Evaluating the druggability of TrmD, a potential antibacterial target, through design and microbiological profiling of a series of potent TrmD inhibitors.

The post-transcriptional modifier tRNA-(N1G37) methyltransferase (TrmD) has been proposed to be essential for growth in many Gram-negative and Gram-positive pathogens, however previously reported inhibitors show only weak antibacterial activity. In this work, optimisation of fragment hits resulted in compounds with low nanomolar TrmD inhibition incorporating features designed to enhance bacterial permeability and covering a range of physicochemical space. The resulting lack of significant antibacterial activity suggests that whilst TrmD is highly ligandable, its essentiality and druggability are called into question.

Restoring carbapenem efficacy: a novel carbapenem companion targeting metallo-ß-lactamases in carbapenem-resistant Enterobacterales.

BACKGROUND: The dissemination of MBLs compromises effective use of many ß-lactams in the treatment of patients with life-threatening bacterial infections. Predicted global increases in the prevalence of MBL-producing carbapenem-resistant Enterobacterales (CRE) are being realized, yielding infections that are untreatable with existing therapies including newly approved ß-lactam/ß-lactamase inhibitor combinations. Developing MBL inhibitors (MBLIs) now is essential to address the growing threat that MBL-producing CRE pose to patients. METHODS: A novel MBLI series was assessed by susceptibility testing and time-kill assays. Target activity and selectivity was evaluated using bacterial NDM, VIM and IMP enzyme assays and human matrix metallopeptidase enzyme assays, respectively, and cytotoxicity was assessed in HepG2 cells. In vivo efficacy of meropenem/MBLI combinations was evaluated in a mouse thigh infection model using an NDM-1-producing Escherichia coli strain. RESULTS: Combination of MBLIs with carbapenems reduced MICs for NDM/IMP/VIM-producing Enterobacterales by up to 128-fold compared with the carbapenems alone. Supplementation of meropenem with the promising compound 272 reduced the MIC90 from 128 to 0.25 mg/L in a panel of MBL-producing CRE clinical isolates (n = 115). Compound 272 restored the bactericidal activity of meropenem and was non-cytotoxic, potentiating the antimicrobial action of meropenem through specific inhibition of NDM, IMP and VIM. In vivo efficacy was achieved in a mouse thigh infection model with meropenem/272 dosed subcutaneously. CONCLUSIONS: We have developed a series of rationally designed MBLIs that restore activity of carbapenems against NDM/IMP/VIM-producing Enterobacterales. This series warrants further development towards a novel combination therapy that combats antibiotic-resistant organisms, which pose a critical threat to human health.

Emotion in animal contests.

Emotions encompass cognitive and behavioural responses to reward and punishment. Using contests as a case-study, we propose that short-term emotions underpin animals' assessments, decision-making and behaviour. Equating contest assessments to emotional 'appraisals', we describe how contestants appraise more than resource value and outcome probability. These appraisals elicit the cognition, drive and neurophysiology that governs aggressive behaviour. We discuss how recent contest outcomes induce long-term moods, which impact subsequent contest behaviour. Finally, we distinguish between integral (objectively relevant) and incidental (objectively irrelevant) emotions and moods (affective states). Unlike existing ecological models, our approach predicts that incidental events influence contest dynamics, and that contests become incidental influences themselves, potentially causing maladaptive decision-making. As affective states cross contexts, a more holistic ethology (incorporating emotions and moods) would illuminate animal cognition and behaviour.

Potential for repurposing the personal care product preservatives bronopol and bronidox as broad-spectrum antibiofilm agents for topical application.

