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Abstract Background Mesenchymal stromal cells (MSCs) can be used intra-articularly to quell inflammation and promote cartilage healing; however, mechanisms by which MSCs mitigate joint disease remain poorly understood. Galectins, a family of β-galactoside binding proteins, regulate inflammation, adhesion and cell migration in diverse cell types. Galectin-1 and galectin-3 are proposed to be important intra-articular modulators of inflammation in both osteoarthritis and rheumatoid arthritis. Here, we asked whether equine bone marrow-derived MSCs (BMSCs) express higher levels of galectin-1 and -3 relative to synovial fibroblasts and chondrocytes and if an inflammatory environment affects BMSC galectin expression and motility. Methods Equine galectin-1 and -3 gene expression was quantified using qRT-PCR in cultured BMSCs, synoviocytes and articular chondrocytes, in addition to synovial membrane and articular cartilage tissues. Galectin gene expression, protein expression, and protein secretion were measured in equine BMSCs following exposure to inflammatory cytokines (IL-1β 5 and 10 ng/mL, TNF-α 25 and 50 ng/mL, or LPS 0.1, 1, 10 and 50 μg/mL). BMSC focal adhesion formation was assessed using confocal microscopy, and BMSC motility was quantified in the presence of inflammatory cytokines (IL-1β or TNF-α) and the pan-galectin inhibitor β-lactose (100 and 200 mM). Results Equine BMSCs expressed 3-fold higher galectin-1 mRNA levels as compared to cultured synovial fibroblasts (p = 0.0005) and 30-fold higher galectin-1 (p

Galectins are potent regulators of cell adhesion, growth and apoptosis in diverse cell types, including chondrocytes and synovial fibroblasts. Elevations in synovial fluid galectin-3 have been observed in rheumatoid arthritis, juvenile idiopathic arthritis and experimental inflammatory arthritis in animal models, whereas galectin-1 is thought to be protective. Less is known about galectins-1 and-3 in osteoarthritis (OA). Therefore, the purpose of this study was: (1) to determine whether galectin-1 and-3 synovial fluid concentrations and synovial membrane and cartilage histochemical staining were altered following osteochondral injury in an experimental equine osteoarthritis (OA) model and (2) to measure galectin-1 and-3 mRNA expression and synovial fluid concentrations in naturally occurring equine carpal OA. Synovial fluid galectin-1 and-3 concentrations were quantified using custom ELISAs in two research horse cohorts undergoing experimental OA induction (n = 5 and 4) and in a cohort of horses with naturally occurring carpal OA (n = 57). Galectin mRNA expression in synovial membrane and cartilage tissue obtained from carpal joints of horses with naturally occurring OA was measured using RT-qPCR, and galectin immunostaining was assessed in synovial membrane and osteochondral tissues in the experimental model (n = 5). Synovial fluid galectin-1 and-3 concentrations increased following experimental carpal osteochondral fragmentation. Cartilage galectin-1 mRNA expression increased with OA severity in naturally occurring disease. The superficial zone of healthy articular cartilage stained intensely for galectin-3 in sham-operated joints, whereas galectin-1 staining was nearly absent. Chondrocyte galectin-1 and-3 immunoreactivity increased following cartilage injury, particularly in galectin-1 positive chondrones. Galectins-1 and-3 are present in healthy equine synovial fluid and increase following post-traumatic OA. Healthy superficial zone chondrocytes express galectin-3, whereas galectin-1 chondrocyte staining is limited predominantly to chondrones and injured cartilage. Further work is needed to clarify the functions of galectins-1 and-3 in healthy and OA joints.

