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Abstract The term supramolecular polymer has been applied to polymeric materials in which the individual units, i.e., building blocks—are bound to each other via noncovalent interactions, including electrostatic or hydrogen bonding, as well as metal–ligand conjugation. The building blocks are generally low molecular weight amphiphiles. Methods for preparing biopolymers based on non-toxic, metal–ligand conjugation have been little studied; however, they offer significant potential for tuning the response of biologically relevant macromolecules. In this communication, we characterize the assembly and morphology of supramolecular biopolymers in which the building blocks are low- or medium-molecular weight globular proteins—ubiquitin and Cas9-interacting via metal–ligand conjugation. In each case, the protein gene was expressed in cell culture with the addition of hexa-His/linkers at both the N and C termini. Divalent cations investigated were Zn2+ and Ni2+. We observe in cryo-TEM imaging an absolute requirement for divalent cations for the formation of supramolecular biopolymers. In the presence of Ni2+, 1D assembled fibers are predominant, while with Zn2+, the more frequently detected structures are sheet-like. We use gel electrophoresis and CD spectroscopy to monitor possible secondary and tertiary structural changes in the protein building blocks during conjugation.

Abstract One of the family patterns crucial for mental and physical well-being is differentiation of self. In this study, our aim was to investigate its impact on the phenomenon of phubbing—where individuals prioritize smartphones over in-person interactions. The prevalence of phubbing behavior has risen substantially in tandem with the increased adoption of smartphones. The study investigated familial, interpersonal, and personal factors that could potentially contribute to the escalation of phubbing behaviors. It was hypothesized that differentiation of self would be associated with phubbing through the mediation of fear of missing out, romantic relationship satisfaction, and loneliness in intimate relationships. We also expected gender differences in the study variables. A sample of 431 young adults, with an average age of 29 (M = 29.05, SD = 9.14), completed the DSI-R, Phubbing, FoMO, ENRICH, and LIRS questionnaires online. Results indicated that fear of missing out mediated the relationship between three dimensions of differentiation of self—emotional reactivity, emotional cutoff, and fusion with others—and phubbing behavior. However, the expected mediation by romantic relationship satisfaction and loneliness in intimate relationships did not reach significance, although these variables were found to be associated with differentiation of self. Women reported higher levels of phubbing behavior, emotional reactivity, and fusion with others, whereas men reported higher levels of I-position. We conclude that fear of missing out may serve as a catalyst, triggering anxiety in individuals, which in turn drives them to adopt phubbing as a coping mechanism. Moreover, individuals with lower levels of differentiation of self appear to be at increased risk of engaging in phubbing behaviors through the mediation of fear of missing out.

Abstract Many nocturnally active fireflies use precisely timed bioluminescent patterns to identify mates, making them especially vulnerable to light pollution. As urbanization continues to brighten the night sky, firefly populations are under constant stress, and close to half of the species are now threatened. Ensuring the survival of firefly biodiversity depends on a large-scale conservation effort to monitor and protect thousands of populations. While species can be identified by their flash patterns, current methods require expert measurement and manual classification and are infeasible given the number and geographic distribution of fireflies. Here we present the application of a recurrent neural network (RNN) for accurate automated firefly flash pattern classification. Using recordings from commodity cameras, we can extract flash trajectories of individuals within a swarm and classify their species with an accuracy of approximately seventy percent. In addition to its potential in population monitoring, automated classification provides the means to study firefly behavior at the population level. We employ the classifier to measure and characterize the variability within and between swarms, unlocking a new dimension of their behavior. Our method is open source, and deployment in community science applications could revolutionize our ability to monitor and understand firefly populations.

Introduction Critically ill patients are at risk of suboptimal beta-lactam antibiotic (beta-lactam) exposure due to the impact of altered physiology on pharmacokinetics. Suboptimal concentrations can lead to treatment failure or toxicity. Therapeutic drug monitoring (TDM) involves adjusting doses based on measured plasma concentrations and individualising dosing to improve the likelihood of improving exposure. Despite its potential benefits, its adoption has been slow, and data on implementation, dose adaptation and safety are sparse. The aim of this trial is to assess the feasibility and fidelity of implementing beta-lactam TDM-guided dosing in the intensive care unit setting.Methods and analysis A beta-lactam antibiotic Dose AdaPtation feasibility randomised controlled Trial using Therapeutic Drug Monitoring (ADAPT-TDM) is a single-centre, unblinded, feasibility randomised controlled trial aiming to enroll up to 60 critically ill adult participants (≥18 years). TDM and dose adjustment will be performed daily in the intervention group; the standard of care group will undergo plasma sampling, but no dose adjustment. The main outcomes include: (1) feasibility of recruitment, defined as the number of participants who are recruited from a pool of eligible participants, and (2) fidelity of TDM, defined as the degree to which TDM as a test is delivered as intended, from accurate sample collection, sample processing to result availability. Secondary outcomes include target attainment, uptake of TDM-guided dosing and incidence of neurotoxicity, hepatotoxicity and nephrotoxicity.Ethics and dissemination This study has been approved by the Alfred Hospital human research ethics committee, Office of Ethics and Research Governance (reference: Project No. 565/22; date of approval: 22/11/2022). Prospective consent will be obtained and the study will be conducted in accordance with the Declaration of Helsinki. The finalised manuscript, including aggregate data, will be submitted for publication in a peer reviewed journal. ADAPT-TDM will determine whether beta-lactam TDM-guided dose adaptation is reproducible and feasible and provide important information required to implement this intervention in a phase III trial.Trial registration number Australian New Zealand Clinical Trials Registry, ACTRN12623000032651.

