Relationship between trauma, psychological distress and help-seeking among corrective service workers.

BACKGROUND: Corrective service workers (CSWs) are at high risk of post-traumatic stress disorder (PTSD) and other mental health problems. Prevalence rates and help-seeking behaviours are under-researched within this population. AIMS: To assess rates of PTSD and distress, and identify predictors of intention to seek help, among workers at an Australian corrective service agency. METHODS: A cross-sectional online survey was used to collect data on staff demographics, employment, PTSD symptoms and current distress. Participants received a tailored feedback report including referral to relevant mental health services (where applicable) and were asked to indicate their likelihood of seeking help. Prevalence data are reported. Binary logistic regression was used to examine relationships between participant characteristics and help-seeking for those with probable PTSD and/or high psychological distress. RESULTS: Participants (n=1001) were predominantly men (56.8%) with a mean age of 46.72 (SD=11.00). Over half (58.0%) were classified as probable PTSD cases, and one-third (33.0%) were experiencing high psychological distress. Around a third (34.3%) of participants with probable PTSD and/or elevated distress indicated they were likely to seek help. Older age and fewer years of service were associated with increased help-seeking intentions. CONCLUSIONS: CSWs were found to be experiencing probable PTSD at higher rates than reported in previous studies. Relatively few intended to seek help from mental health services, despite being provided with personalised screening and feedback along with access to specialised care. Future research should investigate the potential role of organisational support as a facilitator of help-seeking within this population.

Job Loss Due to COVID-19: A Longitudinal Study of Mental Health, Protective and Risk Factors.

The COVID-19 pandemic had a devastating impact on unemployment, which-compounded by the additional stressors associated with the pandemic-had considerable mental health impact. The current study examined the trajectory of mental health amongst those experiencing pandemic-related job loss, alongside the impact of risk and protective factors. Data were obtained from 374 Australian participants who were allocated to a waitlist control arm of a randomised control trial. The outcome variables assessed at baseline and six-month follow-up consisted of depression, anxiety, and suicidality. The assessed risk and protective factors were age, gender, relationship status, education, exercise frequency, COVID-related stress, dispositional resilience, and coping self-efficacy. Re-employment by follow-up was used as a covariate. Overall, there were decreases in depression and anxiety symptoms, and partial evidence of decreased suicidality, demonstrating an apparent capacity for individuals to better cope with their circumstances over time. Demographics and exercise had no effect on changes in mental health. Those with high COVID-related stress, low resilience, and low coping self-efficacy had worse mental health at baseline, although exhibited significantly greater improvements in mental health over time. Obtaining re-employment by follow-up did not predict changes in mental health. The present results offer an optimistic picture of recovery for those experiencing pandemic-related job loss, even for those with the most substantial risk and severity. The likely protective role played by Australian social welfare policies over the course of the study is explored. Stress around one's broader sociocultural or economic circumstances, perceived resilience, and coping self-efficacy are valuable targets for intervention.

Flood-irrigated agriculture mediates climate-induced wetland scarcity for summering sandhill cranes in western North America.

Information about species distributions is lacking in many regions of the world, forcing resource managers to answer complex ecological questions with incomplete data. Information gaps are compounded by climate change, driving ecological bottlenecks that can act as new demographic constraints on fauna. Here, we construct greater sandhill crane (Antigone canadensis tabida) summering range in western North America using movement data from 120 GPS-tagged individuals to determine how landscape composition shaped their distributions. Landscape variables developed from remotely sensed data were combined with bird locations to model distribution probabilities. Additionally, land-use and ownership were summarized within summer range as a measure of general bird use. Wetland variables identified as important predictors of bird distributions were evaluated in a post hoc analysis to measure long-term (1984-2022) effects of climate-driven surface water drying. Wetlands and associated agricultural practices accounted for 1.2% of summer range but were key predictors of occurrence. Bird distributions were structured by riparian floodplains that concentrated wetlands, and flood-irrigated agriculture in otherwise arid and semi-arid landscapes. Findings highlighted the role of private lands in greater sandhill crane ecology as they accounted for 78% of predicted distributions. Wetland drying observed in portions of the range from 1984 to 2022 represented an emerging ecological bottleneck that could limit future greater sandhill crane summer range. Study outcomes provide novel insight into the significance of ecosystem services provided by flood-irrigated agriculture that supported nearly 60% of wetland resources used by birds. Findings suggest greater sandhill cranes function as a surrogate species for agroecology and climate change adaptation strategies seeking to reduce agricultural water use through improved efficiency while also maintaining distinct flood-irrigation practices supporting greater sandhill cranes and other wetland-dependent wildlife. We make our wetland and sandhill crane summering distributions available as interactive web-based mapping tools to inform conservation design.

