Veterinary Services'use of the Global Burden of Animal Diseases to prioritise interventions, monitor impact and develop critical competencies.

Attracting and sustaining investment in Veterinary Services and animal health programmes from national government budgets, development aid and grants, and philanthropic donors requires economic rationale using relevant, reliable and validated analytical approaches. The complex interwoven relationships between animal health, livestock husbandry systems, national food security, global health security and environmental sustainability emphasise the importance of improving data governance and stewardship and applying economic analysis to understand animal disease burdens. These efforts should enable prioritised investment of limited resources and effective monitoring of the impact of programmes over time. Data governance and stewardship capacities are fundamental to development, implementation and performance monitoring of evidence-based policies in animal health. There are challenges in data availability for national and subnational livestock populations in different sectors, for disease incidence and prevalence, and for animal health expenditure in support of optimised allocation of scarce resources, be they finance, land, labour, or management attention and policy focus. Animal health data systems governance and stewardship and economic analysis are core skills for Veterinary Services in developing and applying evidence-based policy, but capability probably varies among World Organisation for Animal Health (WOAH) Members. The WOAH Performance of Veterinary Services programme has several critical competencies that are relevant to economics of animal health and to data governance and stewardship, but these have not yet been targeted for coordinated capacity development. Implementation of publicâ€"private partnership approaches for animal health programmes creates increasing expectations of robust data and methods for prioritisation, options analysis, and assessing impacts and costs. Experience and examples from national systems in New Zealand, Australia, Ethiopia and Indonesia illustrate current challenges associated with prioritisation of animal health programmes using economic analysis. The Global Burden of Animal Diseases programme intends to support WOAH Members and partners to develop capacities for and standardise approaches to economic analysis and prioritisation in animal health programmes.

Les investissements dans les Services vétérinaires et dans les programmes de santé animale à partir des budgets publics nationaux, des aides et subventions au développement et des fonds alloués par des donateurs philanthropiques peuvent être encouragés et pérennisés au moyen d'une argumentation économique solide fondée sur des méthodes analytiques pertinentes, fiables et validées. La complexité et l'imbrication des relations entre la santé animale, les systèmes d'élevage, la sécurité de l'approvisionnement alimentaire à l'échelle nationale, la sécurité sanitaire mondiale et la durabilité environnementale imposent d'améliorer la gouvernance et la gestion des données et de recourir à des analyses économiques pour mieux comprendre l'impact des maladies animales. Ces efforts devraient permettre de définir les investissements prioritaires dans un contexte de ressources limitées et d'assurer un suivi efficace de l'impact des programmes dans le temps. L'existence de capacités de gouvernance et de gestion des données est donc une condition essentielle pour concevoir et mettre en oeuvre des politiques de santé animale fondées sur des données factuelles et pour suivre leurs performances. Les données disponibles sur les populations d'animaux d'élevage des différentes filières aux niveaux national ou infranational, sur l'incidence et la prévalence des maladies ou sur les dépenses de santé animale sont parfois insuffisantes pour étayer une utilisation optimale de ressources limitées, qu'il s'agisse de moyens financiers, des terres, de la main-d'oeuvre, voire des efforts de gestion ou de la volonté politique. La gouvernance et la gestion des systèmes de données de santé animale et la conduite d'analyses économiques sont des compétences cruciales des Services vétérinaires, que ceux-ci mobilisent pour concevoir et mettre en oeuvre des politiques fondées sur des données factuelles ; il est néanmoins peu probable que ces capacités soient d'un niveau homogène parmi tous les Membres de l'Organisation mondiale de la santé animale (OMSA). Le Processus d'évaluation de la Performance des Services vétérinaires mis en place par l'OMSA définit un certain nombre de compétences critiques dans le domaine de l'économie de la santé animale et de la gouvernance et gestion des données, mais ces compétences n'ont pas encore été intégrées dans un effort coordonné de renforcement des capacités. Les stratégies consistant à confier la mise en oeuvre de programmes de santé animale à des partenariats public-privé suscitent des besoins accrus en données et en méthodes robustes pour l'établissement des priorités, l'analyse des options et l'évaluation des impacts et des coûts. Les auteurs mentionnent les expériences et exemples de systèmes nationaux en Nouvelle-Zélande, en Australie, en Ethiopie et en Indonésie pour illustrer les enjeux actuels liés à l'utilisation des analyses économiques pour définir les priorités des programmes de santé animale. Le programme " Impact mondial des maladies animales " vise à aider les Membres et les partenaires de l'OMSA à renforcer leurs capacités dans le domaine de l'analyse économique et de la définition des priorités des programmes de santé animale et à normaliser leurs approches en la matière.

