The nucleotide-binding domain of NRC-dependent disease resistance proteins is sufficient to activate downstream helper NLR oligomerization and immune signaling.

Nucleotide-binding domain and leucine-rich repeat (NLR) proteins with pathogen sensor activities have evolved to initiate immune signaling by activating helper NLRs. However, the mechanisms underpinning helper NLR activation by sensor NLRs remain poorly understood. Although coiled coil (CC) type sensor NLRs such as the Potato virus X disease resistance protein Rx have been shown to activate the oligomerization of their downstream helpers NRC2, NRC3 and NRC4, the domains involved in sensor-helper signaling are not known. Here, we used Agrobacterium tumefaciens-mediated transient expression in Nicotiana benthamiana to show that the nucleotide-binding (NB) domain within the NB-ARC of Rx is necessary and sufficient for oligomerization and immune signaling of downstream helper NLRs. In addition, the NB domains of the disease resistance proteins Gpa2 (cyst nematode resistance), Rpi-amr1, Rpi-amr3 (oomycete resistance) and Sw-5b (virus resistance) are also sufficient to activate their respective downstream NRC helpers. Using transient expression in the lettuce (Lactuca sativa), we show that Rx (both as full length or as NB domain truncation) and its helper NRC2 form a minimal functional unit that can be transferred from solanaceous plants (lamiids) to Campanulid species. Our results challenge the prevailing paradigm that NLR proteins exclusively signal via their N-terminal domains and reveal a signaling activity for the NB domain of NRC-dependent sensor NLRs. We propose a model in which helper NLRs can perceive the status of the NB domain of their upstream sensors.

Contrasting Objective and Perceived Risk: Predicting COVID-19 Health Behaviors in a Nationally Representative U.S. Sample.

BACKGROUND: Individuals confronting health threats may display an optimistic bias such that judgments of their risk for illness or death are unrealistically positive given their objective circumstances. PURPOSE: We explored optimistic bias for health risks using k-means clustering in the context of COVID-19. We identified risk profiles using subjective and objective indicators of severity and susceptibility risk for COVID-19. METHODS: Between 3/18/2020-4/18/2020, a national probability sample of 6,514 U.S. residents reported both their subjective risk perceptions (e.g., perceived likelihood of illness or death) and objective risk indices (e.g., age, weight, pre-existing conditions) of COVID-19-related susceptibility and severity, alongside other pandemic-related experiences. Six months later, a subsample (N = 5,661) completed a follow-up survey with questions about their frequency of engagement in recommended health protective behaviors (social distancing, mask wearing, risk behaviors, vaccination intentions). RESULTS: The k-means clustering procedure identified five risk profiles in the Wave 1 sample; two of these demonstrated aspects of optimistic bias, representing almost 44% of the sample. In OLS regression models predicting health protective behavior adoption at Wave 2, clusters representing individuals with high perceived severity risk were most likely to report engagement in social distancing, but many individuals who were objectively at high risk for illness and death did not report engaging in self-protective behaviors. CONCLUSIONS: Objective risk of disease severity only inconsistently predicted health protective behavior. Risk profiles may help identify groups that need more targeted interventions to increase their support for public health policy and health enhancing recommendations more broadly.

As we move into an endemic stage of the COVID-19 pandemic, understanding engagement in health behaviors to curb the spread of disease remains critically important to manage COVID-19 and other health threats. However, peoples' perceptions about their risk of getting sick and having severe outcomes if they do fall ill are subject to bias. We studied a nationally representative probability sample of over 6,500 U.S. residents who completed surveys immediately after the COVID-19 pandemic began and approximately 6 months later. We used a computer processing (i.e., machine learning) approach to categorize participants based on both their actual risk factors for COVID-19 and their subjective understanding of that risk. Our analysis identified groups of individuals whose subjective perceptions of risk did not align with their actual risk characteristics. Specifically, almost 44% of our sample demonstrated an optimistic bias: they did not report higher risk of death from COVID-19 despite having one or more well-known risk factors for poor disease outcomes (e.g., older age, obesity). Six months later, membership in these risk groups prospectively predicted engagement in health protective and risky behaviors, as well as vaccine intentions, demonstrating how early risk perceptions may influence health behaviors over time.