Analysis of SARS-CoV-2 Variant-Specific Serum Antibody Post-Vaccination Utilizing Immortalized Human Hepatocyte-Like Cells (HLC) to Assess Development of Immunity.

Background: Our previous studies demonstrated that SARS-CoV-2 spike protein could bind to primary hepatocytes and immortalized Hepatocyte-like cells (HLC) via the asialoglycoprotein receptor-1 (ASGR-1). The binding of biotinylated spike protein could be inhibited by Spike-neutralizing monoclonal antibodies, anti-ASGR-1 antibodies and unlabeled spike protein. The cells were unable to bind Spike S1 and Spike S1 was incapable of blocking labeled Spike protein, suggesting that the Receptor Binding Domain (RBD) was not involved in the binding event. This study was done to investigate the utility of these cells and immortalized alveolar type 2-like (AT-2) cells in studying the development of variant-specific antibodies post-vaccination. Methods: Serum was collected from 10 individuals pre- and post-vaccination with the J&J, Moderna or Pfizer vaccines. The serum samples were quantified for variant-specific antibodies in a flow cytometry-based immunofluorescent assay utilizing beads coated with biotinylated variant spike proteins. Inhibition of spike protein binding to HLC and AT-2 cells by donor serum was analyzed by immunofluorescent confocal analysis. Results: All variant spike proteins bound to HLC and AT-2 cells. Post-vaccination serum samples demonstrated increases of SARS-CoV-2 antibody levels from 2 weeks to 2.5 months post-vaccination with associated increased spike-blocking capacity. It was also demonstrated that vaccination with all the available vaccines stimulated antibodies that inhibited binding of all the available variant spike proteins to both HLC and AT-2 cells. Conclusion: HLC, along with AT-2 cells, provides a useful platform to study the development of neutralizing antibodies post-vaccination. Vaccination with the 3 available vaccines all elicited neutralizing serum antibodies that inhibited binding of each of the variant spike proteins to both AT-2 and HLC cells. This study suggests that inhibition of spike binding to target cells may be a more useful technique to assess immunity than gross quantitation of antibody.

A mixed-method survey to understand the role of dog welfare organisations in Ireland, including reported challenges and potential solutions.

BACKGROUND: This novel study forms part of a larger research programme seeking an improved understanding of aspects of the owned dog population in Ireland. Dog welfare organisations (DWOs) in Ireland are recognised as an instrumental pillar of the animal welfare sector with some receiving substantial public funding. We conducted a survey of DWOs in Ireland (n = 39) to gain a better understanding of their role and function, including their policies and procedures and the rehoming of dogs to other regions. In addition, we wanted to get a better understanding of the challenges experienced by DWOs in fulfilling their role and their perspectives on potential solutions to these challenges. The survey questions consisted of closed and open-ended items. Closed items were analysed quantitively; open-ended items were analysed thematically. RESULTS: Most DWOs (> 80%) had written protocols for important welfare actions including rehoming procedures, assessment of owner suitability and euthanasia. DWOs sent dogs to Northern Ireland (13%), Great Britain (38.5%) and to other countries outside the United Kingdom (36%, including Germany, Sweden, Italy, the Netherlands and Czechia). Reported challenges included a general lack of funding, limited public awareness of the importance of dog welfare and insufficient capacity to handle dog numbers. To address these challenges, the DWOs highlighted the potential contribution of subsidised programmes and access to resources to educate potential owners. In a further qualitative evaluation to capture perceptions of appropriate solutions by DWOs, several themes emerged, relating to legislation, education, an overwhelmed workforce, and funding. CONCLUSIONS: This study provides important insights into the roles and functions of DWOs and challenges they experience in Ireland. It is hoped that the findings from this research will inform future research investigating potential solutions to these challenges as well as the development of policy in Ireland.