Copper is an essential micronutrient and the ability to scavenge tightly bound or trace levels of copper ions at the host–pathogen interface is vital for fungal proliferation in animal hosts. Recent studies suggest that trace metal ion acquisition is critical for the establishment and propagation of Pseudogymnoascus destructans, the fungal pathogen responsible for white-nose syndrome (WNS), on their bat host. However, little is known about these metal acquisition pathways in P. destructans. In this study, we report the characterization of the P. destructans high-affinity copper transporter VC83_00191 (PdCTR1a), which is implicated as a virulence factor associated with the WNS disease state. Using Saccharomyces cerevisiae as a recombinant expression host, we find that PdCTR1a can efficiently traffic Cu ions into the yeast cytoplasm. Complementary studies in the native P. destructans fungus provide evidence that PdCTR1a transcripts and protein levels are dictated by Cu-bioavailability in the growth media. Our study demonstrates that PdCTR1a is a functional high-affinity copper transporter and is relevant to Cu homeostasis pathways in P. destructans.

Abstract Identification of long‐term calcium channel blocker (CCB) responders with acute vasodilator challenge is critical in the evaluation of patients with pulmonary arterial hypertension. Currently there is no standardized approach for use of supplemental oxygen during acute vasodilator challenge. In this retrospective analysis of patients identified as acute vasoresponders, treated with CCBs, all patients had hemodynamic measurements in three steps: (1) at baseline; (2) with 100% fractional inspired oxygen; and (3) with 100% fractional inspired oxygen plus inhaled nitric oxide (iNO). Those meeting the definition of acute vasoresponsiveness only after first normalizing for the effects of oxygen in step 2 were labeled “iNO Responders.” Those who met the definition of acute vasoresponsiveness from a combination of the effects of 100% FiO2 and iNO were labeled “oxygen responders.” Survival, hospitalization for decompensated right heart failure, duration of CCB monotherapy, and functional data were collected. iNO responders, when compared to oxygen responders, had superior survival (100% vs. 50.1% 5‐year survival, respectively), fewer hospitalizations for acute decompensated right heart failure (0% vs. 30.4% at 1 year, respectively), longer duration of CCB monotherapy (80% vs. 52% at 1 year, respectively), and superior 6‐min walk distance. Current guidelines for acute vasodilator testing do not standardize oxygen coadministration with iNO. This study demonstrates that adjusting for the effects of supplemental oxygen before assessing for acute vasoresponsiveness identifies a cohort with superior functional status, tolerance of CCB monotherapy, and survival while on long‐term CCB therapy.

Abstract Molecular markers of autoimmunity, such as antibodies to citrullinated protein antigens (ACPA), are detectable prior to inflammatory arthritis (IA) in rheumatoid arthritis (RA) and may define a state that is ‘at-risk’ for future RA. Here we present a cross-sectional comparative analysis among three groups that include ACPA positive individuals without IA (At-Risk), ACPA negative individuals and individuals with early, ACPA positive clinical RA (Early RA). Differential methylation analysis among the groups identifies non-specific dysregulation in peripheral B, memory and naïve T cells in At-Risk participants, with more specific immunological pathway abnormalities in Early RA. Tetramer studies show increased abundance of T cells recognizing citrullinated (cit) epitopes in At-Risk participants, including expansion of T cells reactive to citrullinated cartilage intermediate layer protein I (cit-CILP); these T cells have Th1, Th17, and T stem cell memory-like phenotypes. Antibody-antigen array analyses show that antibodies targeting cit-clusterin, cit-fibrinogen and cit-histone H4 are elevated in At-Risk and Early RA participants, with the highest levels of antibodies detected in those with Early RA. These findings indicate that an ACPA positive at-risk state is associated with multifaceted immune dysregulation that may represent a potential opportunity for targeted intervention.

