High expression of oleoyl-ACP hydrolase underpins life-threatening respiratory viral diseases.

Respiratory infections cause significant morbidity and mortality, yet it is unclear why some individuals succumb to severe disease. In patients hospitalized with avian A(H7N9) influenza, we investigated early drivers underpinning fatal disease. Transcriptomics strongly linked oleoyl-acyl-carrier-protein (ACP) hydrolase (OLAH), an enzyme mediating fatty acid production, with fatal A(H7N9) early after hospital admission, persisting until death. Recovered patients had low OLAH expression throughout hospitalization. High OLAH levels were also detected in patients hospitalized with life-threatening seasonal influenza, COVID-19, respiratory syncytial virus (RSV), and multisystem inflammatory syndrome in children (MIS-C) but not during mild disease. In olah-/- mice, lethal influenza infection led to survival and mild disease as well as reduced lung viral loads, tissue damage, infection-driven pulmonary cell infiltration, and inflammation. This was underpinned by differential lipid droplet dynamics as well as reduced viral replication and virus-induced inflammation in macrophages. Supplementation of oleic acid, the main product of OLAH, increased influenza replication in macrophages and their inflammatory potential. Our findings define how the expression of OLAH drives life-threatening viral disease.

Influenza Virus Z-RNAs Induce ZBP1-Mediated Necroptosis.

Influenza A virus (IAV) is a lytic RNA virus that triggers receptor-interacting serine/threonine-protein kinase 3 (RIPK3)-mediated pathways of apoptosis and mixed lineage kinase domain-like pseudokinase (MLKL)-dependent necroptosis in infected cells. ZBP1 initiates RIPK3-driven cell death by sensing IAV RNA and activating RIPK3. Here, we show that replicating IAV generates Z-RNAs, which activate ZBP1 in the nucleus of infected cells. ZBP1 then initiates RIPK3-mediated MLKL activation in the nucleus, resulting in nuclear envelope disruption, leakage of DNA into the cytosol, and eventual necroptosis. Cell death induced by nuclear MLKL was a potent activator of neutrophils, a cell type known to drive inflammatory pathology in virulent IAV disease. Consequently, MLKL-deficient mice manifest reduced nuclear disruption of lung epithelia, decreased neutrophil recruitment into infected lungs, and increased survival following a lethal dose of IAV. These results implicate Z-RNA as a new pathogen-associated molecular pattern and describe a ZBP1-initiated nucleus-to-plasma membrane "inside-out" death pathway with potentially pathogenic consequences in severe cases of influenza.

Human CD8+ T cell cross-reactivity across influenza A, B and C viruses.

Influenza A, B and C viruses (IAV, IBV and ICV, respectively) circulate globally and infect humans, with IAV and IBV causing the most severe disease. CD8+ T cells confer cross-protection against IAV strains, however the responses of CD8+ T cells to IBV and ICV are understudied. We investigated the breadth of CD8+ T cell cross-recognition and provide evidence of CD8+ T cell cross-reactivity across IAV, IBV and ICV. We identified immunodominant CD8+ T cell epitopes from IBVs that were protective in mice and found memory CD8+ T cells directed against universal and influenza-virus-type-specific epitopes in the blood and lungs of healthy humans. Lung-derived CD8+ T cells displayed tissue-resident memory phenotypes. Notably, CD38+Ki67+CD8+ effector T cells directed against novel epitopes were readily detected in IAV- or IBV-infected pediatric and adult subjects. Our study introduces a new paradigm whereby CD8+ T cells confer unprecedented cross-reactivity across all influenza viruses, a key finding for the design of universal vaccines.

Necroptosis blockade prevents lung injury in severe influenza.

Severe influenza A virus (IAV) infections can result in hyper-inflammation, lung injury and acute respiratory distress syndrome1-5 (ARDS), for which there are no effective pharmacological therapies. Necroptosis is an attractive entry point for therapeutic intervention in ARDS and related inflammatory conditions because it drives pathogenic lung inflammation and lethality during severe IAV infection6-8 and can potentially be targeted by receptor interacting protein kinase 3 (RIPK3) inhibitors. Here we show that a newly developed RIPK3 inhibitor, UH15-38, potently and selectively blocked IAV-triggered necroptosis in alveolar epithelial cells in vivo. UH15-38 ameliorated lung inflammation and prevented mortality following infection with laboratory-adapted and pandemic strains of IAV, without compromising antiviral adaptive immune responses or impeding viral clearance. UH15-38 displayed robust therapeutic efficacy even when administered late in the course of infection, suggesting that RIPK3 blockade may provide clinical benefit in patients with IAV-driven ARDS and other hyper-inflammatory pathologies.

