Metagenome diversity illuminates the origins of pathogen effectors.

Recent metagenome-assembled genome (MAG) analyses have profoundly impacted Rickettsiology systematics. The discovery of basal lineages (novel families Mitibacteraceae and Athabascaceae) with predicted extracellular lifestyles exposed an evolutionary timepoint for the transition to host dependency, which seemingly occurred independent of mitochondrial evolution. Notably, these basal rickettsiae carry the Rickettsiales vir homolog (rvh) type IV secretion system and purportedly use rvh to kill congener microbes rather than parasitize host cells as described for later-evolving rickettsial pathogens. MAG analysis also substantially increased diversity for the genus Rickettsia and delineated a sister lineage (the novel genus Tisiphia) that stands to inform on the emergence of human pathogens from protist and invertebrate endosymbionts. Herein, we probed Rickettsiales MAG and genomic diversity for the distribution of Rickettsia rvh effectors to ascertain their origins. A sparse distribution of most Rickettsia rvh effectors outside of Rickettsiaceae lineages illuminates unique rvh evolution from basal extracellular species and other rickettsial families. Remarkably, nearly every effector was found in multiple divergent forms with variable architectures, indicating profound roles for gene duplication and recombination in shaping effector repertoires in Rickettsia pathogens. Lateral gene transfer plays a prominent role in shaping the rvh effector landscape, as evinced by the discovery of many effectors on plasmids and conjugative transposons, as well as pervasive effector gene exchange between Rickettsia and Legionella species. Our study exemplifies how MAGs can yield insight into pathogen effector origins, particularly how effector architectures might become tailored to the discrete host cell functions of different eukaryotic hosts.IMPORTANCEWhile rickettsioses are deadly vector-borne human diseases, factors distinguishing Rickettsia pathogens from the innumerable bevy of environmental rickettsial endosymbionts remain lacking. Recent metagenome-assembled genome (MAG) studies revealed evolutionary timepoints for rickettsial transitions to host dependency. The rvh type IV secretion system was likely repurposed from congener killing in basal extracellular species to parasitizing host cells in later-evolving pathogens. Our analysis of MAG diversity for over two dozen rvh effectors unearthed their presence in some non-pathogens. However, most effectors were found in multiple divergent forms with variable architectures, indicating gene duplication and recombination-fashioned effector repertoires of Rickettsia pathogens. Lateral gene transfer substantially shaped pathogen effector arsenals, evinced by the discovery of effectors on plasmids and conjugative transposons, as well as pervasive effector gene exchanges between Rickettsia and Legionella species. Our study exemplifies how MAGs yield insight into pathogen effector origins and evolutionary processes tailoring effectors to eukaryotic host cell biology.

Using healthcare systems data for outcomes in clinical trials: issues to consider at the design stage.

BACKGROUND: Healthcare system data (HSD) are increasingly used in clinical trials, augmenting or replacing traditional methods of collecting outcome data. This study, PRIMORANT, set out to identify, in the UK context, issues to be considered before the decision to use HSD for outcome data in a clinical trial is finalised, a methodological question prioritised by the clinical trials community. METHODS: The PRIMORANT study had three phases. First, an initial workshop was held to scope the issues faced by trialists when considering whether to use HSDs for trial outcomes. Second, a consultation exercise was undertaken with clinical trials unit (CTU) staff, trialists, methodologists, clinicians, funding panels and data providers. Third, a final discussion workshop was held, at which the results of the consultation were fed back, case studies presented, and issues considered in small breakout groups. RESULTS: Key topics included in the consultation process were the validity of outcome data, timeliness of data capture, internal pilots, data-sharing, practical issues, and decision-making. A majority of consultation respondents (n = 78, 95%) considered the development of guidance for trialists to be feasible. Guidance was developed following the discussion workshop, for the five broad areas of terminology, feasibility, internal pilots, onward data sharing, and data archiving. CONCLUSIONS: We provide guidance to inform decisions about whether or not to use HSDs for outcomes, and if so, to assist trialists in working with registries and other HSD providers to improve the design and delivery of trials.

Are Welsh primary schools Sunproofed? Results of a national survey, part 1: scoping the landscape of sun safety policies in Wales.