Despite the central role of T cells in tumor immunity, attempts to harness their cytotoxic capacity as a therapy have met limited efficacy, partially as a result of the suppressive microenvironment which limits their migration and activation. In contrast, myeloid cells massively infiltrate tumors and are well adapted to survive these harsh conditions. While they are equipped with cell-killing abilities, they often adopt an immunosuppressive phenotype upon migration to tumors. Therefore, the questions of how to modify their activation programming against cancer, and what signaling cascades should be activated in myeloid cells to elicit their cytotoxicity have remained unclear. Here, we found that activation of IgM-induced signaling in murine myeloid cells results in secretion of lytic granules and massive tumor cell death. These findings open venues for designing novel immunotherapy by equipping monocytes with chimeric receptors that target tumor antigens and consequently, signal through IgM receptor. Nonetheless, we found that myeloid cells do not express the antibody-derived portion used to recognize the tumor antigen due to the induction of an ER stress response. To overcome this limitation, we designed chimeric receptors that are based on the high-affinity FcγRI for IgG. Incubation of macrophages expressing these receptors along with tumor-binding IgG induced massive tumor cell killing and secretion of reactive oxygen species and Granzyme B. Overall, this work highlights the challenges involved in genetically reprogramming the signaling in myeloid cells and provides a framework for endowing myeloid cells with antigen-specific cytotoxicity.

Abstract Patients with chronic hepatitis B (CHB) are regularly monitored for HBV DNA and liver enzymes in order to assess disease progression and the need for antiviral therapy. Identifying patients with a stable course of disease can potentially prolong the intervals between visits, withhold unnecessary tests and save money. Accordingly, we aimed to find predictors for a stable disease course in patients with CHB. 579 patients with CHB, who were followed in a tertiary referral center between January 2004–December 2018, were retrospectively analyzed. Patients with low and steady viral load titer (

Photobacterium damselae subspecies damselae, an abundant, generalist marine pathogen, has been reported in various cetaceans worldwide. We report a bottlenose dolphin in the eastern Mediterranean Sea that was found stranded and dead. The dolphin had a severe case of chronic suppurative pneumonia and splenic lymphoid depletion caused by this pathogen.

Biological aging can be described as accumulative, prolonged metabolic stress and is the major risk factor for cognitive decline and Alzheimer’s disease (AD). Recently, we identified and described a quinone reductase 2 (QR2) pathway in the brain, in which QR2 acts as a removable memory constraint and metabolic buffer within neurons. QR2 becomes overexpressed with age, and it is possibly a novel contributing factor to age-related metabolic stress and cognitive deficit. We found that, in human cells, genetic removal of QR2 produced a shift in the proteome opposing that found in AD brains while simultaneously reducing oxidative stress. We therefore created highly specific QR2 inhibitors (QR2is) to enable evaluation of chronic QR2 inhibition as a means to reduce biological age–related metabolic stress and cognitive decline. QR2is replicated results obtained by genetic removal of QR2, while local QR2i microinjection improved hippocampal and cortical-dependent learning in rats and mice. Continuous consumption of QR2is in drinking water improved cognition and reduced pathology in the brains of AD-model mice (5xFAD), with a noticeable between-sex effect on treatment duration. These results demonstrate the importance of QR2 activity and pathway function in the healthy and neurodegenerative brain and what we believe to be the great therapeutic potential of QR2is as first-in-class drugs.

BackgroundSince treatment with immune checkpoint inhibitors (ICIs) is becoming standard therapy for patients with high-risk and advanced melanoma, an increasing number of patients experience treatment-related adverse events such as fatigue. Until now, studies have demonstrated the benefits of using eHealth tools to provide either symptom monitoring or interventions to reduce treatment-related symptoms such as fatigue. However, an eHealth tool that facilitates the combination of both symptom monitoring and symptom management in patients with melanoma treated with ICIs is still needed. ObjectiveIn this pilot study, we will explore the use of the CAPABLE (Cancer Patients Better Life Experience) app in providing symptom monitoring, education, and well-being interventions on health-related quality of life (HRQoL) outcomes such as fatigue and physical functioning, as well as patients’ acceptance and usability of using CAPABLE. MethodsThis prospective, exploratory pilot study will examine changes in fatigue over time in 36 patients with stage III or IV melanoma during treatment with ICI using CAPABLE (a smartphone app and multisensory smartwatch). This cohort will be compared to a prospectively collected cohort of patients with melanoma treated with standard ICI therapy. CAPABLE will be used for a minimum of 3 and a maximum of 6 months. The primary endpoint in this study is the change in fatigue between baseline and 3 and 6 months after the start of treatment. Secondary end points include HRQoL outcomes, usability, and feasibility parameters. ResultsStudy inclusion started in April 2023 and is currently ongoing. ConclusionsThis pilot study will explore the effect, usability, and feasibility of CAPABLE in patients with melanoma during treatment with ICI. Adding the CAPABLE system to active treatment is hypothesized to decrease fatigue in patients with high-risk and advanced melanoma during treatment with ICIs compared to a control group receiving standard care. The Medical Ethics Committee NedMec (Amsterdam, The Netherlands) granted ethical approval for this study (reference number 22-981/NL81970.000.22). Trial RegistrationClinicalTrials.gov NCT05827289; https://clinicaltrials.gov/study/NCT05827289 International Registered Report Identifier (IRRID)DERR1-10.2196/49252