Para atraer y mantener las inversiones en los Servicios Veterinarios y los programas de sanidad animal procedentes de los presupuestos de los gobiernos nacionales, la ayuda para el desarrollo y las subvenciones, así como de donantes filántropos, se requiere un razonamiento económico en el que se utilicen enfoques analíticos pertinentes, fiables y validados. Las complejas relaciones entre la sanidad animal, los sistemas de ganadería, la seguridad alimentaria nacional, la seguridad sanitaria mundial y la sostenibilidad ambiental ponen de relieve la importancia de mejorar la gobernanza y la gestión de datos y de aplicar el análisis económico para comprender el impacto de las enfermedades animales. Estos esfuerzos deberían permitir establecer prioridades para la inversión de los limitados recursos y realizar un seguimiento eficaz de las repercusiones de los programas a lo largo del tiempo. Las capacidades de gobernanza y gestión de datos son fundamentales para el desarrollo y la implementación de políticas de sanidad animal con una base empírica y para el seguimiento de sus resultados. Existen dificultades en cuanto a la disponibilidad de datos sobre las cabañas ganaderas nacionales y subnacionales de los distintos sectores, la incidencia y prevalencia de las enfermedades y el gasto en sanidad animal que plantean problemas a la hora de optimizar la asignación de unos recursos que son escasos, ya sean los recursos financieros, las tierras, la mano de obra o la atención a la gestión y la orientación de las políticas. La gobernanza y la gestión de los sistemas de datos zoosanitarios y el análisis económico son competencias esenciales para que los Servicios Veterinarios elaboren y apliquen políticas con base empírica, pero es probable que la capacidad varíe entre los Miembros de la Organización Mundial de Sanidad Animal (OMSA). El Proceso de Prestaciones de los Servicios Veterinarios de la OMSA abarca varias competencias esenciales que son relevantes para la economía de la sanidad animal y para la gobernanza y la gestión de datos, pero que aún no han sido objeto de actividades coordinadas de desarrollo de capacidades. La aplicación de enfoques de asociación público-privada para los programas de sanidad animal aumenta aún más las expectativas de datos y métodos sólidos para el establecimiento de prioridades, el análisis de opciones y la evaluación de las repercusiones y los costos. La experiencia y los ejemplos de los sistemas nacionales de Nueva Zelanda, Australia, Etiopía e Indonesia ilustran los retos actuales asociados al establecimiento de prioridades en los programas de sanidad animal mediante el análisis económico. El programa sobre el impacto global de las enfermedades animales pretende ayudar a los Miembros y socios de la OMSA a desarrollar capacidades y armonizar enfoques para el análisis económico y el establecimiento de prioridades en los programas de sanidad animal.

Romosozumab in patients who experienced an on-study fracture: post hoc analyses of the FRAME and ARCH phase 3 trials.