BACKGROUND: Schools with formal sun safety polices generally show better sun safety practices than schools without. OBJECTIVES: To understand the extent to which Welsh primary schools have sun safety policies; to identify the key characteristics of policies; to assess whether policy adoption varies by school characteristics; and to consider what support schools need to develop sun safety policies. METHODS: An online multiple-choice survey on sun safety was distributed to all 1241 primary schools in Wales. RESULTS: In total, 471 (38.0%) schools responded. Of these, 183 (39.0%) reported having a formal sun safety policy. Welsh medium schools (P = 0.036) and schools in North Wales (P = 0.008) were more likely to report having a policy. Schools with a higher percentage of pupils receiving free school meals (P = 0.046) and with lower attendance rates (P = 0.008) were less likely to report having a sun safety policy. The primary reasons for schools not having a policy included being 'not aware of the need' (34.6%); 'need assistance with policy or procedure development' (30.3%); and 'not got around to it just yet' (26.8%). CONCLUSIONS: With less than half of schools reporting a sun safety policy and variation in the presence/absence of a policy by school characteristics, our survey revealed inconsistency in formal sun safety provision in Welsh schools. The findings also suggest that schools are unaware of the importance of sun safety and need support to develop and implement policies. This snapshot of the current situation in primary schools in Wales provides a basis upon which the comprehensiveness, effectiveness and implementation of sun safety policies can be further evaluated.

Mediport use as an acceptable standard for CAR T cell infusion.

Introduction: Mediport use as a clinical option for the administration of chimeric antigen receptor T cell (CAR T cell) therapy in patients with B-cell malignancies has yet to be standardized. Concern for mediport dislodgement, cell infiltration, and ineffective therapy delivery to systemic circulation has resulted in variable practice with intravenous administration of CAR T cell therapy. With CAR T cell commercialization, it is important to establish practice standards for CAR T cell delivery. We conducted a study to establish usage patterns of mediports in the clinical setting and provide a standard of care recommendation for mediport use as an acceptable form of access for CAR T cell infusions. Methods: In this retrospective cohort study, data on mediport use and infiltration rate was collected from a survey across 34 medical centers in the Pediatric Real-World CAR Consortium, capturing 504 CAR T cell infusion routes across 489 patients. Data represents the largest, and to our knowledge sole, report on clinical CAR T cell infusion practice patterns since FDA approval and CAR T cell commercialization in 2017. Results: Across 34 sites, all reported tunneled central venous catheters, including Broviac® and Hickman® catheters, as accepted standard venous options for CAR T cell infusion. Use of mediports as a standard clinical practice was reported in 29 of 34 sites (85%). Of 489 evaluable patients with reported route of CAR T cell infusion, 184 patients were infused using mediports, with no reported incidences of CAR T cell infiltration. Discussion/Conclusion: Based on current clinical practice, mediports are a commonly utilized form of access for CAR T cell therapy administration. These findings support the safe practice of mediport usage as an accepted standard line option for CAR T cell infusion.

Projection of high temperature-related burden of kidney disease in Australia under different climate change, population and adaptation scenarios: population-based study.

Background: The dual impacts of a warming climate and population ageing lead to an increasing kidney disease prevalence, highlighting the importance of quantifying the burden of kidney disease (BoKD) attributable to high temperature, yet studies on this subject are limited. The study aims to quantify the BoKD attributable to high temperatures in Australia across all states and territories, and project future BoKD under climatic, population and adaptation scenarios. Methods: Data on disability-adjusted-life-years (DALYs) due to kidney disease, including years of life lost (YLL), and years lived with disability (YLD), were collected during 2003-2018 (baseline) across all states and territories in Australia. The temperature-response association was estimated using a meta-regression model. Future temperature projections were calculated using eight downscaled climate models to estimate changes in attributable BoKD centred around 2030s and 2050s, under two greenhouse gas emissions scenarios (RCP4.5 and RCP8.5), while considering changes in population size and age structure, and human adaptation to climate change. Findings: Over the baseline (2003-2018), high-temperature contributed to 2.7% (Standard Deviation: 0.4%) of the observed BoKD in Australia. The future population attributable fraction and the attributable BoKD, projected using RCP4.5 and RCP8.5, showed a gradually increasing trend when assuming no human adaptation. Future projections were most strongly influenced by the population change, with the high temperature-related BoKD increasing by 18.4-67.4% compared to the baseline under constant population and by 100.2-291.2% when accounting for changes in population size and age structure. However, when human adaptation was adopted (from no to partial to full), the high temperature-related BoKD became smaller. Interpretation: It is expected that increasing high temperature exposure will substantially contribute to higher BoKD across Australia, underscoring the urgent need for public health interventions to mitigate the negative health impacts of a warming climate on BoKD. Funding: Australian Research Council Discovery Program.

