Production of the baculovirus-expressed dengue virus glycoprotein NS1 can be improved dramatically with optimised regimes for fed-batch cultures and the addition of the insect moulting hormone, 20-Hydroxyecdysone

A perennial problem in recombinant protein expression is low yield of the product of interest. A strategy which has been shown to increase the production of baculovirus-expressed proteins is to utilise fed-batch cultures. One disadvantage of this approach is the time-consuming task of optimising the feeding strategy. Previously, a statistical optimisation routine was applied to develop a feeding strategy that increased the yield of beta-Galactosidase (beta-Gal) by 2.4-fold (Biotechnol. Bioeng, 59 (1998) 178). This involves the single addition of nutrient concentrates (amino acids, lipids. glucose and yeastolate ultrafiltrate) into Sf9 cell cultures grown in SF900II medium. In this study, it is demonstrated that this optimised fed-batch strategy developed for a high-yielding intracellular product beta-Gal could be applied successfully to a relatively low-yielding glycosylated and secreted product such as the dengue virus glycoprotein NS1. Optimised batch infections yielded 4 mug/ml of NS1 at a peak cell density of 4.2 x 10(6) cells/ml. In contrast. optimised fed-batch infections exhibited a 3-fold improvement in yield, with 12 mug ml of NS1 produced at a peak cell density of 11.3 x 10(6) cells/ml. No further improvements in yield were recorded when the feed volumes were doubled and the peak cell density was increased to 23 x 10(6) cells/ml, unless the cultures were stimulated by the addition of 4 mug/ml of 20-Hydroxyecdysone (an insect moulting hormone). In this case, the NS1 yield was increased to 20 mug/ml. which was nearly 5-fold higher than optimised batch cultures. (C) 2002 Elsevier Science B.V. All rights reserved.

Epidemiology and strain characteristics of Echinococcus granulosus in the Benghazi area of eastern Libya

The incidence of surgically confirmed cystic echinococcosis in eastern Libya was estimated to be at least 4.2 cases/100,000, with significantly more female cases than male. The prevalences of infection with Echinococcus granulosus among 1087 sheep, 881 goats, 428 camels and 614 cattle from the same region, determined postmortem in abattoirs, were 20%, 3.4%, 13.6% and 11%, respectively. Infection in the livestock was age-dependent and, generally, the female animals were more often infected than the male. The measurements of rostellar hooks on protoscoleces collected from sheep and cattle were similar but significantly different from the corresponding measurements of parasites of human or camel origin. However, when a portion of the cytochrome c-oxidase subunit I (coxl) gene from each of 30 protoscolex samples from Libya (12 from cattle, three from humans, five from camels and 10 from sheep) was sequenced, the sequences were all found to be identical to that published for the common sheep strain of E. granulosus.

Changes in postural stability in women aged 20 to 80 years

Background. A study of postural stability was undertaken to identify the relationship between vision and support surface across age decades. Understanding when reliance on vision for postural stability emerges and the support conditions contributing to this instability may provide the evidence required to introduce falls-prevention strategies in younger age decades. Methods. We measured postural stability in 453 women aged 20 to 80 years using the Balance Master force-plate system while the women performed the modified Clinical Test for the Sensory Interaction and Balance (firm and foam surfaces, eyes open and closed) and the Single-Limb Stance Test (eyes open and closed). Results. Women in their 60s and 70s were more unstable than younger women in bilateral stance on a firm surface with the eyes closed. This instability was evident from the 50s when a foam surface was introduced and from the 40s when single-limb stance was tested with eyes closed. A further decline in stability was demonstrated for each subsequent decade when the eyes were closed in single-limb stance. Conclusions. Age, visual condition, and support surface were significant variables influencing postural stability in women. Reliance on vision for postural stability was evident for women from the 40s when single-limb stance was tested, from the 50s when bilateral stance on foam was tested, and from the 60s when a firm surface was used. The cause(s) of this decline in stability requires further investigation, and screening for postural instability between the ages of 40 and 60 is advocated.