Abstract To survive during colony reproduction, bees create dense clusters of thousands of suspended individuals. How does this swarm, which is orders of magnitude larger than the size of an individual, maintain mechanical stability? We hypothesize that the internal structure in the bulk of the swarm, about which there is little prior information, plays a key role in mechanical stability. Here, we provide the first-ever 3D reconstructions of the positions of the bees in the bulk of the swarm using x-ray computed tomography. We find that the mass of bees in a layer decreases with distance from the attachment surface. By quantifying the distribution of bees within swarms varying in size (made up of 4000–10,000 bees), we find that the same power law governs the smallest and largest swarms, with the weight supported by each layer scaling with the mass of each layer to the $$\approx 1.5$$ ≈ 1.5 power. This arrangement ensures that each layer exerts the same fraction of its total strength, and on average a bee supports a lower weight than its maximum grip strength. This illustrates the extension of the scaling law relating weight to strength of single organisms to the weight distribution within a superorganism made up of thousands of individuals.

Objective To conduct a systematic review of the literature on the association between fibromyalgia and mortality and to pool the results in a meta-analysis.Methods The authors searched the PubMed, Scopus, and Web of Science databases using the key words ‘fibromyalgia’ and ‘mortality’ to identify studies that addressed an association between fibromyalgia and mortality. Original papers that assessed associations between fibromyalgia and mortality (all or specific causes) and provided an effect measure (hazard ratio (HR), standardised mortality ratio (SMR), odds ratio (OR)) quantifying the relationship between fibromyalgia and mortality were included in the systematic review. Of 557 papers that were initially identified using the search words, 8 papers were considered eligible for the systematic review and meta-analysis. We used a Newcastle-Ottawa scale to assess the risk of bias in the studies.Results The total fibromyalgia group included 188 751 patients. An increased HR was found for all-cause mortality (HR 1.27, 95% CI 1.04 to 1.51), but not for the subgroup diagnosed by the 1990 criteria. There was a borderline increased SMR for accidents (SMR 1.95, 95% CI 0.97 to 3.92), an increased risk for mortality from infections (SMR 1.66, 95% CI 1.15 to 2.38), and suicide (SMR 3.37, 95% CI 1.52 to 7.50), and a decreased mortality rate for cancer (SMR 0.82, 95% CI 0.69 to 0.97). The studies showed significant heterogeneity.Conclusions These potential associations indicate that fibromyalgia should be taken seriously, with a special focus on screening for suicidal ideation, accident prevention, and the prevention and treatment of infections.

Here, we present the first analysis of the connectome of a small volume of the Octopus vulgaris vertical lobe (VL), a brain structure mediating the acquisition of long-term memory in this behaviorally advanced mollusk. Serial section electron microscopy revealed new types of interneurons, cellular components of extensive modulatory systems, and multiple synaptic motifs. The sensory input to the VL is conveyed via~1.8 × 106 axons that sparsely innervate two parallel and interconnected feedforward networks formed by the two types of amacrine interneurons (AM), simple AMs (SAMs) and complex AMs (CAMs). SAMs make up 89.3% of the~25 × 106VL cells, each receiving a synaptic input from only a single input neuron on its non-bifurcating primary neurite, suggesting that each input neuron is represented in only~12 ± 3.4SAMs. This synaptic site is likely a ‘memory site’ as it is endowed with LTP. The CAMs, a newly described AM type, comprise 1.6% of the VL cells. Their bifurcating neurites integrate multiple inputs from the input axons and SAMs. While the SAM network appears to feedforward sparse ‘memorizable’ sensory representations to the VL output layer, the CAMs appear to monitor global activity and feedforward a balancing inhibition for ‘sharpening’ the stimulus-specific VL output. While sharing morphological and wiring features with circuits supporting associative learning in other animals, the VL has evolved a unique circuit that enables associative learning based on feedforward information flow.