Metagenome diversity illuminates origins of pathogen effectors.

Recent metagenome assembled genome (MAG) analyses have profoundly impacted Rickettsiology systematics. Discovery of basal lineages (Mitibacteraceae and Athabascaceae) with predicted extracellular lifestyles reveals an evolutionary timepoint for the transition to host dependency, which occurred independent of mitochondrial evolution. Notably, these basal rickettsiae carry the Rickettsiales vir homolog (rvh) type IV secretion system (T4SS) and purportedly use rvh to kill congener microbes rather than parasitize host cells as described for derived rickettsial pathogens. MAG analysis also substantially increased diversity for genus Rickettsia and delineated a basal lineage (Tisiphia) that stands to inform on the rise of human pathogens from protist and invertebrate endosymbionts. Herein, we probed Rickettsiales MAG and genomic diversity for the distribution of Rickettsia rvh effectors to ascertain their origins. A sparse distribution of most Rickettsia rvh effectors outside of Rickettsiaceae lineages indicates unique rvh evolution from basal extracellular species and other rickettsial families. Remarkably, nearly every effector was found in multiple divergent forms with variable architectures, illuminating profound roles for gene duplication and recombination in shaping effector repertoires in Rickettsia pathogens. Lateral gene transfer plays a prominent role shaping the rvh effector landscape, as evinced by the discover of many effectors on plasmids and conjugative transposons, as well as pervasive effector gene exchange between Rickettsia and Legionella species. Our study exemplifies how MAGs can provide incredible insight on the origins of pathogen effectors and how their architectural modifications become tailored to eukaryotic host cell biology.

Expert consensus guidance on respiratory physiotherapy and rehabilitation of patients with rib fractures: An international, multidisciplinary e-Delphi study.

BACKGROUND: There is limited research supporting optimal respiratory physiotherapy or physical rehabilitation strategies for patients with rib fractures. The aim of this study was to develop key recommendations for the physiotherapy management of patients with rib fractures. METHODS: A three-round modified e-Delphi survey design, using an international Delphi panel including physiotherapy clinicians, researchers and lecturers, physician associates, trauma surgeons, and intensivists, was used in this study. The draft recommendations were developed by the Steering Group, based on available research. Over three rounds, panelists rated their agreement (using a Likert scale) with regard to recommendation for physiotherapists delivering respiratory physiotherapy and physical rehabilitation to patients following rib fractures. Recommendations were retained if they achieved consensus (defined as ≥70% of panelists ≥5/7) at the end of each round. RESULTS: A total of 121 participants from 18 countries registered to participate in the study, with 87 (72%), 77 (64%), and 79 (65%) registrants completing the three rounds, respectively. The final guidance document included 18 respiratory physiotherapy and rehabilitation recommendations, mapped over seven clinical scenarios for patients (1) not requiring mechanical ventilation, (2) requiring mechanical ventilation, (3) with no concurrent fracture of the shoulder girdle complex, (4) with a concurrent fracture of the shoulder girdle complex, (5) with/without concurrent upper limb orthopedic injuries, (6) undergoing surgical stabilization of rib fractures, and (7) at hospital discharge. CONCLUSION: This guidance provides key recommendations for respiratory physiotherapy and physical rehabilitation of patients with rib fractures. It could also be used to inform future research priorities in the field. LEVEL OF EVIDENCE: Therapeutic/Care Management; Level IV.

Estimating the burden of disease attributable to high ambient temperature across climate zones: methodological framework with a case study.

BACKGROUND: With high temperature becoming an increasing health risk due to a changing climate, it is important to quantify the scale of the problem. However, estimating the burden of disease (BoD) attributable to high temperature can be challenging due to differences in risk patterns across geographical regions and data accessibility issues. METHODS: We present a methodological framework that uses Köppen-Geiger climate zones to refine exposure levels and quantifies the difference between the burden observed due to high temperatures and what would have been observed if the population had been exposed to the theoretical minimum risk exposure distribution (TMRED). Our proposed method aligned with the Australian Burden of Disease Study and included two parts: (i) estimation of the population attributable fractions (PAF); and then (ii) estimation of the BoD attributable to high temperature. We use suicide and self-inflicted injuries in Australia as an example, with most frequent temperatures (MFTs) as the minimum risk exposure threshold (TMRED). RESULTS: Our proposed framework to estimate the attributable BoD accounts for the importance of geographical variations of risk estimates between climate zones, and can be modified and adapted to other diseases and contexts that may be affected by high temperatures. CONCLUSIONS: As the heat-related BoD may continue to increase in the future, this method is useful in estimating burdens across climate zones. This work may have important implications for preventive health measures, by enhancing the reproducibility and transparency of BoD research.

