Molecular Lego of Human Cytochrome P450: The Key Role of Heme Domain Flexibility for the Activity of the Chimeric Proteins

The cytochrome P450 superfamily are heme-thiolate enzymes able to carry out monooxygenase reactions. Several studies have demonstrated the feasibility of using a soluble bacterial reductase from Bacillus megaterium, BMR, as an artificial electron transfer partner fused to the human P450 domain in a single polypeptide chain in an approach known as ‘molecular Lego’. The 3A4-BMR chimera has been deeply characterized biochemically for its activity, coupling efficiency, and flexibility by many different biophysical techniques leading to the conclusion that an extension of five glycines in the loop that connects the two domains improves all the catalytic parameters due to improved flexibility of the system. In this work, we extend the characterization of 3A4-BMR chimeras using differential scanning calorimetry to evaluate stabilizing role of BMR. We apply the ‘molecular Lego’ approach also to CYP19A1 (aromatase) and the data show that the activity of the chimeras is very low (<0.003 min−1) for all the constructs tested with a different linker loop length: ARO-BMR, ARO-BMR-3GLY, and ARO-BMR-5GLY. Nevertheless, the fusion to BMR shows a remarkable effect on thermal stability studied by differential scanning calorimetry as indicated by the increase in Tonset by 10 °C and the presence of a cooperative unfolding process driven by the BMR protein domain. Previously characterized 3A4-BMR constructs show the same behavior of ARO-BMR constructs in terms of thermal stabilization but a higher activity as a function of the loop length. A comparison of the ARO-BMR system to 3A4-BMR indicates that the design of each P450-BMR chimera should be carefully evaluated not only in terms of electron transfer, but also for the biophysical constraints that cannot always be overcome by chimerization

New insights into structural and functional roles of indole-3-acetic acid (IAA) : Changes in DNA topology and gene expression in bacteria

Indole-3-acetic acid (IAA) is a major plant hormone that affects many cellular processes in plants, bacteria, yeast, and human cells through still unknown mechanisms. In this study, we demonstrated that the IAA-treatment of two unrelated bacteria, the Ensifer meliloti 1021 and Escherichia coli, harboring two different host range plasmids, influences the supercoiled state of the two plasmid DNAs in vivo. Results obtained from in vitro assays show that IAA interacts with DNA, leading to DNA conformational changes commonly induced by intercalating agents. We provide evidence that IAA inhibits the activity of the type IA topoisomerase, which regulates the DNA topological state in bacteria, through the relaxation of the negative supercoiled DNA. In addition, we demonstrate that the treatment of E. meliloti cells with IAA induces the expression of some genes, including the ones related to nitrogen fixation. In contrast, these genes were significantly repressed by the treatment with novobiocin, which reduces the DNA supercoiling in bacterial cells. Taking into account the overall results reported, we hypothesize that the IAA action and the DNA structure/function might be correlated and involved in the regulation of gene expression. This work points out that checking whether IAA influences the DNA topology under physiological conditions could be a useful strategy to clarify the mechanism of action of this hormone, not only in plants but also in other unrelated organisms

Attention Capture by Direct Gaze is Robust to Context and Task Demands

The final publication is available at Springer via https://dx.doi.org/10.1007/s10919-011-0128-z.Eye-tracking was used to investigate whether gaze direction would influence the visual scanning of faces, when presented in the context of a full character, in different social settings, and with different task demands. Participants viewed individual computer agents against either a blank background or a bar scene setting, during both a free-viewing task and an attractiveness rating task for each character. Faces with a direct gaze were viewed longer than faces with an averted gaze regardless of body context, social settings, and task demands. Additionally, participants evaluated characters with a direct gaze as more attractive than characters with an averted gaze. These results, obtained with pictures of computer agents rather than real people, suggest that direct gaze is a powerful attention grabbing stimulus that is robust to background context or task demands.103305-1/Canadian Institutes of Health Research89822-1/Canadian Institutes of Health Research103305-1/PHS HHS/United States89822-1/PHS HHS/United State

The use of different adhesive filling material and mass combinations to restore class II cavities under loading and shrinkage effects: a 3D-FEA

3D tooth models were virtually restored: flowable composite resin + bulk-fill composite (A), glass ionomer cement + bulk-fill composite (B) or adhesive + bulk-fill composite (C). Polymerization shrinkage and masticatory loads were simulated. All models exhibited the highest stress concentration at the enamel–restoration interfaces. A and C showed similar pattern with lower magnitude in A in comparison to C. B showed lower stress in dentine and C the highest cusps displacement. The use of glass ionomer cement or flowable composite resin in combination with a bulk-fill composite improved the biomechanical behavior of deep class II MO cavities

Assessing the effects of Bt maize on the non-target pest Rhopalosiphum maidis by demographic and life-history measurement endpoints