Post hoc analysis of FRAME and ARCH revealed that on-study nonvertebral and vertebral fractures by Month 12 were less common in women initially treated with romosozumab versus placebo or alendronate. Recurrent fracture risk was also lower in romosozumab­treated patients, and there were no fracture­related complications. Results support continuing romosozumab treatment post­fracture. PURPOSE: Post hoc analysis evaluating efficacy and safety of romosozumab, administered in the immediate post­fracture period, in the FRAME and ARCH phase 3 trials. METHODS: In FRAME (NCT01575834) and ARCH (NCT01631214), postmenopausal women with osteoporosis were randomized 1:1 to romosozumab 210 mg monthly or comparator (FRAME, placebo; ARCH, alendronate 70 mg weekly) for 12 months, followed by antiresorptive therapy (FRAME, denosumab; ARCH, alendronate). In patients who experienced on-study nonvertebral or new/worsening vertebral fracture by Month 12, we report the following: fracture and treatment­emergent adverse event (TEAE) incidence through 36 months, bone mineral density changes (BMD), and romosozumab timing. Due to the sample sizes employed, meaningful statistical comparisons between treatments were not possible. RESULTS: Incidence of on-study nonvertebral and vertebral fractures by Month 12 was numerically lower in romosozumab- versus comparator-treated patients (FRAME, 1.6% and 0.5% versus 2.1% and 1.6%; ARCH, 3.4% and 3.3% versus 4.6% and 4.9%, respectively). In those who experienced on-study nonvertebral fracture by Month 12, recurrent nonvertebral and subsequent vertebral fracture incidences were numerically lower in patients initially treated with romosozumab versus comparator (FRAME, 3.6% [2/56] and 1.8% [1/56] versus 9.2% [7/76] and 3.9% [3/76]; ARCH, 10.0% [7/70] and 5.7% [4/70] versus 12.6% [12/95] and 8.4% [8/95], respectively). Among those with on-study vertebral fracture by Month 12, recurrent vertebral and subsequent nonvertebral fracture incidences were numerically lower with romosozumab versus comparator (FRAME, 0.0% [0/17] and 0.0% [0/17] versus 11.9% [7/59] and 8.5% [5/59]; ARCH, 9.0% [6/67] and 7.5% [5/67] versus 15.0% [15/100] and 16.0% [16/100], respectively). In patients with fracture by Month 12, no fracture­related complications were reported in romosozumab-treated patients. BMD gains were numerically greater with romosozumab than comparators. CONCLUSION: Data suggest support for the efficacy and safety of continuing romosozumab treatment following fracture. TRIAL REGISTRATIONS: NCT01575834; NCT01631214.

The impact of face coverings on audio-visual contributions to communication with conversational speech.

The use of face coverings can make communication more difficult by removing access to visual cues as well as affecting the physical transmission of speech sounds. This study aimed to assess the independent and combined contributions of visual and auditory cues to impaired communication when using face coverings. In an online task, 150 participants rated videos of natural conversation along three dimensions: (1) how much they could follow, (2) how much effort was required, and (3) the clarity of the speech. Visual and audio variables were independently manipulated in each video, so that the same video could be presented with or without a superimposed surgical-style mask, accompanied by one of four audio conditions (either unfiltered audio, or audio-filtered to simulate the attenuation associated with a surgical mask, an FFP3 mask, or a visor). Hypotheses and analyses were pre-registered. Both the audio and visual variables had a statistically significant negative impact across all three dimensions. Whether or not talkers' faces were visible made the largest contribution to participants' ratings. The study identifies a degree of attenuation whose negative effects can be overcome by the restoration of visual cues. The significant effects observed in this nominally low-demand task (speech in quiet) highlight the importance of the visual and audio cues in everyday life and that their consideration should be included in future face mask designs.

Direct assessment of nitrative stress in lipid environments: Applications of a designer lipid-based biosensor for peroxynitrite.

Lipid membranes and lipid-rich organelles are targets of peroxynitrite (ONOO-), a highly reactive species generated under nitrative stress. We report a membrane-localized phospholipid (DPPC-TC-ONOO-) that allows the detection of ONOO- in diverse lipid environments: biomimetic vesicles, mammalian cell compartments, and within the lung lining. DPPC-TC-ONOO- and POPC self-assemble to membrane vesicles that fluorogenically and selectively respond to ONOO-. DPPC-TC-ONOO-, delivered through lipid nanoparticles, allowed for ONOO- detection in the endoplasmic reticulum upon cytokine-induced nitrative stress in live mammalian cells. It also responded to ONOO- within lung tissue murine models upon acute lung injury. We observed nitrative stress around bronchioles in precision cut lung slices exposed to nitrogen mustard and in pulmonary macrophages following intratracheal bleomycin challenge. Results showed that DPPC-TC-ONOO- functions specifically toward iNOS, a key enzyme modulating nitrative stress, and offers significant advantages over its hydrophilic analog in terms of localization and signal generation.