OBJECTIVES: Bacterial biofilms represent a major impediment to healing in chronic wounds and are largely refractory to the antibacterial agents currently used in wound management. From a repurposing screen of compounds considered safe for topical application in humans, we report the identification of the personal care product preservatives bronopol and bronidox as broad-spectrum antibiofilm agents and potential candidates for reducing biofilm burden in chronic wounds. METHODS: Antibiofilm activity was assessed by viable counting against single-species biofilms of Staphylococcus aureus and Pseudomonas aeruginosa in the Calgary Biofilm Device, and against mixed-species biofilms of the two organisms growing on nitrocellulose discs. RESULTS: Bronopol and bronidox exhibited broad-spectrum antibiofilm activity that encompassed the two major wound pathogens, S. aureus and P. aeruginosa. When impregnated into gauze dressings at their existing maximum authorized concentrations for safe use and placed onto an established mixed-species biofilm, bronopol and bronidox completely eradicated P. aeruginosa and achieved an ∼5 log10 reduction in the S. aureus population. The antibiofilm action of bronopol and bronidox was attributed to their ability to kill slow- or non-growing bacteria found in biofilms, and both compounds exhibited synergistic antibiofilm effects in combination with established wound-treatment agents. CONCLUSIONS: Bronopol and bronidox kill bacteria regardless of growth state, a property that endows them with broad-spectrum antibiofilm activity. As this effect is observed at concentrations authorized for use on human skin, these compounds represent promising candidates for the treatment of chronic wounds.

On the Stability of Polymeric Nanoparticles Fabricated through Rapid Solvent Mixing.

We study the stability of polymeric nanoparticles fabricated through the rapid mixing of polymers in a good solvent with a poor solvent that is miscible with the good solvent. In previous experiments where water was used as the poor solvent, a negative surface charge was measured on the precipitated nanoparticles, which led to the long-time stability of the dispersion. It was argued that these charges originate presumably from either water or hydroxide adsorption at the hydrophobic nanoparticle surface or from impurities in the feed streams that preferentially adsorb on the precipitated nanoparticles. To elucidate the origin of this stabilization mechanism, we performed experiments wherein we replaced water with a nonpolar poor solvent. The polymers aggregated into stable nanoparticles for a range of processing parameters. We investigated theoretically three possible explanations for this stability, i.e., electrostatic stabilization, conditional thermodynamic equilibrium, and steric stabilization. Our experiments and considerations suggest that steric stabilization is the most likely candidate.

Scalable Platform for Structured and Hybrid Soft Nanocolloids by Continuous Precipitation in a Confined Environment.

Geometrically structured polymer nanocolloids, including Janus nanocolloids, have been widely investigated for their unique properties, which are derived from their anisotropy. Controlled surface decoration with inorganic nanoparticles could induce another level of functionality into structured nanocolloids that could enable applications in fields ranging from rewriteable electronics to biphasic catalysis. Here, we demonstrate flash nanoprecipitation (FNP) as a one-step, scalable process platform for manufacturing hybrid polymer-inorganic nanocolloids in which one phase is selectively decorated with a metal nanocatalyst by tuning the molecular interactions between the feed ingredients during the process. For instance, by modifying the polymer end-group functionality, we document the ability to tune the location of the metal nanocatalyst, including placement at the nanocolloid circumference. Moreover, the addition of molecular additives is shown to transform the Janus nanocolloid structure from spherical to dumbbell or snowman while maintaining the ability to control the nanocatalyst location. In considering the flexibility and continuous nature of the FNP process, it offers an industrial-scale platform for the manufacturing of nanomaterials that are anticipated to impact many technologies.

Combining Precipitation and Vitrification to Control the Number of Surface Patches on Polymer Nanocolloids.

