A theoretical model of the tridimensional structure of Bacillus thuringiensis subsp. medellin Cry 11Bb toxin deduced by homology modelling

Cry11Bb is an insecticidal crystal protein produced by Bacillus thuringiensis subsp. medellin during its stationary phase; this ¶-endotoxin is active against dipteran insects and has great potential for mosquito borne disease control. Here, we report the first theoretical model of the tridimensional structure of a Cry11 toxin. The tridimensional structure of the Cry11Bb toxin was obtained by homology modelling on the structures of the Cry1Aa and Cry3Aa toxins. In this work we give a brief description of our model and hypothesize the residues of the Cry11Bb toxin that could be important in receptor recognition and pore formation. This model will serve as a starting point for the design of mutagenesis experiments aimed to the improvement of toxicity, and to provide a new tool for the elucidation of the mechanism of action of these mosquitocidal proteins.

Monitoring resistance to Bacillus thuringiensis subsp. israelensis in the field by performing bioassays with each Cry toxin separately

Bacillus thuringiensis subsp. israelensis (Bti) is increasingly used worldwide for mosquito control and is the only larvicide used in the French Rhône-Alpes region since decades. The artificial selection of mosquitoes with field-persistent Bti collected in breeding sites from this region led to a moderate level of resistance to Bti, but to relatively high levels of resistance to individual Bti Cry toxins. Based on this observation, we developed a bioassay procedure using each Bti Cry toxin separately to detect cryptic Bti-resistance evolving in field mosquito populations. Although no resistance to Bti was detected in none of the three mosquito species tested (Aedes rusticus, Aedes sticticus and Aedes vexans), an increased tolerance to Cry4Aa (3.5-fold) and Cry11Aa toxins (8-fold) was found in one Ae. sticticus population compared to other populations of the same species, suggesting that resistance to Bti may be arising in this population. This study confirms previous works showing a lack of Bti resistance in field mosquito populations treated for decades with this bioinsecticide. It also provides a first panorama of their susceptibility status to individual Bti Cry toxins. In combination with bioassays with Bti, bioassays with separate Cry toxins allow a more sensitive monitoring of Bti-resistance in the field.

CAVIN-3 regulates circadian period length and PER:CRY protein abundance and interactions.

In mammals, transcriptional autorepression by Period (PER) and Cryptochrome (CRY) protein complexes is essential for the generation of circadian rhythms. We have identified CAVIN-3 as a new, cytoplasmic PER2-interacting protein influencing circadian clock properties. Thus, CAVIN-3 loss- and gain-of-function shortened and lengthened, respectively, the circadian period in fibroblasts and affected PER:CRY protein abundance and interaction. While depletion of protein kinase Cδ (PKCδ), a known partner of CAVIN-3, had little effect on circadian gene expression, CAVIN-3 required the PKCδ-binding site to exert its effect on period length. This suggests the involvement of yet uncharacterized protein kinases. Finally, CAVIN-3 activity in circadian gene expression was independent of caveolae.

Respostas da lagarta‑do‑cartucho a milho geneticamente modificado expressando a toxina Cry 1A(b)

O objetivo deste trabalho foi avaliar parâmetros biológicos da lagarta-do-cartucho do milho (Spodoptera frugiperda), alimentada com híbridos de milho Bt, que expressam a toxina Cry 1A(b), e com seus respectivos isogênicos não Bt. Os experimentos foram realizados no laboratório da Embrapa Milho e Sorgo, em Sete Lagoas, MG. Os parâmetros avaliados foram: sobrevivência de larvas após 48 horas, sobrevivência da fase larval e pré-imaginal, biomassa de larvas aos 14 dias de idade, biomassa de pupas, período de desenvolvimento larval, e não preferência alimentar de larvas do primeiro ínstar. Larvas de S. frugiperda apresentam menor sobrevivência nas primeiras 48 horas de alimentação e durante toda a fase larval, na maioria dos híbridos de milho Bt, em comparação ao milho não Bt. A biomassa de larvas e pupas foi sempre menor no milho Bt, e o período larval e o pré-imaginal, maior. Houve interação entre a toxina Cry 1A(b) e a base genética dos híbridos transgênicos, quanto à sobrevivência e à biomassa larval. Larvas recém-eclodidas de S. frugiperda apresentam preferência pela alimentação em milho não Bt.