We developed a novel machine learning (ML) algorithm with the goal of producing transparent models (i.e., understandable by humans) while also flexibly accounting for nonlinearity and interactions. Our method is based on ranked sparsity, and it allows for flexibility and user control in varying the shade of the opacity of black box machine learning methods. The main tenet of ranked sparsity is that an algorithm should be more skeptical of higher-order polynomials and interactions a priori compared to main effects, and hence, the inclusion of these more complex terms should require a higher level of evidence. In this work, we put our new ranked sparsity algorithm (as implemented in the open source R package, sparseR) to the test in a predictive model “bakeoff” (i.e., a benchmarking study of ML algorithms applied “out of the box”, that is, with no special tuning). Algorithms were trained on a large set of simulated and real-world data sets from the Penn Machine Learning Benchmarks database, addressing both regression and binary classification problems. We evaluated the extent to which our human-centered algorithm can attain predictive accuracy that rivals popular black box approaches such as neural networks, random forests, and support vector machines, while also producing more interpretable models. Using out-of-bag error as a meta-outcome, we describe the properties of data sets in which human-centered approaches can perform as well as or better than black box approaches. We found that interpretable approaches predicted optimally or within 5% of the optimal method in most real-world data sets. We provide a more in-depth comparison of the performances of random forests to interpretable methods for several case studies, including exemplars in which algorithms performed similarly, and several cases when interpretable methods underperformed. This work provides a strong rationale for including human-centered transparent algorithms such as ours in predictive modeling applications.

Abstract Background Temporary, voluntary storage of firearms away from the home is a recommended option for individuals with risk of suicide, but it may also be used in other situations (e.g., long trips). Prior work has explored the availability of storage options and the views of storage locations. Little is known about out-of-home storage practices among those who live in homes with firearms (including owners). Methods We surveyed English-speaking adults (18 or older) in two states (Colorado and Washington) living in a home with a firearm (June–July 2021). Results Among the final sample of 1029, most respondents were white (88.1%) and non-Hispanic (85.0%); half were female (50.8%), and the most common age group was ages 35–44 (25.5%). Just over one quarter (27.3%) of respondents indicated they had stored a firearm away from their home/car/garage in the last 5 years. The place most respondents said they were somewhat or very likely to consider was at a family members home (62.7%) or at a self-storage facility (52.5%). Conclusion Out-of-home firearm storage is a relatively common practice and endorsed by many gun-owners, suggesting out-of-home storage is feasible for firearm owners as an approach to suicide prevention.

Abstract Background Temporary, voluntary storage of firearms away from the home during times of risk is a recommended strategy for suicide prevention. Law enforcement agencies (LEAs) are often suggested as storage sites, and online maps in Colorado and Washington display LEAs willing to consider storage. Questions remain about the experiences and views of LEAs, including barriers to providing storage. Methods LEAs in Colorado and Washington were invited to complete a survey via mail or online from June to July 2021; invitations were sent by email and mail, with telephone calls to non-responders. Survey data were analyzed using descriptive statistics, with testing between states and other subgroups using Fisher’s exact tests. Results Overall, 168 LEAs in Colorado (n = 91) or Washington (n = 77) participated (40% participation rate). Of those, 53% provided temporary, voluntary storage upon request by community members at the time of the survey. More LEAs said they had ever provided storage when the requester was under a court order (74% overall). Over half (60%) of responding LEAs had received at least one storage request in the prior 12 months. Many (41%) said they had declined to return a firearm after temporary storage due to safety concerns. Most LEAs supported engagement in suicide prevention (89%) and provision of community services (77%), but they simultaneously preferred being a storage option of last resort (73%). Factors negatively influencing storage provision included liability and funding concerns. Conclusions In Colorado and Washington, half of LEAs currently offer temporary, voluntary firearm storage upon request. While LEAs support suicide prevention and community engagement, broader provision of storage and participation in online maps may be limited by logistic, liability, and financial concerns. Addressing these barriers may facilitate broader suicide prevention efforts.