Effect of stretching on inflammation in a subcutaneous carrageenan mouse model analyzed at single-cell resolution.

Understanding the factors that influence the biological response to inflammation is crucial, due to its involvement in physiological and pathological processes, including tissue repair/healing, cancer, infections, and autoimmune diseases. We have previously demonstrated that in vivo stretching can reduce inflammation and increase local pro-resolving lipid mediators in rats, suggesting a direct mechanical effect on inflammation resolution. Here we aimed to explore further the effects of stretching at the cellular/molecular level in a mouse subcutaneous carrageenan-inflammation model. Stretching for 10 min twice a day reduced inflammation, increased the production of pro-resolving mediator pathway intermediate 17-HDHA at 48 h postcarrageenan injection, and decreased both pro-resolving and pro-inflammatory mediators (e.g., PGE2 and PGD2 ) at 96 h. Single-cell RNA sequencing analysis of inflammatory lesions at 96 h showed that stretching increased the expression of both pro-inflammatory (Nos2) and pro-resolution (Arg1) genes in M1 and M2 macrophages at 96 h. An intercellular communication analysis predicted specific ligand-receptor interactions orchestrated by neutrophils and M2a macrophages, suggesting a continuous neutrophil presence recruiting immune cells such as activated macrophages to contain the antigen while promoting resolution and preserving tissue homeostasis.

Protein Adsorption on Mixed Self-Assembled Monolayers: Influence of Chain Length and Terminal Group.

Mixed self-assembled monolayers (SAMs) are often used as highly tunable substrates for biomedical and biosensing applications. It is well documented, however, that mixed SAMs can be highly disordered at the molecular level and do not pack as closely or homogeneously as single-component SAMs, particularly when the chain lengths and head groups of the SAM thiol components are significantly different. In this study, we explore the impact of SAM structure and mixing ratio (-OH and -CH3 termini) on the weak physisorption behavior of bovine serum albumin (BSA), which adsorbs more readily to hydrophobic, methyl-terminated SAMs. Our results suggest that once the mixture includes 50% or more of the methyl terminus, the mixing ratio alone is a relatively good predictor of adsorption, regardless of the relative chain lengths of the thiols used in the mixture. This trend persists at any mixing ratio for SAMs where methyl- and hydroxyl-terminated groups are the same length or where the hydroxyl-terminated thiol is longer. The only variance observed is at low mixing ratios (<50% methyl-terminated) for a mixed SAM where the methyl-terminated component has a longer chain length. Relative protein adsorption increases on these mixtures, perhaps due to the disordered exposure of the excess alkane backbone. Taken together, however, we do not find significant evidence that varying chain lengths for mixed SAMs prepared on polycrystalline substrates and analyzed in air have an outsized influence on nanoscopic adsorption behavior, despite molecular-level disorder in the SAM itself.

Cryo-EM structure of SKP1-SKP2-CKS1 in complex with CDK2-cyclin A-p27KIP1.

p27KIP1 (cyclin-dependent kinase inhibitor 1B, p27) is a member of the CIP/KIP family of CDK (cyclin dependent kinase) regulators that inhibit cell cycle CDKs. p27 phosphorylation by CDK1/2, signals its recruitment to the SCFSKP2 (S-phase kinase associated protein 1 (SKP1)-cullin-SKP2) E3 ubiquitin ligase complex for proteasomal degradation. The nature of p27 binding to SKP2 and CKS1 was revealed by the SKP1-SKP2-CKS1-p27 phosphopeptide crystal structure. Subsequently, a model for the hexameric CDK2-cyclin A-CKS1-p27-SKP1-SKP2 complex was proposed by overlaying an independently determined CDK2-cyclin A-p27 structure. Here we describe the experimentally determined structure of the isolated CDK2-cyclin A-CKS1-p27-SKP1-SKP2 complex at 3.4 Å global resolution using cryogenic electron microscopy. This structure supports previous analysis in which p27 was found to be structurally dynamic, transitioning from disordered to nascent secondary structure on target binding. We employed 3D variability analysis to further explore the conformational space of the hexameric complex and uncovered a previously unidentified hinge motion centred on CKS1. This flexibility gives rise to open and closed conformations of the hexameric complex that we propose may contribute to p27 regulation by facilitating recognition with SCFSKP2. This 3D variability analysis further informed particle subtraction and local refinement approaches to enhance the local resolution of the complex.

