An Expert Elicitation of Public Acceptance of Renewable Energy in Kenya

This article reports evidence for substantial public support for the large-scale deployment of three renewable energy options in Kenya: wind, solar PV, and geothermal energy. With these renewable technologies, the government of Kenya could make a large contribution to reaching its national commitment under the Paris Agreement. Prices, infrastructural needs, and land-use requirements importantly contribute to shaping public opinion about these renewable energy alternatives, in different ways and directions for wind, PV, and geothermal energy. While overall the evaluation of these technologies is positive, public authorities should be wary of the possible inconveniences and drawbacks associated with them. Anticipating and, where possible, mitigating these shortcomings in national climate and energy development plans could preclude some of them becoming possible hindrances for broad-scale adoption of wind, PV, and geothermal energy. Furthering quantitative public acceptance studies, like the one presented here based on (semi-)expert elicitation and information-choice questionnaires, can assist in Kenya fully reaching its national climate and energy ambitions. More generally, we argue that the establishment of affordable, clean, and secure energy systems, as well as the mitigation of global climate change, can benefit from stakeholder engagement and public survey analysis like the one performed in our study – in developing countries as much as in the developed part of the world

Spotlight on the Roles of Whitefly Effectors in Insect–Plant Interactions

The Bemisia tabaci species complex (whitefly) causes enormous agricultural losses. These phloem-feeding insects induce feeding damage and transmit a wide range of dangerous plant viruses. Whiteflies colonize a broad range of plant species that appear to be poorly defended against these insects. Substantial research has begun to unravel how phloem feeders modulate plant processes, such as defense pathways, and the central roles of effector proteins, which are deposited into the plant along with the saliva during feeding. Here, we review the current literature on whitefly effectors in light of what is known about the effectors of phloem-feeding insects in general. Further analysis of these effectors may improve our understanding of how these insects establish compatible interactions with plants, whereas the subsequent identification of plant defense processes could lead to improved crop resistance to insects. We focus on the core concepts that define the effectors of phloem-feeding insects, such as the criteria used to identify candidate effectors in sequence-mining pipelines and screens used to analyze the potential roles of these effectors and their targets in planta. We discuss aspects of whitefly effector research that require further exploration, including where effectors localize when injected into plant tissues, whether the effectors target plant processes beyond defense pathways, and the properties of effectors in other insect excretions such as honeydew. Finally, we provide an overview of open issues and how they might be addressed

A FRET-based biosensor for measuring G alpha 13 activation in single cells

Contains fulltext : 187833.pdf (publisher's version ) (Open Access

An Expert Elicitation of Public Acceptance of Renewable Energy in Kenya

This article reports evidence for substantial public support for the large-scale deployment of three renewable energy options in Kenya: wind, solar PV, and geothermal energy. With these renewable technologies, the government of Kenya could make a large contribution to reaching its national commitment under the Paris Agreement. Prices, infrastructural needs, and land-use requirements importantly contribute to shaping public opinion about these renewable energy alternatives, in different ways and directions for wind, PV, and geothermal energy. While overall the evaluation of these technologies is positive, public authorities should be wary of the possible inconveniences and drawbacks associated with them. Anticipating and, where possible, mitigating these shortcomings in national climate and energy development plans could preclude some of them becoming possible hindrances for broad-scale adoption of wind, PV, and geothermal energy. Furthering quantitative public acceptance studies, like the one presented here based on (semi-)expert elicitation and information-choice questionnaires, can assist in Kenya fully reaching its national climate and energy ambitions. More generally, we argue that the establishment of affordable, clean, and secure energy systems, as well as the mitigation of global climate change, can benefit from stakeholder engagement and public survey analysis like the one performed in our study – in developing countries as much as in the developed part of the world

mScarlet: a bright monomeric red fluorescent protein for cellular imaging.

International audienceWe report the engineering of mScarlet, a truly monomeric red fluorescent protein with record brightness, quantum yield (70%) and fluorescence lifetime (3.9 ns). We developed mScarlet starting with a consensus synthetic template and using improved spectroscopic screening techniques; mScarlet's crystal structure reveals a planar and rigidified chromophore. mScarlet outperforms existing red fluorescent proteins as a fusion tag, and it is especially useful as a Förster resonance energy transfer (FRET) acceptor in ratiometric imaging