The effect of shear-dependent flocculation on the multimodality of effective particle size distributions in a gravel-bed river during high flows.

Purpose: Multimodal effective particle size distributions (EPSDs) develop as flocculation and particle breakage occur dynamically in a fluid shear and such distributions have been previously reported in coastal and estuarine waters to understand flocculation processes. Here, we use time varying multimodal EPSDs and hydraulic parameters (discharge and bed shear stress) to assess freshwater flocculation in a gravel-bed river in southern Alberta, Canada. Methods: Instantaneous discharge, volume concentration (VC), and EPSD of suspended solids were measured during three high discharge events at four study sites in a 10 km reach of the Crowsnest River. The EPSD and VC of suspended solids (< 500 µm) were measured in the centroid of flow with a LISST-200x. Bed shear stress for measured discharge was obtained using a flow model, MOBED. Results: Multimodal EPSDs consisted of primary particles, flocculi, microflocs, and macroflocs. Shear dependent flocculation was consistently observed for all sites and events, due to low and high shear stress flocculation, particle breakage, and mobilization of tributary sub-catchment derived particles. Higher shear stress limited flocculation to smaller floc sizes, while lower bed shear stress conditions created higher volumes of macroflocs. Conclusion: Flocculation and particle breakage processes based on relationships between particle size and hydraulic properties presented herein have implications for advancing fine sediment transport models by a variable cohesion factor as a function of floc size class.

Efficacy of urban road salt reduction strategies on public supply well quality.

Long-term road salt application has increased chloride (Cl-) concentrations in public drinking water wells in many cold climate communities. A range of Best Management Practices (BMPs) have been adopted to mitigate the impact of road deicing compounds on groundwater quality. Chronic increases in chloride levels have been observed in several municipal well fields within the southern Ontario Regional Municipality of Waterloo (RMOW). In response, the RMOW and City of Kitchener implemented a plan to reduce salt application on roads by 25% within the local capture zones of one of the impacted well fields, the Greenbrook Well Field. Here the influence of salt reduction BMPs on subsurface water quality are examined by documenting changes in pore water Cl- concentrations and stored salt mass in vadose zone core samples collected at sites near the well field both before and after the implementation of the BMPs. The data indicate that ~6 years after salt reduction measures were initiated, average pore water Cl- concentration and average cumulative stored chloride mass in the vadose zone had decreased by approximately 60% and 40%, respectively. Groundwater samples collected from shallow monitoring wells installed at each field site showed similar post-BMP reductions in shallow groundwater Cl- concentration (~35%). Long-term (1973-2022) trends in raw water Cl- concentration data from the deeper public drinking water supply wells clearly demonstrate a slow, time-lagged response of the municipal supply wells to the salt reduction BMPs. The combined results suggest that controlled reductions in road salt applications within vulnerable, capture zone regions of public supply wells can reduce the impact of road salt deicing practices on municipal groundwater supplies over time.

Caregiver perspectives on the continued impact of the COVID-19 pandemic on children with intellectual/developmental disabilities.

The COVID-19 pandemic has significantly impacted caregivers, especially those raising a child with an intellectual/developmental disability (IDD). While research has shown substantial disruption to the family, school, and occupational lives of the IDD community, little is known about the long-term impacts of COVID-19. To address this question, 249 caregivers were surveyed via an online questionnaire, between April and August of 2022 (more than 2 years into the pandemic) about potential impacts of the COVID-19 pandemic on their child's access to health- and school-based therapeutic services, caregiver mental health, and family life. The majority of caregivers reported disruptions in access to and quality of school-based therapeutic services for their child as well as a reduction in educational accommodations in the 2021-2022 academic year. Nearly half of caregivers reported feeling anxious and almost a quarter reported feeling depressed for the majority of their days. More than half of respondents reported decreased social support, and one-fifth reported employment disruptions and decreased access to food. These findings suggest that families of children with IDD are still experiencing ongoing negative impacts of the pandemic, emphasizing the critical need for continued support in the wake of the initial and more obvious disruptions caused by the COVID-19 outbreak.