Lung γδ T Cells Mediate Protective Responses during Neonatal Influenza Infection that Are Associated with Type 2 Immunity.

Compared to adults, infants suffer higher rates of hospitalization, severe clinical complications, and mortality due to influenza infection. We found that γδ T cells protected neonatal mice against mortality during influenza infection. γδ T cell deficiency did not alter viral clearance or interferon-γ production. Instead, neonatal influenza infection induced the accumulation of interleukin-17A (IL-17A)-producing γδ T cells, which was associated with IL-33 production by lung epithelial cells. Neonates lacking IL-17A-expressing γδ T cells or Il33 had higher mortality upon influenza infection. γδ T cells and IL-33 promoted lung infiltration of group 2 innate lymphoid cells and regulatory T cells, resulting in increased amphiregulin secretion and tissue repair. In influenza-infected children, IL-17A, IL-33, and amphiregulin expression were correlated, and increased IL-17A levels in nasal aspirates were associated with better clinical outcomes. Our results indicate that γδ T cells are required in influenza-infected neonates to initiate protective immunity and mediate lung homeostasis.

Exuberant fibroblast activity compromises lung function via ADAMTS4.

Severe respiratory infections can result in acute respiratory distress syndrome (ARDS)1. There are no effective pharmacological therapies that have been shown to improve outcomes for patients with ARDS. Although the host inflammatory response limits spread of and eventually clears the pathogen, immunopathology is a major contributor to tissue damage and ARDS1,2. Here we demonstrate that respiratory viral infection induces distinct fibroblast activation states, which we term extracellular matrix (ECM)-synthesizing, damage-responsive and interferon-responsive states. We provide evidence that excess activity of damage-responsive lung fibroblasts drives lethal immunopathology during severe influenza virus infection. By producing ECM-remodelling enzymes-in particular the ECM protease ADAMTS4-and inflammatory cytokines, damage-responsive fibroblasts modify the lung microenvironment to promote robust immune cell infiltration at the expense of lung function. In three cohorts of human participants, the levels of ADAMTS4 in the lower respiratory tract were associated with the severity of infection with seasonal or avian influenza virus. A therapeutic agent that targets the ECM protease activity of damage-responsive lung fibroblasts could provide a promising approach to preserving lung function and improving clinical outcomes following severe respiratory infections.

Publisher Correction: Exuberant fibroblast activity compromises lung function via ADAMTS4.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

A qualitative study of military service members undergoing medical separation.

OBJECTIVE: Little research explores military perspectives on medical disability-related transition. A qualitative study sought to understand transition experiences of United States military Service members found unfit for duty following medical and physical evaluation boards (MEBs and PEBs). METHODS: Confidential telephone interviews were conducted with 25 current and prior Service members. Participants were asked to share their experiences before, during, and after the MEB and PEB processes. Interview questions explored (1) health conditions that prompted the medical disability evaluation, (2) reactions to being recommended for separation, (3) transition-related stress and challenges, and (4) coping strategies. Salient themes were identified across chronological narratives. RESULTS: Participants expressed that debilitating physical (e.g., injury) and/or mental (e.g., post-traumatic stress disorder) illnesses prompted their medical evaluation. In response to the unfit for duty notice, some participants reported emotional distress (e.g., anxiety, anger) connected to uncertainty about the future. Other participants reported relief connected to a sense of progression toward their medical disability claim status. Transition stress included the length of the MEB/PEB process, impact of the COVID-19 pandemic on the process, financial stress, impact on family life, and compounded effect of these stressors on emotional distress, including depression and suicidal thoughts. Participants reported using adaptive (e.g., psychotherapy) and maladaptive (e.g., excessive drinking) strategies to cope with stress. CONCLUSION: Preliminary reports of emotional distress and transition stress following unfit for duty notices highlight the need for increased support and interventions to facilitate adaptive coping strategies during this vulnerable period.