Selective Prevention of Depression in Workers Using a Smartphone App: Randomized Controlled Trial.

BACKGROUND: There is increasing evidence that depression can be prevented; however, universal approaches have had limited success. Appropriate targeting of interventions to at-risk populations has been shown to have potential, but how to selectively determine at-risk individuals remains unclear. Workplace stress is a risk factor for depression and a target for intervention, but few interventions exist to prevent depression among workers at risk due to heightened stress. OBJECTIVE: This trial aimed to evaluate the efficacy of a smartphone-based intervention in reducing the onset of depression and improving related outcomes in workers experiencing at least moderate levels of stress. METHODS: A randomized controlled trial was conducted with participants who were currently employed and reported no clinically significant depression and at least moderate stress. The intervention group (n=1053) were assigned Anchored, a 30-day self-directed smartphone app-based cognitive behavioral- and mindfulness-based intervention. The attention-control group (n=1031) were assigned a psychoeducation website. Assessment was performed via web-based self-report questionnaires at baseline and at 1-, 3-, and 6-month postbaseline time points. The primary outcome was new depression caseness aggregated over the follow-up period. The secondary outcomes included depressive and anxiety symptoms, stress, well-being, resilience, work performance, work-related burnout, and quality of life. Analyses were conducted within an intention-to-treat framework using mixed modeling. RESULTS: There was no significant between-group difference in new depression caseness (z score=0.69; P=.49); however, those in the Anchored arm had significantly greater depressive symptom reduction at 1 month (Cohen d=0.02; P=.049) and 6 months (Cohen d=0.08; P=.03). Anchored participants also showed significantly greater reduction in anxiety symptoms at 1 month (Cohen d=0.07; P=.04) and increased work performance at 1 month (Cohen d=0.07; P=.008) and 6 months (Cohen d=0.13; P=.01), compared with controls. Notably, for Anchored participants completing at least two-thirds of the intervention, there was a significantly lower rate of depression onset (1.1%, 95% CI 0.0%-3.7%) compared with controls (9.0%, 95% CI 6.8%-12.3%) at 1 month (z score=4.50; P<.001). Significant small to medium effect sizes for most secondary outcomes were seen in the highly engaged Anchored users compared with controls, with effects maintained at the 6-month follow-up for depressive symptoms, well-being, stress, and quality of life. CONCLUSIONS: Anchored was associated with a small comparative reduction in depressive symptoms compared with controls, although selective prevention of case-level depression was not observed in the intention-to-treat analysis. When users adequately engaged with the app, significant findings pertaining to depression prevention, overall symptom reduction, and functional improvement were found, compared with controls. There is a need for a greater focus on engagement techniques in future research. TRIAL REGISTRATION: Australian New Zealand Clinical Trials Registry (ANZCTR) ACTRN12620000178943; https://www.anzctr.org.au/Trial/Registration/TrialReview.aspx?id=378592.

Mental health and unemployment: A systematic review and meta-analysis of interventions to improve depression and anxiety outcomes.

