Dominant Negative Mutants of Bacillus thuringiensis Cry1Ab Toxin Function as Anti-Toxins: Demonstration of the Role of Oligomerization in Toxicity

BACKGROUND:Bacillus thuringiensis Cry toxins, that are used worldwide in insect control, kill insects by a mechanism that depends on their ability to form oligomeric pores that insert into the insect-midgut cells. These toxins are being used worldwide in transgenic plants or spray to control insect pests in agriculture. However, a major concern has been the possible effects of these insecticidal proteins on non-target organisms mainly in ecosystems adjacent to agricultural fields. METHODOLOGY/PRINCIPAL FINDINGS:We isolated and characterized 11 non-toxic mutants of Cry1Ab toxin affected in different steps of the mechanism of action namely binding to receptors, oligomerization and pore-formation. These mutant toxins were analyzed for their capacity to block wild type toxin activity, presenting a dominant negative phenotype. The dominant negative phenotype was analyzed at two levels, in vivo by toxicity bioassays against susceptible Manduca sexta larvae and in vitro by pore formation activity in black lipid bilayers. We demonstrate that some mutations located in helix alpha-4 completely block the wild type toxin activity at sub-stoichiometric level confirming a dominant negative phenotype, thereby functioning as potent antitoxins. CONCLUSIONS/SIGNIFICANCE:This is the first reported case of a Cry toxin dominant inhibitor. These data demonstrate that oligomerization is a fundamental step in Cry toxin action and represent a potential mechanism to protect special ecosystems from the possible effect of Cry toxins on non-target organisms

Applying Bayesian model averaging for uncertainty estimation of input data in energy modelling

Background Energy scenarios that are used for policy advice have ecological and social impact on society. Policy measures that are based on modelling exercises may lead to far reaching financial and ecological consequences. The purpose of this study is to raise awareness that energy modelling results are accompanied with uncertainties that should be addressed explicitly. Methods With view to existing approaches of uncertainty assessment in energy economics and climate science, relevant requirements for an uncertainty assessment are defined. An uncertainty assessment should be explicit, independent of the assessor’s expertise, applicable to different models, including subjective quantitative and statistical quantitative aspects, intuitively understandable and be reproducible. Bayesian model averaging for input variables of energy models is discussed as method that satisfies these requirements. A definition of uncertainty based on posterior model probabilities of input variables to energy models is presented. Results The main findings are that (1) expert elicitation as predominant assessment method does not satisfy all requirements, (2) Bayesian model averaging for input variable modelling meets the requirements and allows evaluating a vast amount of potentially relevant influences on input variables and (3) posterior model probabilities of input variable models can be translated in uncertainty associated with the input variable. Conclusions An uncertainty assessment of energy scenarios is relevant if policy measures are (partially) based on modelling exercises. Potential implications of these findings include that energy scenarios could be associated with uncertainty that is presently neither assessed explicitly nor communicated adequately

The Linear Response of ENSO to the Madden–Julian Oscillation

Abstract The possibility that the tropical Pacific coupled system linearly amplifies perturbations produced by the Madden–Julian oscillation (MJO) is explored. This requires an estimate of the low-frequency tail of the MJO. Using 23 yr of NCEP–NCAR reanalyses of surface wind and Reynolds SST, we show that the spatial structure that dominates the intraseasonal band (i.e., the MJO) also dominates the low-frequency band once the anomalies directly related to ENSO have been removed. This low-frequency contribution of the intraseasonal variability is not included in most ENSO coupled models used to date. Its effect in a coupled model of intermediate complexity has, therefore, been studied. It is found that this “MJO forcing” (τMJO) can explain a large fraction of the interannual variability in an asymptotically stable version of the model. This interaction is achieved via linear dynamics. That is, it is the cumulative effect of individual events that maintains ENSOs in this model. The largest coupled wind anomalies are initiated after a sequence of several downwelling Kelvin waves of the same sign have been forced by τMJO. The cumulative effect of the forced Kelvin waves is to persist the (small) SST anomalies in the eastern Pacific just enough for the coupled ocean–atmosphere dynamics to amplify the anomalies into a mature ENSO event. Even though τMJO explains just a small fraction of the energy contained in the stress not associated with ENSO, a large fraction of the modeled ENSO variability is excited by this forcing. The characteristics that make τMJO an optimal stochastic forcing for the model are discussed. The large zonal extent is an important factor that differentiates the MJO from other sources of stochastic forcing

Sensitivity of Hybrid ENSO Models to Unresolved Atmospheric Variability

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Maturation and fertilization in vitro of Pteria sterna (Gould, 1851, Mollusca)

 P. sterna oocytes cannot be fertilized in vitro unless they are obtained by inducing spawning. The present work was undertaken to develop a simple method to induce germinal vesicle breakdown that would enable the oocytes to be fertilized without prior spawning. We found that Pteria sterna oocytes removed from the ovaries are induced to undergo germinal vesicle breakdown (GVBD) by 10 mM NH4Cl in seawater, and proceed to metaphase I of meiosis, at which stage they can be fertilized and will develop at least as far as "D" larvae. On the other hand, ovarian oocytes exposed to 2 × 10–5 M serotonin in seawater remained in prophase and did not undergo GVBD. Ammonium chloride treatment provides a simple, inexpensive procedure that can be useful in studies of fertilization and development, and for attempts to repopulate this exploited species