Cell-free expression and SMA copolymer encapsulation of a functional receptor tyrosine kinase disease variant, FGFR3-TACC3

Despite their high clinical relevance, obtaining structural and biophysical data on transmembrane proteins has been hindered by challenges involved in their expression and extraction in a homogeneous, functionally-active form. The inherent enzymatic activity of receptor tyrosine kinases (RTKs) presents additional challenges. Oncogenic fusions of RTKs with heterologous partners represent a particularly difficult-to-express protein subtype due to their high flexibility, aggregation propensity and the lack of a known method for extraction within the native lipid environment. One such protein is the fibroblast growth factor receptor 3 fused with transforming acidic coiled-coil-containing protein 3 (FGFR3-TACC3), which has failed to express to sufficient quality or functionality in traditional expression systems. Cell-free protein expression (CFPE) is a burgeoning arm of synthetic biology, enabling the rapid and efficient generation of recombinant proteins. This platform is characterised by utilising an optimised solution of cellular machinery to facilitate protein synthesis in vitro. In doing so, CFPE can act as a surrogate system for a range of proteins that are otherwise difficult to express through traditional host cell-based approaches. Here, functional FGFR3-TACC3 was expressed through a novel cell-free expression system in under 48 h. The resultant protein was reconstituted using SMA copolymers with a specific yield of 300 µg/mL of lysate. Functionally, the protein demonstrated significant kinase domain phosphorylation (t < 0.0001). Currently, there is no published, high-resolution structure of any full-length RTK. These findings form a promising foundation for future research on oncogenic RTKs and the application of cell-free systems for synthesising functional membrane proteins

Impact of terminal group on temperature-dependent excited state relaxation in cationic dyes

Cationic organic dyes carry a positive charge distributed along the molecule, and the localization of this charge significantly affects their symmetry and optical properties. Depending on the different factors (topology of the terminal groups, the polarity of the solvent, and the temperature) the polyene, polymethine, or donor-acceptor structure form in such dyes, and excited state relaxation for such systems is not fully explored, particularly at low temperatures. At room temperature, the studied cationic dyes, regardless of symmetry in the ground state, are mostly symmetrical in the excited state. At low temperatures, charge localization effects become evident, leading to symmetry breaking in both ground and excited states. In this paper, we distinguish how terminal groups at the end of the cationic dyes impact the relaxation of excited states by analyzing experimental low-temperature time-resolved spectra combined with quantum-chemical calculations. Distinctive emission (690 nm) in the anti-Stokes range of polymethine band (700–730 nm) features polyene structures forming depending on the temperature, solvent polarity, and charge-donating properties of the dye's terminal groups. Furthermore, in low-temperature time-resolved photoluminescence, a 760 nm band is distinguished and associated with intramolecular charge transfer. Our calculations revealed unequal distribution of total positive charge in different molecular fragments (polymethine chain and terminal groups) and formation of negative charge on polymethine chain. We propose a model of excited state relaxation transitions for linear cationic molecular systems that enable donor-acceptor features. This model offers valuable insights for designing new functional materials with tunable properties or efficient energy transfer systems for artificial photosynthesis

Next-Generation Ultrasound Propagation in Suspended Core Optical Fibers

We demonstrate the temporal propagation of high-frequency ultrasonic pulses inside of a suspended core fiber (SCF) for the first time. An SCF composed of a 5 μm core suspended by nano bridges and four air holes is analytically and numerically investigated using the 2D and 3D finite element method. We reveal the propagation of ultrasonic modes in the SCF and describe the variation of their properties with increasing frequency from 1 to 100 MHz. Ultrasonic Gaussian-modulated pulses traveling with distinct frequencies (25 and 72 MHz) and bandwidths (56 and 10 MHz) are evaluated along the SCF length and time. The induced strain, pressure, and sensing responsivity are reviewed and discussed. Analytical and numerical results show highly confined effective single-mode propagation in the SCF core, amplifying strains along the fiber (up to 18 times) and pressures at the fiber output (up to 3 times). Compared to previous studies, the demonstrated SCF provides the widest 86 MHz high-responsivity bandwidth (with a -218 dB average and - 75 dB maximum responsivity at 93 MHz). These achievements are promising for developing smaller, faster, and more efficient acousto-optic modulators for fiber lasers, enhancing simultaneous light-sound guidance and spatial resolution of fiber-based optoacoustic imaging sensors and neurostimulators in biomedicine

Renegotiating HRM Practices and Delivering Service Transformation of Frontline Workers: Insights From the UK 's NHS Ambulance Trust During a Significant Disruption

We note that while the institutional environment determines the actions and structures of constituent organizations, there are still limited insights available in the literature on the nature and process of interactions between key stakeholders in ambulance services, especially as they respond during a significant crisis. This study specifically examines how a positive social exchange between senior leadership, managers, and frontline staff, accompanied by changes to the leadership style and HR practices, resulted in significant service transformation. An in‐depth qualitative case study design was used to interview 52 frontline staff and leaders from the UK National Health Service's (NHS) Ambulance Service over a prolonged period of 8 months (October 2020 to May 2021). Interviews and secondary data were abductively analyzed, showing how changes to HRM and leadership practices shaped service transformation's content, process, and logic. The findings suggest that an iterative social exchange between the key social actors in the Ambulance Trust led to adaptations to the nature of HRM and leadership practices, which positively impacted the changes to service delivery, leading to service transformation and improved health and well‐being of paramedics. Our conceptual framework contributes to the literature by integrating how the use of agency, new HRM, and leadership practices in the context of institutionalized structures can result in service transformation. Implications for theory and practice are also discussed

Innovation and internationalisation in a digital world: the focal role of innovation incubators