BACKGROUND: Unemployment is associated with substantially greater depression and anxiety, constituting a considerable public health concern. The current review provides the most comprehensive synthesis to date, and first meta-analysis, of controlled intervention trials aimed at improving depression and anxiety outcomes during unemployment. METHODS: Searches were conducted within PsycInfo, Cochrane Central, PubMed and Embase from their inception to September 2022. Included studies conducted controlled trials of interventions focused on improving mental health within unemployed samples, and reported on validated measures of depression, anxiety, or distress (mixed depression and anxiety). Narrative syntheses and random effects meta-analyses were conducted among prevention- and treatment-level interventions for each outcome. RESULTS: A total of 39 articles reporting on 33 studies were included for review (sample sizes ranging from 21 to 1801). Both prevention and treatment interventions tended to be effective overall, with treatment interventions producing larger effect sizes than prevention interventions. The clearest evidence for particular intervention approaches emerged for prevention-level Cognitive Therapy/CBT, followed by prevention-level work-related interventions, although neither produced entirely consistent effects. LIMITATIONS: Risk of bias was generally high across studies. Low numbers of studies within subgroups precluded any comparisons between long-term and short-term unemployment, limited comparisons among treatment studies, and reduced the power of meta-analyses. CONCLUSIONS: Both prevention- and treatment-level mental health-focused interventions have merit for reducing symptoms of anxiety and depression among those experiencing unemployment. Cognitive Therapy/CBT and work-related interventions hold the most robust evidence base, which can inform both prevention and treatment strategies implemented by clinicians, employment services providers, and governments.

The development of a smartphone app to enhance post-traumatic stress disorder treatment in high-risk workers.

Objective: Post-traumatic stress disorder (PTSD) is highly prevalent in certain populations. However, evidence indicates that many individuals do not respond to treatment. Digital supports hold promise for increasing service provision and engagement but there is a lack of evidence on blended care options and still less research guiding the development of such tools. This study details the development and overarching framework used to build a smartphone app to support PTSD treatment. Methods: The app was developed in line with the Integrate, Design, Assess, and Share (IDEAS) framework for the development of digital health interventions and involved clinicians (n=3), frontline worker clients (n=5) and trauma-exposed frontline workers (n=19). Integrated iterative rounds of testing were conducted via in-depth interviews, surveys, prototype testing and workshops, alongside app and content development. Results: Clinicians and frontline workers both expressed a clear preference for the app to augment but not replace face-to-face therapy, with the aim of increasing between-session support, and facilitating homework completion. Content was adapted for app delivery from manualised therapy (trauma-focused cognitive behavioural therapy (CBT). Prototype versions of the app were well received, with both clinicians and clients reporting the app was easy to use, understandable, appropriate and highly recommendable. System Usability Scale (SUS) scores were on average in the excellent range (82/100). Conclusions: This is one of the first studies to document the development of a blended care app designed specifically to augment clinical care for PTSD, and the first within a frontline worker population. Through a systematic framework with active end user consultation, a highly usable app was built to undergo subsequent evaluation.

Joint use of location and acceleration data reveals influences on transitions among habitats in wintering birds.

Quantifying relationships between animal behavior and habitat use is essential to understanding animal decision-making. High-resolution location and acceleration data allows unprecedented insights into animal movement and behavior. These data types allow researchers to study the complex linkages between behavioral plasticity and habitat distribution. We used a novel Markov model in a Bayesian framework to quantify the influence of behavioral state frequencies and environmental variables on transitions among landcover types through joint use of location and tri-axial accelerometer data. Data were collected from 56 greater white-fronted geese (Anser albifrons frontalis) across seven ecologically distinct winter regions over two years in midcontinent North America. We showed that goose decision-making varied across landcover types, ecoregions, and abiotic conditions, and was influenced by behavior. We found that time spent in specific behaviors explained variation in the probability of transitioning among habitats, revealing unique behavioral responses from geese among different habitats. Combining GPS and acceleration data allowed unique study of potential influences of an ongoing large-scale range shift in the wintering distribution of a migratory bird across midcontinent North America. We anticipate that behavioral adaptations among variable landscapes is a likely mechanism explaining goose use of highly variable ecosystems during winter in ways which optimize their persistence.

Pilot Trial of Workable: A Therapist-Supported Digital Program for Injured Workers.