Repetitive brief ischemia: Intermittent reperfusion during ischemia ameliorates the extent of injury in the perfused kidney

Acute renal failure commonly follows reduced renal perfusion or ischemia. Reperfusion is essential for recovery but can itself cause functional and structural injury to the kidney. The separate contributions of ischemia and of reperfusion were examined in the isolated perfused rat kidney. Three groups were studied: brief (5 min) ischemia, 20 min ischemia, and repetitive brief ischemia (4 periods of 5 min) with repetitive intervening reperfusion of 5 min. A control group had no intervention, the three ischemia groups were given a baseline perfusion of 30 min before intervention and all groups were perfused for a total of 80 min. In addition, the effects of exogenous (NO)-N-. from sodium nitroprusside and xanthine oxidase inhibition by allopurinol were assessed in the repetitive brief ischemia-reperfusion model. Brief ischemia produced minimal morphological injury with near normal functional recovery. Repetitive brief ischemia reperfusion caused less functional and morphological injury than an equivalent single period of ischemia (20 min) suggesting that intermittent reperfusion is less injurious than ischemia alone over the time course of study. Pretreatment with allopurinol improved renal function after repetitive brief ischemia-reperfusion compared with the allopurinol-untreated repetitive brief ischemia-reperfusion group. Similarly, sodium nitroprusside reduced renal vascular resistance but did not improve the glomerular filtration rate or sodium reabsorption in the repetitive brief ischemia-reperfusion model. Thus, these studies show that the duration of uninterrupted ischemia is more critical than reperfusion in determining the extent of renal ischemia-reperfusion injury and that allopurinol, in particular, counteracts the oxidative stress of reperfusion.

Amino acids 1-20 of the hepatitis C virus (HCV) core protein specifically inhibit HCV IRES- dependent translation in Hep G2 cells, and inhibit both HCV IRES- and cap-dependent translation in HuH7 and CV-1 cells.

A self-modulating mechanism by the hepatitis C virus (HCV) core protein has been suggested to influence the level of HCV replication, but current data on this subject are contradictory. We examined the effect of wild-type and mutated core protein on HCV IRES- and cap-dependent translation. The wild-type core protein was shown to inhibit both IRES- and cap-dependent translation in an in vitro system. This effect was duplicated in a dose-dependent manner with a synthetic peptide representing amino acids 1-20 of the HCV core protein. This peptide was able to bind to the HCV IRES as shown by a mobility shift assay. In contrast, a peptide derived from the hepatitis B virus (HBV) core protein that contained a similar proportion of basic residues was unable to inhibit translation or bind the HCV IRES. A recombinant vaccinia-HCV core virus was used to examine the effect of the HCV core protein on HCV IRES-dependent translation in cells and this was compared with the effects of an HBV core-recombinant vaccinia virus. In CV-1 and HuH7 cells, the HCV core protein inhibited translation directed by the IRES elements of HCV, encephalomyocarditis virus and classical swine fever virus as well as cap-dependent translation, whereas in HepG2 cells, only HCV IRES-dependent translation was affected. Thus, the ability of the HCV core protein to selectively inhibit HCV IRES-dependent translation is cell-specific. N-terminal truncated (aa 1-20) HCV core protein that was expressed from a novel recombinant vaccinia virus in cells abrogated the inhibitory phenotype of the core protein in vivo, consistent with the above in vitro data.

Synthesis of optically complex core-shell colloidal suspensions: Pathways to multiplexed biological screening

The ability to generate enormous random libraries of DNA probes via split-and-mix synthesis on solid supports is an important biotechnological application of colloids that has not been fully utilized to date. To discriminate between colloid-based DNA probes each colloidal particle must be 'encoded' so it is distinguishable from all other particles. To this end, we have used novel particle synthesis strategies to produce large numbers of optically encoded particle suitable for DNA library synthesis. Multifluorescent particles with unique and reproducible optical signatures (i.e., fluorescence and light-scattering attributes) suitable for high-throughput flow cytometry have been produced. In the spectroscopic study presented here, we investigated the optical characteristics of multi-fluorescent particles that were synthesized by coating silica 'core' particles with up to six different fluorescent dye shells alternated with non-fluorescent silica 'spacer' shells. It was observed that the diameter of the particles increased by up to 20% as a result of the addition of twelve concentric shells and that there was a significant reduction in fluorescence emission intensities from inner shells as an increasing number of shells were deposited.

Bridgehead arylation: A direct route to advanced intermediates for the synthesis of C-20 diterpene alkaloids

[GRAPHICS] Rapid access to the ABCE ring system of the C-20 diterpene alkaloids was achieved by silver(I)-promoted intramolecular Friedel-Crafts arylation of a functional group-specific 5-bromo-3-azabicyclo[3.3.1]nonane derivative.