A retrospective validation study of the STUMBL score for emergency department patients with blunt thoracic trauma.

INTRODUCTION: Blunt thoracic trauma (BTT) is a leading cause of emergency department (ED) trauma-related attendance. Risk prediction tools are commonly to predict patients' outcomes and assign them to the most appropriate care setting. The STUMBL score is a prognostic model for BTT, derived and validated in the United Kingdom; items comprising the score are age, number of rib fractures, use of pre-injury anticoagulants, chronic lung disease and oxygen saturation levels. This study's aim was to validate the STUMBL score in an Italian ED. METHODS: This single-centre retrospective validation study was conducted in the ED of Santa Croce and Carle hub hospital in Cuneo, north-western Italy. All patients with an ED attendance for isolated BTT from 2018 to 2021 were included. Exclusion criteria were an age of under eighteen and the presence of any immediately life-threatening lesion. The primary outcome was the development of trauma-related complications, defined by the occurrence of one or more of the following: in-hospital mortality, pulmonary complications (infection, pleural effusion, haemothorax, pneumothorax, pleural empyema), need for intensive care unit admission, hospital length of stay equal to or greater than seven days. The performance of the STUMBL score was analysed in terms of discrimination with the evaluation of the receiver operating characteristic curve and calibration with the Hosmer-Lemeshow test and with the calibration belt. RESULTS: 745 patients were enroled (median age 64 [25th;75th percentile: 50;78], male/female ratio 1:4, median Charlson comorbidity index 2 [1;4], median STUMBL score 11 [6;17]). 65.2% of patients were discharged home after ED evaluation. 203 patients (27.2%) developed the primary outcome. The STUMBL score was significantly different in patients with complications compared to those without complications (9 [5;13] vs 21 [17;25], p < 0.001). The C index of the score for the primary outcome was 0.90 (95% CI 0.88-0.93), and the result of the Hosmer-Lemeshow test was 9.01 (p = 0.34). STUMBL score = 16 has a sensitivity of 0.80 (95% CI 0.75-0.85), specificity of 0.87 (95% CI 0.84-0.90), a positive predictive value of 0.70 (95% CI 0.64-0.76), and a negative predictive value of 0.92 (95% CI 0.90-0.94). CONCLUSION: In this validation study, the STUMBL score demonstrated excellent discrimination and calibration in predicting the outcome of patients attending the ED with a BTT.

Factors Influencing the Use of Control Measures to Reduce Occupational Exposure to Welding Fume in Australia: A Qualitative Study.

Background: Exposure to welding fume is associated with adverse effects on worker health. The use of various control measures can reduce levels of exposure and the resulting health effects. However, little is known about the factors that may influence workers' use of control measures in the workplace and their perceived intervention needs. This study aimed to investigate workers' and other stakeholders' views on ways to improve the use of welding fume control measures in Australian workplaces. Methods: We conducted a series of online focus group discussions and individual interviews with participants who have some occupational involvement in welding, whether as workers, employers or industry representatives, union representatives, or regulators. A semi-structured question guide was used, and all discussions and interviews were recorded and transcribed for analysis. Results: Five focus group discussions and five individual interviews were conducted with a total of 21 participants. Three major themes emerged. The first addressed the current awareness of welding fume harms and concern about exposure; the second focussed on the current use of control measures, and barriers and facilitators to their use; and the last centred around intervention needs and the contents of a potential effective intervention. Conclusion: Improving the use of control measures to prevent exposure to welding fume requires knowledge around the barriers and facilitators of control, use, and the intervention needs of stakeholders. This study has provided such knowledge, which will facilitate the design and implementation of an intervention to reduce welding fume exposure and ultimately protect the health of workers.

Reduced death rates of elite Australian Rules footballers compared to age-matched general population.