Abstract Background Missing data prove troublesome in data analysis; at best they reduce a study’s statistical power and at worst they induce bias in parameter estimates. Multiple imputation via chained equations is a popular technique for dealing with missing data. However, techniques for combining and pooling results from fitted generalized additive models (GAMs) after multiple imputation have not been well explored. Methods We simulated missing data under MCAR, MAR, and MNAR frameworks and utilized random forest and predictive mean matching imputation to investigate a variety of rules for combining GAMs after multiple imputation with binary and normally distributed outcomes. We compared multiple pooling procedures including the “D2” method, the Cauchy combination test, and the median p-value (MPV) rule. The MPV rule involves simply computing and reporting the median p-value across all imputations. Other ad hoc methods such as a mean p-value rule and a single imputation method are investigated. The viability of these methods in pooling results from B-splines is also examined for normal outcomes. An application of these various pooling techniques is then performed on two case studies, one which examines the effect of elevation on a six-minute walk distance (a normal outcome) for patients with pulmonary arterial hypertension, and the other which examines risk factors for intubation in hospitalized COVID-19 patients (a dichotomous outcome). Results In comparison to the results from generalized additive models fit on full datasets, the median p-value rule performs as well as if not better than the other methods examined. In situations where the alternative hypothesis is true, the Cauchy combination test appears overpowered and alternative methods appear underpowered, while the median p-value rule yields results similar to those from analyses of complete data. Conclusions For pooling results after fitting GAMs to multiply imputed datasets, the median p-value is a simple yet useful approach which balances both power to detect important associations and control of Type I errors.

BackgroundFirearm safety among individuals with Alzheimer disease and related dementias (ADRD) is an underdiscussed and underresearched concern in the United States, especially given the growing population of community-dwelling adults with ADRD. The “Safety in Dementia” (SiD) web-based decision aid was developed to support caregivers in addressing firearm access; the efficacy of SiD is unknown. ObjectiveThrough the SiD decision aid, the Safe at Home (S@H) study aims to support caregivers in making decisions about home safety that align with their goals and values, and behaviors regarding firearm access for persons with ADRD and firearm access. MethodsThe S@H study is a 2-armed randomized controlled trial to test the effect of the SiD decision aid on caregivers of community-dwelling adults with ADRD who have firearm access. S@H aims to recruit 500 ADRD caregivers (age ≥18 years, fluent in English or Spanish, and in the United States) through online or social media advertisements and through relevant organizations. Participants are randomized to view SiD or a control website at their own pace; all participants complete web-based questionnaires at baseline, 2 weeks, 2 months, and 6 months. The primary outcome is immediate preparation for decision-making; secondary outcomes include longitudinal decision outcomes and self-reported modifications to firearm access. The relative reach and effectiveness of each recruitment method (online/social media and through relevant organizations) will be assessed by examining differences in caregiver participation, retention rates, and relative cost. ResultsThe study enrollment began in May 2022. As of December 2022, a total of 117 participants had enrolled. ConclusionsThe S@H study is the first randomized trial of a firearm safety decision aid for ADRD caregivers. The results from this study will inform how best to support caregivers in decision-making regarding firearm safety. Further, results may guide approaches for recruiting caregivers and for dissemination of resources. Trial RegistrationClinicalTrials.gov NCT05173922; https://clinicaltrials.gov/ct2/show/NCT05173922 International Registered Report Identifier (IRRID)DERR1-10.2196/43702

Purpose – Building on tourism crisis studies and behavioral economics, this study describes a national survey conducted among 439 Aruban tourism and nontourism employees. Design/methodology/approach – Regression analysis was subsequently conducted to analyze the relationship between experienced well-being, crisis duration and tourism and nontourism employee sentiments. Findings – The findings indicate that tourism employee sentiments are generally, and significantly, more negative and their concerns about the future are significantly more pessimistic than nontourism employees. The results show that the experienced well-being and expected duration of the COVID-19 crisis have a significant negative effect on tourism employees' sentiments. The paper provides several policies and industry recommendations for strengthening tourism employee well-being and economic resilience. Several avenues for future research are presented. Originality/value – The current study contributes to this literature by showing that the increased pessimism and negativity of the tourism employees as compared to nontourism employees during the current pandemic influence their thoughts about future income and earnings as well as future purchases.