Synthesis of vancomycin fluorescent probes that retain antimicrobial activity, identify Gram-positive bacteria, and detect Gram-negative outer membrane damage.

Antimicrobial resistance is an urgent threat to human health, and new antibacterial drugs are desperately needed, as are research tools to aid in their discovery and development. Vancomycin is a glycopeptide antibiotic that is widely used for the treatment of Gram-positive infections, such as life-threatening systemic diseases caused by methicillin-resistant Staphylococcus aureus (MRSA). Here we demonstrate that modification of vancomycin by introduction of an azide substituent provides a versatile intermediate that can undergo copper-catalysed azide-alkyne cycloaddition (CuAAC) reaction with various alkynes to readily prepare vancomycin fluorescent probes. We describe the facile synthesis of three probes that retain similar antibacterial profiles to the parent vancomycin antibiotic. We demonstrate the versatility of these probes for the detection and visualisation of Gram-positive bacteria by a range of methods, including plate reader quantification, flow cytometry analysis, high-resolution microscopy imaging, and single cell microfluidics analysis. In parallel, we demonstrate their utility in measuring outer-membrane permeabilisation of Gram-negative bacteria. The probes are useful tools that may facilitate detection of infections and development of new antibiotics.

Geometrical frustration versus Kitaev interactions in BaCo2(AsO4)2.

Recently, Co-based honeycomb magnets have been proposed as promising candidate materials to host the Kitaev spin liquid (KSL) state. One of the front-runners is BaCo2(AsO4)2 (BCAO), where it was suggested that the exchange processes between Co2+ ions via the surrounding edge-sharing oxygen octahedra could give rise to bond-dependent Kitaev interactions. In this work, we present and analyze a comprehensive inelastic neutron scattering (INS) study of BCAO with fields in the honeycomb plane. Combining the constraints from the magnon excitations in the high-field polarized state and the inelastic spin structure factor measured in zero magnetic field, we examine two leading theoretical models: the Kitaev-type [Formula: see text] model and the XXZ[Formula: see text]model. We show that the existing experimental data can be consistently accounted for by the XXZ[Formula: see text]model but not by the [Formula: see text] model, and we discuss the implications of these results for the realization of a spin liquid phase in BCAO and more generally for the realization of the Kitaev model in cobaltates.

Bioorthogonal Functionalization of Material Surfaces with Bioactive Molecules.

The functionalization of material surfaces with biologically active molecules is crucial for enabling technologies in life sciences, biotechnology, and medicine. However, achieving biocompatibility and bioorthogonality with current synthetic methods remains a challenge. We report herein a novel surface functionalization method that proceeds chemoselectively and without a free transition metal catalyst. In this method, a coating is first formed via the tyrosinase-catalyzed putative polymerization of a tetrazine-containing catecholamine (DOPA-Tet). One or more types of molecule of interest containing trans-cyclooctene are then grafted onto the coating via tetrazine ligation. The entire process proceeds under physiological conditions and is suitable for grafting bioactive molecules with diverse functions and structural complexities. Utilizing this method, we functionalized material surfaces with enzymes (alkaline phosphatase, glucose oxidase, and horseradish peroxidase), a cyclic peptide (cyclo[Arg-Gly-Asp-D-Phe-Lys], or c(RGDfK)), and an antibiotic (vancomycin). Colorimetric assays confirmed the maintenance of the biocatalytic activities of the grafted enzymes on the surface. We established the mammalian cytocompatibility of the functionalized materials with fibroblasts. Surface functionalization with c(RGDfK) showed improved fibroblast cell morphology and cytoskeletal organization. Microbiological studies with Staphylococcus aureus indicated that surfaces coated using DOPA-Tet inhibit the formation of biofilms. Vancomycin-grafted surfaces additionally display significant inhibition of planktonic S. aureus growth.

Feasibility testing of a standardised virtual clinic for follow-up of patients after hip and knee arthroplasty.

