GM-CSF suppresses antioxidant signaling and drives IL-1ß secretion through NRF2 downregulation.

GM-CSF is a potent inflammatory cytokine regulating myeloid cell differentiation, hematopoiesis, and various other functions. It is functionally associated with a number of inflammatory pathologies including rheumatoid arthritis and inflammatory bowel disease. GM-CSF has been found to promote NLRP3-dependent IL-1ß secretion, which may have a significant role in driving inflammatory pathologies. However, the molecular mechanisms remain unknown. Here, we show that GM-CSF induces IL-1ß secretion through a ROS-dependent pathway. TNF is required for reactive oxygen species (ROS) generation that strikingly does not promote NLRP3 activation, but instead drives ubiquitylation of IL-1ß, promoting its cleavage through basal NRLP3 activity. GM-CSF regulates this pathway through suppression of antioxidant responses via preventing upregulation of NRF2. Thus, the pro-inflammatory effect of GM-CSF on IL-1ß is through suppression of antioxidant responses, which leads to ubiquitylation of IL-1ß and enhanced processing. This study highlights the role of metabolic regulation of inflammatory signaling and reveals a novel mechanism for GM-CSF to promote inflammation.

A non-death function of the mitochondrial apoptosis apparatus in immunity.

Apoptosis is a frequent form of programmed cell death, but the apoptotic signaling pathway can also be engaged at a low level, in the absence of cell death. We here report that such sub-lethal engagement of mitochondrial apoptosis signaling causes the secretion of cytokines from human epithelial cells in a process controlled by the Bcl-2 family of proteins. We further show that sub-lethal signaling of the mitochondrial apoptosis pathway is initiated by infections with all tested viral, bacterial, and protozoan pathogens and causes damage to the genomic DNA. Epithelial cells infected with these pathogens secreted cytokines, and this cytokine secretion upon microbial infection was substantially reduced if mitochondrial sub-lethal apoptosis signaling was blocked. In the absence of mitochondrial pro-apoptotic signaling, the ability of epithelial cells to restrict intracellular bacterial growth was impaired. Triggering of the mitochondrial apoptosis apparatus thus not only causes apoptosis but also has an independent role in immune defense.

Pattern Recognition Receptors of Nucleic Acids Can Cause Sublethal Activation of the Mitochondrial Apoptosis Pathway during Viral Infection.

The mitochondrial apoptosis pathway has the function to kill the cell, but recent work shows that this pathway can also be activated to a sublethal level, where signal transduction can be observed but the cell survives. Intriguingly, this signaling has been shown to contribute to inflammatory activity of epithelial cells upon infection with numerous agents. This suggests that microbial recognition can generate sublethal activity in the mitochondrial apoptosis pathway. Because this recognition is achieved by pattern recognition receptors (PRRs), it also implies that PRR signals are linked to the mitochondrial apoptosis apparatus. We here test this hypothesis during infection of epithelial cells with modified vaccinia virus Ankara (MVA). MVA recognition is achieved through receptors specific for nucleic acids, and we present evidence that the three receptors, Toll-like receptor 3 (TLR3), RIG-I/MDA5, and cyclic GMP-AMP synthase (cGAS)/stimulator of interferon genes (STING), are involved in this signaling. When stimulated directly by specific ligands, all three receptors could trigger sublethal apoptosis signals. During infection with MVA, sublethal apoptosis signals were unmasked in X-linked IAP (XIAP)-deficient cells, where apoptosis induction was observed. Deletion of any of the three signaling adapters, TRIF, MAVS, and STING, reduced the DNA damage response, a sensitive measure of sublethal apoptosis signals. Our results suggest that PRRs signal via mitochondria, where they generate sublethal signals through the BCL-2-family, which may contribute to the response to infectious agents. IMPORTANCE A contribution of the mitochondrial apoptosis apparatus, in the absence of cell death, to the reaction of nonprofessional immune cells to viruses is suggested to play a role as a broad alert system of an infected cell: the apoptosis system can be activated by many upstream signals and could therefore act as a central coordinator of viral recognition. The proapoptotic activity of PRRs has been documented in multiple situations, but this activity seems too low to be meaningful, and a physiological significance of such activity is not immediately obvious. This work suggests the alternative interpretation that PRRs do not have the primary function to induce apoptosis but to trigger sublethal signals in the apoptosis system. A number of lines of recent research suggest that mitochondria contribute to cellular reactions, and this pathway may be a way of triggering an early host response.