In an effort to incorporate increasingly higher levels of functionality into soft nanoparticles, heterogeneously structured particles stand out as a simple means to enhance functionality by tailoring only particle architecture. Various means exist for the fabrication of particles with specific structural configurations; however, the tunability of particle morphology is still a challenging and often laborious task, especially in self-assembled systems where a single equilibrium configuration dominates. Improved strategies for multipatch particle assembly are therefore needed to allow for the tailoring of particle structure via a single, continuous assembly route. One means of accomplishing this is through kinetic trapping of particle morphologies along the path to the final equilibrium configuration in precipitation-induced, phase-separating polymer blends. Here, we demonstrate this capability by using rapid nanoprecipitation to control the overall size, composition, and patch distribution of soft colloids. In particular, we illustrate that polymer feed concentration, blend ratio, and polymer molecular weight can all serve as functional handles with which to consistently alter particle patch distributions in a self-assembling homopolymer system without redesigning the starting materials. We furthermore delineate the role of polymer vitrification in the determination of particle structure.

Social behavior in farm animals: Applying fundamental theory to improve animal welfare.

A fundamental understanding of behavior is essential to improving the welfare of billions of farm animals around the world. Despite living in an environment managed by humans, farm animals are still capable of making important behavioral decisions that influence welfare. In this review, we focus on social interactions as perhaps the most dynamic and challenging aspects of the lives of farm animals. Social stress is a leading welfare concern in livestock, and substantial variation in social behavior is seen at the individual and group level. Here, we consider how a fundamental understanding of social behavior can be used to: (i) understand agonistic and affiliative interactions in farm animals; (ii) identify how artificial environments influence social behavior and impact welfare; and (iii) provide insights into the mechanisms and development of social behavior. We conclude by highlighting opportunities to build on previous work and suggest potential fundamental hypotheses of applied relevance. Key areas for further research could include identifying the welfare benefits of socio-positive interactions, the potential impacts of disrupting important social bonds, and the role of skill in allowing farm animals to navigate competitive and positive social interactions. Such studies should provide insights to improve the welfare of farm animals, while also being applicable to other contexts, such as zoos and laboratories.

Tuning Morphologies and Reactivities of Hybrid Organic-Inorganic Nanoparticles.

Hybrid nanoparticles (hNPs), or nanoparticles composed of both organic and inorganic components, hold promise for diverse energy and environmental applications due to their ability to stabilize reactive nanomaterials against aggregation, enhancing their ability to pervade tortuous spaces and travel long distances to degrade contaminants in situ. Past studies have investigated the use of polymer or surfactant coatings to stabilize nanomaterials against aggregation. However, fabrication of these materials often requires multiple steps and lacks specificity in the control of their morphologies and reactivities. Here, we demonstrated a method of producing stable hNPs with tunable morphologies by incubating polystyrene nanoparticles formed via Flash NanoPrecipitation with citrate-stabilized gold nanocatalysts. Using this simple fabrication technique, we found that gold adsorption to polystyrene nanoparticles was enabled by the presence of a good solvent for polystyrene. Furthermore, changing process parameters, such as gold incubation time, and molecular parameters, such as polymer molecular weight and end-group functionality, provided control over the resultant nanocatalyst loading and dispersal atop hNPs. We classified these morphologies into three distinct regimes─aggregated, dispersed, or internalized─and we showed that the emergence of these regimes has key implications for controlling reaction rates in applications such as heterogeneous catalysis or groundwater remediation. Specifically, we found that hNPs with gold nanocatalysts embedded below the surfaces of polystyrene nanoparticles exhibited slower bulk catalytic reduction capacity than their disperse, surface-decorated counterparts. Taken together, our work demonstrates a simple way by which hNPs can be fabricated and presents a method to control catalytic reactions using reactive nanomaterials.

Animal cognition in an urbanised world.

