Laryngeal Vibrotactile Stimulation Is Feasible, Acceptable To People With Unexplained Chronic Cough.

OBJECTIVES: Unexplained chronic cough (UCC) is common and has significant impacts on quality of life. Ongoing cough can sensitize the larynx, increasing the urge to cough and perpetuating the cycle of chronic cough. Vibrotactile stimulation (VTS) of the larynx is a noninvasive stimulation technique that can modulate laryngeal somatosensory and motor activity. Study objectives were to assess feasibility and acceptability of VTS use by people with UCC. Secondarily, changes in cough-related quality of life measures were assessed. METHODS: Adults with UCC recorded cough measures at baseline and after completing 2 weeks of daily VTS. Feasibility and acceptability were assessed through participant-reported device use and structured feedback. Cough-related quality of life measures were the Leicester Cough Questionnaire (LCQ) and the Newcastle Laryngeal Hypersensitivity Questionnaire (NLHQ). RESULTS: Nineteen adults participated, with mean age 67 years and cough duration 130 months. Notably, 93% of planned VTS sessions were logged, 94% of participants found the device comfortable to wear, 89% found it easy to operate and 79% would recommend it to others. Pre-post LCQ change achieved a minimal important difference (MID) (mean 1.3 [SD 2.4, p = 0.015]). NLHQ scores improved, but did not reach an MID. CONCLUSIONS: Laryngeal VTS use was feasible and acceptable for use by patients with UCC and was associated with a meaningful improvement in cough-related quality of life. Future studies will include VTS dose refinement and the inclusion of a comparison arm to further assess the potential for laryngeal VTS as a novel treatment modality for UCC. LEVEL OF EVIDENCE: 4 Laryngoscope, 2024.

"I knew the effects the whole time." Examination of facilitators and barriers to quit vaping among adolescents.

Despite declines in adolescent nicotine vaping during the COVID-19 pandemic, vaping continues to be a public health issue for many adolescents. Most studies on teen reasons for vaping and on barriers and facilitators of vaping were conducted prior to the pandemic. Because teen culture changes so rapidly, and because the pandemic had wide-reaching effects on teens and mental health, it is critical to have a current understanding what helps adolescents to quit vaping (facilitators), as well as what prevents them from quitting (barriers) in order to design effective and engaging interventions. The objective of this qualitative study was to examine reasons for vaping, and barriers and facilitators to quitting vaping among high school age (14-18 years old) students. Students (n = 28; 60.7 % female; 50 % White, 10.7 % Black or African American, 25 % Asian or Asian American, 3.6 % other non-Hispanic; 7.2 % Hispanic; 21.4 % former vapers, 78.6 % current vapers) were recruited online from four regions in the US and participated in one of five online focus groups. We conducted a content analysis of the focus groups using a team-based coding approach. The most frequently cited reasons for vaping were mood, peer influence, and boredom. Results also showed that the stigma of seeking treatment acted as a barrier to quitting vaping, while self-reflection acted as a facilitator. In addition, two factors, peer influence and health effects, served as both barriers and facilitators, depending on the context. Program developers can use this information to design engaging vaping cessation programs intended for adolescents.

Methods used to quit vaping among adolescents and associations with perceived risk, addiction, and socio-economic status.

Despite widespread evidence suggesting that e-cigarettes are harmful to youth, the prevalence of adolescent vaping continues to rise. The first aim of this cross-sectional study was to determine which methods adolescents have previously used to quit vaping and their interest in using various methods to quit vaping in the future. The second aim was to investigate the associations between vaping-related characteristics and (1) motivation to quit vaping and (2) previous use of pharmacological methods to quit vaping or previous use of unassisted quitting. Participants were 185 current or former vapers, aged 14-19 (M age = 16.9 years, SD = 1.1; 52% female), recruited online from different U.S. regions. Data were collected through online questionnaires. Most participants reported at least one prior attempt to quit vaping (81.3%). Unassisted quitting was the most prevalent method to quit vaping (78.4%), followed by "advice from a friend" (51.9%). Participants were interested in using a variety of methods for quitting vaping in the future, including medication (28.8%), and mobile apps (34.6%). Greater perceived harm of vaping was significantly associated with motivation to quit vaping, while greater perceived risk of addiction and higher SES were significantly associated with lifetimeuse of nicotine replacement to quit vaping. Adolescents may be open to trying methods to quit that are different from what they have used previously. These findings could help direct resources toward the development of vaping cessation programs that are acceptable to adolescents.