The impact of film deposition and annealing on the nanostructure and dielectric constant of organic semiconductor thin films.

The strategy of using a bulk-heterojunction light-absorbing layer has led to the most efficient organic solar cells. However, optimising the blend morphology to maximise light absorption, charge generation and extraction can be challenging. Homojunction devices containing a single component have the potential to overcome the challenges associated with bulk heterojunction films. A strategy towards this goal is to increase the dielectric constant of the organic semiconductor to ≈10, which in principle would lead to free charge carrier generation upon photoexcitation. However, the factors that affect the thin film dielectric constants are still not well understood. In this work we report an organic semiconductor material that can be solution processed or vacuum evaporated to form good quality thin films to explore the effect of chromophore structure and film morphology on the dielectric constant and other optoelectronic properties. 2,2'-[(4,4,4',4'-Tetrakis{2-[2-methoxyethoxy]ethyl}-4H,4'H-{2,2'-bi[cyclo-penta[2,1-b:3,4-b']dithiophene]}-6,6'-diyl)bis(methaneylylidene)]dimalononitrile [D(CPDT-DCV)] was designed to have high electron-affinity end groups and low ionisation-potential central moieties. It can be processed from solution or be thermally evaporated, with the film morphology changing from face-on to a herringbone arrangement upon solvent or thermal annealing. The glycol solubilising groups led to the static dielectric constant (taken from capacitance measurements) of the films to be between 6 and 7 (independent of processing conditions), while the optical frequency dielectric constant depended on the processing conditions. The less ordered solution processed film was found to have the lowest optical frequency dielectric constant of 3.6 at 2.0 × 1014 Hz, which did not change upon annealing. In contrast, the more ordered evaporated film had an optical frequency dielectric constant 20% higher at 4.2 and thermal annealing further increased it to 4.5, which is amongst the highest reported for an organic semiconductor at that frequency. Finally, the more ordered evaporated films had more balanced charge transport, which did not change upon annealing.

A Deeper Understanding of Metal Nucleation and Growth in Rechargeable Metal Batteries Through Theory and Experiment.

Lithium and sodium metal batteries continue to occupy the forefront of battery research. Their exceptionally high energy density and nominal voltages are highly attractive for cutting-edge energy storage applications. Anode-free metal batteries are also coming into the research spotlight offering improved safety and even higher energy densities than conventional metal batteries. However, uneven metal nucleation and growth which leads to dendrites continues to limit the commercialisation of conventional and anode-free metal batteries alike. This review connects models and theories from well-established fields in metallurgy and electrodeposition to both conventional and anode-free metal batteries. These highly applicable models and theories explain the driving forces of uneven metal growth and can inform future experiment design. Finally, the models and theories that are most relevant to each anode-related cell component are identified. Keeping these specific models and theories in mind will assist with rational design for these components.

Solvent-derived Fluorinated Secondary Interphase for Reversible Zn-graphite Dual-ion Batteries.

The irreversibility of anion intercalation-deintercalation is a fundamental issue in determining the cycling stability of a dual-ion battery (DIB). In this work, we demonstrate that using a partially fluorinated carbonate solvent can drive a beneficial fluorinated secondary interphase layer formation. Such layer facilitates reversible anion (de-)intercalation processes by impeding solvent molecule co-intercalation and the associated graphite exfoliation. The enhanced reversibility of anion transport contributes to the overall cycling stability for a Zn-graphite DIB-a high Coulombic efficiency of 98.5 % after 800 cycles, with an attractive discharge capacity of 156 mAh g-1 and a mid-point discharge voltage of ≈1.7 V (at 0.1 A g-1 ). In addition, the formed fluorinated secondary interphase suppresses the self-discharge behavior, preserving 29 times of the capacity retention rate compared to the battery with a commonly used carbonate solvent, after standing for 24 hours. This work provides a simple and effective strategy for addressing the critical challenges in graphite-based DIBs and contributes to fundamental understanding to help accelerate their practical application.