Genome-wide association study of chronic sputum production implicates loci involved in mucus production and infection.

BACKGROUND: Chronic sputum production impacts on quality of life and is a feature of many respiratory diseases. Identification of the genetic variants associated with chronic sputum production in a disease agnostic sample could improve understanding of its causes and identify new molecular targets for treatment. METHODS: We conducted a genome-wide association study (GWAS) of chronic sputum production in UK Biobank. Signals meeting genome-wide significance (p<5×10-8) were investigated in additional independent studies, were fine-mapped and putative causal genes identified by gene expression analysis. GWASs of respiratory traits were interrogated to identify whether the signals were driven by existing respiratory disease among the cases and variants were further investigated for wider pleiotropic effects using phenome-wide association studies (PheWASs). RESULTS: From a GWAS of 9714 cases and 48 471 controls, we identified six novel genome-wide significant signals for chronic sputum production including signals in the human leukocyte antigen (HLA) locus, chromosome 11 mucin locus (containing MUC2, MUC5AC and MUC5B) and FUT2 locus. The four common variant associations were supported by independent studies with a combined sample size of up to 2203 cases and 17 627 controls. The mucin locus signal had previously been reported for association with moderate-to-severe asthma. The HLA signal was fine-mapped to an amino acid change of threonine to arginine (frequency 36.8%) in HLA-DRB1 (HLA-DRB1*03:147). The signal near FUT2 was associated with expression of several genes including FUT2, for which the direction of effect was tissue dependent. Our PheWAS identified a wide range of associations including blood cell traits, liver biomarkers, infections, gastrointestinal and thyroid-associated diseases, and respiratory disease. CONCLUSIONS: Novel signals at the FUT2 and mucin loci suggest that mucin fucosylation may be a driver of chronic sputum production even in the absence of diagnosed respiratory disease and provide genetic support for this pathway as a target for therapeutic intervention.

Pandemic-Induced Telework Challenges and Strategies.

Prior to the COVID-19 pandemic, telework was an established discretionary practice with a considerable amount of research. However, the COVID-19 pandemic forced people who had never worked from home before to do so. Our two-wave descriptive investigation provides a historical snapshot of what approximately 400 teleworkers experienced in the first two to three months of the pandemic. We explored how this experience differed for those who had previously teleworked, those who had children in their home, and those who had supervisory responsibilities. The data exposed telework challenges and pandemic-specific challenges. The results support job crafting theories that teleworkers proactively implement strategies to adjust their boundaries and relationships to meet their need (Biron et al., Personnel Review, 2022). The data also revealed that employees were still struggling two months later, despite implementing strategies like self-care, taking breaks, and psychological reframing. This research provides detailed evidence of how pandemic-induced telework is not the same as traditional telework and some initial evidence of the pandemic-induced telework adjustment time period. Supplementary Information: The online version contains supplementary material available at 10.1007/s41542-023-00151-1.

Radiation induced changes in profibrotic markers in the submental muscles and their correlation with tongue movement.