Associations Between Work-Related Factors and Psychological Distress Among Construction Workers.

OBJECTIVE: Identify work-related factors associated with the mental health and well-being of construction workers. METHODS: We completed eight key informant interviews, six worker focus groups, and a survey, informed by the interviews and focus groups, of 259 construction workers on five construction sites. Negative binomial regressions examined associations between psychological distress and work-related factors including safety climate, work-to-family conflict, psychological demands, social support, harassment, and job security. RESULTS: Three themes emerged from the interviews and focus groups, job demands and structure, social support and workplace relations, and job precarity. From the survey higher psychological demands, higher work-to-family conflict, lower supervisor support, higher discrimination, and higher likelihood of losing a job were associated with higher psychological distress. When combined into a single model job demands and work-to-family conflict remained significant. CONCLUSIONS: Work-related factors were associated with high levels of distress.

Mental Health Stigma and Wellbeing Among Commercial Construction Workers: A Mixed Methods Study.

OBJECTIVE: The aim of this study was to understand the relationship between mental health stigma and wellbeing in a sample of commercial construction workers. METHODS: A sequential quantitative to qualitative mixed methods study was conducted. First, 264 workers were surveyed and logistic regression examined the associations between mental health stigma and psychological distress, substance abuse, and sleep; then, data from eight key informant interviews and six focus groups with workers were analyzed thematically. RESULTS: Stigma was significantly associated with psychological distress and impaired sleep, but not with substance abuse. Key informants and workers discussed the prominence of stigma on worksites and how the masculine culture of the industry and job insecurity contributed to stigma. CONCLUSION: Stigma poses a barrier to addressing mental health among workers; peer support and improved human resources may be promising.

Associations between a safety prequalification survey and worker safety experiences on commercial construction sites.

BACKGROUND: While assessment of subcontractors' safety performance during project bidding processes are common in commercial construction, the validation of organizational surveys used in these processes is largely absent. METHODS: As part of a larger research project called Assessment of Contractor Safety (ACES), we designed and tested through a cross-sectional study, a 63-item organizational survey assessing subcontractors' leading indicators of safety performance. We administered the ACES Survey to 43 subcontractors on 24 construction sites. Concurrently, we captured the safety climate of 1426 workers on these sites through worker surveys, as well as injury rates, for the duration of the project. RESULTS: At the worksite level, higher average ACES scores were associated with higher worker safety climate scores (P < .01) and lower rates of injury involving days away (P < .001). Within subcontracting companies, no associations were observed between ACES and worker safety climate scores and injuries. CONCLUSIONS: These results suggest the overall and collective importance of the construction project and its worksite in mediating worker experiences, perhaps somewhat independent of the individual subcontractor level.

Tbx2 mediates dorsal patterning and germ layer suppression through inhibition of BMP/GDF and Activin/Nodal signaling.

BACKGROUND: Members of the T-box family of DNA-binding proteins play a prominent role in the differentiation of the three primary germ layers. VegT, Brachyury, and Eomesodermin function as transcriptional activators and, in addition to directly activating the transcription of endoderm- and mesoderm-specific genes, serve as regulators of growth factor signaling during induction of these germ layers. In contrast, the T-box gene, tbx2, is expressed in the embryonic ectoderm, where Tbx2 functions as a transcriptional repressor and inhibits mesendodermal differentiation by the TGFß ligand Activin. Tbx2 misexpression also promotes dorsal ectodermal fate via inhibition of the BMP branch of the TGFß signaling network. RESULTS: Here, we report a physical association between Tbx2 and both Smad1 and Smad2, mediators of BMP and Activin/Nodal signaling, respectively. We perform structure/function analysis of Tbx2 to elucidate the roles of both Tbx2-Smad interaction and Tbx2 DNA-binding in germ layer suppression. CONCLUSION: Our studies demonstrate that Tbx2 associates with intracellular mediators of the Activin/Nodal and BMP/GDF pathways. We identify a novel repressor domain within Tbx2, and have determined that Tbx2 DNA-binding activity is required for repression of TGFß signaling. Finally, our data also point to overlapping yet distinct mechanisms for Tbx2-mediated repression of Activin/Nodal and BMP/GDF signaling.