Workplace sickness absence is a major public health and economic problem, and common mental disorders (CMDs) such as anxiety and depression are associated with particularly high rates of long-term sickness absence. Effective return-to-work (RTW) interventions are required. This pilot study investigates the feasibility, acceptability, and potential effectiveness of a new therapist-assisted Web-based RTW intervention (Workable) for injured workers on sick leave for a psychological or physical injury. A single-group open pilot trial design was used, with assessments at pre-treatment and post-treatment. The intervention consisted of 6 weeks of online modules and 6 coaching calls from a psychologist. A total of 13 participants were recruited and 9 completed all questionnaires. Program adherence was high, with 92% of participants completing the 6-week intervention. Participants reported high levels of intervention satisfaction and ease of use. There were large and significant reductions between pre- and post-treatment on measures of depression, anxiety, stress, and workdays missed over the past four weeks, along with a significant increase in self-reported work ability. These results suggest that Workable is a feasible and acceptable intervention for injured workers, with the potential to improve mental health and RTW outcomes. A randomized controlled trial is required to determine the efficacy of the intervention.

Harnessing synthetic biology to enhance ocean health.

Ocean health is faltering, its capability for regeneration and renewal being eroded by a steady pulse of anthropomorphic impacts. Plastic waste has infiltrated all ocean biomes, climate change threatens coral reefs with extinction, and eutrophication has unleashed vast algal blooms. In the face of these challenges, synthetic biology approaches may hold untapped solutions to mitigate adverse effects, repair ecosystems, and put us on a path towards sustainable stewardship of our planet. Leveraging synthetic biology tools would enable innovative engineering approaches to augment the natural adaptive capacity of ocean biological systems to cope with the swiftness of human-induced change. Here, we present a framework for developing synthetic biology solutions for the challenges of plastic pollution, coral bleaching, and harmful algal blooms.

A Qualitative Exploration of Challenges and Opportunities for Dog Welfare in Ireland Post COVID-19, as Perceived by Dog Welfare Organisations.

This novel qualitative study identifies challenges and opportunities to improve dog welfare in Ireland, as perceived by dog welfare organisations (DWOs), a previously underutilised stakeholder. This study sought the views of this predominantly voluntary sector of the next steps for policy and action in dog welfare, in light of the effects of the "puppy pandemic", increased costs and COVID-19 restrictions. An integrated online focus group and interview design involving DWOs was analysed using inductive thematic analysis. Thematic analysis identified 2 key themes: (1) Key challenges and solutions in general dog welfare and (2) Challenges and opportunities in the welfare organisation sector. DWOs perceived poor public awareness of appropriate dog-husbandry, inadequate legislation enforcement, negative impact of puppy farms, and increased financial and volunteer burden. DWOs helped construct a best practice rehoming pathway, identified how overall standards could be improved and recommendations to enhance dog welfare. The DWOs perceived an increased numbers of households acquiring dogs, difficulties in rehoming, and financial challenges as threatening their viability as organisations and Irish dog welfare. Greater enforcement of legislation, greater communication between organisations and the government, and more media awareness were seen as helpful by the DWOs to improve dog welfare standards and their organisations.

Apprentices' Attitudes Toward Using a Mental Health Mobile App to Support Healthy Coping: Mixed Methods Study.