A legal assessment of private land conservation in South America.

Privately protected areas (PPAs) are a potentially innovative conservation tool. Legal recognition is necessary for their success, especially where there are institutional challenges to nature conservation, such as in South America. Although PPAs have increased in South America since the early 2000s, there is a critical information gap pertaining to their legal frameworks. We analyzed the level of landowner commitment to and governmental support for PPAs across countries in South America that officially recognize PPAs. We analyzed the legal framework governing PPAs and reviewed literature on them. This process was done in English and Spanish. The information we gathered was validated by 16 conservation experts from 10 South American countries. Because Peru is 1 of only 2 South American countries where local communities create and manage PPAs, we studied Peruvian PPAs in more detail by examining official creation documents and interviewing 13 local conservation professionals. We found inadequate minimum duration of PPAs and vague guidelines for conducting economic activities within them and a lack of governmental support (e.g., financial and technical support) for PPAs. Support was limited to the exemption from rural property taxes, which are relatively low compared with countries outside South America. In Peru, PPAs run by individuals and communities needed different legal frameworks because they were created with different objectives and had different sizes and duration of commitments. The prompt improvement of legal frameworks across South America is necessary for PPAs to achieve their aim of being places for enduring nature conservation in the region.

Una revisión legal de la conservación voluntaria entierras privadas de América del Sur Resumen Las áreas protegidas privadas (APP) son una herramienta de conservación con potencial innovador. Para ser exitosas, las APP necesitan reconocimiento legal, especialmente cuando existen obstáculos institucionales para la conservación de la naturaleza, como sucede en América del Sur. Aunque las APP han aumentado en esta zona desde principios de la década del 2000, existe un vacío de información con respecto a sus marcos legales. Analizamos el nivel de compromiso de los terratenientes y el apoyo gubernamental hacia las APP en los países de América del Sur que reconocen de forma oficial las APP. Analizamos el encuadre legal que rige a las APP y revisamos la literatura existente sobre ellas; realizamos este proceso en inglés y en español. Dieciséis expertos de la conservación de diez países sudamericanos validaron la información recopilada. Ya que Perú es uno de los dos países de la zona en donde las comunidades locales crean y manejan las APP, nos enfocamos en sus APP y examinamos a detalle los documentos oficiales de creación y entrevistamos a 13 profesionales de la conservación locales. Encontramos una duración mínima inadecuada de las APP y directrices vagas para la realización de actividades dentro de ellas, así como una falta de apoyo gubernamental (p. ej.: apoyo económico y técnico). Este apoyo se limitaba a la exención de los impuestos sobre la propiedad rural, los cuales son relativamente bajos en comparación con los países fuera de América del Sur. En Perú, las APP a cargo de individuos y comunidades necesitaban diferentes encuadres legales porque fueron creados con diferentes objetivos y tenían diferentes tamaños y plazos para los compromisos. Se necesita una rápida mejora de los marcos legales en América del Sur para que las APP logren el objetivo de ser sitios para que perdure la conservación de la naturaleza en la región.

Association Between Patient Race and Ethnicity and Outcomes With COVID-19: A Retrospective Analysis From a Large Mid-Atlantic Health System.

BACKGROUND: Members of racial and ethnic minority groups have been disproportionately impacted by coronavirus-2019 (COVID-19). The objective of the study is to describe associations between race and ethnicity on clinical outcomes such as need for mechanical ventilation and mortality. METHODS: Retrospective cohort study of patients with severe COVID-19 infection admitted within a large, not-for-profit healthcare system in the mid-Atlantic region between March and July, 2020. Patient demographic data and clinical outcomes were abstracted from the electronic health record. Logistic regressions were performed to estimate associations between race and ethnicity and the clinical outcomes. RESULTS: The study population (N = 2931) was stratified into 1 of 3 subgroups: non-Hispanic White (n = 466), non-Hispanic Black (n = 1611), and Hispanic (n = 654). The average age of White, Black, and Hispanic patients was 69 ± 17.06, 64 ± 15.9, and 50 ± 15.53 years old, respectively (P < .001). Compared to White patients, Black and Hispanic patients were at increased odds of needing mechanical ventilation due to COVID-19 pneumonia (odds ratio [OR] Black = 1.35, 95% confidence interval [CI] = 1.04 to 1.75, P < .05; OR Hispanic = 1.43, 95% CI = 1.06 to 1.93, P < .05). When compared to White patients, Hispanic patients were at decreased odds of death (OR = 0.45, 95% CI = 0.32 to 0.63, P < .001). However, when adjusting for age, there were no statistically significant differences in the odds of death between these groups (adjusted OR [aOR] Black = 1.05, 95% CI = 0.80 to 1.38, P = .71; aOR Hispanic = 1.10, 95% CI = 0.76 to 1.60, P = .62). CONCLUSION: Our analysis demonstrated that Hispanic patients were more likely require mechanical ventilation but had lower mortality when compared to White patients, with lower average age likely mediating this association. These findings emphasize the importance of outreach efforts to communities of color to increase prevention measures and vaccination uptake to reduce infection with COVID-19.

