Cell homeostasis in a Leishmania major mutant overexpressing the spliced leader RNA is maintained by an increased proteolytic activity

Although several stage-specific genes have been identified in Leishmania, the molecular mechanisms governing developmental gene regulation in this organism are still not well understood. We have previously reported an attenuation of virulence in Leishmania major and L braziliensis carrying extra-copies of the spliced leader RNA gene. Here, we surveyed the major differences in proteome and transcript expression profiles between the spliced leader RNA overexpressor and control lines using two-dimensional gel electrophoresis and differential display reverse transcription PCR, respectively. Thirty-nine genes related to stress response, cytoskeleton, proteolysis, cell cycle control and proliferation, energy generation, gene transcription, RNA processing and post-transcriptional regulation have abnormal patterns of expression in the spliced leader RNA overexpressor line. The evaluation of proteolytic pathways in the mutant revealed a selective increase of cysteine protease activity and an exacerbated ubiquitin-labeled protein population. Polysome profile analysis and measurement of cellular protein aggregates showed that protein translation in the spliced leader RNA overexpressor line is increased when compared to the control line. We found that L major promastigotes maintain homeostasis in culture when challenged with a metabolic imbalance generated by spliced leader RNA surplus through modulation of intracellular proteolysis. However, this might interfere with a fine-tuned gene expression control necessary for the amastigote multiplication in the mammalian host. (c) 2010 Elsevier Ltd. All rights reserved.

The macrophage-derived neutrophil chemotactic factor, MNCF: A lectin with TNF-alpha-like activities on neutrophils

The macro phage-derived neutrophil chemotactic factor (MNCF) is an alpha-galactoside-binding lectin, known to induce dexamethasone-insensitive neutrophil recruitment. We further characterized MNCF effects on neutrophils and showed that it shares with TNF-alpha the ability to delay apoptosis and to trigger degranulation. MNCF and TNF-alpha effects show similar kinetics and involve Src kinases and MAPKinases dependent pathways. They were, however, clearly distinguished, since the soluble TNF-receptor etanercept prevented TNF but not MNCF effects, while melibiose disaccharide inhibited MNCF but not TNF effects. Absorption of MNCF on detoxi-gel did not alter its properties, precluding an LPS contamination effect. By contrast, galectin-3 required LPS to activate neutrophils. Specific antibodies allowed to further demonstrate that MNCF and galectin-3 are two distinct molecules. Finally, MNCF- and IL-8-induced neutrophil activation differed by their kinetic and sensitivity to pertussis toxin. In conclusion, MNCF is a distinct neutrophil agonist, with pro-inflammatory activities involving its carbohydrate recognition domain. (C) 2008 Elsevier Inc. All rights reserved.

Antimicrobial Activities of Ethanol Extract and Coumestans from Eclipta alba (L.) Hassk (Asteraceae)

Ethanol extract and fractions from aerial parts of Eclipta alba (L.) Hassk (Asteraceae) were screened for the antibacterial and antifungal activities against different species of human pathogenic bacterial ATCC, antibiotic-resistant clinical isolates and strains of the dermatophyte Trichophyton rubrum (wild and mutant for TruMDR2 gene) using a microdilution method. Demethylwedelolactone/wedelolactone (DWL/WL) and only wedelolactone (WL), both in a high homogeneity degree, were efficient to inhibit the ATCC strains of Staphylococus aureus (Minimal Inhibitory Concentration MIC = 75 mu g/mL), Staphylococcus epidemidis (MIC = 125 mu g/mL) and Escherichia coli (MIC = 125 mu g/mL) as well as antibiotic-resistant clinical isolates of Enterococcus spp (MIC = 250 mu g/mL) and S. aureus (MIC = 125 mu g/mL). Ethanol extract was more effective than the purified fractions against Trichophyton rubrum strains (MIC = 125 mu g/mL), suggesting that anti-fungal activity is not only related to demethylwedelolactone and wedelolactone, but also to a synergistic action between these coumestans and other compounds found in that extract. Thus, this work suggests that E. alba possesses a significant antimicrobial activity, including that against multi-drug resistant microorganisms, which could be of relevance for the treatment of infectious diseases.