OBJECTIVES: To determine age-matched death rates of current and retired elite male Australian football players (Australian and/or Victorian Football League) with the general population. DESIGN: Analysis of publicly-available birth, debut and death data for all Australian Victorian Football/Victorian Football League players who debuted prior to (and were still alive at) the start of 1971 or debuted 1971-2020. METHODS: Wikipedia was used to source the dates of death (or record that the player was alive in 2021) for the cohort. New players became part of the cohort for analysis on debut and existing cohort members left it at death. Actual death rates (per year and per decade) were then compared to expected deaths, based on age-specific population death rates. RESULTS: There were 5400 players and ex-players in the cohort at the start of 1971 (average age 47.3) and a further 4532 players debuted between 1971 and 2020 inclusive (a total of 9932 players). The expected deaths for the cohort in this 50-year period were 4955, but only 3914 deaths occurred (Standardized Mortality Ratio 0.79, 95 %CI 0.76-0.82). For younger members of the cohort (age <50) the discrepancy between expected (222) and actual (98) deaths was also significant (Standardized Mortality Ratio 0.44, 95 %CI 0.35-0.56). CONCLUSIONS: We conclude that the death rates of elite male Australian footballers are lower than the reference general population, similar to other studies of elite athletes. Some of this may be explained by "healthy cohort" selection bias, a limitation which affects almost all studies in this genre.

Heat exposure and cardiovascular health outcomes: a systematic review and meta-analysis.

BACKGROUND: Heat exposure is an important but underappreciated risk factor contributing to cardiovascular disease. Warming temperatures might therefore pose substantial challenges to population health, especially in a rapidly aging population. To address a potential increase in the burden of cardiovascular disease, a better understanding of the effects of ambient heat on different types of cardiovascular disease and factors contributing to vulnerability is required, especially in the context of climate change. This study reviews the current epidemiological evidence linking heat exposures (both high temperatures and heatwaves) with cardiovascular disease outcomes, including mortality and morbidity. METHODS: In this systematic review and meta-analysis, we searched PubMed, Embase, and Scopus for literature published between Jan 1, 1990, and March 10, 2022, and evaluated the quality of the evidence following the Navigation Guide Criteria. We included original research on independent study populations in which the exposure metric was high temperatures or heatwaves, and observational studies using ecological time series, case crossover, or case series study designs comparing risks over different exposures or time periods. Reviews, commentaries, grey literature, and studies that examined only seasonal effects without explicitly considering temperature were excluded. The risk estimates were derived from included articles and if insufficient data were available we contacted the authors to provide clarification. We did a random-effects meta-analysis to pool the relative risk (RR) of the association between high temperatures and heatwaves and cardiovascular disease outcomes. The study protocol was registered with PROSPERO (CRD42021232601). FINDINGS: In total, 7360 results were returned from our search of which we included 282 articles in the systematic review, and of which 266 were eligible for the meta-analysis. There was substantial heterogeneity for both mortality (high temperatures: I2=93·6%, p<0·0001; heatwaves: I2=98·9%, p<0·0001) and morbidity (high temperatures: I2=98·8%, p<0·0001; heatwaves: I2=83·5%, p<0·0001). Despite the heterogeneity in environmental conditions and population dynamics among the reviewed studies, results showed that a 1°C increase in temperature was positively associated with cardiovascular disease-related mortality across all considered diagnoses. The overall risk of cardiovascular disease-related mortality increased by 2·1% (RR 1·021 [95%CI 1·020-1·023]), with the highest specific disease risk being for stroke and coronary heart disease. A 1°C temperature rise was also associated with a significant increase in morbidity due to arrhythmias and cardiac arrest and coronary heart disease. Our findings suggest heat exposure leads to elevated risk of morbidity and mortality for women, people 65 years and older, individuals living in tropical climates, and those in countries of lower-middle income. Heatwaves were also significantly associated with a 17% increase in risk of mortality (RR 1·117 [95% CI 1·093-1·141]), and increasing heatwave intensity with an increasing risk (RR 1·067 [95% CI 1·056-1·078] for low intensity, 1·088 [1·058-1·119] for middle intensity, and 1·189 [1·109-1·269] for high intensity settings). INTERPRETATION: This review strengthens the evidence on the increase in cardiovascular disease risk due to ambient heat exposures in different climate zones. The widespread prevalence of exposure to hot temperatures, in conjunction with an increase in the proportion of older people in the population, might result in a rise in poor cardiovascular disease health outcomes associated with a warming climate. Evidence-based prevention measures are needed to attenuate peaks in cardiovascular events during hot spells, thereby lowering the worldwide total heat-related burden of cardiovascular disease-related morbidity and death. FUNDING: Australian Research Council Discovery Program.