Swallowing impairment is a major complication of radiation treatment for oropharyngeal cancers. Developing targeted therapies that improve swallowing outcomes relies on an understanding of the mechanisms that influence motor function after radiation treatment. The purpose of this study was to determine whether there is a correlation between radiation induced changes in tongue movement and structural changes in irradiated submental muscles, as well as assess other possible causes for dysfunction. We hypothesized that a clinically relevant total radiation dose to the submental muscles would result in: a) quantifiable changes in tongue strength and displacement during drinking two months post treatment; and b) a profibrotic response and/or fiber type transition in the irradiated tissue. Sprague-Dawley adult male rats received radiation to the submental muscles at total dose-volumes known to provoke dysphagia in humans. A clinical linear accelerator administered 8 fractions of 8Gy for a total of 64Gy. Comparisons were made to sham-treated rats that received anesthesia only. Swallowing function was assessed using videofluoroscopy and tongue strength was analyzed via force lickometer. TGFß1 expression was analyzed via ELISA. The amount of total collagen was analyzed by picrosirius red staining. Immunofluorescence was used to assess fiber type composition and size. Significant changes in licking function during drinking were observed at two months post treatment, including a slower lick rate and reduced tongue protrusion during licking. In the mylohyoid muscle, significant increases in TGFß1 protein expression were found post radiation. Significant increases in the percentage of collagen content were observed in the irradiated geniohyoid muscle. No changes in fiber type expression were observed. Results indicate a profibrotic transition within the irradiated swallowing muscles that contributes to tongue dysfunction post-radiation treatment.

Idiosyncratic media exposures during a pandemic and their link to well-being, cognition, and behavior over time.

Over the past two decades of research, increased media consumption in the context of collective traumas has been cross-sectionally and longitudinally linked to negative psychological outcomes. However, little is known about the specific information channels that may drive these patterns of response. The current longitudinal investigation uses a probability-based sample of 5,661 Americans measured at the onset of the COVID-19 pandemic to identify a) distinct patterns of information-channel use (i.e., dimensions) for COVID-related information, b) demographic correlates of these patterns, and c) prospective associations of these information channel dimensions with distress (i.e., worry, global distress, and emotional exhaustion), cognition (e.g., beliefs about the seriousness of COVID-19, response efficacy, and dismissive attitudes), and behavior (e.g., engaging in health-protective behaviors and risk-taking behaviors) 6 mo later. Four distinct information-channel dimensions emerged: journalistic complexity; ideologically focused news; domestically focused news; and nonnews. Results indicate that journalistic complexity was prospectively associated with more emotional exhaustion, belief in the seriousness of the coronavirus, response efficacy, engaging in health-protective behaviors, and less dismissiveness of the pandemic. A reliance on conservative-leaning media was prospectively associated with less psychological distress, taking the pandemic less seriously, and engaging in more risk-taking behaviors. We discuss the implications of this work for the public, policy makers, and future research.

Prox2 and Runx3 vagal sensory neurons regulate esophageal motility.

Vagal sensory neurons monitor mechanical and chemical stimuli in the gastrointestinal tract. Major efforts are underway to assign physiological functions to the many distinct subtypes of vagal sensory neurons. Here, we use genetically guided anatomical tracing, optogenetics, and electrophysiology to identify and characterize vagal sensory neuron subtypes expressing Prox2 and Runx3 in mice. We show that three of these neuronal subtypes innervate the esophagus and stomach in regionalized patterns, where they form intraganglionic laminar endings. Electrophysiological analysis revealed that they are low-threshold mechanoreceptors but possess different adaptation properties. Lastly, genetic ablation of Prox2 and Runx3 neurons demonstrated their essential roles for esophageal peristalsis in freely behaving mice. Our work defines the identity and function of the vagal neurons that provide mechanosensory feedback from the esophagus to the brain and could lead to better understanding and treatment of esophageal motility disorders.

Plantar Grasp sign as a screening tool for Orthostatic Tremor (OT).

