Safety of mapping the motor networks in the spinal cord using penetrating microelectrodes in Yucatan minipigs.

OBJECTIVE: The goal of this study was to assess the safety of mapping spinal cord locomotor networks using penetrating stimulation microelectrodes in Yucatan minipigs (YMPs) as a clinically translational animal model. METHODS: Eleven YMPs were trained to walk up and down a straight line. Motion capture was performed, and electromyographic (EMG) activity of hindlimb muscles was recorded during overground walking. The YMPs underwent a laminectomy and durotomy to expose the lumbar spinal cord. Using an ultrasound-guided stereotaxic frame, microelectrodes were inserted into the spinal cord in 8 animals. Pial cuts were made to prevent tissue dimpling before microelectrode insertion. Different locations within the lumbar enlargement were electrically stimulated to map the locomotor networks. The remaining 3 YMPs served as sham controls, receiving the laminectomy, durotomy, and pial cuts but not microelectrode insertion. The Porcine Thoracic Injury Behavioral Scale (PTIBS) and hindlimb reflex assessment results were recorded for 4 weeks postoperatively. Overground gait kinematics and hindlimb EMG activity were recorded again at weeks 3 and 4 postoperatively and compared with preoperative measures. The animals were euthanized at the end of week 4, and the lumbar spinal cords were extracted and preserved for immunohistochemical analysis. RESULTS: All YMPs showed transient deficits in hindlimb function postoperatively. Except for 1 YMP in the experimental group, all animals regained normal ambulation and balance (PTIBS score 10) at the end of weeks 3 and 4. One animal in the experimental group showed gait and balance deficits by week 4 (PTIBS score 4). This animal was excluded from the kinematics and EMG analyses. Overground gait kinematic measures and EMG activity showed no significant (p > 0.05) differences between preoperative and postoperative values, and between the experimental and sham groups. Less than 5% of electrode tracks were visible in the tissue analysis of the animals in the experimental group. There was no statistically significant difference in damage caused by pial cuts between the experimental and sham groups. Tissue damage due to the pial cuts was more frequently observed in immunohistochemical analyses than microelectrode tracks. CONCLUSIONS: These findings suggest that mapping spinal locomotor networks in porcine models can be performed safely, without lasting damage to the spinal cord.

Vascular adhesion protein-1 blockade in primary sclerosing cholangitis: Open-label, multicenter, single-arm, phase II trial.

BACKGROUND: Primary sclerosing cholangitis is a progressive inflammatory liver disease characterized by biliary and liver fibrosis. Vascular adhesion protein-1 (VAP-1) is important in the inflammatory process driving liver fibrosis. We evaluated the safety and efficacy of VAP-1 blockade with a monoclonal antibody (timolumab, BTT1023) in patients with primary sclerosing cholangitis. METHODS: BUTEO was a prospective, single-arm, open-label, multicenter, phase II trial, conducted in 6 centers in the United Kingdom. Patients with primary sclerosing cholangitis aged 18-75 years had an alkaline phosphatase value of >1.5 times the upper limit of normal. The dose-confirmatory stage aimed to confirm the safety of timolumab through the incidence of dose-limiting toxicity and sufficient trough levels of circulating antibody to block VAP-1 function. The primary outcome of the dose-expansion portion of the trial was patient's response to timolumab at day 99, as measured by a reduction in serum alkaline phosphatase by 25% or more from baseline to day 99. RESULTS: Twenty-three patients were recruited: 7 into the initial dose-confirmatory stage and a further 16 into an expansion stage. Timolumab (8 mg/kg) was confirmed to be safe for the duration of administration with sufficient circulating levels. Only 2 of the 18 evaluable patients (11.1%) achieved a reduction in alkaline phosphatase levels of 25% or more, and both the proportion of circulating inflammatory cell populations and biomarkers of fibrosis remained unchanged from baseline. CONCLUSIONS: The BUTEO trial confirmed 8 mg/kg timolumab had no short-term safety signals and resulted in sufficient circulating levels of VAP-1 blocking timolumab. However, the trial was stopped after an interim assessment due to a lack of efficacy as determined by no significant change in serum liver tests.

Effects of cannabidiol on psychosocial stress, situational anxiety and nausea in a virtual reality environment: a protocol for a single-centre randomised clinical trial.