Abstract Background The training of near-peer (NP) teachers and junior faculty instructors received major attention as a possible solution for the shortage of experienced anatomy instructors in faculties of medicine and health professions. Several studies described the training of NP teachers and junior instructors (≤ 2 years of teaching experience) using various methods. However, few publications include On the Job Training (OJT), which enables reflection and performance evaluation and encourages professionals to cope with their blind spots. Previous publications describing OJT did not include formal observation of the NP teacher or junior instructor. Therefore, this study aimed to present a novel approach to OJT inclusion during prosection laboratories based on the Lewinian experiential model. Methods Eight physical therapy (PT) graduates were recruited as junior anatomy instructors into the prosection laboratories. All participated in a unique training program during two consecutive academic years (2017, 2018) and received OJT during the teaching sessions. Two questionnaires were filled out to evaluate the educational impact of the training program. Eighty-three first-year PT students participated in prosection laboratories in anatomy taught by junior instructors, and filled out a questionnaire evaluating the performance of both junior and senior instructors. In addition, we compared the final grades in anatomy obtained by students taught by senior instructors to the grades of those taught by junior instructors. Results Each junior anatomy instructor participated in four OJT sessions. Based on self-reported measures, all professional and didactic aspects of the training program received a median score of 4.5 or higher on a five-point Likert scale. Students obtained similar grades in anatomy when taught by junior instructors compared with senior ones, and were similarly satisfied from the teaching performance of both senior and junior anatomy instructors. Conclusions OJT is applicable in a small-sized PT program facing a shortage of anatomy instructors. Including junior anatomy instructors in prosection laboratories for PT students is a viable solution to the shortage of experienced anatomy instructors. Further study, involving a larger cohort with a longer follow up will strengthen the preliminary results presented here.

Reading is considered a non-intuitive, cognitively demanding ability requiring synchronization between several neural networks supporting visual, language processing and higher-order abilities. With the involvement of technology in our everyday life, reading from a screen has become widely used. Several studies point to challenges in processing written materials from the screen due to changes in attention allocation when reading from a screen compared to reading from a printed paper. The current study examined the differences in brain activation when reading from a screen compared to reading from a printed paper focusing on spectral power related to attention in fifteen 6-8-year-old children. Using an electroencephalogram, children read two different age-appropriate texts, without illustrations, presented randomly on the screen and on a printed paper. Data were analyzed using spectral analyses in brain regions related to language, visual processing, and cognitive control, focusing on theta vs. beta waveforms. Results indicated that while reading from a printed paper was accompanied by higher energy in high-frequency bands (beta, gamma), reading from the screen was manifested by a higher power in the lower frequency bands (alpha, theta). Higher theta compared to the beta ratio, representing challenges in allocating attention to a given task, was found for the screen reading compared to the printed paper reading condition. Also, a significant negative correlation was found between differences in theta/beta ratio for screen vs paper reading and accuracy level in the age-normalized Sky-Search task measuring attention and a positive correlation with performance time. These results provide neurobiological support for the greater cognitive load and reduced focused attention during screen-based compared to print-based reading and suggest a different reliance on attention resources for the two conditions in children.

Introduction There has been renewed interest in the therapeutic use of bacteriophages (phages); however, standardised therapeutic protocols are lacking, and there is a paucity of rigorous clinical trial data assessing efficacy.Methods and analysis We propose an open-label, single-arm trial investigating a standardised treatment and monitoring protocol for phage therapy. Patients included will have exhausted other therapeutic options for control of their infection and phage therapy will be administered under Australia’s Therapeutic Goods Administration Special Access Scheme. A phage product with high in vitro activity against the targeted pathogen(s) must be available in line with relevant regulatory requirements. We aim to recruit 50–100 patients over 5 years, from any public or private hospitals in Australia. The standardised protocol will specify clinical assessments and biological sampling at scheduled time points. The primary outcome is safety at day 29, assessed by the frequency of adverse events, and overseen by an independent Data Safety Monitoring Board. Secondary outcomes include long-term safety (frequency of adverse events until at least 6 months following phage therapy), and feasibility, measured as the proportion of participants with>80% of minimum data available for analysis. Additional endpoints assessed include clinical response, patient/guardian reported quality of life measures, phage pharmacokinetics, human host immune responses and microbiome analysis. All trial outcomes will be summarised and presented using standard descriptive statistics.Ethics and dissemination Participant inclusion will be dependent on obtaining written informed consent from the patient or guardian. The trial protocol was approved by the Sydney Children’s Hospitals Network Human Research Ethics Committee in December 2021 (Reference 2021/ETH11861). In addition to publication in a peer-reviewed scientific journal, a lay summary of study outcomes will be made available for participants and the public on the Phage Australia website (https://www.phageaustralia.org/).Trial registration number Registered on ANZCTR, 10 November 2021 (ACTRN12621001526864; WHO Universal Trial Number: U1111-1269-6000).

Despite the remarkable successes of cancer immunotherapies, the majority of patients will experience only partial response followed by relapse of resistant tumors. While treatment resistance has frequently been attributed to clonal selection and immunoediting, comparisons of paired primary and relapsed tumors in melanoma and breast cancers indicate that they share the majority of clones. Here, we demonstrate in both mouse models and clinical human samples that tumor cells evade immunotherapy by generating unique transient cell-in-cell structures, which are resistant to killing by T cells and chemotherapies. While the outer cells in this cell-in-cell formation are often killed by reactive T cells, the inner cells remain intact and disseminate into single tumor cells once T cells are no longer present. This formation is mediated predominantly by IFNγ-activated T cells, which subsequently induce phosphorylation of the transcription factors signal transducer and activator of transcription 3 (STAT3) and early growth response-1 (EGR-1) in tumor cells. Indeed, inhibiting these factors prior to immunotherapy significantly improves its therapeutic efficacy. Overall, this work highlights a currently insurmountable limitation of immunotherapy and reveals a previously unknown resistance mechanism which enables tumor cells to survive immune-mediated killing without altering their immunogenicity.