Texto para discuss??o 20: a reforma administrativa francesa: da crise da fun????o p??blica a uma nova racionalidade da a????o coletiva, uma dif??cil transi????o

Um dos marcos da reforma francesa ?? a crise da gest??o p??blica. Sabemos que a crise da gest??o p??blica ?? um fen??meno hoje universal que se tornou vis??vel a partir da d??cada de 70, especificamente com a crise do petr??leo.Com o fim do conflito Leste/Oeste em 1989, esta crise torna-se mais evidente e v??m-se refor??adas suas conota????es ideol??gicas. No caso franc??s, ao lado desta crise mundial, os analistas identificam uma crise espec??fica, a saber, a fal??ncia do modelo de Estado. Dentre os pa??ses industrializados do Ocidente, o Estado franc??s ??, sem d??vida, um dos que mais interv??m na vida social, econ??mica e pol??tica

ENAP 20 anos: caminhos de uma escola de governo

Apresenta????o da trajet??ria da ENAP Escola Nacional de Administra????o P??blica desde sua funda????o, em 1996. Pe??a importante na forma????o de um servi??o p??blico competente, a ENAP tem sua atua????o focada no desenvolvimento de compet??ncias dos servidores para aumentar a capacidade de governo na gest??o de pol??ticas p??blicas

Os 20 anos da carreira de Especialista em Pol??ticas P??blicas e Gest??o Governamental do governo federal e seu papel estrat??gico para o aprimoramento da gest??o e das pol??ticas p??blicas

O trabalho visa apresentar o atual est??gio de desenvolvimento dos processos de gest??o da carreira de Especialista em Pol??ticas P??blicas e Gest??o Governamental do governo federal. Assim, discorrer-se-?? sobre o modelo de gest??o adotado ?? luz dos estudos e discuss??es realizados pela Secretaria de Gest??o e consultores externos, com a participa????o dos pr??prios gestores. Essas reflex??es, realizadas ao longo do ??ltimo ano, permitiram identificar as estrat??gias que devem orientar todos os processos relacionados ?? carreira. O trabalho perpassa os temas: defini????o do perfil do ocupante do cargo de Especialista em Pol??ticas P??blicas e Gest??o Governamental; diretrizes para aprimorar o concurso p??blico; o processo de identifica????o das compet??ncias a serem desenvolvidas pelos seus membros, estrat??gias utilizadas para definir a aloca????o inicial; o acompanhamento do est??gio probat??rio; e algumas quest??es relacionadas ?? mobilidade funcional dos membros desta carreira. Constata-se que estas iniciativas refor??am a presen??a dos membros da carreira na condu????o das principais pol??ticas p??blicas e intensificam sua atua????o em rede, o que resulta em impactos positivos para a formula????o e execu????o de pol??ticas complexas, que demandam o envolvimento de m??ltiplos atores.

Alianças e coalizões internacionais do governo Lula: o Ibas e o G-20

O objetivo deste artigo é demonstrar que o protagonismo internacional do governo Lula busca ressuscitar a questão do desenvolvimento tendo como base tanto o regionalismo, quanto a recuperação da clivagem Norte/Sul na política internacional em benefício das novas demandas dos países em desenvolvimento. Baseado nessa constatação, indagamos se as estratégias internacionais e o diálogo Sul-Sul presente na retórica e na ação da política externa do governo Lula representam alianças duradouras ou coalizões efêmeras nas relações internacionais contemporâneas entre os países em desenvolvimento e menor desenvolvidos.

A Governança econômica global e os desafios do G-20 pós-crise financeira: análise das posições de Estados Unidos, China, Alemanha e Brasil

A crise financeira de 2008 teve impactos significativos no capitalismo global, sendo um de seus reflexos na estrutura da governança global a constituição e evolução do G-20. Neste contexto, o objetivo do artigo é analisar tais mudanças e, em especial, as posições de quatro dos principais atores nas cúpulas do G-20, a saber: Estados Unidos, China, Alemanha e Brasil.

China's proposing behavior in Global Governance: the cases of the WTO Doha Round negotiation and G-20 process

This article examines China's proposals on the reform of global governance, and discusses the main features of China's proposing behavior in the cases of the WTO Doha Round negotiation and G-20 Process. The main findings are: (1) in the critical junctures of global governance reform, China engaged the reform of the global governance institutions proactively, and put forward a series of reform proposals; (2) in proposing behavior, China argued the global governance institutions should be properly adjusted without intention to change the basic principles, refrained from playing a leadership role while proposing jointly with other countries, and upheld the principled idea of pro-development.