentum), and various species of scarab grubs are summarized. The determination of enantiomeric excesses (ee's) for component epoxides, lactones, spiroacetals, and allenes are described. Stereochemical and related aspects of the biosynthesis of spiroacetals in certain fruit-fly species (Bactrocerae sp.) are also presented.

Localization of the Epstein-Barr virus protein LMP 1 to exosomes

The Epstein-Barr virus latent membrane protein (LMP 1) functions as a constitutively active signalling molecule and associates in lipid rafts clustered with other signalling molecules. Using immunofluorescent confocal microscopy, LMP 1 was shown to have an heterogeneous distribution among individual cells which was not related to the cell cycle stage. LMP 1 was shown to localize to intracellular compartments in cells other than the plasma membrane, Co-labelling of cells with both an LIMP 1 antibody and an antibody to the Golgi protein GS15 revealed that the intracellular LMP 1 partly co-localized with the Golgi apparatus. Further confirmation of intracellular LMP 1 localization was obtained by immunoelectron microscopy with rabbit polyclonal LIMP 1 antibodies and cryosectioning. As well as being present in intracellular foci, LMP 1 co-localized in part with MHC-II and was present on exosomes derived from a lymphoblastoid cell line. Preparations of LMP 1 containing exosomes were shown to inhibit the proliferation of peripheral blood mononuclear cells, suggesting that LIMP 1 could be involved in immune regulation. This may be of particular relevance in EBV-associated tumours such as nasopharyngeal carcinoma and Hodgkin's disease, as LMP 1-containing exosomes may be taken up by infiltrating T-lymphocytes, where LMP 1 could exert an anti-proliferative effect, allowing the tumour cells to evade the immune system.

The t-SNARE syntaxin 4 is regulated during macrophage activation to function in membrane traffic and cytokine secretion

Activation of macrophages with lipopolysaccharide (LPS) induces the rapid synthesis and secretion of proinflammatory cytokines, such as tumor necrosis factor (TNFalpha), for priming the immune response [1, 2]. TNFalpha plays a key role in inflammatory disease [3]; yet, little is known of the intracellular trafficking events leading to its secretion. In order to identify molecules involved in this secretory pathway, we asked whether any of the known trafficking proteins are regulated by LPS. We found that the levels of SNARE proteins were rapidly and significantly up- or downregulated during macrophage activation. A subset of t-SNAREs (Syntaxin 4/SNAP23/Munc18c) known to control regulated exocytosis in other cell types [4, 5] was substantially increased by LPS in a temporal pattern coinciding with peak TNFalpha secretion. Syntaxin 4 formed a complex with Munc18c at the cell surface of macrophages. Functional studies involving the introduction of Syntaxin 4 cDNA or peptides into macrophages implicate this t-SNARE in a rate-limiting step of TNFalpha secretion and in membrane ruffling during macrophage activation. We conclude that in macrophages, SNAREs are regulated in order to accommodate the rapid onset of cytokine secretion and for membrane traffic associated with the phenotypic changes of immune activation. This represents a novel regulatory role for SNAREs in regulated secretion and in macrophage-mediated host defense.

An approach to verifying and debugging simulation models governed by ordinary differential equations: Part 1. Methodology for residual generation

For dynamic simulations to be credible, verification of the computer code must be an integral part of the modelling process. This two-part paper describes a novel approach to verification through program testing and debugging. In Part 1, a methodology is presented for detecting and isolating coding errors using back-to-back testing. Residuals are generated by comparing the output of two independent implementations, in response to identical inputs. The key feature of the methodology is that a specially modified observer is created using one of the implementations, so as to impose an error-dependent structure on these residuals. Each error can be associated with a fixed and known subspace, permitting errors to be isolated to specific equations in the code. It is shown that the geometric properties extend to multiple errors in either one of the two implementations. Copyright (C) 2003 John Wiley Sons, Ltd.

Recombinant expression of Munc18c in a baculovirus system and interaction with syntaxin4

Two protein families that are critical for vesicle transport are the Syntaxin and Munc18/Sec1. families of proteins. These two molecules form a high affinity complex and play an essential role in vesicle docking and fusion. Munc18c was expressed as an N-terminally His-tagged fusion protein from recombinant baculovirus in Sf9 insect cells. His-tagged Munc18c was purified to homogeneity using both cobalt-chelating affinity chromatography and gel filtration chromatography. With this simple two-step protocol, 3.5 mg of purified Munc18c was obtained from a 1 L culture. Further, the N-terminal His-tag could be removed by thrombin cleavage while the tagged protein was bound to metal affinity resin. Recombinant Munc18c produced in this way is functional, in that it forms a stable complex with the SNARE interacting partner, syntaxin4. Thus we have developed a method for producing and purifying large amounts of functional Munc18c-both tagged and detagged-from a baculovirus expression system. We have also developed a method to purify the Munc18c:syntaxin4 complex. These methods will be employed for future functional and structural studies. Crown copyright (C) 2003 Published by Elsevier Inc. All rights reserved.