Introduction: Orthostatic tremor (OT) is a rare neurological disorder characterized by a sensation of instability while standing. Very few clinical signs have been described for OT to date. Finding other symptoms and signs could prove valuable for this hard-to-recognized disease. Methods: This protocol is part of the University of Nebraska Medical Center Orthostatic Tremor longitudinal study. It was noted that OT patients flex their toes and sometimes the foot arch while standing (Plantar Grasp). They reported doing this to "grab" the floor and improve stability. This paper analyses the diagnostic test characteristics of the patient-self-reported Plantar Grasp, a new sign in OT. Results: There were 34 OT patients (88% females), and 20 controls (65% females). Eighty-eight percent of patients with OT reported the plantar grasp sign and none of the controls. The Plantar Grasp Sign was found to be very sensitive (88%), and extremely specific (100%) in our cohort. Non-weighted Negative Likelihood Ratio (NLR) was 0.12. And the 3% prevalence-weighted NLR was so low that the negative post-test probability was close to zero. Conclusion: Due to its high sensitivity, specificity, and ideal likelihood ratio, we propose that the Plantar Grasp sign could be considered to screen patients with possible OT. Further studies are needed to determine the specificity of this sign in OT versus other balance disorders.

High Resolution Membrane Structures within Hybrid Lipid-Polymer Vesicles Revealed by Combining X-Ray Scattering and Electron Microscopy.

Hybrid vesicles consisting of phospholipids and block-copolymers are increasingly finding applications in science and technology. Herein, small angle X-ray scattering (SAXS) and cryo-electron tomography (cryo-ET) are used to obtain detailed structural information about hybrid vesicles with different ratios of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and poly(1,2-butadiene-block-ethylene oxide) (PBd22 -PEO14 , Ms  = 1800 g mol-1 ). Using single particle analysis (SPA) the authors are able to further interpret the information gained from SAXS and cryo-ET experiments, showing that increasing PBd22 -PEO14 mole fraction increases the membrane thickness from 52 Å for a pure lipid system to 97 Å for pure PBd22 -PEO14 vesicles. Two vesicle populations with different membrane thicknesses in hybrid vesicle samples are found. As these lipids and polymers are reported to homogeneously mix, bistability is inferred between weak and strong interdigitation regimes of PBd22 -PEO14 within the hybrid membranes. It is hypothesized that membranes of intermediate structure are not energetically favorable. Therefore, each vesicle exists in one of these two membrane structures, which are assumed to have comparable free energies. The authors conclude that, by combining biophysical methods, accurate determination of the influence of composition on the structural properties of hybrid membranes is achieved, revealing that two distinct membranes structures can coexist in homogeneously mixed lipid-polymer hybrid vesicles.

Multi-ancestry genome-wide association analyses improve resolution of genes and pathways influencing lung function and chronic obstructive pulmonary disease risk.

Lung-function impairment underlies chronic obstructive pulmonary disease (COPD) and predicts mortality. In the largest multi-ancestry genome-wide association meta-analysis of lung function to date, comprising 580,869 participants, we identified 1,020 independent association signals implicating 559 genes supported by ≥2 criteria from a systematic variant-to-gene mapping framework. These genes were enriched in 29 pathways. Individual variants showed heterogeneity across ancestries, age and smoking groups, and collectively as a genetic risk score showed strong association with COPD across ancestry groups. We undertook phenome-wide association studies for selected associated variants as well as trait and pathway-specific genetic risk scores to infer possible consequences of intervening in pathways underlying lung function. We highlight new putative causal variants, genes, proteins and pathways, including those targeted by existing drugs. These findings bring us closer to understanding the mechanisms underlying lung function and COPD, and should inform functional genomics experiments and potentially future COPD therapies.

The predictive validity of the DSM-5 alternative model for borderline personality disorder: Associations with coping strategies, general distress, rumination, and suicidal ideation across one year.