The deubiquitinase Usp27x as a novel regulator of cFLIPL protein expression and sensitizer to death-receptor-induced apoptosis.

Death receptors are transmembrane proteins that can induce the activation of caspase-8 upon ligand binding, initiating apoptosis. Recent work has highlighted the great molecular complexity of death receptor signalling, in particular through ubiquitination/deubiquitination. We have earlier defined the deubiquitinase Ubiquitin-Specific Protease 27x (Usp27x) as an enzyme capable of stabilizing the pro-apoptotic Bcl-2 family member Bim. Here, we report that enhanced expression of Usp27x in human melanoma cells leads to the loss of cellular FLICE-like inhibitory protein (cFLIP) and sensitizes to Tumor necrosis factor receptor 1 (TNF-R1) or Toll-like receptor 3 (TLR3)-induced extrinsic apoptosis through enabling enhanced processing of caspase-8. The loss of cFLIPL upon overexpression of Usp27x was not due to reduced transcription, could be partially counteracted by blocking the ubiquitin proteasome system and was independent of the known cFLIPL destabilizing ubiquitin E3-ligases Itch and DTX1. Instead, Usp27x interacted with the E3-ligase TRIM28 and reduced ubiquitination of TRIM28. Reduction of cFLIPL protein levels by Usp27x-induction depended on TRIM28, which was also required for polyI:C-induced cell death. This work defines Usp27x as a novel regulator of cFLIPL protein expression and a deubiquitinase in fine tuning death receptor signalling pathways to execute apoptosis.

Inhibitor of apoptosis proteins limit RIP3 kinase-dependent interleukin-1 activation.

Interleukin-1ß (IL-1ß) is a potent inflammatory cytokine that is usually cleaved and activated by inflammasome-associated caspase-1. To determine whether IL-1ß activation is regulated by inhibitor of apoptosis (IAP) proteins, we treated macrophages with an IAP-antagonist "Smac mimetic" compound or genetically deleted the genes that encode the three IAP family members cIAP1, cIAP2, and XIAP. After Toll-like receptor priming, IAP inhibition triggered cleavage of IL-1ß that was mediated not only by the NLRP3-caspase-1 inflammasome, but also by caspase-8 in a caspase-1-independent manner. In the absence of IAPs, rapid and full generation of active IL-1ß by the NLRP3-caspase-1 inflammasome, or by caspase-8, required the kinase RIP3 and reactive oxygen species production. These results demonstrate that activation of the cell death-inducing ripoptosome platform and RIP3 can generate bioactive IL-1ß and implicate them as additional targets for the treatment of pathological IL-1-driven inflammatory responses.

A statistical approach for modelling the physical process of bacterial attachment to abiotic surfaces.

A statistical approach using a polynomial linear model in combination with a probability distribution model was developed to mathematically represent the process of bacterial attachment and study its mechanism. The linear deterministic model was built based on data from experiments investigating bacterial and substratum surface physico-chemical factors as predictors of attachment. The prediction results were applied to a normal-approximated binomial distribution model to probabilistically predict attachment. The experimental protocol used mixtures of Streptococcus salivarius and Escherichia coli, and mixtures of porous poly(butyl methacrylate-co-ethyl dimethacrylate) and aluminum sec-butoxide coatings, at varying ratios, to allow bacterial attachment to substratum surfaces across a range of physico-chemical properties (including the surface hydrophobicity of bacterial cells and the substratum, the surface charge of the cells and the substratum, the substratum surface roughness and cell size). The model was tested using data from independent experiments. The model indicated that hydrophobic interaction was the most important predictor while reciprocal interactions existed between some of the factors. More importantly, the model established a range for each factor within which the resultant attachment is unpredictable. This model, however, considers bacterial cells as colloidal particles and accounts only for the essential physico-chemical attributes of the bacterial cells and substratum surfaces. It is therefore limited by a lack of consideration of biological and environmental factors. This makes the model applicable only to specific environments and potentially provides a direction to future modelling for different environments.

Infection of epithelial cells with Chlamydia trachomatis inhibits TNF-induced apoptosis at the level of receptor internalization while leaving non-apoptotic TNF-signalling intact.