BackgroundPrescription of psychostimulants has significantly increased in most countries worldwide for both preschool and school-aged children. Understanding the trends of chronic medication use among children in different age groups and from different sociodemographic backgrounds is essential. It is essential to distinguish between selected therapy areas to help decision-makers evaluate not only the relevant expected medication costs but also the specific services related to these areas. ObjectiveThis study will analyze differences in trends regarding medications considered psychobehavioral treatments and medications considered nonpsychobehavioral treatments and will identify risk factors and predictors for chronic medication use among children. MethodsThis is a retrospective study. Data will be extracted from the Clalit Health Services data warehouse. For each year between 2010 and 2019, there are approximately 1,500,000 children aged 0-18 years. All medication classes will be identified using the Anatomical Therapeutic Chemical code. A time-trend analysis will be performed to investigate if there is a significant difference between the trends of children’s psychobehavioral and nonpsychobehavioral medication prescriptions. A logistic regression combined with machine learning models will be developed to identify variables that may increase the risk for specific chronic medication types and identify children likely to get such treatment. ResultsThe project was funded in 2019. Data analysis is currently underway, and the results are expected to be submitted for publication in 2022. Understanding trends regarding medications considered psychobehavioral treatments and medications considered nonpsychobehavioral treatments will support the identification of risk factors and predictors for chronic medication use among children. ConclusionsAnalyzing the response of the patient (and their parents or caregivers) population over time will hopefully help improve policies for prescriptions and follow-up of chronic treatments in children. International Registered Report Identifier (IRRID)DERR1-10.2196/36756

Summary: Background: Clinical phage therapy is often delivered alongside antibiotics. However, the phenomenon of phage-antibiotic synergy has been mostly studied in vitro. Here, we assessed the in vivo bactericidal effect of a phage-antibiotic combination on Acinetobacter baumannii AB900 using phage øFG02, which binds to capsular polysaccharides and leads to antimicrobial resensitisation in vitro. Methods: We performed a two-stage preclinical study using a murine model of severe A. baumannii AB900 bacteraemia. In the first stage, with an endpoint of 11 h, mice (n = 4 per group) were treated with either PBS, ceftazidime, phage øFG02, or the combination of phage and ceftazidime. The second stage involved only the latter two groups (n = 5 per group), with a prolonged endpoint of 16 h. The primary outcome was the average bacterial burden from four body sites (blood, liver, kidney, and spleen). Bacterial colonies from phage-treated mice were retrieved and screened for phage-resistance. Findings: In the first stage, the bacterial burden (CFU/g of tissue) of the combination group (median: 4.55 × 105; interquartile range [IQR]: 2.79 × 105–2.81 × 106) was significantly lower than the PBS (median: 2.42 × 109; IQR: 1.97 × 109–3.48 × 109) and ceftazidime groups (median: 3.86 × 108; IQR: 2.15 × 108–6.35 × 108), but not the phage-only group (median: 1.28 × 107; IQR: 4.71 × 106–7.13 × 107). In the second stage, the combination treatment (median: 1.72 × 106; IQR: 5.11 × 105–4.00 × 106) outperformed the phage-only treatment (median: 7.46 × 107; IQR: 1.43 × 107–1.57 × 108). Phage-resistance emerged in 96% of animals receiving phages, and all the tested isolates (n = 11) had loss-of-function mutations in genes involved in capsule biosynthesis and increased sensitivity to ceftazidime. Interpretation: øFG02 reliably drives the in vivo evolution of A. baumannii AB900 towards a capsule-deficient, phage-resistant phenotype that is resensitised to ceftazidime. This mechanism highlights the clinical potential of using phage therapy to target A. baumannii and restore antibiotic activity. Funding: National Health and Medical Research Council (Australia).

Abstract Sinonasal teratocarcinosarcoma is a rare, highly aggressive tumor of the anterior skull base composed of malignant epithelial, mesenchymal, and neural tissue. Examination of cases in patients in minority populations is important in order to better understand the behavior of this neoplasm and outcomes of treatment in our nation's diverse population.