Repression of Inappropriate Gene Expression in the Vertebrate Embryonic Ectoderm.

During vertebrate embryogenesis, precise regulation of gene expression is crucial for proper cell fate determination. Much of what we know about vertebrate development has been gleaned from experiments performed on embryos of the amphibian Xenopus laevis; this review will focus primarily on studies of this model organism. An early critical step during vertebrate development is the formation of the three primary germ layers-ectoderm, mesoderm, and endoderm-which emerge during the process of gastrulation. While much attention has been focused on the induction of mesoderm and endoderm, it has become clear that differentiation of the ectoderm involves more than the simple absence of inductive cues; rather, it additionally requires the inhibition of mesendoderm-promoting genes. This review aims to summarize our current understanding of the various inhibitors of inappropriate gene expression in the presumptive ectoderm.

Tbx2 is required for the suppression of mesendoderm during early Xenopus development.

BACKGROUND: T-box family proteins are DNA-binding transcriptional regulators that play crucial roles during germ layer formation in the early vertebrate embryo. Well-characterized members of this family, including the transcriptional activators Brachyury and VegT, are essential for the proper formation of mesoderm and endoderm, respectively. To date, T-box proteins have not been shown to play a role in the promotion of the third primary germ layer, ectoderm. RESULTS: Here, we report that the T-box factor Tbx2 is both sufficient and necessary for ectodermal differentiation in the frog Xenopus laevis. Tbx2 is expressed zygotically in the presumptive ectoderm, during blastula and gastrula stages. Ectopic expression of Tbx2 represses mesoderm and endoderm, while loss of Tbx2 leads to inappropriate expression of mesoderm- and endoderm-specific genes in the region fated to give rise to ectoderm. Misexpression of Tbx2 also promotes neural tissue in animal cap explants, suggesting that Tbx2 plays a role in both the establishment of ectodermal fate and its dorsoventral patterning. CONCLUSIONS: Our studies demonstrate that Tbx2 functions as a transcriptional repressor during germ layer formation, and suggest that this activity is mediated in part through repression of target genes that are stimulated, in the mesendoderm, by transactivating T-box proteins. Taken together, our results point to a critical role for Tbx2 in limiting the potency of blastula-stage progenitor cells during vertebrate germ layer differentiation. Developmental Dynamics 247:903-913, 2018. © 2018 Wiley Periodicals, Inc.

Pitx1 regulates cement gland development in Xenopus laevis through activation of transcriptional targets and inhibition of BMP signaling.

The cement gland in Xenopus laevis has long been used as a model to study the interplay of cell signaling and transcription factors during embryogenesis. It has been shown that an intermediate level of Bone Morphogenetic Protein (BMP) signaling is essential for cement gland formation. In addition, several transcription factors have been linked to cement gland development. One of these, the homeodomain-containing protein Pitx1, can generate ectopic cement gland formation; however, the mechanisms underlying this process remain obscure. We report here, for the first time, a requirement for Pitx proteins in cement gland formation, in vivo: knockdown of both pitx1 and the closely related pitx2c inhibit endogenous cement gland formation. Pitx1 transcriptionally activates cement gland differentiation genes through both direct and indirect mechanisms, and functions as a transcriptional activator to inhibit BMP signaling. This inhibition, required for the expression of pitx genes, is partially mediated by Pitx1-dependent follistatin expression. Complete suppression of BMP signaling inhibits induction of cement gland markers by Pitx1; furthermore, we find that Pitx1 physically interacts with Smad1, an intracellular transducer of BMP signaling. We propose a model of cement gland formation in which Pitx1 limits local BMP signaling within the cement gland primordium, and recruits Smad1 to activate direct downstream targets.

Cloning and spatiotemporal expression of Xenopus laevis Apolipoprotein CI.