Identificação e caracterização de genes vip e cry coleóptero‑específicos em isolados de Bacillus thuringiensis

O objetivo deste trabalho foi identificar e caracterizar os genes cry3, vip1, vip2 e vip1/vip2 em uma coleção de 1.078 isolados de Bacillus thuringiensis potencialmente tóxicos para larvas de coleópteros. Foram utilizados pares de oligonucleotídeos iniciadores gerais obtidos a partir de regiões conservadas dos genes e do alinhamento de sequências consenso. Posteriormente, os isolados positivos foram caracterizados por meio da técnica de PCR‑RFLP, tendo-se utilizado enzimas de restrição específicas, para identificar novas subclasses de genes nos isolados. Cento e cinquenta e um isolados foram positivos para os genes avaliados, com maior frequência para o gene vip1/vip2 (139 isolados). Pela técnica de PCR‑RFLP, foram observados 14 perfis polimórficos, o que indica a presença de diferentes alelos e, consequentemente, de distintas subclasses desses genes.

Weep and cry

Soitinnus: trumpetti, bassoklarinetti, piano, kontrabasso, rummut.

Sleeping with half open eyes: dreams and realities in The Cry of the Owl

This article is a close analysis of The Cry of the Owl (Thraves, 2009). It is also part of larger project to bring together traditions of detailed criticism with those of production history, which culminates in second article on the film due to be published in 2011. The detail of the argument concerns analysing a range of the film’s key signifying systems, with a particular interest in the way the film explores the gap between images / impressions and characters’ realities; engages in a complex way with generic traditions and modes of address; establishes complex patterns of connection and contrast through blocking, camera strategies and narrative structure.

The cry of the owl: investigating decision-making in a contemporary feature film

This article extends the traditions of style-based criticism through an encounter with the insights that can be gained from engaging with filmmakers at work. By bringing into relationship two things normally thought of as separate: production history and disinterested critical analysis, the discussion aims to extend the subjects which criticism can appreciate as well as providing some insights on the creative process. Drawing on close analysis, on observations made during fieldwork and on access to earlier cuts of the film, this article looks at a range of interrelated decision-making anchored by the reading of a particular sequence. The article examines changes the film underwent in the different stages of production, and some of the inventions deployed to ensure key themes and ideas remained in play, as other elements changed. It draws conclusions which reveal perspectives on the filmmaking process, on collaboration, and on the creative response to material realities. The article reveals elements of the complexity of the process of the construction of image and soundtrack, and extends the range of filmmakers’ choices which are part of a critical dialogue. Has a relationship to ‘Sleeping with half open eyes: dreams and realities in The Cry of the Owl’, Movie: A Journal of Film Criticism, 1, (2010) which provides a broader interpretative context for the enquiry.

Voorhees, A Far Cry From the Old School

Clipping from unidentified student publication of Voorhees Institute of Technology and the Voorhees Campus of New York City Community College.

Identificação e caracterização de genes vip e cry coleóptero-específicos em isolados de Bacillus thuringiensis

O objetivo deste trabalho foi identificar e caracterizar os genes cry3, vip1, vip2 e vip1/vip2 em uma coleção de 1.078 isolados de Bacillus thuringiensis potencialmente tóxicos para larvas de coleópteros. Foram utilizados pares de oligonucleotídeos iniciadores gerais obtidos a partir de regiões conservadas dos genes e do alinhamento de sequências consenso. Posteriormente, os isolados positivos foram caracterizados por meio da técnica de PCR‑RFLP, tendo-se utilizado enzimas de restrição específicas, para identificar novas subclasses de genes nos isolados. Cento e cinquenta e um isolados foram positivos para os genes avaliados, com maior frequência para o gene vip1/vip2 (139 isolados). Pela técnica de PCR‑RFLP, foram observados 14 perfis polimórficos, o que indica a presença de diferentes alelos e, consequentemente, de distintas subclasses desses genes.

Isolation of Bacillus thuringiensis strains that contain Dipteran-specific cry genes from Ilha Bela (São Paulo, Brazil) soil samples

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)