Impact of increased alcohol consumption during the COVID-19-related lockdowns on admissions with liver disease, gastrointestinal bleeding and pancreatitis in Melbourne, Victoria.

This audit collates data on alcohol-related gastrointestinal (GI) admissions at Monash Health, Victoria, during the prolonged, coronavirus disease 2019 (COVID-19)-related lockdown July to October 2020 compared with the same periods in 2019 and 2021. We found a 58% increase in admissions in 2020 and a 16% increase in 2021, which also increased disproportionately to overall health service emergency presentations. Self-reported alcohol consumption increased by 2.5-fold and was greatest in 2020. Clinical severity was unchanged and cirrhosis was the only factor associated with severe disease. This study suggests an association between the pandemic-related lockdown, alcohol consumption and alcohol-related GI hospitalisation. Our study provides support for resourcing and adapting alcohol and other drug services during and beyond the COVID-19 lockdown.

Selecting Data Analytic and Modeling Methods to Support Air Pollution and Environmental Justice Investigations: A Critical Review and Guidance Framework.

Given the serious adverse health effects associated with many pollutants, and the inequitable distribution of these effects between socioeconomic groups, air pollution is often a focus of environmental justice (EJ) research. However, EJ analyses that aim to illuminate whether and how air pollution hazards are inequitably distributed may present a unique set of requirements for estimating pollutant concentrations compared to other air quality applications. Here, we perform a scoping review of the range of data analytic and modeling methods applied in past studies of air pollution and environmental injustice and develop a guidance framework for selecting between them given the purpose of analysis, users, and resources available. We include proxy, monitor-based, statistical, and process-based methods. Upon critically synthesizing the literature, we identify four main dimensions to inform method selection: accuracy, interpretability, spatiotemporal features of the method, and usability of the method. We illustrate the guidance framework with case studies from the literature. Future research in this area includes an exploration of increasing data availability, advanced statistical methods, and the importance of science-based policy.

Household Transmission of Carbapenemase-producing Enterobacterales in Ontario, Canada.

BACKGROUND: Data on household transmission of carbapenemase-producing Enterobacterales (CPE) remain limited. We studied risk of CPE household co-colonization and transmission in Ontario, Canada. METHODS: We enrolled CPE index cases (identified via population-based surveillance from January 2015 to October 2018) and their household contacts. At months 0, 3, 6, 9, and 12, participants provided rectal and groin swabs. Swabs were cultured for CPE until September 2017, when direct polymerase chain reaction (PCR; with culture of specimens if a carbapenemase gene was detected) replaced culture. CPE risk factor data were collected by interview and combined with isolate whole-genome sequencing to determine likelihood of household transmission. Risk factors for household contact colonization were explored using a multivariable logistic regression model with generalized estimating equations. RESULTS: Ninety-five households with 177 household contacts participated. Sixteen (9%) household contacts in 16 (17%) households were CPE-colonized. Household transmission was confirmed in 3/177 (2%) cases, probable in 2/177 (1%), possible in 9/177 (5%), and unlikely in 2/177 (1%). Household contacts were more likely to be colonized if they were the index case's spouse (odds ratio [OR], 6.17; 95% confidence interval [CI], 1.05-36.35), if their index case remained CPE-colonized at household enrollment (OR, 7.00; 95% CI, 1.92-25.49), or if they had at least 1 set of specimens processed after direct PCR was introduced (OR, 6.46; 95% CI, 1.52-27.40). CONCLUSIONS: Nine percent of household contacts were CPE-colonized; 3% were a result of household transmission. Hospitals may consider admission screening for patients known to have CPE-colonized household contacts.