Sunproofed study protocol: A mixed-methods scoping study of sun safety policies in primary schools in Wales.

BACKGROUND: Skin cancer, including melanoma and non-melanoma (keratinocyte), is increasing in incidence in the UK. Accounting for half of all cancers in England and Wales, the disease significantly impacts overstretched dermatology services. Research suggests that 86% of melanoma is preventable with modified sun exposure. Educating children about sun safety in schools can help prevent skin cancer and is recommended by major health organisations. In England, teaching sun safety in primary schools is compulsory, while in Wales this is left to school discretion. AIMS: Understand how primary schools in Wales are responding to growing skin cancer rates and explore the effectiveness of sun safety policies in schools on knowledge and behaviour. METHODS: Sunproofed is a mixed-methods scoping study comprising 5 work packages (WP) using survey and routine electronic health record (EHR) data supplemented by qualitative case studies. Objective(s) are to: WP1: Discover if primary schools in Wales have sun safety policies; policy characteristics; determine factors that may influence their presence and identify areas where schools need support. WP2: Determine what EHR data is available regarding the incidence of sunburn in primary school children and the feasibility of using this data to evaluate the impact of sun safety policies. WP3: Understand the impact of sun safety policies on sun-safe knowledge and behaviour amongst children, parents, teachers, and school management; identify barriers and facilitators to schools implementing sun safety policies. WP4: Co-produce guidance regarding sun safety policies and best methods for implementation in schools. WP5: Disseminate guidance and findings widely to ensure impact and uptake. DISCUSSION: Skin cancer rates are increasing in the UK, straining limited resources. Sunproofed has the potential to inform the development of future prevention activities, both in Wales and beyond. This could reduce the number of skin cancer cases in the future and keep people healthier for longer.

Cryptic Genes for Interbacterial Antagonism Distinguish Rickettsia Species Infecting Blacklegged Ticks From Other Rickettsia Pathogens.

Background: The genus Rickettsia (Alphaproteobacteria: Rickettsiales) encompasses numerous obligate intracellular species with predominantly ciliate and arthropod hosts. Notable species are pathogens transmitted to mammals by blood-feeding arthropods. Mammalian pathogenicity evolved from basal, non-pathogenic host-associations; however, some non-pathogens are closely related to pathogens. One such species, Rickettsia buchneri, is prevalent in the blacklegged tick, Ixodes scapularis. While I. scapularis transmits several pathogens to humans, it does not transmit Rickettsia pathogens. We hypothesize that R. buchneri established a mutualism with I. scapularis, blocking tick superinfection with Rickettsia pathogens. Methods: To improve estimates for assessing R. buchneri infection frequency in blacklegged tick populations, we used comparative genomics to identify an R. buchneri gene (REIS_1424) not present in other Rickettsia species present throughout the I. scapularis geographic range. Bioinformatic and phylogenomics approaches were employed to propose a function for the hypothetical protein (263 aa) encoded by REIS_1424. Results: REIS_1424 has few analogs in other Rickettsiales genomes and greatest similarity to non-Proteobacteria proteins. This cohort of proteins varies greatly in size and domain composition, possessing characteristics of Recombination hotspot (Rhs) and contact dependent growth inhibition (CDI) toxins, with similarity limited to proximal C-termini (~145 aa). This domain was named CDI-like/Rhs-like C-terminal toxin (CRCT). As such proteins are often found as toxin-antidote (TA) modules, we interrogated REIS_1423 (151 aa) as a putative antidote. Indeed, REIS_1423 is similar to proteins encoded upstream of CRCT domain-containing proteins. Accordingly, we named these proteins CDI-like/Rhs-like C-terminal toxin antidotes (CRCA). R. buchneri expressed both REIS_1423 and REIS_1424 in tick cell culture, and PCR assays showed specificity for R. buchneri over other rickettsiae and utility for positive detection in three tick populations. Finally, phylogenomics analyses uncovered divergent CRCT/CRCA modules in varying states of conservation; however, only R. buchneri and related Tamurae/Ixodes Group rickettsiae carry complete TA modules. Conclusion: We hypothesize that Rickettsia CRCT/CRCA modules circulate in the Rickettsia mobile gene pool, arming rickettsiae for battle over arthropod colonization. While its functional significance remains to be tested, R. buchneri CRCT/CRCA serves as a marker to positively identify infection and begin deciphering the role this endosymbiont plays in the biology of the blacklegged tick.