Chlamydia trachomatis is an obligate intracellular bacterial pathogen of medical importance. C. trachomatis develops inside a membranous vacuole in the cytosol of epithelial cells but manipulates the host cell in numerous ways. One prominent effect of chlamydial infection is the inhibition of apoptosis in the host cell, but molecular aspects of this inhibition are unclear. Tumour necrosis factor (TNF) is a cytokine with important roles in immunity, which is produced by immune cells in chlamydial infection and which can have pro-apoptotic and non-apoptotic signalling activity. We here analysed the signalling through TNF in cells infected with C. trachomatis. The pro-apoptotic signal of TNF involves the activation of caspase-8 and is controlled by inhibitor of apoptosis proteins. We found that in C. trachomatis-infected cells, TNF-induced apoptosis was blocked upstream of caspase-8 activation even when inhibitor of apoptosis proteins were inhibited or the inhibitor of caspase-8 activation, cFLIP, was targeted by RNAi. However, when caspase-8 was directly activated by experimental over-expression of its upstream adapter Fas-associated protein with death domain, C. trachomatis was unable to inhibit apoptosis. Non-apoptotic TNF-signalling, particularly the activation of NF-κB, initiates at the plasma membrane, while the activation of caspase-8 and pro-apoptotic signalling occur subsequently to internalization of TNF receptor and the formation of a cytosolic signalling complex. In C. trachomatis-infected cells, NF-κB activation through TNF was unaffected, while the internalization of the TNF-TNF-receptor complex was blocked, explaining the lack of caspase-8 activation. These results identify a dichotomy of TNF signalling in C. trachomatis-infected cells: Apoptosis is blocked at the internalization of the TNF receptor, but non-apoptotic signalling through this receptor remains intact, permitting a response to this cytokine at sites of infection.

The structural impact of water sorption on device-quality melanin thin films.

The melanins are a class of pigmentary bio-macromolecules ubiquitous in the biosphere. They possess an intriguing set of physico-chemical properties and have been shown to exhibit hybrid protonic-electronic electrical conductivity, a feature derived from a process termed chemical self-doping driven by the sorption of water. Although the mechanism underlying the electrical conduction has been established, how the sorbed water interacts with the melanin structure at the physical level has not. Herein we use neutron reflectometry to study changes in the structure of synthetic melanin thin films as a function of H2O and D2O vapour pressure. Water is found to be taken up evenly throughout the films, and by employing the contrast effect, the existence of labile protons through reversible deuterium exchange is demonstrated. Finally, we determine a sorption isotherm to enable quantification of the melanin-water interactions.

Elucidating the Spatial Arrangement of Emitter Molecules in Organic Light-Emitting Diode Films.

The effect of varying the emitter concentration on the structural properties of an archetypal phosphorescent blend consisting of 4,4'-bis(N-carbazolyl)biphenyl and tris(2-phenylpyridyl)iridium(III) has been investigated using non-equilibrium molecular dynamics (MD) simulations that mimic the process of vacuum deposition. By comparison with reflectometry measurements, we show that the simulations provide an accurate model of the average density of such films. The emitter molecules were found not to be evenly distributed throughout film, but rather they can form networks that provide charge and/or energy migration pathways, even at emitter concentrations as low as ≈5 weight percent. At slightly higher concentrations, percolated networks form that span the entire system. While such networks would give improved charge transport, they could also lead to more non-radiative pathways for the emissive state and a resultant loss of efficiency.

Inhibitors of apoptosis proteins (IAPs) are required for effective T-cell expansion/survival during antiviral immunity in mice.

Inhibitors of apoptosis proteins (IAPs) were originally described as regulating apoptosis by direct binding to caspases. More recently, IAPs have been identified as important modulators of canonical and noncanonical nuclear factor κB signaling via their ubiquitin-E3 ligase activity. IAPs are therefore, not only gatekeepers of cell death, but are probably also involved in the regulation of inflammation, as well as innate and adaptive immunity. In this study, we analyzed the role of IAPs in T-cell immunity during lymphocytic choriomeningitis virus (LCMV) infection by pharmacological targeting with an IAP antagonist/second mitochondria-derived activator of caspase-mimetic. Expansion of virus-specific CD8 T cells was drastically reduced in LCMV-infected mice exposed to IAP antagonists. Accordingly, virus control was substantially impaired, indicated by high virus titres in the spleen and the spread of LCMV to peripheral organs. The profound negative effect of IAP antagonists on T-cell immunity was partially linked to tumor necrosis factor-mediated cell death of activated T cells and required inhibition of X-linked inhibitor of apoptosis, as well as cellular IAP-1. Thus, IAPs play an important role in T-cell expansion and survival in the context of a highly inflammatory environment such as a virus infection, indicating that IAP antagonists may interfere with immune responses.