The most commercialized Bt maize plants in Europe were transformed with genes which express a truncated form of the insecticidal delta-endotoxin (Cry1Ab) from the soil bacterium Bacillus thuringiensis (Bt) specifically against Lepidoptera. Studies on the effect of transgenic maize on non-target arthropods have mainly converged on beneficial insects. However, considering the worldwide extensive cultivation of Bt maize, an increased availability of information on their possible impact on non-target pests is also required. In this study, the impact of Bt-maize on the non-target corn leaf aphid, Rhopalosiphum maidis, was examined by comparing biological traits and demographic parameters of two generations of aphids reared on transgenic maize with those on untransformed near-isogenic plants. Furthermore, free and bound phenolics content on transgenic and near-isogenic plants were measured. Here we show an increased performance of the second generation of R. maidis on Bt-maize that could be attributable to indirect effects, such as the reduction of defense against pests due to unintended changes in plant characteristics caused by the insertion of the transgene. Indeed, the comparison of Bt-maize with its corresponding near-isogenic line strongly suggests that the transformation could have induced adverse effects on the biosynthesis and accumulation of free phenolic compounds. In conclusion, even though there is adequate evidence that aphids performed better on Bt-maize than on non-Bt plants, aphid economic damage has not been reported in commercial Bt corn fields in comparison to non-Bt corn fields. Nevertheless, Bt-maize plants can be more easily exploited by R. maidis, possibly due to a lower level of secondary metabolites present in their leaves. The recognition of this mechanism increases our knowledge concerning how insect-resistant genetically modified plants impact on non-target arthropods communities, including tritrophic web interactions, and can help support a sustainable use of genetically modified crops

The use of different adhesive filling material and mass combinations to restore class II cavities under loading and shrinkage effects: a 3D-FEA

3D tooth models were virtually restored: flowable composite resin + bulk-fill composite (A), glass ionomer cement + bulk-fill composite (B) or adhesive + bulk-fill composite (C). Polymerization shrinkage and masticatory loads were simulated. All models exhibited the highest stress concentration at the enamel-restoration interfaces. A and C showed similar pattern with lower magnitude in A in comparison to C. B showed lower stress in dentine and C the highest cusps displacement. The use of glass ionomer cement or flowable composite resin in combination with a bulk-fill composite improved the biomechanical behavior of deep class II MO cavities

Characterization of Strombolian events by using independent component analysis

We apply Independent Component Analysis (ICA) to seismic signals recorded at Stromboli volcano. Firstly, we show how ICA works considering synthetic signals, which are generated by dynamical systems. We prove that Strombolian signals, both tremor and explosions, in the high frequency band (>0.5 Hz), are similar in time domain. This seems to give some insights to the organ pipe model generation for the source of these events. Moreover, we are able to recognize in the tremor signals a low frequency component (<0.5 Hz), with a well defined peak corresponding to 30s

The Transformation of the Great Decommissioned Farini Railroad Yard: The Research for a Modern Housing Settlement

In 2016, the Public Company for Social Services of the Municipality of Bologna "ASP City of Bologna" requires the ABC Department to analyze its huge real estate portfolio with the aim to identify efficiency strategies to free up resources to be reinvested in the offer of personal social services. Corporate governance sets up its strategy by coherently matching: the statutory values, the corporate mission and the growing need to respond to the needs expressed by the community in terms of providing personal services. The collaboration found strong foundations of understanding in the intention to pursue the mission of general social interest with environmental and economic sustainability, in a research path that made explicit the values of transparency and accountability of the Company. On the one hand, the research outlined the levers for making management, resources and processes more efficient, and on the other, strategies for enhancing assets in a social and circular economy perspective (see Bilancio Sociale ASP 2017)

High Frequency Peakers: young radio sources or flaring blazars?

We present new, simultaneous, multifrequency observations of 45 out of the 55 candidate High Frequency Peakers (HFP) selected by Dallacasa et al. (2000), carried out 3 to 4 years after a first set of observations. Our sub-sample consists of 10 galaxies, 28 stellar objects (``quasars'') and 7 unidentified sources. Both sets of observations are sensitive enough to allow the detection of variability at the 10% level or lower. While galaxies do not show significant variability, most quasars do. Seven of them no longer show the convex spectrum which is the defining property of Gigahertz Peaked Spectrum (GPS)/HFP sources and are interpreted as blazars caught by Dallacasa et al. (2000) during a flare, when a highly self-absorbed component dominated the emission. In general, the variability properties (amplitude, timescales, correlation between peak luminosity and peak frequency of the flaring component) of the quasar sub-sample resemble those of blazars. We thus conclude that most HFP candidates identified with quasars may well be flaring blazars.Comment: 20 pages, 18 figures, accepted for publication in Astronomy & Astrophysic