BACKGROUND: Apprenticeships are a common pathway for young people transitioning into the workforce. Apprentices often face many employment-related challenges and have high levels of psychological distress, drug and alcohol use, and suicidal ideation. Little is known about the attitudes of apprentices toward using smartphone apps to support their mental health and the content that would engage them. OBJECTIVE: This study explored (1) apprentices' interest in using an app to support their mental health and (2) the healthy coping strategies used to manage their mental well-being in the face of workplace challenges, in order to inform future app content. METHODS: A mixed methods study was conducted with 54 apprentices (50/54 male, 93%) with a mean age of 22.7 (SD 5.7) years. Participants completed a survey on preferred ways of using an app to support mental health. Across 8 focus groups, participants were asked to describe healthy strategies they used to cope with occupational stressors. RESULTS: Only 11% (6/54) of participants currently used a well-being app, but there was high interest in using an app to support their friends (47/54 participants, 87%) and develop self-help strategies to manage or prevent mental health issues (42/54 participants, 78%). Four major types of coping behaviors were identified: (1) social connection for disclosure, advice, and socializing; (2) pleasurable activities, such as engaging in hobbies, time-outs, and developing work-life separation; (3) cognitive approaches, including defusing from thoughts and cognitive reframing; and (4) self-care approaches, including exercise, a healthy diet, and getting adequate sleep. CONCLUSIONS: There is interest among apprentices to use an app with a positive well-being focus that helps them to develop self-management skills and support their friends. Apprentices utilized a range of healthy behaviors to cope with workplace stressors that can be incorporated into mental health apps to improve uptake and engagement. However, many of the preferred coping strategies identified are not those focused on by currently available apps, indicating the need for more targeted digital interventions for this group.

Caught in the act: Monitoring OO bond cleavage in Acylperoxoferric cytochrome P450cam to form compound I in real time.

While monitoring the reaction of ferric cytochrome P450cam (Cyp101) with substituted peroxybenzoic acids using rapid-scanning, stopped-flow (RSSF) spectroscopy, an intermediate appears en route to formation of the high-valent moiety known as Compound I [Fe(IV)=O/porphyrin radical cation] that is thought to be the key catalytic species for O-atom transfer to substrate. We have previously suggested (Spolitak, T., Dawson, J.H., Ballou, D.P., J. Biol. Chem.2005, 280, 20,300-20,309) that this species is an acylperoxo-ferric heme adduct that subsequently undergoes OO bond cleavage to generate Compound I. Singular value decomposition analysis of the RSSF data for formation of this intermediate shows that the energy of its Soret absorption peak is sensitive to the electron donor properties of the aryl substituents on the peracid. A linear Hammett correlation plot is seen for the energy of the Soret absorption peak vs. the Hammett σ constant. This correlation requires that the aryl substituents remain as part of the ligand bound to the heme iron, providing direct evidence that the adduct is indeed a ferric acylperoxo derivative. Linear Hammett correlation plots are also seen for both the rate of formation of the intermediate as well as for its conversion to Compound I. It is proposed that the electron donating/withdrawing properties of the aryl-bound substituents affect the electrophilic nature for binding substrate, changing the observed rate of formation for the acylperoxo intermediate, as well as the propensity and stability of the substituted benzoic acid to serve as the leaving group during OO bond cleavage yielding Compound I.

Understanding the dog population in the Republic of Ireland: insight from existing data sources?

BACKGROUND: Reliable information about national pet dog populations is an important contributor to informed decision-making, both by governments and national dog welfare organisations. In some countries, there is an improved understanding of aspects of the national pet dog population, but as yet limited published information is available in Ireland. The current study reviews the utility of existing data to inform our understanding of recent changes to the pet dog population in Ireland, including both biological and organisational processes. RESULTS: Based on national data on dog licencing and microchipping registration, pet dog numbers have remained relatively stable in recent years (ie prior to the COVID-19 pandemic). Since 2015, there has been a substantial decrease in the number of dogs managed through dog control centres. Although the completeness of the data are likely variable, there appears to be substantial, and increasing, number of dogs moving from Ireland to other countries, including UK, Sweden, Italy, Germany and Singapore. We also note an increase (albeit much smaller) in the number of dogs being moved into Ireland. CONCLUSIONS: This study highlights the challenges faced when using existing national data to gain insights into the dog population of Ireland. The linking of existing national databases (individual dog identification, dog licencing, dog control statistics) has the potential to improve both the representativeness and accuracy of information about the Irish pet dog population. In the next phases of our work, we will focus on the work of dog welfare organisations, given both the increased role played by these organisations and the substantial public funding that has been committed in this sector.