INTRODUCTION: Over 200,000 hip and knee total joint arthroplasties (TJAs) are performed annually in England and Wales. UK guidelines recommend regular follow-up because missed early failure can result in complex revision surgery, which places additional burden on overstretched orthopaedic services. This study evaluated the feasibility and acceptability of an expert, consensus-based, standardised virtual clinic (VC) approach for TJA follow-up. METHODS: Five UK secondary care orthopaedic centres implemented a standardised VC. Feedback was obtained through patient satisfaction questionnaires and telephone interviews with arthroplasty care practitioners. Key stakeholders subsequently attended an expert discussion forum to achieve consensus on the final VC format and to address obstacles identified during testing. RESULTS: From 19 June 2018 to 11 December 2018, 561 TJA patients [mean age (SD) 70 (9.4) years, 57.8% female, 69.0% hip TJA, 1-28 years postsurgery (median 5 years)] completed a VC. Of these 561 patients, 82.2% were discharged without attending an outpatient appointment and 46 (8.8%) required early face-to-face consultant review. Patient satisfaction with the VC was high (156/188; 83.0%); over 70% of patients indicated a preference for the VC. DISCUSSION: This feasibility study suggested significant resource savings, including time spent by consultant orthopaedic surgeons in outpatient clinics, hospital transport and an estimated saving of up to two-thirds of usual clinic-allotted time. The expert discussion forum provided helpful feedback for supporting more efficient implementation of the VC. CONCLUSIONS: A standardised VC is a feasible alternative to outpatient clinics for the follow-up of hip and knee TJA patients, and is acceptable to key stakeholders, including patients.

Quantum wake dynamics in Heisenberg antiferromagnetic chains.

Traditional spectroscopy, by its very nature, characterizes physical system properties in the momentum and frequency domains. However, the most interesting and potentially practically useful quantum many-body effects emerge from local, short-time correlations. Here, using inelastic neutron scattering and methods of integrability, we experimentally observe and theoretically describe a local, coherent, long-lived, quasiperiodically oscillating magnetic state emerging out of the distillation of propagating excitations following a local quantum quench in a Heisenberg antiferromagnetic chain. This "quantum wake" displays similarities to Floquet states, discrete time crystals and nonlinear Luttinger liquids. We also show how this technique reveals the non-commutativity of spin operators, and is thus a model-agnostic measure of a magnetic system's "quantumness."

"Are You Doing Any Sport Science?" A Brief Editorial.

This brief opinion-based editorial addresses what the authors perceive to be a fundamental issue in the application of sport science, and these issues are reflected by the question "Are you doing any sport science?" As sport science has grown within the United States, organizational sport science budgets have grown, with increasing interest in developing various sport science initiatives. While it is indeed an exciting time for sport science, the authors suggest that, too often, sport science pursuits are driven by commercially available technologies and viewed as an "add-on" instead of pursuing an integrated systematic approach to informing the training process.

CHESS: The future direct geometry spectrometer at the second target station.

CHESS, chopper spectrometer examining small samples, is a planned direct geometry neutron chopper spectrometer designed to detect and analyze weak signals intrinsic to small cross sections (e.g., small mass, small magnetic moments, or neutron absorbing materials) in powders, liquids, and crystals. CHESS is optimized to enable transformative investigations of quantum materials, spin liquids, thermoelectrics, battery materials, and liquids. The broad dynamic range of the instrument is also well suited to study relaxation processes and excitations in soft and biological matter. The 15 Hz repetition rate of the Second Target Station at the Spallation Neutron Source enables the use of multiple incident energies within a single source pulse, greatly expanding the information gained in a single measurement. Furthermore, the high flux grants an enhanced capability for polarization analysis. This enables the separation of nuclear from magnetic scattering or coherent from incoherent scattering in hydrogenous materials over a large range of energy and momentum transfer. This paper presents optimizations and technical solutions to address the key requirements envisioned in the science case and the anticipated uses of this instrument.

Fast bacterial growth reduces antibiotic accumulation and efficacy.