Explaining how animals respond to an increasingly urbanised world is a major challenge for evolutionary biologists. Urban environments often present animals with novel problems that differ from those encountered in their evolutionary past. To navigate these rapidly changing habitats successfully, animals may need to adjust their behaviour flexibly over relatively short timescales. These behavioural changes, in turn, may be facilitated by an ability to acquire, store and process information from the environment. The question of how cognitive abilities allow animals to avoid threats and exploit resources (or constrain their ability to do so) is attracting increasing research interest, with a growing number of studies investigating cognitive and behavioural differences between urban-dwelling animals and their non-urban counterparts. In this review we consider why such differences might arise, focusing on the informational challenges faced by animals living in urban environments, and how different cognitive abilities can assist in overcoming these challenges. We focus largely on birds, as avian taxa have been the subject of most research to date, but discuss work in other species where relevant. We also address the potential consequences of cognitive variation at the individual and species level. For instance, do urban environments select for, or influence the development of, particular cognitive abilities? Are individuals or species with particular cognitive phenotypes more likely to become established in urban habitats? How do other factors, such as social behaviour and individual personality, interact with cognition to influence behaviour in urban environments? The aim of this review is to synthesise current knowledge and identify key avenues for future research, in order to improve our understanding of the ecological and evolutionary consequences of urbanisation.

The Role of Animal Cognition in Human-Wildlife Interactions.

Humans have a profound effect on the planet's ecosystems, and unprecedented rates of human population growth and urbanization have brought wild animals into increasing contact with people. For many species, appropriate responses toward humans are likely to be critical to survival and reproductive success. Although numerous studies have investigated the impacts of human activity on biodiversity and species distributions, relatively few have examined the effects of humans on the behavioral responses of animals during human-wildlife encounters, and the cognitive processes underpinning those responses. Furthermore, while humans often present a significant threat to animals, the presence or behavior of people may be also associated with benefits, such as food rewards. In scenarios where humans vary in their behavior, wild animals would be expected to benefit from the ability to discriminate between dangerous, neutral and rewarding people. Additionally, individual differences in cognitive and behavioral phenotypes and past experiences with humans may affect animals' ability to exploit human-dominated environments and respond appropriately to human cues. In this review, we examine the cues that wild animals use to modulate their behavioral responses toward humans, such as human facial features and gaze direction. We discuss when wild animals are expected to attend to certain cues, how information is used, and the cognitive mechanisms involved. We consider how the cognitive abilities of wild animals are likely to be under selection by humans and therefore influence population and community composition. We conclude by highlighting the need for long-term studies on free-living, wild animals to fully understand the causes and ecological consequences of variation in responses to human cues. The effects of humans on wildlife behavior are likely to be substantial, and a detailed understanding of these effects is key to implementing effective conservation strategies and managing human-wildlife conflict.

In Silico Design Enables the Rapid Production of Surface-Active Colloidal Amphiphiles.

A new technology platform built on the integration of theory and experiments to enable the design of Janus colloids with precision control of surface anisotropy and amphiphilicity could lead to a disruptive transformation in the next generation of surfactants, photonic or phononic materials, and coatings. Here, we exploit molecular dynamics (MD) simulations to guide the rational design of amphiphilic polymer Janus colloids by Flash NanoPrecipitation (FNP), a method capable of the production of colloids with complex structure without the compromise of reduced scalability. Aided by in silico design, we show in experiments that amphiphilic Janus colloids can be produced using a unique blend of hydrophobic homopolymers and the addition of an amphiphilic block copolymer. The final structure of the colloids depends on the mass fraction of each homopolymer as well as the concentration and composition of the block copolymer additive. To confirm the surface activity of the colloids, we demonstrate their potential to stabilize Pickering emulsions. This hybrid approach of simulations and experiments provides a pathway to designing and manufacturing complex polymeric colloids on an industrial scale.

Responsive Polymers as Smart Nanomaterials Enable Diverse Applications.

Responsive polymers undergo reversible or irreversible physical or chemical modifications in response to a change in environment or stimulus, e.g., temperature, pH, light, and magnetic or electric fields. Polymeric nanoparticles (NPs), which constitute a diverse set of morphologies, including micelles, vesicles, and core-shell geometries, have been successfully prepared from responsive polymers and have shown great promise in applications ranging from drug delivery to catalysis. In this review, we summarize pH, thermo-, photo-, and enzymatic responsiveness for a selection of polymers. We then discuss the formation of NPs made from responsive polymers. Finally, we highlight how NPs and other nanomaterials are enabling a wide range of smart applications with improved efficiency, as well as improved sustainability and recyclability of polymeric systems.

