The actin regulators Enabled and Diaphanous direct distinct protrusive behaviors in different tissues during Drosophila development

Actin-based protrusions are important for signaling and migration during development and homeostasis. Gain- and loss-of-function and quantitative approaches are used to define differential roles for the actin elongation factors Diaphanous and Enabled in regulating distinct protrusive behaviors in different tissues during Drosophila morphogenesis.Actin-based protrusions are important for signaling and migration during development and homeostasis. Defining how different tissues in vivo craft diverse protrusive behaviors using the same genomic toolkit of actin regulators is a current challenge. The actin elongation factors Diaphanous and Enabled both promote barbed-end actin polymerization and can stimulate filopodia in cultured cells. However, redundancy in mammals and Diaphanous’ role in cytokinesis limited analysis of whether and how they regulate protrusions during development. We used two tissues driving Drosophila dorsal closure—migratory leading-edge (LE) and nonmigratory amnioserosal (AS) cells—as models to define how cells shape distinct protrusions during morphogenesis. We found that nonmigratory AS cells produce filopodia that are morphologically and dynamically distinct from those of LE cells. We hypothesized that differing Enabled and/or Diaphanous activity drives these differences. Combining gain- and loss-of-function with quantitative approaches revealed that Diaphanous and Enabled each regulate filopodial behavior in vivo and defined a quantitative “fingerprint”—the protrusive profile—which our data suggest is characteristic of each actin regulator. Our data suggest that LE protrusiveness is primarily Enabled driven, whereas Diaphanous plays the primary role in the AS, and reveal each has roles in dorsal closure, but its robustness ensures timely completion in their absence

Contextual Simulated Annealing Q-Learning for Pre-negotiation of Agent-Based Bilateral Negotiations

Electricity markets are complex environments, which have been suffering continuous transformations due to the increase of renewable based generation and the introduction of new players in the system. In this context, players are forced to re-think their behavior and learn how to act in this dynamic environment in order to get as much benefit as possible from market negotiations. This paper introduces a new learning model to enable players identifying the expected prices of future bilateral agreements, as a way to improve the decision-making process in deciding the opponent players to approach for actual negotiations. The proposed model introduces a con-textual dimension in the well-known Q-Learning algorithm, and includes a simulated annealing process to accelerate the convergence process. The proposed model is integrated in a multi-agent decision support system for electricity market players negotiations, enabling the experimentation of results using real data from the Iberian electricity market.This work has received funding from the European Union's Horizon 2020 research and innovation programme under project DOMINOES (grant agreement No 771066) and from FEDER Funds through COMPETE program and from National Funds through FCT under the project UID/EEA/00760/2019.info:eu-repo/semantics/publishedVersio

Abelson kinase acts as a robust, multifunctional scaffold in regulating embryonic morphogenesis

Abelson family kinases (Abl) are key regulators of cell behavior and the cytoskeleton during development and in leukemia. Abl's SH3, SH2, and tyrosine kinase domains are joined via a linker to an F-actin-binding domain (FABD). Research on Abl's roles in cell culture led to several hypotheses for its mechanism of action: 1) Abl phosphorylates other proteins, modulating their activity. 2) Abl directly regulates the cytoskeleton via its cytoskeletal interaction domains, and/or 3) Abl is a scaffold for a signaling complex. The importance of these roles during normal development remains untested. We tested these mechanistic hypotheses during Drosophila morphogenesis using a series of mutants to examine Abl's many cell biological roles. Strikingly, Abl lacking the FABD fully rescued morphogenesis, cell shape change, actin regulation, and viability, while kinase dead Abl, though reduced in function, retained substantial rescuing ability in some but not all Abl functions. We also tested the function of four conserved motifs in the linker region, revealing a key role for a conserved PXXP motif known to bind Crk and Abi. We propose Abl acts as a robust multi-domain scaffold with different protein motifs and activities contributing differentially to diverse cellular behaviors

Preventing acute infection in total hip prostheses implanted after external fixation of the femur: is there a need for a staged procedure?