Phenotypic variations between individual microbial cells play a key role in the resistance of microbial pathogens to pharmacotherapies. Nevertheless, little is known about cell individuality in antibiotic accumulation. Here, we hypothesise that phenotypic diversification can be driven by fundamental cell-to-cell differences in drug transport rates. To test this hypothesis, we employed microfluidics-based single-cell microscopy, libraries of fluorescent antibiotic probes and mathematical modelling. This approach allowed us to rapidly identify phenotypic variants that avoid antibiotic accumulation within populations of Escherichia coli, Pseudomonas aeruginosa, Burkholderia cenocepacia, and Staphylococcus aureus. Crucially, we found that fast growing phenotypic variants avoid macrolide accumulation and survive treatment without genetic mutations. These findings are in contrast with the current consensus that cellular dormancy and slow metabolism underlie bacterial survival to antibiotics. Our results also show that fast growing variants display significantly higher expression of ribosomal promoters before drug treatment compared to slow growing variants. Drug-free active ribosomes facilitate essential cellular processes in these fast-growing variants, including efflux that can reduce macrolide accumulation. We used this new knowledge to eradicate variants that displayed low antibiotic accumulation through the chemical manipulation of their outer membrane inspiring new avenues to overcome current antibiotic treatment failures.

Bacteria can cause an array of diseases ranging from mildly inconvenient to deadly. In fact, every year around the world, five million people succumb to a bacterial infection. Antibiotics can kill bacteria or stop their growth, but many bacterial species are now able to evade these drugs. To be efficient, most antibiotics first need to get inside a bacterium; there, they accumulate until they reach the concentration they need to act. Often, the drugs make their way through channel-like structures ('pores') studded through the external membranes of bacteria and which control the passage of molecules in and out of cells. Resistance usually emerges when genetic changes provide the microorganism with an advantage against antibiotics, or when the microorganism performs the biochemical reactions necessary for life at a slower pace. In contrast, Lapinska, Pagliara et al. decided to examine how genetically similar Escherichia coli bacteria which differed in their growth rate would fare against antibiotics. The drug targeted ribosomes, the machinery that produces proteins in a cell. A combination of techniques was used to follow individual cells, revealing that fast-growing variants better managed to survive. A closer look showed that bacteria which were growing quickly had a surplus of ribosomes, which then produced more pores that could pump the antibiotic out the cell. Next, Lapinska, Pagliara et al. exposed the bacteria to both the antibiotic and a compound that weakens bacterial membrane; this erased the advantage shown by the fast-growing variants. Overall, this work gives a finer understanding of the mechanisms that underlie antibiotic resistance, which could help pave the way to new strategies to combat harmful bacteria.

Application of antibiotic-derived fluorescent probes to bacterial studies.

Fluorescent probes are extensively applied as useful tools for imaging and determining dynamic processes in bacterial cells. In particular, antibiotic-derived fluorescent probes which can visualize the presence or the localization of antibiotics within bacteria through the monitoring of changes in fluorescence signal, are particularly useful. They form an emerging set of tools for studying the mode of action of their parent antibiotics and examining bacterial resistance and persistence, with the long-term goal of developing fresh approaches to the treatment of drug-resistant bacterial infections. In this chapter, we discuss the applications of antibiotic-based fluorescent probes to visualize bacteria, focusing on the techniques we have utilized to study their localization, penetration and efflux. We describe detailed protocols for analysis of bacteria using microscopy, flow cytometry, and plate reader-based methods based on these probes.

Dirac Magnons, Nodal Lines, and Nodal Plane in Elemental Gadolinium.

We investigate the magnetic excitations of elemental gadolinium (Gd) using inelastic neutron scattering, showing that Gd is a Dirac magnon material with nodal lines at K and nodal planes at half integer ℓ. We find an anisotropic intensity winding around the K-point Dirac magnon cone, which is interpreted to indicate Berry phase physics. Using linear spin wave theory calculations, we show the nodal lines have nontrivial Berry phases, and topological surface modes. We also discuss the origin of the nodal plane in terms of a screw-axis symmetry, and introduce a topological invariant characterizing its presence and effect on the scattering intensity. Together, these results indicate a highly nontrivial topology, which is generic to hexagonal close packed ferromagnets. We discuss potential implications for other such systems.