Testing relationship recognition in wild jackdaws (Corvus monedula).

According to the social intelligence hypothesis, understanding the challenges faced by social animals is key to understanding the evolution of cognition. In structured social groups, recognising the relationships of others is often important for predicting the outcomes of interactions. Third-party relationship recognition has been widely investigated in primates, but studies of other species are limited. Furthermore, few studies test for third-party relationship recognition in the wild, where cognitive abilities are deployed in response to natural socio-ecological pressures. Here, we used playback experiments to investigate whether wild jackdaws (Corvus monedula) track changes in their own relationships and the relationships of others. Females were presented with 'infidelity simulations': playbacks of their male partner copulating with a neighbouring female, and their male neighbour copulating with another female, against a congruent control. Our results showed substantial inter-individual variation in responses, but females did not respond more strongly to infidelity playbacks, indicating that jackdaws may not attend and/or respond to relationship information in this experimental context. Our results highlight the need for further study of relationship recognition and other cognitive traits that facilitate group-living in the wild, particularly in non-primates and in a wider range of social systems.

Social learning about dangerous people by wild jackdaws.

For animals that live alongside humans, people can present both an opportunity and a threat. Previous studies have shown that several species can learn to discriminate between individual people and assess risk based on prior experience. To avoid potentially costly encounters, it may also pay individuals to learn about dangerous people based on information from others. Social learning about anthropogenic threats is likely to be beneficial in habitats dominated by human activity, but experimental evidence is limited. Here, we tested whether wild jackdaws (Corvus monedula) use social learning to recognize dangerous people. Using a within-subjects design, we presented breeding jackdaws with an unfamiliar person near their nest, combined with conspecific alarm calls. Subjects that heard alarm calls showed a heightened fear response in subsequent encounters with the person compared to a control group, reducing their latency to return to the nest. This study provides important evidence that animals use social learning to assess the level of risk posed by individual humans.

Testing social learning of anti-predator responses in juvenile jackdaws: the importance of accounting for levels of agitation.

Social learning is often assumed to help young animals respond appropriately to potential threats in the environment. We brought wild, juvenile jackdaws briefly into captivity to test whether short exposures to conspecific vocalizations are sufficient to promote anti-predator learning. Individuals were presented with one of two models-a stuffed fox representing a genuine threat, or a toy elephant simulating a novel predator. Following an initial baseline presentation, juveniles were trained by pairing models with either adult mobbing calls, indicating danger, or contact calls suggesting no danger. In a final test phase with no playbacks, birds appeared to have habituated to the elephant, regardless of training, but responses to the fox remained high throughout, suggesting juveniles already recognized it as a predator before the experiment began. Training with mobbing calls did seem to generate elevated escape responses, but this was likely to be a carry-over effect of the playback in the previous trial. Overall, we found little evidence for social learning. Instead, individuals' responses were mainly driven by their level of agitation immediately preceding each presentation. These results highlight the importance of accounting for agitation in studies of anti-predator learning, and whenever animals are held in captivity for short periods.

Rapid Production of Internally Structured Colloids by Flash Nanoprecipitation of Block Copolymer Blends.

Colloids with internally structured geometries have shown great promise in applications ranging from biosensors to optics to drug delivery, where the internal particle structure is paramount to performance. The growing demand for such nanomaterials necessitates the development of a scalable processing platform for their production. Flash nanoprecipitation (FNP), a rapid and inherently scalable colloid precipitation technology, is used to prepare internally structured colloids from blends of block copolymers and homopolymers. As revealed by a combination of experiments and simulations, colloids prepared from different molecular weight diblock copolymers adopt either an ordered lamellar morphology consisting of concentric shells or a disordered lamellar morphology when chain dynamics are sufficiently slow to prevent defect annealing during solvent exchange. Blends of homopolymer and block copolymer in the feed stream generate more complex internally structured colloids, such as those with hierarchically structured Janus and patchy morphologies, due to additional phase separation and kinetic trapping effects. The ability of the FNP process to generate such a wide range of morphologies using a simple and scalable setup provides a pathway to manufacturing internally structured colloids on an industrial scale.