Apolipoprotein CI (ApoCI) belongs to the Apolipoprotein superfamily, members of which are involved in lipid transport, uptake and homeostasis. Excessive ApoCI has been implicated in atherosclerosis and Alzheimer's disease in humans. In this study we report the isolation of Xenopus laevis apoCI and describe the expression pattern of this gene during early development, using reverse transcription polymerase chain reaction and whole mount in situ hybridization. Xenopus apoCI is enriched in the dorsal ectoderm during gastrulation, and is subsequently expressed in sensory placodes, neural tube and cranial neural crest. These data suggest as yet uncharacterized roles for ApoCI during early vertebrate embryogenesis.

Exercise-associated Excessive Dynamic Airway Collapse in Military Personnel.

RATIONALE: Evaluation of military personnel for exertional dyspnea can present a diagnostic challenge, given multiple unique factors that include wide variation in military deployment. Initial consideration is given to common disorders such as asthma, exercise-induced bronchospasm, and inducible laryngeal obstruction. Excessive dynamic airway collapse has not been reported previously as a cause of dyspnea in these individuals. OBJECTIVES: To describe the clinical and imaging characteristics of military personnel with exertional dyspnea who were found to have excessive dynamic collapse of large airways during exercise. METHODS: After deployment to Afghanistan or Iraq, 240 active U.S. military personnel underwent a standardized evaluation to determine the etiology of persistent dyspnea on exertion. Study procedures included full pulmonary function testing, impulse oscillometry, exhaled nitric oxide measurement, methacholine challenge testing, exercise laryngoscopy, cardiopulmonary exercise testing, and fiberoptic bronchoscopy. Imaging included high-resolution computed tomography with inspiratory and expiratory views. Selected individuals underwent further imaging with dynamic computed tomography. MEASUREMENTS AND MAIN RESULTS: A total of five men and one woman were identified as having exercise-associated excessive dynamic airway collapse on the basis of the following criteria: (1) exertional dyspnea without resting symptoms, (2) focal expiratory wheezing during exercise, (3) functional collapse of the large airways during bronchoscopy, (4) expiratory computed tomographic imaging showing narrowing of a large airway, and (5) absence of underlying apparent pathology in small airways or pulmonary parenchyma. Identification of focal expiratory wheezing correlated with bronchoscopic and imaging findings. CONCLUSIONS: Among 240 military personnel evaluated after presenting with postdeployment exertional dyspnea, a combination of symptoms, auscultatory findings, imaging, and visualization of the airways by bronchoscopy identified six individuals with excessive dynamic central airway collapse as the sole apparent cause of dyspnea. Exercise-associated excessive dynamic airway collapse should be considered in the differential diagnosis of exertional dyspnea.

Group music performance causes elevated pain thresholds and social bonding in small and large groups of singers.

Over our evolutionary history, humans have faced the problem of how to create and maintain social bonds in progressively larger groups compared to those of our primate ancestors. Evidence from historical and anthropological records suggests that group music-making might act as a mechanism by which this large-scale social bonding could occur. While previous research has shown effects of music making on social bonds in small group contexts, the question of whether this effect 'scales up' to larger groups is particularly important when considering the potential role of music for large-scale social bonding. The current study recruited individuals from a community choir that met in both small (n = 20 - 80) and large (a 'megachoir' combining individuals from the smaller subchoirs n = 232) group contexts. Participants gave self-report measures (via a survey) of social bonding and had pain threshold measurements taken (as a proxy for endorphin release) before and after 90 minutes of singing. Results showed that feelings of inclusion, connectivity, positive affect, and measures of endorphin release all increased across singing rehearsals and that the influence of group singing was comparable for pain thresholds in the large versus small group context. Levels of social closeness were found to be greater at pre- and post-levels for the small choir condition. However, the large choir condition experienced a greater change in social closeness as compared to the small condition. The finding that singing together fosters social closeness - even in large contexts where individuals are not known to each other - is consistent with evolutionary accounts that emphasize the role of music in social bonding, particularly in the context of creating larger cohesive groups than other primates are able to manage.