A Framework for National Collaboration to Reduce Documentation Burden and Design for Usable and Reusable Data.

A focused effort is needed to capture the utility and usability of the electronic health record for providing usable and reusable data while reducing documentation burden. A collaborative effort of nurse leaders and experts was able to generate national consensus recommendations on documentation elements related to admission history. The process used in this effort is summarized in a framework that can be used by other groups to develop content that reduces documentation burden while maximizing the creation of usable and reusable data.

Hospital entrance smoking is reduced by broadcasting recorded antitobacco messages from Australian primary school children over entrance public address system.

ISSUE ADDRESSED: Smoking near hospital entrances occurs frequently despite smoke-free policies, resulting in multiple issues including second-hand smoke exposure (SHS) to vulnerable populations. Primary school children were engaged through their health curriculum to produce antismoking audio recordings for broadcast over a hospital entrance loudspeaker system to determine if this reduced smoking. METHOD: Students produced original recordings against hospital grounds smoking during class workshops, from which a collection (n = 16) was selected. Episodes of entrance smoking and total entrance traffic were recorded using security camera infrastructure over a 5-week period. A computer-controlled entrance loudspeaker played a message which was followed by silence until a new (different) message was played. Intensity of messaging was moderate in week 3 (every 5 minutes), increasing to high in week 4 (3 minutely) and compared to no messages (weeks 1-2 preintervention) and week 5 (postintervention). RESULTS: Smokers presented 316 times, smoking 523 cigarettes over 155 hours of observation (patients 70.6%, visitors 29.4%). SHS exposure was high given 172 others used the entrance/hour. Smoking was highest in the preintervention period (weeks 1-2), median five cigarettes/hr [IQR,3-7 (min = 0, max = 12)], falling to four cigarettes/hr during 5-minutely broadcasts [IQR,2-5 (min = 1, max = 14)] (P = .06), but zero cigarettes/hr during 3-minutely broadcasts [IQR,0-1(min = 0, max = 7)] (P < .0001). Postintervention (no broadcasts), smoking increased from zero to 1 cigarette/hr [IQR,0-3 (min = 0, max = 5)]; (P = .052). Nonsmoker movements did not change significantly between each period. CONCLUSION: Intensive (3 minutely) broadcasting of short antismoking messages significantly reduced hospital entrance smoking. SO WHAT?: Health services can positively interact with the health curriculum of primary schools against tobacco use while developing low-cost strategies to effectively deter entrance smoking.

Arrival of the rare carbapenemase OXA-204 in Canada causing a multispecies outbreak over 3 years.

OBJECTIVES: To investigate a persistent multispecies OXA-204 outbreak occurring simultaneously in multiple distant hospitals in the province of Quebec, Canada. METHODS: OXA-204 carbapenemase-producing Enterobacterales (CPE) isolated from multiple hospitals between January 2016 and October 2018 were included in the study. An epidemiological inquiry was conducted in order to elucidate possible transmission routes and a putative source. Isolates were characterized by standardized antibiotic susceptibility testing and by WGS, using Illumina short-read data and MinION long-read data. RESULTS: The outbreak comprised 65 patients and 82 isolates from four hospital sites. Most patients were ≥65 years old, had multiple comorbidities and had received antibiotics recently. The infection to colonization ratio was 1:20. No persistent environmental reservoir was identified. The most frequent organism was Citrobacter freundii (n = 78), followed by Klebsiella spp. (n = 3) and Escherichia coli (n = 1). WGS analysis showed 77/78 C. freundii isolates differing by 0-26 single nucleotide variants (SNVs). Results of WGS analysis showed blaOXA-204 was present on three plasmids types (IncX1, IncA/C2 and IncFII/FIB/A/C2) and on a prophage. All C. freundii isolates harboured multiple copies of blaOXA-204, both on the chromosome and a plasmid. Plasmid IncFII/FIB/A/C2 was observed in all three species. CONCLUSIONS: Transfer of OXA-204 plasmids likely occurred between species within the same patient, highlighting the plasticity of these plasmids and potential for widespread dissemination. OXA-204 carbapenemase has been introduced into Quebec and has rapidly disseminated. Although the infection to colonization ratio was low in this outbreak, this carbapenemase has been associated with severe infection elsewhere.