INTRODUCTION: The non-intoxicating plant-derived cannabinoid, cannabidiol (CBD), has demonstrated therapeutic potential in a number of clinical conditions. Most successful clinical trials have used relatively high (≥300 mg) oral doses of CBD. Relatively few studies have investigated the efficacy of lower (<300 mg) oral doses, typical of those available in over-the-counter CBD products. METHODS: We present a protocol for a randomised, double-blind, placebo-controlled, parallel-group clinical trial investigating the effects of a low oral dose (150 mg) of CBD on acute psychosocial stress, situational anxiety, motion sickness and cybersickness in healthy individuals. Participants (n=74) will receive 150 mg of CBD or a matched placebo 90 min before completing three virtual reality (VR) challenges (tasks) designed to induce transient stress and motion sickness: (a) a 15 min 'Public Speaking' task; (b) a 5 min 'Walk the Plank' task (above a sheer drop); and (c) a 5 min 'Rollercoaster Ride' task. The primary outcomes will be self-reported stress and nausea measured on 100 mm Visual Analogue Scales. Secondary outcomes will include salivary cortisol concentrations, skin conductance, heart rate and vomiting episodes (if any). Statistical analyses will test the hypothesis that CBD reduces nausea and attenuates subjective, endocrine and physiological responses to stress compared with placebo. This study will indicate whether low-dose oral CBD has positive effects in reducing acute psychosocial stress, situational anxiety, motion sickness and cybersickness. ETHICS AND DISSEMINATION: The University of Sydney Human Research Ethics Committee has granted approval (2023/307, version 1.6, 16 February 2024). Study findings will be disseminated in a peer-reviewed journal and at academic conferences. TRIAL REGISTRATION NUMBER: Australian New Zealand Clinical Trials Registry (ACTRN12623000872639).

Wildlife-friendly garden practices increase butterfly abundance and species richness in urban and arable landscapes.

Insect declines are being reported worldwide and private, residential gardens could provide refugia for these species under increasing land use change. Interest in wildlife-friendly gardening has increased, but many management recommendations lack a scientific evidence-base. We used a large citizen science scheme, the Garden Butterfly Survey (GBS), with data from over 600 gardens across Great Britain (2016-2021) to determine how the surrounding landscape influences the abundance and species richness of butterflies in gardens and whether wildlife-friendly gardening practices, such as having long grass and providing nectar plants, benefit butterflies. First, we show that GBS provides reliable estimates of species abundances by comparing with results from standardised, long-term monitoring data. Garden size and surrounding land use had significant effects on butterfly abundance and richness in gardens, including positive relationships with garden size, woodland and arable farmland and negative relationships with urbanisation. Both the presence and area of long grass in gardens were positively related to higher butterfly richness and abundance, with the latter being driven by butterflies that use grasses as larval host plants. These effects differed depending on the surrounding landscape, such that long grass resulted in higher garden butterfly abundance in landscapes dominated by arable farming, and higher abundance and richness in highly urbanised areas. The presence of flowering ivy (Hedera spp.) in gardens resulted in higher abundance of Celastrina argiolus holly blue which uses ivy as a larval host, and of Vanessa atalanta red admiral and Polygonia c-album comma, which favour it as a nectar source. Our work provides evidence that undertaking simple wildlife-friendly garden practices can be beneficial for attracting butterflies, particularly in heavily modified areas. With over 728,000 ha of gardens in Great Britain, the cumulative effect of leaving areas of lawn uncut and providing nectar and larval host plants could be key for helping biodiversity.

Anthropogenic climate and land-use change drive short- and long-term biodiversity shifts across taxa.

Climate change and habitat loss present serious threats to nature. Yet, due to a lack of historical land-use data, the potential for land-use change and baseline land-use conditions to interact with a changing climate to affect biodiversity remains largely unknown. Here, we use historical land use, climate data and species observation data to investigate the patterns and causes of biodiversity change in Great Britain. We show that anthropogenic climate change and land conversion have broadly led to increased richness, biotic homogenization and warmer-adapted communities of British birds, butterflies and plants over the long term (50+ years) and short term (20 years). Biodiversity change was found to be largely determined by baseline environmental conditions of land use and climate, especially over shorter timescales, suggesting that biodiversity change in recent periods could reflect an inertia derived from past environmental changes. Climate-land-use interactions were mostly related to long-term change in species richness and beta diversity across taxa. Semi-natural grasslands (in a broad sense, including meadows, pastures, lowland and upland heathlands and open wetlands) were associated with lower rates of biodiversity change, while their contribution to national-level biodiversity doubled over the long term. Our findings highlight the need to protect and restore natural and semi-natural habitats, alongside a fuller consideration of individual species' requirements beyond simple measures of species richness in biodiversity management and policy.

Linking climate warming and land conversion to species' range changes across Great Britain.