An outbreak of livestock-associated meticillin-resistant Staphylococcus aureus (LA-MRSA) clonal complex 398 in a regional burns centre.

BACKGROUND: Meticillin-resistant Staphylococcus aureus (MRSA) outbreaks have been reported previously in burns centres with resulting mortality and morbidity. This article describes the first human-associated outbreak in the UK caused by a strain of mupirocin-resistant (MuR) livestock-associated MRSA clonal complex 398 (LA-MRSA CC398) in an adult burns centre. The centre historically had a very low prevalence of MRSA infections. AIM: To describe the clinical and epidemiological context of how the outbreak was identified and contained using a range of infection prevention and control (IPC) measures guided by both traditional and genetic methods. METHODS: A cluster of MuR-MRSA led to an outbreak investigation. Cases were detected via retrospective search and real-time laboratory surveillance. Isolates were sent continuously for whole-genome sequencing (WGS). A live timeline of cases and interventions was produced throughout the period. FINDINGS: The outbreak consisted of 12 cases (seven males and five females) aged between 22 and 70 years. Patients were identified between May and October 2020. All patients were colonized rather than infected. The strain acquired the plasmid bearing MupA while colonizing the index case before dissemination. The index case was found to be a chicken farmer. This outbreak was eventually controlled using IPC measures, audits, and blind staff decolonization guided by insight from WGS. CONCLUSION: It was not possible to determine how the strain entered the centre, or if a staff carrier was involved. The outbreak demonstrated the potential for continued transmissions for months despite active surveillance and stringent control measures.

CryoEM structure of the outer membrane secretin channel pIV from the f1 filamentous bacteriophage.

The Ff family of filamentous bacteriophages infect gram-negative bacteria, but do not cause lysis of their host cell. Instead, new virions are extruded via the phage-encoded pIV protein, which has homology with bacterial secretins. Here, we determine the structure of pIV from the f1 filamentous bacteriophage at 2.7 Å resolution by cryo-electron microscopy, the first near-atomic structure of a phage secretin. Fifteen f1 pIV subunits assemble to form a gated channel in the bacterial outer membrane, with associated soluble domains projecting into the periplasm. We model channel opening and propose a mechanism for phage egress. By single-cell microfluidics experiments, we demonstrate the potential for secretins such as pIV to be used as adjuvants to increase the uptake and efficacy of antibiotics in bacteria. Finally, we compare the f1 pIV structure to its homologues to reveal similarities and differences between phage and bacterial secretins.

Serum Complement Activation by C4BP-IgM Fusion Protein Can Restore Susceptibility to Antibiotics in Neisseria gonorrhoeae.

Neisseria gonorrhoeae is the etiological agent of gonorrhea, the second most common bacterial sexually transmitted infection worldwide. Reproductive sequelae of gonorrhea include infertility, ectopic pregnancy and chronic pelvic pain. Most antibiotics currently in clinical use have been rendered ineffective due to the rapid spread of antimicrobial resistance among gonococci. The developmental pipeline of new antibiotics is sparse and novel therapeutic approaches are urgently needed. Previously, we utilized the ability of N. gonorrhoeae to bind the complement inhibitor C4b-binding protein (C4BP) to evade killing by human complement to design a chimeric protein that linked the two N-terminal gonococcal binding domains of C4BP with the Fc domain of IgM. The resulting molecule, C4BP-IgM, enhanced complement-mediated killing of gonococci. Here we show that C4BP-IgM induced membrane perturbation through complement deposition and membrane attack complex pore insertion facilitates the access of antibiotics to their intracellular targets. Consequently, bacteria become more susceptible to killing by antibiotics. Remarkably, C4BP-IgM restored susceptibility to azithromycin of two azithromycin-resistant clinical gonococcal strains because of overexpression of the MtrC-MtrD-MtrE efflux pump. Our data show that complement activation can potentiate activity of antibiotics and suggest a role for C4BP-IgM as an adjuvant for antibiotic treatment of drug-resistant gonorrhea.