Implementation Analysis of a Perioperative Patient Safety Program in Guatemala.

BACKGROUND: The implementation of programs to improve patient safety remains challenging in low- and middle-income countries. The goal of our study was to define the barriers and facilitators to implementation of a perioperative patient safety program in Guatemala. MATERIAL AND METHODS: We conducted semi-structured interviews with 16 staff pre-intervention and a follow-up focus group discussion 1 year later in the perioperative department at the Roosevelt Hospital in Guatemala. We performed qualitative thematic analysis to identify barriers and facilitators to the implementation process, with analysis guided by the Consolidated Framework for Implementation Research. RESULTS: We found several dominant themes affecting implementation of a patient safety program. Implementation facilitators included strong prioritization of patient needs, program compatibility with existing workflow, and staff attributes. Barriers included a lack of knowledge about patient safety, limited resources, limited leadership engagement, and lack of formal implementation leaders. Several program modifications were made to enhance successful implementation iteratively during the implementation process. DISCUSSION: Our analysis highlights several dominant themes which affect the implementation of a perioperative safety program in Guatemala. Understanding the barriers and facilitators to implementation during program deployment allows for program modification and improvement of the implementation process itself.

Assessment of Maya women's knowledge, attitudes, and beliefs on sexually transmitted infections in Guatemala: a qualitative pilot study.

BACKGROUND: Despite Guatemala's large indigenous population, indigenous health is often neglected in reported health data and interventions. Although this data is limited in scope, it shows that indigenous people have poorer health outcomes. Sexually transmitted infections (STIs) are now a growing threat in Guatemala and pose great risk to the wellbeing of its indigenous population. METHODS: This qualitative pilot study assessed the knowledge, attitudes, and beliefs of STIs through semi-structured interviews among a previously unstudied population of indigenous Maya women (n = 35, ages 18-50) in the six municipalities of Santa Cruz La Laguna, Guatemala. RESULTS: Four key themes were identified: 1) indigenous Maya women have limited factual knowledge about sex and STIs; 2) widespread partner infidelity minimizes women's control over preventing STI contraction; 3) close-knit communities and the resulting heightened fear of gossip prevents communication and hinders care seeking; and 4) lack of quality medical care and inaccessibility of biomedical healthcare systems pose barriers to seeking care for potential STIs. CONCLUSIONS: To address these findings, we suggest methods to improve sexual education, combat male infidelity, promote condom use, and improve health services to reduce the incidence of STIs in Maya Guatemala.

People, Pipelines, and Probabilities: Clarifying Significance and Uncertainty in Environmental Impact Assessments.

Determinations of significance play a pivotal role in environmental impact assessments because they point decision makers to the predicted effects of an action most deserving of attention and further study. Impact predictions are always subject to uncertainty because they rely on estimates of future consequences. Yet uncertainty is often neglected or treated in a perfunctory manner as part of the characterization, evaluation, and communication of anticipated consequences and their significance. Proposals to construct fossil fuel pipelines in North America provide a highly visible example; casual treatment of how uncertainty affects significance determinations has resulted in poorly informed stakeholders, frustrated industry proponents, and inconsistent choices on the part of public decision makers. Using environmental assessments for recent pipeline proposals as examples, we highlight five ways in which uncertainty is often neglected when determining impact significance and suggest that a mix of known methods, new guidelines, and appropriate oversight could greatly improve current practices.

Two Hypervirulent Klebsiella pneumoniae Isolates Producing a blaKPC-2 Carbapenemase from a Canadian Patient.

This report describes two hypervirulent Klebsiella pneumoniae isolates that produced K. pneumoniae carbapenemase (KPC), which were identified from a rectal swab and a urine culture upon hospital admission. The patient had recently traveled to Greece, where he was hospitalized. The isolates were sequence type 86 and contained an IncHI1B IncFIBK hypervirulent plasmid and an IncFIIK plasmid harboring KPC.