Inhibition of Snake Venoms and Phospholipases A(2) by Extracts from Native and Genetically Modified Eclipta alba: Isolation of Active Coumestans

We genetically modified Eclipta alba using Agrobacterium rhizogenes LBA 9402, with the aim of producing secondary metabolites with pharmacological properties against phospholipase A(2) and the myotoxic activities of snake venom. Extracts from in natura aerial parts and roots, both native and genetically modified (in vitro), were prepared and analysed by high-performance liquid chromatography. In natura materials showed the coumestan wedelolactone at higher concentration in the aerial parts, while demethylwedelolactone appeared at higher concentration in roots. Among the modified roots, clone 19 showed higher concentrations of these coumestans. Our results show that the in natura extracts of plants collected from Botucatu and Ribeirao Preto were efficient in inhibiting snake venom phospholipase A(2) activity. Regarding in vitro material, the best effect against Crotalus durissus terrificus venom was that of clone 19. Clone 19 and isolated coumestans (wedelolactone and demethylwedelolactone) inhibited the myotoxic activity induced by basic phospholipases A(2) isolated from the venoms of Crotalus durissus terrificus (CB) and Bothrops jararacussu (BthTX-I and II). The search for antivenom is justified by the need of finding active principles that are more efficient in neutralizing snake venoms and also as an attempt to complement serum therapy.

Transferability and characterization of nine microsatellite markers for the tropical tree species Tabebuia roseo-alba

Microsatellite loci that were previously developed in the tropical tree Tabebuia aurea were used for the genetic analysis of Tabebuia roseo-alba populations. Nine of 10 simple sequence repeat markers were amplified, and the polymorphism was assessed in 58 individuals sampled from two stands in southeastern Brazil. All loci were polymorphic with Mendelian inheritance. The allele numbers were high, ranging from 5 to 13 in population I and 3 to 7 in population II, with means of 8.9 and 5.5, respectively. We conclude that these markers can be efficiently used for parentage and gene-flow studies.

Multiple maps and activity-dependent representational plasticity in the anterior Wulst of the adult barn owl (Tyto alba)

In the present study we addressed the issue of somatosensory representation and plasticity in a nonmammalian species, the barn owl. Multiunit mapping techniques were used to examine the representation of the specialized receptor surface of the claw in the anterior Wulst. We found dual somatotopic mirror image representations of the skin surface of the contralateral claw. In addition, we examined both representations 2 weeks after denervation of the distal skin surface of a single digit. In both representations, the denervated digital representation became responsive to stimulation of the adjacent, mutually functional, digit. The mutability and multiple representations indicates that the Wulst provides the owl with sensory processing capabilities analogous to those in mammals.

Pseudotumor inflamatório de seios paranasais

Pseudotumor inflamatório pode ser definido como uma lesão que simula neoplasia clínica e radiologicamente. Não se trata de uma entidade clínico-patológica única, mas um termo genérico para qualquer lesão expansiva inflamatória crônica inespecífica. Há poucos relatos de pseudotumor inflamatório em cavidade nasal e seios paranasais. RELATO DE CASOS: Apresentamos três casos de pseudotumor inflamatório em nariz e seios paranasais atendidos na Divisão de Clínica Otorrinolaringológica do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo. DISCUSSÃO: Pseudotumor inflamatório de seios paranasais apresenta sintomatologia variável conforme o local de origem. Geralmente, segue curso benigno, porém pode ser localmente agressivo.

Las Memorias del duque de Sully (o los avatares del primer proyecto de unión europea)

El presente artículo se consagra al análisis de la redacción, edición y difusión de las Memorias del duque de Sully. Con ese propósito, la prim-era sección refiere el contexto que rodea la preparación de la obra; la segunda presenta las características generales de la confederación y la tercera profundiza en las discrepancias de sus versiones; en la cuarta, quinta y la sexta se estudian la cronología y las etapas de la redacción, los cambios operados en los manuscritos y sus vías de difusión; la séptima examina el debate sobre la primacía histórica de las Memorias. El artículo pretende reconstruir las condiciones que determinan la elaboración y recepción del primer proyecto paneuropeo.

Temas e regiões nas políticas externas de Lula e Fernando Henrique: comparação do discurso dos dois presidentes

Neste artigo, comparamos o discurso sobre a política externa dos dois presidentes que governaram o País nos últimos 16 anos (Fernando Henrique e Lula). Por meio da técnica de análise de conteúdo, foram avaliados mais de mil pronunciamentos realizados em eventos diplomáticos no Brasil e no exterior. Encontramos semelhanças e diferenças importantes, tanto no que diz respeito às temáticas abordadas quanto nos parceiros que buscaram na arena internacional. Ambos os presidentes privilegiaram os temas referentes à economia, às instituições internacionais e ao relacionamento com os países da América do Sul. Fernando Henrique parece ter tido uma preocupação maior com a questão da democracia e aproximou-se mais dos países europeus. Lula estendeu a sua preocupação com o tema da desigualdade para a arena internacional e abriu maior diálogo com os países asiáticos, do Oriente Médio e, principalmente, da África.