Although increased temperatures are known to reinforce the effects of habitat destruction at local to landscape scales, evidence of their additive or interactive effects is limited, particularly over larger spatial extents and longer timescales. To address these deficiencies, we created a dataset of land-use changes over 75 years, documenting the loss of over half (>3000 km2) the semi-natural grassland of Great Britain. Pairing this dataset with climate change data, we tested for relationships to distribution changes in birds, butterflies, macromoths, and plants (n = 1192 species total). We show that individual or additive effects of climate warming and land conversion unambiguously increased persistence probability for 40% of species, and decreased it for 12%, and these effects were reflected in both range contractions and expansions. Interactive effects were relatively rare, being detected in less than 1 in 5 species, and their overall effect on extinction risk was often weak. Such individualistic responses emphasise the importance of including species-level information in policies targeting biodiversity and climate adaptation.

Optimizing the strain engineering process for industrial-scale production of bio-based molecules.

Biomanufacturing could contribute as much as ${\$}$30 trillion to the global economy by 2030. However, the success of the growing bioeconomy depends on our ability to manufacture high-performing strains in a time- and cost-effective manner. The Design-Build-Test-Learn (DBTL) framework has proven to be an effective strain engineering approach. Significant improvements have been made in genome engineering, genotyping, and phenotyping throughput over the last couple of decades that have greatly accelerated the DBTL cycles. However, to achieve a radical reduction in strain development time and cost, we need to look at the strain engineering process through a lens of optimizing the whole cycle, as opposed to simply increasing throughput at each stage. We propose an approach that integrates all 4 stages of the DBTL cycle and takes advantage of the advances in computational design, high-throughput genome engineering, and phenotyping methods, as well as machine learning tools for making predictions about strain scale-up performance. In this perspective, we discuss the challenges of industrial strain engineering, outline the best approaches to overcoming these challenges, and showcase examples of successful strain engineering projects for production of heterologous proteins, amino acids, and small molecules, as well as improving tolerance, fitness, and de-risking the scale-up of industrial strains.

The first report of meningitis in a Greenland shark (Somniosus microcephalus).

The Greenland shark (Somniosus microcephalus) is a large species of shark found in the North Atlantic and Arctic Oceans and is believed to be the longest living vertebrate. Relatively little is known about its biology, abundance, health or diseases. In March 2022, only the third reported UK stranding of this species occurred and it was the first to undergo post-mortem examination. The animal was a sexually immature female, measuring 3.96 m in length and 285 kg in weight, and was in poor nutritional state. Gross findings included haemorrhages in the skin and soft tissues, particularly of the head, and silt in the stomach suggestive of live stranding, bilateral corneal opacity, slightly turbid cerebrospinal fluid (CSF) and patchy congestion of the brain. Histopathological findings included keratitis and anterior uveitis, fibrinonecrotic and lymphohistiocytic meningitis of the brain and proximal spinal cord and fibrinonecrotizing choroid plexitis. A near pure growth of a Vibrio organism was isolated from CSF. This is believed to be the first report of meningitis in this species.

Precipitation buffers temperature-driven local extinctions of moths at warm range margins.

Species' distributions are moving polewards in response to climate change, and although range expansions of relatively warm-adapted species are widely reported, reports of range retractions in cool-adapted species are less common. Here, we analysed species' distribution shifts for 76 cool-adapted moths in Great Britain using citizen science occurrence records from the National Moth Recording Scheme over a 40-year period. Although we find evidence for trailing edge shifts to higher latitudes, shifts in species' range centroids are oriented towards the north-west, and are more closely correlated with directional changes in total precipitation than average temperature. We also found that species' local extinction risk is higher in areas where temperature is high and precipitation is low, but this risk diminishes as precipitation increases. Adaptation efforts should therefore focus on maintaining or increasing water availability as the climate continues to change.

Virtual reality digital surgical planning for jaw reconstruction: a usability study.

BACKGROUND: Digital surgical planning (DSP) has revolutionized the preparation and execution of the management of complex head and neck pathologies. The addition of virtual reality (VR) allows the surgeon to have a three-dimensional experience with six degrees of freedom for visualizing and manipulating objects. This pilot study describes the participants experience with the first head and neck reconstructive VR-DSP platform. METHODS: An original VR-DSP platform has been developed for planning the ablation and reconstruction of head and neck pathologies. A prospective trial utilizing this platform involving reconstructive surgeons was performed. Participants conducted a simulated VR-DSP planning session, pre- and post-questionnaire as well as audio recordings allowing for qualitative analysis. RESULTS: Thirteen consultant reconstructive surgeons representing three surgical backgrounds with varied experience were recruited. The majority of surgeons had no previous experience with VR. Based on the system usability score, the VR-DSP platform was found to have above average usability. The qualitative analysis demonstrated the majority had a positive experience. Participants identified some perceived barriers to implementing the VR-DSP platform. CONCLUSIONS: Virtual reality-digital surgical planning is usable and acceptable to reconstructive surgeons. Surgeons were able to perform the steps in an efficient time despite limited experience. The addition of VR offers additional benefits to current VSP platforms. Based on the results of this pilot study, it is likely that VR-DSP will be of benefit to the reconstructive surgeon.