A FRET-based biosensor for measuring Gα13 activation in single cells

<div><p>Förster Resonance Energy Transfer (FRET) provides a way to directly observe the activation of heterotrimeric G-proteins by G-protein coupled receptors (GPCRs). To this end, FRET based biosensors are made, employing heterotrimeric G-protein subunits tagged with fluorescent proteins. These FRET based biosensors complement existing, indirect, ways to observe GPCR activation. Here we report on the insertion of mTurquoise2 at several sites in the human Gα13 subunit, aiming to develop a FRET-based Gα13 activation biosensor. Three fluorescently tagged Gα13 variants were found to be functional based on i) plasma membrane localization and ii) ability to recruit p115-RhoGEF upon activation of the LPA2 receptor. The tagged Gα13 subunits were used as FRET donor and combined with cp173Venus fused to the Gγ2 subunit, as the acceptor. We constructed Gα13 biosensors by generating a single plasmid that produces Gα13-mTurquoise2, Gβ1 and cp173Venus-Gγ2. The Gα13 activation biosensors showed a rapid and robust response when used in primary human endothelial cells that were exposed to thrombin, triggering endogenous protease activated receptors (PARs). This response was efficiently inhibited by the RGS domain of p115-RhoGEF and from the biosensor data we inferred that this is due to GAP activity. Finally, we demonstrated that the Gα13 sensor can be used to dissect heterotrimeric G-protein coupling efficiency in single living cells. We conclude that the Gα13 biosensor is a valuable tool for live-cell measurements that probe spatiotemporal aspects of Gα13 activation.</p></div

Capacity of the tagged Gα13 variants to recruit p115-RhoGEF.

<p>(A) Confocal images of a representative HeLa cell expressing SYFP1-p115-RhoGEF, Gα13.2-mTurquoise2-Δ9 and LPA2-P2A-mCherry (here only SYFP1-p115-RhoGEF is shown, for the localization of the other constructs see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0193705#pone.0193705.s002" target="_blank">S2 Fig</a>) (before (t = 0s) and after (t = 100s) addition of 3μM LPA). The width of the pictures is 67μm. (B) The mean cytoplasmic fluorescence intensity of SYFP1-p115-RhoGEF over time. After 8s, 3μM LPA was added. All cells transiently expressed LPA2 receptor-P2A-mCherry. The number of cells imaged is p115-RhoGEF <i>n</i> = 5, Gα13 untagged + p115-RhoGEF <i>n</i> = 15, Gα13.1 + p115-RhoGEF <i>n</i> = 27, Gα13.2 + p115-RhoGEF <i>n</i> = 28, Gα13.3 + p115-RhoGEF <i>n</i> = 24, Gα13.5 + p115-RhoGEF <i>n</i> = 20. Data have been derived from three independent experiments. (C) Quantification of the fluorescence intensity at t = 50s for each Gα13 variant, relative to t = 0s. The dots indicate individual cells and the error bars show 95% confidence intervals. The numbers of cells analyzed is the same as in (B).</p

Development and characterization of Gα13 activation FRET based biosensors.

<p>(A) Architecture of the Gα13 biosensor construct, encoding Gβ-2A-cp173Venus-Gγ₂-IRES-Gα13-mTurquoise2-Δ9, under control of the CMV promoter. (B) CFP and YFP emission was measured from individual cells expressing the Gα13.2 sensor from a single plasmid or from cells transfected with separate plasmids that encoded Gα13.2 and cp173Venus-Gγ₂. The r² is the correlation coefficient. (C) Confocal images showing the localization of the Gα13 in the sensor variants (upper, cyan) and cp173Venus-Gγ₂ (lower, yellow) in HeLa cells (for Gα13.2 sensor localization in Hek293T and HUVEC see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0193705#pone.0193705.s003" target="_blank">S3 Fig</a>). The width of the images is 75μm. (D) FRET ratio traces of HUVECs expressing the different Gα13 biosensors, stimulated with Thrombin at t = 100s (dotted lines depict 95% CI). For the corresponding YFP and CFP traces see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0193705#pone.0193705.s004" target="_blank">S4 Fig</a>. The number of cells analyzed is: Gα13.2 sensor <i>n</i> = 16, Gα13.3 sensor <i>n</i> = 11, Gα13.5 sensor <i>n</i> = 16.</p

Insertion of a fluorescent protein at different positions in Gα13.

<p>(A) The protein structure of human Gα13 (PDB ID: 1ZCB). The highlighted residues indicate the amino acid preceding the inserted fluorescent protein. Successful sites for inserting mTurquoise2-Δ9 into Gα13 in pink and unsuccessful sites in orange. (B) A partial protein sequence alignment (full alignment see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0193705#pone.0193705.s001" target="_blank">S1 Fig</a>) of different Gα classes. The highlighted residues indicate the amino acid preceding the inserted fluorescent protein (or luciferase). In bold, the sites that were previously used to insert Rluc [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0193705#pone.0193705.ref026" target="_blank">26</a>]. Insertion of mTurquoise2-Δ9 in Gα13 after residue Q144 (black) was based on homology with previous insertions in Gαq and Gαi (black). Successful sites for inserting mTurquoise2-Δ9 (R128, A129 and R140) in pink and unsuccessful sites (L106 and L143) in orange. The numbers indicated below the alignment correspond with the Gα13 variant numbers, used throughout the manuscript. The colors under the alignment match with the colors of the αHelices shown in (A). (C) Confocal images of the tagged Gα13 variants transiently expressed in HeLa cells. The numbers in the left bottom corner of each picture indicate the number of cells that showed plasma membrane localization out of the total number of cells analyzed. The tagged Gα13 variants also localize to structures inside the cell, which are presumably endomembranes,. The width of the images is 76μm.</p