Early exercise in blunt chest wall trauma: protocol for a mixed-methods, multicentre, parallel randomised controlled trial (ELECT2 trial).

INTRODUCTION: Chronic pain and disability are now well-recognised long-term complications of blunt chest wall trauma. Limited research exists regarding therapeutic interventions that can be used to address these complications. A recent feasibility study was completed testing the methods of a definitive trial. This protocol describes the proposed definitive trial, the aim of which is to investigate the impact of an early exercise programme on chronic pain and disability in patients with blunt chest wall trauma. METHODS/ANALYSIS: This mixed-methods, multicentre, parallel randomised controlled trial will run in four hospitals in Wales and one in England over 12-month recruitment period. Patients will be randomised to either the control group (routine physiotherapy input) or the intervention group (routine physiotherapy input plus a simple exercise programme completed individually by the patient). Baseline measurements including completion of two surveys (Brief Pain Inventory and EuroQol 5-dimensions, 5-Levels) will be obtained on initial assessment. These measures and a client services receipt inventory will be repeated at 3-month postinjury. Analysis of outcomes will focus on rate and severity of chronic pain and disability, cost-effectiveness and acceptability of the programme by patients and clinicians. Qualitative feedback regarding acceptability will be obtained through patient and clinician focus groups. ETHICS/DISSEMINATION: London Riverside Research Ethics Committee (Reference number: 21/LO/0782) and the Health Research Authority granted approval for the trial in December 2021. Patient recruitment will commence in February 2022. Planned dissemination is through publication in a peer-reviewed Emergency Medicine Journal, presentation at appropriate conferences and to stakeholders at professional meetings. TRIAL REGISTRATION NUMBER: ISRCTN65829737; Pre-results.

Genome sequencing of Pseudomonas aeruginosa strain M2 illuminates traits of an opportunistic pathogen of burn wounds.

Pseudomonas aeruginosa is a Gram-negative nosocomial pathogen and one of the most prevalent organisms isolated from burn wounds worldwide. Pseudomonas aeruginosa strain M2 (O5 serotype, type B flagella) is utilized for examining the murine model associated with burns. Pseudomonas aeruginosa M2 is similar in lethality to common laboratory P. aeruginosa strains when infecting CD-1 mice. Conversely, we recently showed that, relative to these strains, P. aeruginosa M2-infected mice are more susceptible to sepsis and demonstrate a 6-log reduction in LD50 from subcutaneous infection at the infection site directly after 10% total body surface area burn. To better understand this striking phenotypic difference from other P. aeruginosa strains employed in burn models, we sequenced the P. aeruginosa M2 genome. A total of 4,136,641 read pairs were obtained, providing an average genome coverage of 97.5X; subsequent assembly yielded a draft genome with 187 contigs comprising 6,360,304 bp with a G + C content of 66.45%. Genome-based phylogeny estimation of 92 P. aeruginosa strains placed P. aeruginosa M2 with P. aeruginosa-12-4-4(59), a nonairway clinical strain isolated from the blood culture of a burn patient. Phylogenomic analyses identified genes shared between P. aeruginosa M2 and P. aeruginosa 14, another strain exhibiting increased lethality in thermal tissues, as well as P. aeruginosa M2 unique genes with diverse functions like degradation of toxic aromatic compounds, iron scavenging, swarming motility and biofilm formation, defense against invasive DNA, and host assault. Predicted lateral gene transfers illuminate proteins heretofore uncharacterized for roles in P. aeruginosa biology. Our work yields a rich resource for assessing P. aeruginosa genes required for increased lethality in burn tissue seroma.

Ruxolitinib for the treatment of steroid refractory pediatric chronic graft-versus-host disease.

Chronic graft-versus-host disease (cGVHD) of the skin is a serious cause of long-term morbidity and mortality among patients who receive hematopoietic stem cell transplants. Systemic corticosteroids remain first-line treatment for cutaneous cGVHD; however, there is currently no consensus on second-line therapy for steroid-refractory disease. We herein present a case of a pediatric patient with severe cGVHD of the skin, nonresponsive to corticosteroids, who was successfully treated with a prolonged course of ruxolitinib with minimal side effects.