Traits data for the butterflies and macro-moths of Great Britain and Ireland.

Butterflies and moths, collectively Lepidoptera, are integral components of ecosystems, providing key services such as pollination and a prey resource for vertebrate and invertebrate predators. Lepidoptera are a relatively well studied group of invertebrates. In Great Britain and Ireland numerous citizen science projects provide data on changes in distribution and abundance. The availability of high-quality monitoring and recording data, combined with the rapid response of Lepidoptera to environmental change, makes them ideal candidates for traits-based ecological studies. Recently, there has been an increase in the number of studies documenting traits-based responses of Lepidoptera, highlighting the demand for a standardized and referenced traits database. There is a wide range of primary and secondary literature sources available regarding the ecology of British and Irish Lepidoptera to support such studies. Currently these sources have not been collated into one central repository that would facilitate and enhance future research. Here, we present a comprehensive traits database for the butterflies and macro-moths of Great Britain and Ireland. The database covers 968 species in 21 families. Ecological traits fall into four main categories: life cycle ecology and phenology, host plant specificity and characteristics, breeding habitat, and morphological characteristics. The database also contains data regarding species distribution, conservation status, and temporal trends for abundance and occupancy. This database can be used for a wide array of purposes including further fundamental research on species and community responses to environmental change, conservation and management studies, and evolutionary biology. There are no copyright restrictions, and this paper must be cited if data are used in publications.

Street lighting has detrimental impacts on local insect populations.

Reported declines in insect populations have sparked global concern, with artificial light at night (ALAN) identified as a potential contributing factor. Despite strong evidence that lighting disrupts a range of insect behaviors, the empirical evidence that ALAN diminishes wild insect abundance is limited. Using a matched-pairs design, we found that street lighting strongly reduced moth caterpillar abundance compared with unlit sites (47% reduction in hedgerows and 33% reduction in grass margins) and affected caterpillar development. A separate experiment in habitats with no history of lighting revealed that ALAN disrupted the feeding behavior of nocturnal caterpillars. Negative impacts were more pronounced under white light-emitting diode (LED) street lights compared to conventional yellow sodium lamps. This indicates that ALAN and the ongoing shift toward white LEDs (i.e., narrow- to broad-spectrum lighting) will have substantial consequences for insect populations and ecosystem processes.

Magnetic resonance imaging analysis of human skull diploic venous anatomy.

BACKGROUND: The skull diploic venous space (DVS) represents a potential route for cerebrospinal fluid (CSF) diversion and absorption in the treatment of hydrocephalus. The goal of this study was to carry out a detailed characterization of the drainage pattern of the DVS of the skull using high-resolution MRI, especially the diploic veins draining to the lacunae laterales (LLs) since the LLs constitute an important channel for the CSF to access the superior sagittal sinus and subsequently the systemic circulation. The objective was to identify those skull regions optimally suited for an intraosseous CSF diversion system. METHODS: High-resolution, T1-weighted MRI scans from 20 adult and 16 pediatric subjects were selected for analysis. Skulls were divided into four regions, that is, frontal, parietal, temporal, and occipital. On each scan, a trained observer counted all diploic veins in every skull region. Each diploic vein was also followed to determine its final drainage pathway (i.e., dural venous sinus, dural vein, LL, or indeterminate). RESULTS: In the adult age group, the frontal and occipital skull regions showed the highest number of diploic veins. However, the highest number of draining diploic veins connecting to the lacunae lateralis was found in the frontal and parietal skull region, just anterior and just posterior to the coronal suture. In the pediatric age group, the parietal skull region, just posterior to the coronal suture, showed the highest overall number of diploic veins and also the highest number of draining diploic veins connecting to the LL. CONCLUSION: This study suggested that diploic venous density across the skull varies with age, with more parietal diploic veins in the pediatric age range, and more occipital and frontal diploic veins in adults. If the DVS is ultimately used for CSF diversion, our anatomical data point to optimal sites for the insertion of specially designed intraosseous infusion devices for the treatment of hydrocephalus. Likely the optimal sites for CSF diversion would be the parietal region just posterior to the coronal suture in children, and in adults, frontal and/or parietal just anterior or just posterior to the coronal suture.