BackgroundExisting mental health apps are largely not aimed at generally healthy young people who may be experimenting with addictive substances and mind-altering experiences. ObjectiveThe aim of this study is to examine the interest and expectations of young people regarding a proposed smartphone app designed to help protect and promote mental health and resilience in the face of risks associated with substance use. MethodsThe study was based on agile system development and had 3 empirical substudies. Our feasibility study (study 1) included an anonymous questionnaire that examined the potential interest of young people in this type of app. It was answered by 339 Israelis aged 18-30 years. The second part of the feasibility study was a pilot study with 1.2% (4/339) of the people who answered the questionnaire and expressed interest in participating in a focus group. They tested and refined the elements planned for the focus groups. Study 2 was a participatory design study involving 7 focus groups of 5 to 7 participants each (young people aged 18-35 years, n=38). Persona development, open discussion, and a Technology Acceptance Model questionnaire were used to elicit user expectations and requirements for the app and to understand the perceived usefulness and usability of the proposed features. Study 3 comprised in-depth interviews with experts in the field of youth mental health and drug use to enlist their professional opinion regarding the value of such an app and recommendations about the features it should include. ResultsThe mock-up for the proposed app had five key features: personalized assessment of risk for a drug-associated mental crisis, support for self-monitoring, useful information (eg, warning signs and first-aid guidelines), resilience-building exercises, and a support center. Participants rated highly the usefulness of all 5 main features and 96% (24/25) of the specific features we proposed within those main categories. The participants also suggested additional features as well as a new user persona we had not considered: the parents or family members of the young person. The focus groups rated highly the perceived usability of the app. Most of the experts saw value in all the main features and suggested specific knowledge sources for the app’s content. Finally, participants of both the feasibility study and the participatory design study expressed moderate to high interest in using the app for self-help and high interest in using the app to help friends. ConclusionsThe findings provide preliminary encouraging support for the 5 main features suggested by the research team and reinforce recommendations for mobile health apps found in the literature. The findings emphasize the insight that this kind of app should be designed primarily for use by individuals seeking to help others.

Development of autoantibodies following BNT162b2 mRNA COVID-19 vaccination and their association with disease flares in adult patients with autoimmune inflammatory rheumatic diseases (AIIRD) and the general population: results of 1-year prospective follow-up study. We conducted a prospective study aimed at investigating the incidence of appearance of autoantibodies (antinuclear, antiphospholipid, and rheumatoid factor) in the sera of 463 adult patients with AIIRD compared to 55 controls from the general population prior to, and following the second and third vaccine doses, and at 1-year of follow-up. Pre- and post-vaccination disease activity indices and the association of autoantibodies with rheumatic disease flares and new onset AIIRD were examined. Autoantibody development of any type in AIIRD patients vs. the controls was 4.0% (vs. 6.7%, p = 0.423) following two vaccine doses and 7.6% (vs. 0%, p = 0.152) after three doses. There was no significant difference in sex, age, or disease-type among individuals with and without autoantibody development, regardless of the immunosuppressant use. More patients developed autoantibodies following the third than the second vaccine dose (p = 0.004). Disease flares occurred in 5.8% and 7.2% of AIIRD patients following second and third vaccine doses, respectively, with autoantibody production increasing the risk of flares following the second (p = 0.002) and third (p = 0.004) vaccine doses. BNT162b2 vaccination resulted in the development of autoantibodies in a minority of AIIRD patients and controls. Autoantibody development was associated with disease flares in patients, but no new-onset autoimmunity was observed.

Introduction Antimicrobial resistance is a growing global health threat, driven by increasing inappropriate use of antimicrobials. High prevalence of unnecessary use of antimicrobials in residential aged care facilities (RACFs) has driven demand for the development and implementation of antimicrobial stewardship (AMS) programmes. The Stepped-wedge Trial to increase antibiotic Appropriateness in Residential aged care facilities and model Transmission of antimicrobial resistance (START) will implement and evaluate the impact of a nurse-led AMS programme on antimicrobial use in 12 RACFs.Methods and analysis The START trial will implement and evaluate a nurse-led AMS programme via a stepped-wedge cluster randomised controlled trial design in 12 RACFs over 16 months. The AMS programme will incorporate education, aged care-specific treatment guidelines, documentation forms, and audit and feedback strategies that will target aged care staff, general practitioners, pharmacists, and residents and their families. The intervention will primarily focus on urinary tract infections, lower respiratory tract infections, and skin and soft tissue infections. RACFs will transition from control to intervention phases in random order, two at a time, every 2 months, with a 2-month transition, wash-in period. The primary outcome is the cumulative proportion of residents within each facility prescribed an antibiotic during each month and total days of antibiotic use per 1000 occupied bed days. Secondary outcomes include the number of courses of systemic antimicrobial therapy, antimicrobial appropriateness, antimicrobial resistant organisms, Clostridioides difficile infection, change in antimicrobial susceptibility profiles, hospitalisations and all-cause mortality. Analyses will be conducted according to the intention-to-treat principle.Ethics and dissemination Ethics approval has been granted by the Alfred Hospital Human Research Ethics Committee (HREC/18/Alfred/591). Research findings will be disseminated through peer-reviewed publications, conferences and summarised reports provided to participating RACFs.Trial registration number NCT03941509.