OBJECTIVES:. Colorado issued a month long statewide lockdown on March 26, 2020, during the initial surge of the COVID-19 pandemic. The impact of this mandate on non–COVID-19 ICU admission rates and outcomes is unclear. DESIGN:. We performed a retrospective analysis of all medical ICU admissions in the University of Colorado Health System in four predefined periods: 1) prepandemic (2 mo prior to lockdown period 1); 2) mandated lockdown from March 26 to April 26, 2020 (period 2); 3) between surges (period 3); and 4) nonmandated lockdown surge (between November 1, 2020, and March 31, 2021, period 4). SETTING:. Nonsurgical ICU admissions at the University of Colorado Health Systems, including 10 hospitals throughout Colorado. SUBJECTS:. All ICU admissions in four predefined time periods. MEASUREMENTS AND MAIN RESULTS:. We included 13,787 patients who were admitted during the four study periods. The 28-day mortality rates for non–COVID-19 ICU admissions following index ICU admission were 13.6%, 18.0%, 13.5%, and 16.0% across periods 1–4, respectively. However, the increased odds in non–COVID-19 ICU mortality during the mandated lockdown period relative to prepandemic 1 (odds ratio [OR], 1.39; 95% CI, 1.11–1.72; p = 0.0.04) was attenuated and nonsignificant after adjustment for demographics, comorbidities, diagnosis flags, and severity (OR, 1.15; 95% CI, 0.89–1.48; p = 0.27). Similar results were found in time-to-event analyses. The most common diagnosis in each time period was acute respiratory failure (ARF), and we found it to have increased during lockdown (p < 0.001), whereas sepsis admissions increased during and decreased after lockdown (p = 0.004). Admissions for alcohol withdrawal syndrome (AWS) increased during lockdown and 6 months afterwards (p = 0.005). CONCLUSIONS:. For non-COVID-19–related ICU admissions, mortality rate was similar before, during, and after Colorado’s month long lockdown after confounder adjustment, including typical ICU admission flags. Primary admission diagnoses shifted throughout the predefined study periods with more admissions for severe critical diagnoses (i.e., ARF, sepsis, AWS) occurring during the mandated lockdown and nonmandated lockdown periods compared with the prepandemic and between surge period. This would suggest that the perceived increase in mortality during the lockdown for non–COVID-19 ICU admissions may be related to a shift inpatient demographics.

Objective To evaluate the prevalence of elevations of anti‐cyclic citrullinated peptide‐3 (anti‐CCP3) antibody, rheumatoid factor IgM (RF‐IgM) and serum calprotectin (sCP) in pre–rheumatoid arthritis (RA) as well as the diagnostic accuracies of these biomarkers for the timing of diagnosis of future RA. Methods A total of 215 RA cases, each with approximately three pre‐RA diagnoses and one post–RA diagnosis serum sample, and controls were identified from the Department of Defense Serum Repository. All case samples and a single sample from each control subject were tested for anti‐CCP3 (IgG), RF‐IgM, and sCP. The diagnostic accuracies of biomarkers for future RA were evaluated. Results Anti‐CCP3, RF‐IgM, and sCP were elevated in pre‐RA, with anti‐CCP3 and sCP significantly elevated compared with RF‐IgM at the earliest time points. Within the cases, the combination of anti‐CCP3 and RF‐IgM positivity had a positive predictive value (PPV) of 35.6% for a diagnosis of RA in 3 years or less, which is significantly higher than the PPV of 18.7% for anti‐CCP3 positivity alone (P < 0.001). A combination of anti‐CCP3, RF‐IgM, and sCP had the highest PPV (53.0%) for a diagnosis of RA in 3 years or less; however, this was not significantly higher than the PPV for anti‐CCP3 and RF‐IgM positivity (P = 0.248). Conclusion Anti‐CCP3, RF‐IgM, and sCP are elevated in pre‐RA; furthermore, combinations of elevations of these biomarkers are more commonly seen in the period of less than or equal to 3 years to diagnosis. This may be considered in creating inclusion criteria in prevention trials in RA. In addition, the biologic relationships of these biomarkers in pre‐RA need exploration.