Reversible precipitation of bovine serum albumin by metal ions and synthesis, structure and reactivity of new tetrathiometallate chelating agents.

Independent research is an important component of any undergraduate chemistry program. This article reports the findings of two of many undergraduate research projects directed by Ed Stiefel in the hopes that the results will be inspiring and useful to the scientific community. The neurological disorders associated with insufficient copper in Menkes disease and an excess of copper in Wilson's disease are well established; however, recent evidence suggests that copper may also be involved in other disorders, such as Alzheimer's, angiogenesis, and prion diseases. The exact role of copper, however, is uncertain. This study examines the role of copper and zinc in the formation of protein deposits and the chelation and removal of the metal ions to reverse the process. The bovine serum albumin (BSA) protein forms a precipitate after the addition of approximately 6 copper(II) atoms or 8 zinc(II) atoms. Other metal ions, such as Ca(II), Al(III), Ni(II), and Co(II), did not precipitate the BSA even when the metal ion to BSA ratios were in excess of 1000. The copper and zinc protein precipitates returned to solution after addition of the chelating agents, ethylenediaminetetraacetic acid (EDTA) or tetrathiometallates [(MS(4)(2-)), where M=Mo, W]. Two new choline and acetylcholine tetrathiomolybdate and tetrathiotungstate chelating agents have been synthesized and characterized. The infrared (IR) and X-ray crystal structures of the complexes revealed that the (MS(4)(2-)) cores had approximate T(d) symmetry in the choline (Ch) salts and C(2v) symmetry in the acetylcholine (AcCh) salts. The AcCh salts hydrolyzed more slowly than the ammonium or Ch salts and the tetrathiotungstate salts hydrolyzed approximately two orders of magnitude more slowly than the tetrathiomolybdate salts. The slower hydrolysis of tetrathiotungstate may make it more useful as an inorganic reagent and therapeutic agent.

Directed Assembly of Soft Colloids through Rapid Solvent Exchange.

We studied the directed assembly of soft nanoparticles through rapid micromixing of polymers in solution with a nonsolvent. Both experiments and computer simulations were performed to elucidate the underlying physics and to investigate the role of various process parameters. In particular, we discovered that no external stabilizing agents or charged end groups are required to keep the colloids separated from each other when water is used as the nonsolvent. Furthermore, the size of the nanoparticles can be reliably tuned through the mixing rate and the ratio between polymer solution and nonsolvent. Our results demonstrate that this mechanism is highly promising for the mass fabrication of uniformly sized colloidal particles, using a wide variety of polymeric feed materials.

Effects of age and experience on contest behavior in the burying beetle, Nicrophorus vespilloides.

Contest behavior forms an important part of reproductive investment. Life-history theory predicts that as individuals age and their residual reproductive value decreases, they should increase investment in contest behavior. However, other factors such as social experience may also be important in determining age-related variation in contest behavior. To understand how selection acts on contest behavior over an individual's lifetime, it is therefore important to tease apart the effects of age per se from other factors that may vary with age. Here, we independently manipulate male age and social experience to examine their effects on male contest behavior in the burying beetle Nicrophorus vespilloides. We found that social experience, but not age, influenced male contest behavior but that these changes in behavior did not alter contest outcomes. Male size (relative to his opponent) was overwhelmingly the most important factor determining contest outcome. Our results suggest that in systems with high variation in fighting ability among males, there may be little opportunity for selection to act on factors that influence contest outcomes by altering motivation to win.