Acyl chain selection couples the consumption and synthesis of phosphoinositides.

Phosphoinositides (PIPn) in mammalian tissues are enriched in the stearoyl/arachidonoyl acyl chain species ("C38:4"), but its functional significance is unclear. We have used metabolic tracers (isotopologues of inositol, glucose and water) to study PIPn synthesis in cell lines in which this enrichment is preserved to differing relative extents. We show that PIs synthesised from glucose are initially enriched in shorter/more saturated acyl chains, but then rapidly remodelled towards the C38:4 species. PIs are also synthesised by a distinct 're-cycling pathway', which utilises existing precursors and exhibits substantial selectivity for the synthesis of C38:4-PA and -PI. This re-cycling pathway is rapidly stimulated during receptor activation of phospholipase-C, both allowing the retention of the C38:4 backbone and the close coupling of PIPn consumption to its resynthesis, thus maintaining pool sizes. These results suggest that one property of the specific acyl chain composition of PIPn is that of a molecular code, to facilitate 'metabolic channelling' from PIP2 to PI via pools of intermediates (DG, PA and CDP-DG) common to other lipid metabolic pathways.

Integrated population modelling reveals potential drivers of demography from partially aligned data: a case study of snowy plover declines under human stressors.

Knowledge of demography is essential for understanding wildlife population dynamics and developing appropriate conservation plans. However, population survey and demographic data (e.g., capture-recapture) are not always aligned in space and time, hindering our ability to robustly estimate population size and demographic processes. Integrated population models (IPMs) can provide inference for population dynamics with poorly aligned but jointly analysed population and demographic data. In this study, we used an IPM to analyse partially aligned population and demographic data of a migratory shorebird species, the snowy plover (Charadrius nivosus). Snowy plover populations have declined dramatically during the last two decades, yet the demographic mechanisms and environmental drivers of these declines remain poorly understood, hindering development of appropriate conservation strategies. We analysed 21 years (1998-2018) of partially aligned population survey, nest survey, and capture-recapture-resight data in three snowy plover populations (i.e., Texas, New Mexico, Oklahoma) in the Southern Great Plains of the US. By using IPMs we aimed to achieve better precision while evaluating the effects of wetland habitat and climatic factors (minimum temperature, wind speed) on snowy plover demography. Our IPM provided reasonable precision for productivity measures even with missing data, but population and survival estimates had greater uncertainty in years without corresponding data. Our model also uncovered the complex relationships between wetland habitat, climate, and demography with reasonable precision. Wetland habitat had positive effects on snowy plover productivity (i.e., clutch size and clutch fate), indicating the importance of protecting wetland habitat under climate change and other human stressors for the conservation of this species. We also found a positive effect of minimum temperature on snowy plover productivity, indicating potential benefits of warmth during night on their population. Based on our results, we suggest prioritizing population and capture-recapture surveys for understanding population dynamics and underlying demographic processes when data collection is limited by time and/or financial resources. Our modelling approach can be used to allocate limited conservation resources for evidence-based decision-making.

Differentiation of immortalized human multi-lineage progenitor to alveolar type 2-like cells: angiotensin-converting enzyme 2 expression and binding of severe acute respiratory syndrome coronavirus 2 spike and spike 1 proteins.