Riding on the back of the country’s maker movement, Chinese central government launched a ‘mass innovation and mass entrepreneurship’ initiative to promote innovation and entrepreneurship across the country. This top-down initiative has contributed to an explosion of innovation incubators focusing on digital technologies and their broader implications for everyday social and cultural life in a globalised era. In this paper, we explore the focal role of these new breed of Chinese innovation incubators in shaping the internationalisation agenda of firms and how their innovation processes play out in the digital innovation environment. Data for the inquiry comes from ethnographic interviews with loosely coupled actors in the Chinese digital innovation ecosystem and two of the most prominent innovation incubators. We found innovation incubators as orchestrators of a new breed of innovation networks that provide safe crucibles for ‘Maker’ ideation, experimentation, and prototyping. Our study suggests that these Chinese innovation incubators, over time, have come to play an intermediary role in linking Chinese traditional manufacturing industries to digital technologies, capital, and talents. These innovation incubators, by virtue of their extended relational networks, provide opportunities for Chinese firms to tap into international markets otherwise overlooked by their competitors

When Do Environmental Regulations Lead to Green Practices? The Role of Resource Commitment and Corporate Entrepreneurship

Environmental regulations increasingly pressure firms to adopt green practices, yet their effectiveness remains debated. Drawing on institutional theory and the resource‐based view, this study investigates the mechanisms linking environmental regulations to green supply chain management (GSCM) practices. We propose and test a moderated mediation model using data from 231 Chinese manufacturers. Results show that circular‐oriented resource commitment mediates the regulation–GSCM relationship, whereas corporate entrepreneurship selectively moderates the path from regulations to resource commitment. These findings extend theory by showing how regulatory pressures shape resource deployment in environmental management and how entrepreneurial orientation enhances firms' ability to transform regulatory requirements into strategic resource commitments

Covalent adaptable networks from functionalised unsaturated polyesters via Diels-Alder cycloaddition

Unsaturated polyesters (UPEs) containing fumarate groups were functionalised via the thiol-Michael addition of furfuryl mercaptan. This “click” reaction occurs rapidly in ambient conditions in the presence of an organic catalyst. The partial addition of the diene moiety allows the polyester chain to participate in Diels-Alder (DA) cycloaddition reactions with the remaining fumarate groups to form a crosslinked network. Such crosslinks dissociate at 90 °C through the retro-Diels-Alder (rDA) reaction and reform upon cooling to ambient temperature, highlighting the reprocessability of these polyester networks. The unsaturated groups in poly(propylene fumarate) were functionalised to varying degrees, producing a range of materials (12–100 % addition of furan) and it was shown via differential scanning calorimetry that 50 % functionalisation produced the highest enthalpy change for the rDA reaction. This composition also produced the highest bond strength in adhesive joints. Similar results were obtained with compositions comprising both 100 % furan functionalised polyester and unfunctionalised fumarate backbone

Advancing Sustainable Glass Manufacturing through Optimized Predictive Maintenance Planning of Critical Forming Components

The glass manufacturing industry is a key contributor to various sectors, including construction, automotive, and packaging. However, it is also energy-intensive and contributes significantly to global carbon emissions. Decarbonizing glass production is essential for aligning industrial practices with global climate goals. This study focuses on advancing sustainability in glass manufacturing through a predictive maintenance planning framework adapted to critical forming components, including Gob Delivery System, Blank Moulds, and Blow Moulds. By optimizing maintenance schedules and minimizing unplanned downtimes, the framework reduces resource wastage, energy inefficiencies, and associated carbon emissions, thereby aligning operational practices with sustainability objectives. The proposed framework integrates reliability analysis, cost evaluation, and advanced optimization techniques to dynamically generate maintenance schedules. A computational tool developed for this purpose simulates degradation and maintenance processes, offering actionable insights into component reliability and cost efficiency. While validated using simulated data, the methodology is adaptable for broader industrial applications, promising significant contributions to the sustainability of glass manufacturing

City-level institutions and perceived entrepreneurial ecosystem’s growth orientation

This study uses both secondary and primary data on perceptions of 1789 ecosystem actors from 17 cities in Europe to perform an empirical analysis of three institutional dimensions: regulatory, cultural values and socio-cultural practices – and tests their association with the entrepreneurial ecosystem’s growth orientation. As a result, we develop a framework for the entrepreneurial ecosystem’s factors and provide policy recommendations for those interested in supporting the entrepreneurial ecosystem’s growth orientation in cities. Among other conclusions, the findings suggest a positive association between the socio-cultural practices of environmental sustainability behaviour in businesses with entrepreneurial ecosystem’s growth orientation

Inferring structure of cortical neuronal networks from activity data: A statistical physics approach

Understanding the relation between cortical neuronal network structure and neuronal activity is a fundamental unresolved question in neuroscience, with implications to our understanding of the mechanism by which neuronal networks evolve over time, spontaneously or under stimulation. It requires a method for inferring the structure and composition of a network from neuronal activities. Tracking the evolution of networks and their changing functionality will provide invaluable insight into the occurrence of plasticity and the underlying learning process. We devise a probabilistic method for inferring the effective network structure by integrating techniques from Bayesian statistics, statistical physics, and principled machine learning. The method and resulting algorithm allow one to infer the effective network structure, identify the excitatory and inhibitory type of its constituents, and predict neuronal spiking activity by employing the inferred structure. We validate the method and algorithm's performance using synthetic data, spontaneous activity of an in silico emulator, and realistic in vitro neuronal networks of modular and homogeneous connectivity, demonstrating excellent structure inference and activity prediction. We also show that our method outperforms commonly used existing methods for inferring neuronal network structure. Inferring the evolving effective structure of neuronal networks will provide new insight into the learning process due to stimulation in general and will facilitate the development of neuron-based circuits with computing capabilities