Detection of Explosive Vapors: The Roles of Exciton and Molecular Diffusion in Real-Time Sensing.

Time-resolved quartz crystal microbalance with in situ fluorescence measurements are used to monitor the sorption of the nitroaromatic (explosive) vapor, 2,4-dinitrotoluene (DNT) into a porous pentiptycene-containing poly(phenyleneethynylene) sensing film. Correlation of the nitroaromatic mass uptake with fluorescence quenching shows that the analyte diffusion follows the Case-II transport model, a film-swelling-limited process, in which a sharp diffusional front propagates at a constant velocity through the film. At a low vapor pressure of DNT of ≈16 ppb, the analyte concentration in the front is sufficiently high to give an average fluorophore-analyte separation of ≈1.5 nm. Hence, a long exciton diffusion length is not required for real-time sensing in the solid state. Rather the diffusion behavior of the analyte and the strength of the binding interaction between the analyte and the polymer play first-order roles in the fluorescence quenching process.

Role of semiconductivity and ion transport in the electrical conduction of melanin.

Melanins are pigmentary macromolecules found throughout the biosphere that, in the 1970s, were discovered to conduct electricity and display bistable switching. Since then, it has been widely believed that melanins are naturally occurring amorphous organic semiconductors. Here, we report electrical conductivity, muon spin relaxation, and electron paramagnetic resonance measurements of melanin as the environmental humidity is varied. We show that hydration of melanin shifts the comproportionation equilibrium so as to dope electrons and protons into the system. This equilibrium defines the relative proportions of hydroxyquinone, semiquinone, and quinone species in the macromolecule. As such, the mechanism explains why melanin at neutral pH only conducts when "wet" and suggests that both carriers play a role in the conductivity. Understanding that melanin is an electronic-ionic hybrid conductor rather than an amorphous organic semiconductor opens exciting possibilities for bioelectronic applications such as ion-to-electron transduction given its biocompatibility.

Time-resolved neutron reflectometry and photovoltaic device studies on sequentially deposited PCDTBT-fullerene layers.

We have used steady-state and time-resolved neutron reflectometry to study the diffusion of fullerene derivatives into the narrow optical gap polymer poly[N-9″-hepta-decanyl-2,7-carbazole-alt-5,5-(4',7'-di-2-thienyl-2',1',3'-benzothiadiazole)] (PCDTBT) to explore the sequential processing of the donor and acceptor for the preparation of efficient organic solar cells. It was found that when [6,6]-phenyl-C61-butyric-acid-methyl-ester (60-PCBM) was deposited onto a thin film of PCDTBT from dichloromethane (DCM), a three-layer structure was formed that was stable below the glass-transition temperature of the polymer. When good solvents for the polymer were used in conjunction with DCM, both 60-PCBM and [6,6]-phenyl-C71-butyric-acid-methyl-ester (70-PCBM) were seen to form films that had a thick fullerene layer containing little polymer and a PCDTBT-rich layer near the interface with the substrate. Devices composed of films prepared by sequential deposition of the polymer and fullerene had efficiencies of up to 5.3%, with those based on 60-PCBM close to optimized bulk heterojunction (BHJ) cells processed in the conventional manner. Sequential deposition of pure components to form the active layer is attractive for large-area device fabrication, and the results demonstrate that this processing method can give efficient solar cells.

Determination of fullerene scattering length density: a critical parameter for understanding the fullerene distribution in bulk heterojunction organic photovoltaic devices.