In Second Year Of Marketplaces, New Entrants, ACA 'Co-Ops,' And Medicaid Plans Restrain Average Premium Growth Rates.

Premiums for health insurance plans offered through the federally facilitated and state-based Marketplaces remained steady or increased only modestly from 2014 to 2015. We used data from the Marketplaces, state insurance departments, and insurer websites to examine patterns of premium pricing and the factors behind these patterns. Our data came from 2,964 unique plans offered in 2014 and 4,153 unique plans offered in 2015 in forty-nine states and the District of Columbia. Using descriptive and multivariate analysis, we found that the addition of a carrier in a rating area lowered average premiums for the two lowest-cost silver plans and the lowest-cost bronze plan by 2.2 percent. When all plans in a rating area were included, an additional carrier was associated with an average decline in premiums of 1.4 percent. Plans in the Consumer Operated and Oriented Plan Program and Medicaid managed care plans had lower premiums and average premium increases than national commercial and Blue Cross and Blue Shield plans. On average, premiums fell by an appreciably larger amount for catastrophic and bronze plans than for gold plans, and premiums for platinum plans increased. This trend of low premium increases overall is unlikely to continue, however, as insurers are faced with mounting medical claims.

ß-Catenin-independent activation of TCF1/LEF1 in human hematopoietic tumor cells through interaction with ATF2 transcription factors.

The role of Wnt signaling in embryonic development and stem cell maintenance is well established and aberrations leading to the constitutive up-regulation of this pathway are frequent in several types of human cancers. Upon ligand-mediated activation, Wnt receptors promote the stabilization of ß-catenin, which translocates to the nucleus and binds to the T-cell factor/lymphoid enhancer factor (TCF/LEF) family of transcription factors to regulate the expression of Wnt target genes. When not bound to ß-catenin, the TCF/LEF proteins are believed to act as transcriptional repressors. Using a specific lentiviral reporter, we identified hematopoietic tumor cells displaying constitutive TCF/LEF transcriptional activation in the absence of ß-catenin stabilization. Suppression of TCF/LEF activity in these cells mediated by an inducible dominant-negative TCF4 (DN-TCF4) inhibited both cell growth and the expression of Wnt target genes. Further, expression of TCF1 and LEF1, but not TCF4, stimulated TCF/LEF reporter activity in certain human cell lines independently of ß-catenin. By a complementary approach in vivo, TCF1 mutants, which lacked the ability to bind to ß-catenin, induced Xenopus embryo axis duplication, a hallmark of Wnt activation, and the expression of the Wnt target gene Xnr3. Through generation of different TCF1-TCF4 fusion proteins, we identified three distinct TCF1 domains that participate in the ß-catenin-independent activity of this transcription factor. TCF1 and LEF1 physically interacted and functionally synergized with members of the activating transcription factor 2 (ATF2) family of transcription factors. Moreover, knockdown of ATF2 expression in lymphoma cells phenocopied the inhibitory effects of DN-TCF4 on the expression of target genes associated with the Wnt pathway and on cell growth. Together, our findings indicate that, through interaction with ATF2 factors, TCF1/LEF1 promote the growth of hematopoietic malignancies in the absence of ß-catenin stabilization, thus establishing a new mechanism for TCF1/LEF1 transcriptional activity distinct from that associated with canonical Wnt signaling.

The mammalian copper transporters CTR1 and CTR2 and their roles in development and disease.

Copper is vital to cell function. The influx of reduced copper ions is controlled by two functionally homologous transmembrane solute carrier transporters CTR1 (encoded by SLC31A1) and CTR2 (encoded by SLC31A2). These copper transporters vary in their expression profiles and intracellular localisation patterns. CTR1 plays roles in the developing embryo as well as regulating homeostasis in the adult mammal. In contrast, the regulation, expression and function of CTR2 is poorly defined. Both are capable of transporting other divalent metal ions and are the primary transporters for platinum-based chemotherapeutic drugs such as cisplatin. This review summarises our current understanding of these two copper transporters and highlights their roles in cellular processes, embryonic development, differentiation, cancer, immunity and disease.

Eif4a3 is required for accurate splicing of the Xenopus laevis ryanodine receptor pre-mRNA.