A cooperação técnica do Brasil com a África: comparando os governos Fernando Henrique Cardoso (1995-2002) e Lula da Silva (2003-2010)

É consensual entre os analistas da Política Externa Brasileira o reconhecimento da priorização do eixo Sul-Sul nas estratégias de inserção internacional do país durante o governo Lula da Silva, bem como o papel de destaque concedido ao continente africano. Com o intuito de dar lastro empírico às análises sobre a retomada e repriorização das relações entre Brasil e África, este trabalho pretende analisar comparativamente a cooperação técnica provida pelo Brasil para os países africanos durante os governos Fernando Henrique Cardoso (1995-2002) e Lula da Silva (2003-2010). Procura-se identificar e comparar o número e os tipos de acordos firmados, a dispersão geográfica desses acordos e a diversidade temática dos projetos de cooperação técnica horizontal desenvolvidos pelo Brasil naquele continente. A fonte principal dos dados analisados, ainda pouco explorada pela literatura acadêmica brasileira, é o banco de dados do Departamento de Atos Internacionais do Ministério das Relações Exteriores.

Eficiência de adubos fosfatados associados ao enxofre elementar na cultura do milho

A eficiência agronômica dos adubos fosfatados pode ser afetada pelas fontes de fósforo, propriedades do solo, modo de aplicação e espécie vegetal. Com o objetivo de comparar a eficiência do fosfato natural reativo (FNR) e do superfosfato triplo (SFT) associado ao enxofre elementar, aplicados no sulco de plantio na cultura do milho, instalouse um experimento em condições de campo no município de Toledo, Paraná, em um Latossolo Vermelho eutroférrico. O delineamento experimental consistiu de blocos inteiramente casualizados, em esquema fatorial 2x4x2, sendo duas fontes de fósforo (FNR e SFT), três doses de adubação (100, 200 e 300 kg ha-1 de P2O5) mais uma testemunha para cada fonte, e adição ou não de enxofre elementar (0 e 30 kg ha-1de S°), aplicados no sulco de plantio em quatro repetições. As parcelas foram constituídas por quatro linhas com 6 m de comprimento espaçadas 0,9 m, utilizando-se como área útil as duas linhas centrais desprezando-se 0,5 m das extremidades. A adubação com N e K e os tratamentos com P e S° foram realizados manualmente no sulco de plantio, sendo posteriormente semeadas com o híbrido Agroceres 8021. O SFT proporcionou maior rendimento de grãos, em comparação com o FNR, no primeiro ano de cultivo. A eficiência agronômica do FNR foi de 43% e 33% na presença e ausência de S, respectivamente. O teor de fósforo no tecido foliar foi inferior com o uso do FNR. A dose de 30 kg ha-1 de S elementar não interferiu no pH do solo, mas aumentou a eficiência da adubação fosfatada.

Spacing, population density and nitrogen fertilization in corn grown in an Oxisoil

The objective of this study was to evaluate yield components, leaf nitrogen content and grain yield in corn as affected by row spacing, plant density and nitrogen topdressing. The experiment was conducted with the single-cross hybrid AG 8021, in the municipality of Toledo-PR, in an Oxisoil under no-tillage system, in the crop year 2005/ 2006. The experiment was arranged in a randomized block design and treatments in split-split-plots, with four replications. The two row spacings (0.45 and 0.90 m) were allocated in the main plots, the two plant densities (60,000 and 80,000 plants ha-1) were allocated in the subplots and the three nitrogen rates (80, 100, 120 and 140 kg ha-1 N) were allocated in the sub-subplots. Topdress nitrogen was applied using urea as N source. The rise of the plant population from 60,000 to 80,000 plants ha-1 and the application of topdress nitrogen resulted in increased production components. The application of topdress fertilization provided increase in leaf N content and grain yield for the spacings 0.45 m and 0.90 m. Yield was higher in the spacing 0.45 m than 0.90 m. Yield was higher with 60,000 plants than with 80,000 plants at 0.90 m, while at 0.45 m there was no difference in relation to the plant density.