Along with the nasal epithelium, the lung epithelium is a portal of entry for sudden acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and many other respiratory viruses. In the case of SARS-CoV-2, the virus surface spike proteins bind to the angiotensin-converting enzyme 2 (ACE-2) receptor to facilitate entry into the respiratory epithelium. Alveolar type 2 (AT2) cells are committed respiratory progenitor cells responsible for the integrity and regeneration of the respiratory epithelium and production of respiratory surfactant proteins. AT2 cells express high levels of surface ACE-2 and thus are a leading target for primary infection by SARS-CoV-2. This study describes a method for directly differentiating telomerase reverse transcriptase-immortalized human cord blood-derived multi-lineage progenitor cells (MLPCs) to AT2-like cells for the purpose of generating an in vitro cellular platform for viral studies. Differentiation was confirmed with the acquisition of AT2 and absence of alveolar type 1 (AT1) specific markers by confocal microscopy. Expression of the ACE-2 receptor was confirmed by immunofluorescence antibody staining, quantitative reverse transcription polymerase chain reaction and binding of biotinylated SARS-CoV-2 spike and spike 1 proteins. The binding of biotinylated spike proteins was specifically blocked by unlabeled spike proteins and neutralizing antibodies. Additionally, it was demonstrated that the spike protein was internalized after binding to the surface membrane of the cells. The authors defined the culture conditions that enabled AT2-like cells to be repeatedly passaged and cryopreserved without further differentiation to AT1. The authors' method provides a stable and renewable source of AT2 cells for respiratory viral binding, blocking and uptake studies.

Binding of the SARS-CoV-2 Spike Protein to the Asialoglycoprotein Receptor on Human Primary Hepatocytes and Immortalized Hepatocyte-Like Cells by Confocal Analysis.

BACKGROUND: The SARS-CoV-2 virus may have direct or indirect effects on other human organs beyond the respiratory system and including the liver, via binding of the spike protein. This study investigated the potential direct interactions with the liver by comparing the binding of SARS-CoV-2 spike proteins to human AT2-like cells, primary human hepatocytes and immortalized hepatocyte-like hybrid cells. Receptors with binding specificity for SARS-CoV-2 spike protein on AT2 cells and hepatocytes were identified. METHODS: The specific binding of biotinylated spike and spike 1 proteins to undifferentiated human E12 MLPC (E12), E12 differentiated alveolar type 2 (AT2) cells, primary human hepatocytes (PHH) and E12 human hepatocyte-like hybrid cells (HLC) was studied by confocal microscopy. We investigated the expression of ACE-2, binding of biotinylated spike protein, biotinylated spike 1 and inhibition of binding by unlabeled spike protein, two neutralizing antibodies and an antibody directed against the hepatocyte asialoglycoprotein receptor 1 (ASGr1). RESULTS: E12 MLPC did not express ACE-2 and did not bind either of spike or spike 1 proteins. AT2-like cells expressed ACE-2 and bound both spike and spike 1. Both PHH and HLC did not express ACE-2 and did not bind spike 1 protein. However, both PHH and HLC actively bound the spike protein. Biotinylated spike protein binding was inhibited by unlabeled spike but not spike 1 protein on PHH and HLC. Two commercial neutralizing antibodies blocked the binding of the spike to PHH and HLC but only one blocked binding to AT2. An antibody to the hepatocyte ASGr1 blocked the binding of the spike protein to PHH and HLC. CONCLUSION: The absence of ACE-2 receptors and inhibition of spike binding by an antibody to the ASGr1 on both PHH and HLC suggested that the spike protein interacts with the ASGr1. The differential antibody blocking of spike binding to AT2, PHH and HLC indicated that neutralizing activity of SARS-CoV-2 binding might involve additional mechanisms beyond RBD binding to ACE-2.

CRISPR/Cas9-Based Lateral Flow and Fluorescence Diagnostics.

Clustered regularly interspaced short palindromic repeat (CRISPR/Cas) proteins can be designed to bind specified DNA and RNA sequences and hold great promise for the accurate detection of nucleic acids for diagnostics. We integrated commercially available reagents into a CRISPR/Cas9-based lateral flow assay that can detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequences with single-base specificity. This approach requires minimal equipment and represents a simplified platform for field-based deployment. We also developed a rapid, multiplex fluorescence CRISPR/Cas9 nuclease cleavage assay capable of detecting and differentiating SARS-CoV-2, influenza A and B, and respiratory syncytial virus in a single reaction. Our findings provide proof-of-principle for CRISPR/Cas9 point-of-care diagnosis as well as a scalable fluorescent platform for identifying respiratory viral pathogens with overlapping symptomology.