Fullerene derivatives are commonly used as electron acceptors in combination with (macro)molecular electron donors in bulk heterojunction (BHJ) organic photovoltaic (OPV) devices. Understanding the BHJ structure at different electron donor/acceptor ratios is critical to the continued improvement and development of OPVs. The high neutron scattering length densities (SLDs) of the fullerenes provide effective contrast for probing the distribution of the fullerene within the blend in a nondestructive way. However, recent neutron scattering studies on BHJ films have reported a wide range of SLDs ((3.6-4.4) × 10(-6) Å(-2)) for the fullerenes 60-PCBM and 70-PCBM, leading to differing interpretations of their distribution in thin films. In this article, we describe an approach for determining more precisely the scattering length densities of the fullerenes within a polymer matrix in order to accurately quantify their distribution within the active layers of OPV devices by neutron scattering techniques.

Reconstructing the 3D Coordinates of Guest:Host OLED Blends with Single Atom Resolution.

The performance of electronic and semiconductor devices is critically dependent on the distribution of guest molecules or atoms in a host matrix. One prominent example is that of organic light-emitting diode (OLED) displays containing phosphorescent emitters, now ubiquitous in handheld devices and high-end televisions. In such OLEDs the phosphorescent guest [normally an iridium(III)-based complex] is typically blended into a host matrix, and charge injection and transport, exciton formation and decay, and hence overall device performance are governed by the distribution of the emissive guest in the host. Here high-angle annular dark-field scanning transmission electron microscopy (HAADF-STEM) is used with depth sectioning to reconstruct the 3D distribution of emissive iridium(III) complexes, fac-tris(2-phenylpyridine)iridium(III) [Ir(ppy)3], blended into the amorphous host material, tris(4-carbazoyl-9-ylphenyl)amine (TCTA), by resolving the position of each single iridium(III) ion. It is found that most Ir(ppy)3 complexes are clustered with at least one other, even at low concentrations, and that for films of 20 wt.% Ir(ppy)3 essentially all the complexes are interconnected. The results validate the morphology of blend films created using molecular dynamics simulations which mimic the evaporation film-forming process and are also consistent with the experimentally measured charge transport and photophysical properties.

Differentiating Between V- and G-Series Nerve Agent and Simulant Vapours Using Fluorescent Film Responses.

In-field rapid and reliable identification of nerve agents is critical for the protection of Defence and National Security personnel as well as communities. Fluorescence-based detectors can be portable and provide rapid detection of chemical threats. However, most current approaches cannot differentiate between dilute vapors of nerve agent classes and are susceptible to false positives due to the presence of common acids. Here a fluorescence-based method is shown for rapid differentiation between the V-series and phosphonofluoridate G-series nerve agents and avoids false positives due to common acids. Differentiation is achieved through harnessing two different mechanisms. Detection of the V-series is achieved using photoinduced hole transfer whereby the fluorescence of the sensing material is quenched in the presence of the V-series agent. The G-series is detected using a turn-on mechanism in which a silylated excited state intramolecular proton transfer sensing molecule is selectively deprotected by hydrogen fluoride, which is typically found as a contaminant and/or breakdown product in G-series agents such as sarin. The strategy provided discrimination between classes, as the sensor for the G-series agent class is insensitive to the V-series agent, and vice versa, and neither responded to common acids.

Dilute Donor Organic Solar Cells Based on Non-fullerene Acceptors.

The advent of small molecule non-fullerene acceptor (NFA) materials for organic photovoltaic (OPV) devices has led to a series of breakthroughs in performance and device lifetime. The most efficient OPV devices have a combination of electron donor and acceptor materials that constitute the light absorbing layer in a bulk heterojunction (BHJ) structure. For many BHJ-based devices reported to date, the weight ratio of donor to acceptor is near equal. However, the morphology of such films can be difficult to reproduce and manufacture at scale. There would be an advantage in developing a light harvesting layer for efficient OPV devices that contains only a small amount of either the donor or acceptor. In this work we explore low donor content OPV devices composed of the polymeric donor PM6 blended with high performance NFA materials, Y6 or ITIC-4F. We found that even when the donor:acceptor weight ratio was only 1:10, the OPV devices still have good photoconversion efficiencies of around 6% and 5% for Y6 and ITIC-4F, respectively. It was found that neither charge mobility nor recombination rates had a strong effect on the efficiency of the devices. Rather, the overall efficiency was strongly related to the film absorption coefficient and maintaining adequate interfacial surface area between donor and acceptor molecules/phases for efficient exciton dissociation.