The DSM-5 Alternative Model for Personality Disorders (AMPD) characterizes borderline personality disorder (BPD) in part as a constellation of maladaptive personality trait facets including emotional lability, anxiousness, separation insecurity, depressivity, impulsivity, risk-taking, and hostility. Previous studies have supported the construct validity of AMPD-BPD; however, research examining its predictive validity in relation to theoretically and clinically relevant constructs remains needed. The present study investigates the longitudinal relationships between AMPD-BPD and general distress, rumination, and suicidal ideation, as well as adaptive and maladaptive coping targeted in Dialectical Behavior Therapy (DBT) in a sample of participants with elevated BPD symptomology. We also examined if dysfunctional coping skill use at 9-month follow-up explained the relationship between baseline BPD traits and outcomes at 1-year. There were significant correlations between baseline trait BPD with dysfunctional coping skill use at 9-month follow-up and psychological distress and rumination at 1-year follow-up. Dysfunctional skill use exhibited a significant indirect effect in the association between trait BPD and rumination after 1 year. The findings of this study support the construct validity of AMPD-BPD that can inform treatment and research.

Stem Cell Transplant Experiences Among Hispanic/Latinx Patients: A Qualitative Analysis.

BACKGROUND: Hispanic/Latinx (H/L) patients with cancer treated with stem cell transplant are vulnerable to adverse outcomes, including higher mortality. This study explored their unmet transplant needs, barriers, and facilitators. METHODS: Eighteen English- or Spanish-speaking H/L patients (M age = 59.2) who had a transplant in the past year were interviewed about their transplant experience and rated their interest in receiving information about transplant topics (0 = not at all to 10 = extremely). RESULTS: Content analysis revealed five main themes: (1) pre-transplant barriers and concerns; (2) complex relationships with medical teams; (3) informational mismatch; (4) impacts on daily life after transplant; and (5) methods of coping. Participants were most interested in information about ways of coping with transplant (M = 9.11, SD = 1.45) and words of hope and encouragement (M = 9.05, SD = 1.80). At just above the scale's midpoint, they were least interested in information about side effects and unintended consequences of transplant (M = 5.61, SD = 3.85). CONCLUSIONS: Cultural factors, social determinants, and structural inequalities give rise to unique needs in this growing patient population. Healthcare team members and researchers can better meet the needs of H/L transplant recipients through attention to described considerations, such as financial barriers, communication difficulties, family dynamics, and coping styles.

Adoptive parents' finances and employment status: a 5-year longitudinal study.

We investigated risk and facilitating factors related to families' change in finances and employment over 5 years following adoption of a child from local authority care in a prospective, longitudinal study of children placed for adoption between 2014 and 2015 (N = 96). Parents completed questionnaires at approximately 5, 21, 36, 48 and 60 months post-placement. We used time series analysis to examine the impact of child (e.g. pre-placement experiences, mental health), family structure (e.g. number of siblings, parent relationship status), and parent (e.g. mental health) factors on change in household income and parent employment status after adoption. We also examined the tendency for parents to comment on employment and finances and the emotional valence of their comments to gauge their concern about their circumstances. Children's mental health problems were associated with primary caregivers reducing their time spent in employment and parents' tendency to comment on their financial and work circumstances. Children who experienced more moves in care were more likely to have a primary caregiver not in full-time work, as were children with higher prosocial behaviour scores. Being in full-time work was associated with parents' symptoms of anxiety. We also detected associations between structural features of the family and changes in income and employment. This study represents one of the first empirical investigations of factors associated with the socioeconomic features of adoptive families' lives and informs ongoing discussion regarding the support needs of families and the timing, nature, and delivery of post-adoption professional services.

Expanding access to cancer peer support: development of a website to deliver written peer support that meets patients' individual emotional and informational needs while reducing potential harms.

Cancer patients often want information from "peers" with the same diagnosis or treatment. To increase access to this valuable resource, we developed a website to deliver written peer support to cancer patients undergoing stem cell transplant. Because little evidence describes how to optimize benefits or reduce potential harms of written peer support, we gathered multiple forms of stakeholder feedback to inform the website's ethical approach, personalization, design, function, and content: a Community Advisory Board; a longitudinal study of patients' written peer support needs and motivations; focus groups; semi-structured interviews; and usability testing. Findings provide a rich foundation for website development.