Population Bottlenecks Strongly Affect the Evolutionary Dynamics of Antibiotic Persistence.

Bacterial persistence is a potential cause of antibiotic therapy failure. Antibiotic-tolerant persisters originate from phenotypic differentiation within a susceptible population, occurring with a frequency that can be altered by mutations. Recent studies have proven that persistence is a highly evolvable trait and, consequently, an important evolutionary strategy of bacterial populations to adapt to high-dose antibiotic therapy. Yet, the factors that govern the evolutionary dynamics of persistence are currently poorly understood. Theoretical studies predict far-reaching effects of bottlenecking on the evolutionary adaption of bacterial populations, but these effects have never been investigated in the context of persistence. Bottlenecking events are frequently encountered by infecting pathogens during host-to-host transmission and antibiotic treatment. In this study, we used a combination of experimental evolution and barcoded knockout libraries to examine how population bottlenecking affects the evolutionary dynamics of persistence. In accordance with existing hypotheses, small bottlenecks were found to restrict the adaptive potential of populations and result in more heterogeneous evolutionary outcomes. Evolutionary trajectories followed in small-bottlenecking regimes additionally suggest that the fitness landscape associated with persistence has a rugged topography, with distinct trajectories toward increased persistence that are accessible to evolving populations. Furthermore, sequencing data of evolved populations and knockout libraries after selection reveal various genes that are potentially involved in persistence, including previously known as well as novel targets. Together, our results do not only provide experimental evidence for evolutionary theories, but also contribute to a better understanding of the environmental and genetic factors that guide bacterial adaptation to antibiotic treatment.

Cabin air quality on non-smoking commercial flights: A review of published data on airborne pollutants.

We reviewed 47 documents published 1967-2019 that reported measurements of volatile organic compounds (VOCs) on commercial aircraft. We compared the measurements with the air quality standards and guidelines for aircraft cabins and in some cases buildings. Average levels of VOCs for which limits exist were lower than the permissible levels except for benzene with average concentration at 5.9 ± 5.5 µg/m3 . Toluene, benzene, ethylbenzene, formaldehyde, acetaldehyde, limonene, nonanal, hexanal, decanal, octanal, acetic acid, acetone, ethanol, butanal, acrolein, isoprene and menthol were the most frequently measured compounds. The concentrations of semi-volatile organic compounds (SVOCs) and other contaminants did not exceed standards and guidelines in buildings except for the average NO2 concentration at 12 ppb. Although the focus was on VOCs, we also retrieved the data on other parameters characterizing cabin environment. Ozone concentration averaged 38 ppb below the upper limit recommended for aircraft. The outdoor air supply rate ranged from 1.7 to 39.5 L/s per person and averaged 6.0 ± 0.8 L/s/p (median 5.8 L/s/p), higher than the minimum level recommended for commercial aircraft. Carbon dioxide concentration averaged 1315 ± 232 ppm, lower than what is permitted in aircraft and close to what is permitted in buildings. Measured temperatures averaged 23.5 ± 0.8°C and were generally within the ranges recommended for avoiding thermal discomfort. Relative humidity averaged 16% ± 5%, lower than what is recommended in buildings.

Outcomes of patients with metastatic cutaneous squamous cell carcinoma to the axilla: a multicentre cohort study.

BACKGROUND: Metastatic cutaneous squamous cell carcinoma to the axilla is uncommon, with limited data to guide management. We sought to assess the outcomes of patients with this condition after surgery and radiotherapy. METHODS: A retrospective cohort study of patients treated at two Australian hospitals from 1994 through 2016 was performed. RESULTS: A total of 74 patients were identified, including 48 treated curatively with surgery-plus-radiotherapy and 15 with surgery alone. Compared with patients treated with surgery alone, a higher proportion of patients treated with surgery-plus-radiotherapy had lymph nodes larger than 6 cm (53% versus 8%, P = 0.012) and multiple adverse histopathological features (75% versus 47%, P = 0.04). The groups had similar 5-year disease-free survival (45% versus 46%) and overall survival (51% versus 48%). Presence of multiple positive lymph nodes was associated with reduced disease-free survival (hazard ratio 4.57, P = 0.01) and overall survival (hazard ratio 3.53, P = 0.02). Regional recurrence was higher in patients treated with surgery alone (38% versus 22%, P = 0.22) and patients with lymph nodes larger than 6 cm (34% versus 10%, P = 0.03). All recurrences occurred within 2 years following treatment. CONCLUSION: Combined-modality therapy for metastatic cutaneous squamous cell carcinoma to the axilla is recommended for high-risk patients, although outcomes remain modest. The key period for recurrence is within 2 years following treatment.