A hist??ria de Henrique: quando as habilidades de l??der s??o testadas ap??s a ascens??o profissional

Henrique, funcion??rio antigo de um importante programa governamental, ?? promovido ao mais alto cargo do programa. Devido a algumas posi????es e a????es ortodoxas, colegas e superiores come??aram a questionar o estilo e as iniciativas de Henrique. Ele, por sua vez, ao identificar as cr??ticas, voltou-se para fora da institui????o, transformando-se em ???embaixador do programa???, com viagens semanais e cada vez mais ausente. Seus subordinados tamb??m come??aram a se queixar e o Ministro finalmente mostrou insatisfa????o a respeito. Ao final do caso, prop??e-se um exerc??cio de avalia????o sobre as habilidades de Henrique

Flexibilidade na gest??o de pessoal na administra????o p??blica

Textos diversos, elaborados por t??cnicos do governo e especialistas de pa??ses-membros da OCDE Organiza????o para Coopera????o e Desenvolvimento Econ??mico, analisando a evolu????o recente da gest??o de pessoal no setor p??blico ao mesmo tempo em que s??o colocados problemas levantados pelas pol??ticas de recursos humanos ent??o adotadas.

Desenvolvimento gerencial no setor p??blico: tend??ncias internacionais e organiza????es-l??deres

Pesquisa comparada realizada por iniciativa do Canadian Centre for Management Development que re??ne nove institui????es em diferentes pa??ses, incluindo a ENAP, direcionadas ao desenvolvimento gerencial no setor p??blico. A vers??o desta pesquisa oferece a possibilidade de ampliar o debate em torno das necessidades de apoiar o desenvolvimento de gerentes e dirigentes p??blicos

Colet??nea planejamento e or??amento governamental: volume 1: pref??cio

Pref??cio da colet??nea, estabelece seu objetivo de fornecer material did??tico focado no desenvolvimento profissional da administra????o p??blica brasileira.

Colet??nea planejamento e or??amento governamental: volume 1: introdu????o

Esta introdu????o, al??m de apresentar e contextualizar cada tem??tica, objetiva ser um texto did??tico b??sico para os cursos da ENAP. Nesse sentido, traz, adicionalmente, uma s??ntese dos temas desenvolvidos nos respectivos textos da colet??nea.

Colet??nea planejamento e or??amento governamental: volume 1: o plano como aposta

O texto sintetiza os principais fundamentos do m??todo PES. Esse m??todo contrap??e-se ao planejamento tradicional e prop??e uma abordagem tecnopol??tica do planejamento.

Colet??nea planejamento e or??amento governamental: volume 1: a experi??ncia brasileira em planejamento econ??mico - uma s??ntese hist??rica

O texto faz uma s??ntese did??tica dos planos de desenvolvimento e de estabiliza????o econ??mica implementados no Brasil, nas ??ltimas seis d??cadas. O relato abrange desde os primeiros exerc??cios de planejamento, no imediato p??s-Segunda Guerra Mundial, at?? os ??ltimos PPAs e, ainda, faz refer??ncias ??s tentativas de planejamento de longo prazo com o ???Brasil 2020???, no governo Fernando Henrique.

Colet??nea planejamento e or??amento governamental: volume 1: a retomada do planejamento governamental no Brasil e seus desafios

O texto apresenta um cen??rio dos avan??os recentes no planejamento governamental. Em sua primeira parte, procura referenciar o debate atual acerca das possibilidades, das limita????es e dos avan??os introduzidos pelo PPA, especialmente a partir de 2000, com destaque para a integra????o ???plano-or??amento???, por meio de programas formulados com origem em problemas ou demandas da sociedade; um plano e um or??amento orientados para resultado; e o novo sistema de planejamento e or??amento como pe??a fundamental para a implanta????o da gest??o por resultados no Pa??s. A segunda parte do texto refere-se ao modelo de gest??o do plano. S??o apresentados os fundamentos do modelo de sua gest??o e os avan??os propostos para o PPA 2004-2007. Em s??ntese, a gest??o do plano ?? fundamentada no princ??pio da responsabiliza????o e nos processos de monitoramento, avalia????o e retroalimenta????o do ciclo do gasto p??blico

Colet??nea planejamento e or??amento governamental: volume 1: a metodologia de gest??o estrat??gica do NAE

O texto incorpora uma diversidade de ferramentas de planejamento estrat??gico. Sua principal caracter??stica ?? a de ver o planejamento estrat??gico como um processo e n??o apenas como um projeto que se finda com a antevis??o de cen??rios prospectivos e suas poss??veis solu????es estrat??gicas.