Background/PurposeIn rheumatoid arthritis (RA) autoantibodies including antibodies to citrullinated protein antigens (ACPA) and rheumatoid factor (RF) can be predictive of incident clinical RA. However, there is limited understanding of how antibody changes over time impact prediction of the likelihood and timing of future clinical RA.Materials and MethodsWe evaluated relationships between ACPA, the shared epitope (SE), RF isotypes and incident RA in a prospective cohort of 90 ACPA(+) individuals without baseline arthritis identified through health-fair testing (i.e. Healthfair). We also evaluated ACPA and RF isotypes and time-to-diagnosis of RA in a retrospective cohort of 215 individuals with RA from the Department of Defense Serum Repository (DoDSR).ResultsTwenty-six of 90 (29%) of ACPA(+) Healthfair participants developed incident RA. Baseline or incident dual RF-IgA and RF-IgM positivity was associated with increased risk for incident RA (HR 3.09; 95% CI 1.15 to 8.29) although RFs were negative in ~50% of individuals with incident RA. SE was associated with increased risk of RA (HR 2.87, 95% CI 1.22-6.76). In the DoDSR cohort, triple positivity for ACPA, RF-IgA and RF-IgM was present a median of 1-2 years prior to RA diagnosis, with some sex-specific differences.ConclusionThese findings can be used to counsel individuals at-risk for future RA and to design clinical trials for RA prevention. The findings also suggest that RF could be a surrogate outcome as a success of an immunologic intervention in RA prevention. Additional studies are needed to understand the biologic of different patterns of autoantibody elevations in RA evolution.

Background and purpose: Effective communication of radiologic findings is crucial in the acute evaluation of stroke. Conventional methods (CM) of communication can present challenges leading to delays in reporting, ultimately impacting patient outcomes. The goal of this retrospective study was to assess the utility of a HIPAA-compliant messaging system (MS) for expediting communication of CT findings by radiologists to members of the stroke team. Materials and methods: Multimodal stroke protocol CT examinations performed at a high-volume stroke center between January and December 2019 were identified as part of this IRB-approved retrospective study. Read-back times were determined based on templated documentation within individual radiologic reports. The time interval between identification of findings and notification was calculated for each report, and usage of CM versus MS was documented. In addition, an online voluntary survey was disseminated to staff to assess user satisfaction with the MS. Results and conclusions: In total, 442 CTs were included, and 126 (29%) documented use of the MS. The MS was associated with significant reductions in median read-back times, both for all reported findings (1.5 versus 16 minutes, p

Acute respiratory distress syndrome (ARDS) is a common cause of respiratory failure yet has few pharmacologic therapies, reflecting the mechanistic heterogeneity of lung injury. We hypothesized that damage to the alveolar epithelial glycocalyx, a layer of glycosaminoglycans interposed between the epithelium and surfactant, contributes to lung injury in patients with ARDS. Using mass spectrometry of airspace fluid noninvasively collected from mechanically ventilated patients, we found that airspace glycosaminoglycan shedding (an index of glycocalyx degradation) occurred predominantly in patients with direct lung injury and was associated with duration of mechanical ventilation. Male patients had increased shedding, which correlated with airspace concentrations of matrix metalloproteinases. Selective epithelial glycocalyx degradation in mice was sufficient to induce surfactant dysfunction, a key characteristic of ARDS, leading to microatelectasis and decreased lung compliance. Rapid colorimetric quantification of airspace glycosaminoglycans was feasible and could provide point-of-care prognostic information to clinicians and/or be used for predictive enrichment in clinical trials.