Treatment with rituximab (RTX) blunts SARS-CoV-2 vaccination-induced humoral response. We sought to identify predictors of a positive immunogenic response to the BNT162b2 mRNA vaccine in patients with autoimmune inflammatory rheumatic diseases (AIIRD) treated with RTX (AIIRD-RTX). We analyzed 108 AIIRD-RTX patients and 122 immunocompetent controls vaccinated with BNT162b2 mRNA participating in a multicenter vaccination study. Immunogenicity was defined by positive anti-SARS-CoV-2 S1/S2 IgG. We used a stepwise backward multiple logistic regression to identify predicting factors for a positive immunogenic response to vaccination and develop a predicting calculator, further validated in an independent cohort of AIIRD-RTX BNT162b2 mRNA vaccinated patients (n = 48). AIIRD-RTX patients who mounted a seropositive immunogenic response significantly differed from patients who did not by a lower number of RTX courses (median (range) 3 (1–10) vs. 5 (1–15), p = 0.007; lower cumulative RTX dose (mean ± SD) 6943.11 ± 5975.74 vs. 9780.95 ± 7240.12 mg, p = 0.033; higher IgG level prior to last RTX course (mean ± SD), 1189.78 ± 576.28 vs. 884.33 ± 302.31 mg/dL, p = 0.002), and extended interval between RTX treatment and vaccination, 469.82 ± 570.39 vs. 162.08 ± 160.12 days, p = 0.0009, respectively. Patients with ANCA-associated vasculitis and inflammatory myositis had a low likelihood of a seropositive immunogenic response compared to patients with rheumatoid arthritis, odds ratio (OR) 0.209, 95% confidence interval (CI) 0.046–0.96, p = 0.044 and OR 0.189, 95% CI 0.036–0.987, p = 0.048, respectively. Based on these findings, we constructed a calculator predicting the probability of a seropositive immunogenic response following BNT162b2 mRNA vaccination which performed with 90.5% sensitivity, 59.3% specificity, and 63.3% positive and 88.9% negative predictive values. In summary, the predicting calculator could guide clinicians for optimal timing of BNT162b2 mRNA vaccination in AIIRD-RTX patients.

Abstract Mutation acquisition is a major mechanism of bacterial antibiotic resistance that remains insufficiently characterised. Here we present RM-seq, a new amplicon-based deep sequencing workflow based on a molecular barcoding technique adapted from Low Error Amplicon sequencing (LEA-seq). RM-seq allows detection and functional assessment of mutational resistance at high throughput from mixed bacterial populations. The sensitive detection of very low-frequency resistant sub-populations permits characterisation of antibiotic-linked mutational repertoires in vitro and detection of rare resistant populations during infections. Accurate quantification of resistance mutations enables phenotypic screening of mutations conferring pleiotropic phenotypes such as in vivo persistence, collateral sensitivity or cross-resistance. RM-seq will facilitate comprehensive detection, characterisation and surveillance of resistant bacterial populations (https://github.com/rguerillot/RM-seq).

Brain tumors: uncovering genomic disruption in meningiomas Meningiomas, which arise from the tissue surrounding the brain and spinal cord, are the most common primary brain tumor in adults. The majority of these are slow-growing and amenable to surgical resection, if treatment is indicated. However, a subset of aggressive meningiomas are considered high-grade, producing significantly worse mortality. In a first study of its kind, Drs. Wenya Linda Bi, Ian Dunn, Sandro Santagata, Rameen Beroukhim, and colleagues at Harvard Medical School sequenced the genomes of 134 high-grade meningiomas and compared their makeup with lower-grade meningiomas. They found that aggressive tumors were more likely to harbor mutations in the NF2 gene and exhibit widespread genomic disruption. They also harbored an elevated rate of predicted immunogenic mutations, with implications for the use of immuno-modulatory therapies.

IntroductionA large proportion of patients with COVID-19 develop acute kidney injury (AKI). While the most severe of these cases require renal replacement therapy (RRT), little is known about their clinical course.MethodsWe describe the clinical characteristics of COVID-19 patients in the ICU with AKI requiring RRT at an academic medical center in New York City and followed patients for outcomes of death and renal recovery using time-to-event analyses.ResultsOur cohort of 115 patients represented 23% of all ICU admissions at our center, with a peak prevalence of 29%. Patients were followed for a median of 29 days (2542 total patient-RRT-days; median 54 days for survivors). Mechanical ventilation and vasopressor use were common (99% and 84%, respectively), and the median Sequential Organ Function Assessment (SOFA) score was 14. By the end of follow-up 51% died, 41% recovered kidney function (84% of survivors), and 8% still needed RRT (survival probability at 60 days: 0.46 [95% CI: 0.36-0.56])). In an adjusted Cox model, coronary artery disease and chronic obstructive pulmonary disease were associated with increased mortality (HRs: 3.99 [95% CI 1.46-10.90] and 3.10 [95% CI 1.25-7.66]) as were angiotensin-converting-enzyme inhibitors (HR 2.33 [95% CI 1.21-4.47]) and a SOFA score >15 (HR 3.46 [95% CI 1.65-7.25).Conclusions and relevanceOur analysis demonstrates the high prevalence of AKI requiring RRT among critically ill patients with COVID-19 and is associated with a high mortality, however, the rate of renal recovery is high among survivors and should inform shared-decision making.