We report two cases of acute infection of an uncemented femoral component in a hip prosthesis implanted after external fixation of a femoral fracture. In both cases, the surgical access did not cross over the pin scars. When the prosthesis was implanted the stem crossed one or more pin tracts. The preoperative clinical examination, laboratory tests and bone scintigraphy with marked granulocytosis did not show signs of local infection in either case. We suggest that every patient destined to receive a prosthesis after external fixation should be treated with a staged procedure, the first step being excision of the soft tissues around the pin tracts and curettage or drilling of the bony holes, followed by prosthesis implant

Electricity Exchange: Demand Side Unit performance monitoring

Demand Side Response management encourages elec- tricity demand reduction during peak hours. One avenue for achieving this is through Demand Side Units (DSUs). These are large electricity consumers who can afford to reduce their demand on the electricity grid when required. Issues with DSUs revolve around verification that the correct demand reduction takes place, with limited monitoring capabilities from the electrical grid operator Eir- Grid. This issue is studied here with the current methods thoroughly analysed and new methods proposed. In this report six different forecasting methods are presented, and their accuracy is compared using two different error metrics. Due to inherent stochasticity in demand it is found that there is no one fore- casting method which is unequivocally best, but the ‘Keep it simple’ weekly and the temperature dependent models are identified as the most promising models to pursue. Initial investigations suggest that a ‘proxy day’ mechanism may be preferable to the current method of verifying that the correct demand reduction takes place

Enabled and Capping protein play important roles in shaping cell behavior during Drosophila oogenesis

During development, cells craft an impressive array of actin-based structures, mediating events as diverse as cytokinesis, apical constriction, and cell migration. One challenge is to determine how cells regulate actin assembly and disassembly to carry out these cell behaviors. During Drosophila oogenesis diverse cell behaviors are seen in the soma and germline. We used oogenesis to explore developmental roles of two important actin regulators: Enabled/VASP proteins and Capping protein. We found that Enabled plays an important role in cortical integrity of nurse cells, formation of robust bundled actin filaments in late nurse cells that facilitate nurse cell dumping, and migration of somatic border cells. During nurse cell dumping, Enabled localizes to barbed ends of the nurse cell actin filaments, suggesting its mechanism of action. We further pursued this mechanism using mutant Enabled proteins, each affecting one of its protein domains. These data suggest critical roles for the EVH2 domain and its tetramerization subdomain, while the EVH1 domain appears less critical. Enabled appears to be negatively regulated during oogenesis by Abelson kinase. We also explored the function of Capping protein. This revealed important roles in oocyte determination, nurse cell cortical integrity and nurse cell dumping, and support the idea that Capping protein and Enabled act antagonistically during dumping. Together these data reveal places these actin regulators shape oogenesis

Uncertainty analysis using Bayesian Model Averaging: a case study of input variables to energy models and inference to associated uncertainties of energy scenarios

Background Energy models are used to illustrate, calculate and evaluate energy futures under given assumptions. The results of energy models are energy scenarios representing uncertain energy futures. Methods The discussed approach for uncertainty quantification and evaluation is based on Bayesian Model Averaging for input variables to quantitative energy models. If the premise is accepted that the energy model results cannot be less uncertain than the input to energy models, the proposed approach provides a lower bound of associated uncertainty. The evaluation of model-based energy scenario uncertainty in terms of input variable uncertainty departing from a probabilistic assessment is discussed. Results The result is an explicit uncertainty quantification for input variables of energy models based on well-established measure and probability theory. The quantification of uncertainty helps assessing the predictive potential of energy scenarios used and allows an evaluation of possible consequences as promoted by energy scenarios in a highly uncertain economic, environmental, political and social target system. Conclusions If societal decisions are vested in computed model results, it is meaningful to accompany these with an uncertainty assessment. Bayesian Model Averaging (BMA) for input variables of energy models could add to the currently limited tools for uncertainty assessment of model-based energy scenarios

Filopodyan: An open-source pipeline for the analysis of filopodia

Filopodia have important sensory and mechanical roles in motile cells. The recruitment of actin regulators, such as ENA/ VASP proteins, to sites of protrusion underlies diverse molecular mechanisms of filopodia formation and extension. We developed Filopodyan (filopodia dynamics analysis) in Fiji and R to measure fluorescence in filopodia and at their tips and bases concurrently with their morphological and dynamic properties. Filopodyan supports high-throughput phenotype characterization as well as detailed interactive editing of filopodia reconstructions through an intuitive graphical user interface. Our highly customizable pipeline is widely applicable, capable of detecting filopodia in four different cell types in vitro and in vivo. We use Filopodyan to quantify the recruitment of ENA and VASP preceding filopodia formation in neuronal growth cones, and uncover a molecular heterogeneity whereby different filopodia display markedly different responses to changes in the accumulation of ENA and VASP fluorescence in their tips over time.J.L. Gallop and V. Urbančič are supported by the Wellcome Trust (WT095829AIA). J. Mason and B. Richier are supported by the European Research Council (281971). C.E. Holt is supported by the Wellcome Trust (program grant 085314) and the European Research Council (advanced grant 322817). The Gurdon Institute is funded by the Wellcome Trust (203144) and Cancer Research UK (C6946/A24843)