The Exon Junction Complex (EJC) plays a critical role in multiple posttranscriptional events, including RNA subcellular localization, nonsense-mediated decay (NMD), and translation. We previously reported that knockdown of the EJC core component Eukaryotic initiation factor 4a3 (Eif4a3) results in full-body paralysis of embryos of the frog, Xenopus laevis. Here, we explore the cellular and molecular mechanisms underlying this phenotype. We find that cultured muscle cells derived from Eif4a3 morphants do not contract, and fail to undergo calcium-dependent calcium release in response to electrical stimulation or treatment with caffeine. We show that ryr (ryanodine receptor) transcripts are incorrectly spliced in Eif4a3 morphants, and demonstrate that inhibition of Xenopus Ryr function similarly results in embryonic paralysis. These results suggest that the EJC mediates muscle cell function via regulation of pre-mRNA splicing during early vertebrate embryogenesis.

Rab11 regulates planar polarity and migratory behavior of multiciliated cells in Xenopus embryonic epidermis.

BACKGROUND: Xenopus embryonic skin is composed of the superficial layer with defined apicobasal polarity and the inner layer lacking the apical domain. Multiciliated cells (MCCs) originate in the inner layer of the epidermal ectoderm and subsequently migrate to the surface. How MCCs acquire the apicobasal polarity and intercalate into the superficial layer during neurulation is largely unknown. As Rab11-dependent vesicle trafficking has been implicated in ciliary membrane assembly and in apical domain formation in epithelial cells, we assessed the involvement of Rab11 in MCC development. RESULTS: Here we report that Rab11 is specifically enriched and becomes apically polarized in skin MCCs. Interference with Rab11 function by overexpression of a dominant negative mutant or injection of a specific morpholino oligonucleotide inhibited MCC intercalation into the superficial layer. Dominant negative Rab11-expressing MCC precursors revealed intrinsic apicobasal polarity, characterized by the apical domain, which is not normally observed in inner layer cells. Despite the presence of the apical domain, the cells with inhibited Rab11 function were randomly oriented relative to the plane of the tissue, thereby demonstrating a defect in planar polarity. CONCLUSIONS: These results establish a requirement for Rab11 in MCC development and support a two-step model, in which the initial polarization of MCC precursors is critical for their integration into the superficial cell layer.

Xmab21l3 mediates dorsoventral patterning in Xenopus laevis.

Specification of the dorsoventral (DV) axis is critical for the subsequent differentiation of regional fate in the primary germ layers of the vertebrate embryo. We have identified a novel factor that is essential for dorsal development in embryos of the frog Xenopus laevis. Misexpression of Xenopus mab21-like 3 (Xmab21l3) dorsalizes gastrula-stage mesoderm and neurula-stage ectoderm, while morpholino-mediated knockdown of Xmab21l3 inhibits dorsal differentiation of these embryonic germ layers. Xmab21l3 is a member of a chordate-specific subclass of a recently characterized gene family, all members of which contain a conserved, but as yet ill-defined, Mab21 domain. Our studies suggest that Xmab21l3 functions to repress ventralizing activity in the early vertebrate embryo, via regulation of BMP/Smad and Ras/ERK signaling.

Regulation of vertebrate embryogenesis by the exon junction complex core component Eif4a3.

The establishment and maintenance of cellular identity are ultimately dependent upon the accurate regulation of gene expression, the process by which genetic information is used to synthesize functional gene products. The post-transcriptional, pre-translational regulation of RNA constitutes RNA processing, which plays a prominent role in the modulation of gene expression in differentiated animal cells. The multi-protein Exon Junction Complex (EJC) serves as a critical signaling hub within the network that underlies many RNA processing events. Here, we identify a requirement for the EJC during early vertebrate embryogenesis. Knockdown of the EJC component Eukaryotic initiation factor 4a3 (Eif4a3) in embryos of the frog Xenopus laevis results in full-body paralysis, with defects in sensory neuron, pigment cell, and cardiac development; similar phenotypes are seen following knockdown of other "core" EJC protein constituents. Our studies point to an essential role for the EJC in the development of neural plate border derivatives.