Introduction Resistant Gram-positive organisms, such as methicillin-resistant staphylococci, account for a significant proportion of infections following joint replacement surgery. Current surgical antimicrobial prophylaxis guidelines recommend the use of first-generation or second-generation cephalosporin antibiotics, such as cefazolin. Cefazolin, however, does not prevent infections due to these resistant organisms; therefore, new prevention strategies need to be examined. One proposed strategy is to combine a glycopeptide antibiotic with cefazolin for prophylaxis. The clinical benefit and cost-effectiveness of this combination therapy compared with usual therapy, however, have not been established.Methods and analysis This randomised, double-blind, parallel, superiority, placebo-controlled, phase 4 trial will compare the incidence of all surgical site infections (SSIs) including superficial, deep and organ/space (prosthetic joint) infections, safety and cost-effectiveness of surgical prophylaxis with cefazolin plus vancomycin to that with cefazolin plus placebo. The study will be performed in patients undergoing joint replacement surgery. In the microbiological sub-studies, we will examine the incidence of SSIs in participants with preoperative staphylococci colonisation (Sub-Study 1) and incidence of VRE acquisition (Sub-Study 2). The trial will recruit 4450 participants over a 4-year period across 13 orthopaedic centres in Australia. The primary outcome is the incidence of SSI at 90 days post index surgery. Secondary outcomes include the incidence of SSI according to joint and microorganism and other healthcare associated infections. Safety endpoints include the incidence of acute kidney injury, hypersensitivity reactions and all-cause mortality. The primary and secondary analysis will be a modified intention-to-treat analysis consisting of all randomised participants who undergo eligible surgery. We will also perform a per-protocol analysis.Ethics and dissemination The study protocol was reviewed and approved by The Alfred Hospital Human Research Ethics Committee (HREC/18/Alfred/102) on 9 July 2018. Study findings will be disseminated in the printed media, and learnt forums.Trial registration number ACTRN12618000642280

BACKGROUND:Liver fibrosis predicts liver-related morbidity and mortality in patients with non-alcoholic fatty liver disease (NAFLD). Non-invasive scores correlate with the degree of liver fibrosis in these patients. AIMS AND METHODS:To investigate the accuracy of noninvasive scoring systems in predicting long-term outcomes and cancer incidence of patients with NAFLD, we performed a single-center retrospective study of patients with biopsy proven NAFLD. Mean follow up period was 100 months. Outcomes included liver-related complications, hospitalizations, overall mortality and the development of any malignancies. RESULTS:32 patients had advanced fibrosis (F3-F4) per biopsy at baseline and 121 patients had mild to moderate fibrosis (F0-F2). Both advanced histologic fibrosis stage as well as higher non-invasive scores predicted repeated hospitalizations and longer hospitalization stays. In a multivariate analysis, liver fibrosis (p = 0.002), FIB-4 score (p

BACKGROUND:Fatty acid synthase 1 (FAS I) from Mycobacterium tuberculosis (Mtb) is an essential protein and a promising drug target. FAS I is a multi-functional, multi-domain protein that is organized as a large (1.9 MDa) homohexameric complex. Acyl intermediates produced during fatty acid elongation are attached covalently to an acyl carrier protein (ACP) domain. This domain is activated by the transfer of a 4'-Phosphopantetheine (4'-PP, also termed P-pant) group from CoA to ACP catalyzed by a 4'-PP transferase, termed acyl carrier protein synthase (AcpS). METHODS:In order to obtain an activated FAS I in E. coli, we transformed E. coli with tagged Mtb fas1 and acpS genes encoded by a separate plasmid. We induced the expression of Mtb FAS I following induction of AcpS expression. FAS I was purified by Strep-Tactin affinity chromatography. RESULTS:Activation of Mtb FAS I was confirmed by the identification of a bound P-pant group on serine at position 1808 by mass spectrometry. The purified FAS I displayed biochemical activity shown by spectrophotometric analysis of NADPH oxidation and by CoA production, using the Ellman reaction. The purified Mtb FAS I forms a hexameric complex shown by negative staining and cryo-EM. CONCLUSION:Purified hexameric and active Mtb FAS I is required for binding and drug inhibition studies and for structure-function analysis of this enzyme. This relatively simple and short procedure for Mtb FAS I production should facilitate studies of this enzyme.

A common two-tier structure for social networks is based on partitioning society into two parts, referred to as the elite and the periphery, where the "elite" is the relatively small but well-connected and highly influential group of powerful individuals around which the society is centered, and the "periphery" consists of the rest of society. It is observed that the relative sizes of economic and social elites in various societies appear to be continually declining. One possible explanation is that this is a natural social phenomenon, resembling Price's "square root" law for the fraction of good scientists in the scientific community. We try to assess the validity of this explanation by studying the elite-periphery structure via introducing a novel axiom-based model for representing and measuring the influence between the elite and the periphery. The model is accompanied by a set of axioms that capture the elite's dominance, robustness and density, as well as a compactness property. Relying on the model and the accompanying axioms, we are able to draw a number of insightful conclusions about the elite-periphery structure. In particular, we show that in social networks that respect our axioms, the size of a compact elite is sublinear in the network size. This agrees with Price's principle but appears to contradict the common belief that the elite size tends to a linear fraction of society (recently claimed to be around 1%). We propose a natural method to create partitions with nice properties, based on the key observation that an elite-periphery partition is at what we call a 'balance point', where the elite and the periphery maintain a balance of powers. Our method is based on setting the elite to be the k most influential nodes in the network and suggest the balance point as a tool for choosing k and therefore the size of the elite. When using nodes degrees to order the nodes, the resulting k-rich club at the balance point is the elite of a partition we refer to as the balanced edge-based partition. We accompany these findings with an empirical study on 32 real-world social networks, which provides evidence that balanced edge-based partitions which satisfying our axioms commonly exist.