Droplet microfluidics for time-resolved serial crystallography.

Serial crystallography requires large numbers of microcrystals and robust strategies to rapidly apply substrates to initiate reactions in time-resolved studies. Here, we report the use of droplet miniaturization for the controlled production of uniform crystals, providing an avenue for controlled substrate addition and synchronous reaction initiation. The approach was evaluated using two enzymatic systems, yielding 3 µm crystals of lysozyme and 2 µm crystals of Pdx1, an Arabidopsis enzyme involved in vitamin B6 biosynthesis. A seeding strategy was used to overcome the improbability of Pdx1 nucleation occurring with diminishing droplet volumes. Convection within droplets was exploited for rapid crystal mixing with ligands. Mixing times of <2 ms were achieved. Droplet microfluidics for crystal size engineering and rapid micromixing can be utilized to advance time-resolved serial crystallography.

Associations Between Risk Perception and HPV Knowledge and Vaccine Uptake: Highlighting the LGBTQ+ Population.

INTRODUCTION: Gay and bisexual males and other LGBTQ+ communities are more frequently exposed to factors associated with an increased risk of human papillomavirus (HPV) acquisition. Vaccination is critical to protect against HPV+ head and neck cancer (HNC). We characterized the association of perceived level of risk of contraction with HPV knowledge, and vaccine decision-making. STUDY DESIGN: Cross-sectional cohort. SETTING: LGBTQ and general survey Reddit forums (control). METHODS: A survey was shared amongst the online forums. Descriptive statistics characterized the data. Multivariable logistic regression was used to understand factors associated with vaccination, self-perceived high risk, and knowledge of HPV + HNC. RESULTS: Of 718 respondents, most were female (41.09%), Caucasian (59.89%), college-educated (33.01%), and insured (77.15%) with a mean age of 30.75 years. Half were vaccinated (49.16%), with most unvaccinated endorsing interest (60.58%). Few dependents were vaccinated (25.91%), with interest in vaccination among parents of unvaccinated children (38.58%). Knowledge of HIV's association with HPV (62.95%), HPV causing HNC (55.57%), and the vaccine's efficacy against HNC (55.57%) was also moderate. Identifying female (P = .042), a self-perceived high-risk (P < .001), and having vaccinated children (P < .001) increased vaccination likelihood; transgender (P = .021), or lesbian or gay sexual identity (P < .001) decreased likelihood. Personal HNC diagnosis (P < .001), self-vaccination (P < .001), having vaccinated children (P < .001), having anal sex (P = .001) or no knowledge of past HPV status (P < .001) increased likelihood of high self-perceived risk. CONCLUSION: Efforts to improve public education regarding the association between HPV and HNC and vaccination efficacy are required to better inform vaccine decision-making among individuals at risk for HPV infection.

Assessing Laryngectomy Patient Education on YouTube: Investigating Quality and Reliability.

Objective: This study aimed to characterize the quality of laryngectomy-related patient education on YouTube and understand factors impacting video content quality. Study Design: Cross-sectional cohort analysis. Setting: Laryngectomy-related videos on YouTube. Methods: YouTube was anonymously queried for various laryngectomy procedure search terms. Video quality was evaluated using the validated DISCERN instrument which assesses treatment-related information quality. Descriptive statistics were used to characterize our cohort. Univariate and multivariable linear regression were used to assess factors associated with increased DISCERN score. Significance was set at P < .05. Results: Our 78-video cohort exhibited moderate levels of engagement, averaging 13,028.40 views (SD = 24,246.93), 69.79 likes (SD = 163.75), and 5.27 comments (SD = 18.81). Videos were most frequently uploaded to accounts belonging to physicians (43.59%) or health care groups (41.03%) and showcased operations (52.56%) or physician-led education (20.51%). Otolaryngologists were featured in most videos (85.90%), and most videos originated outside the United States (67.95%). Laryngectomy videos demonstrated poor reliability (mean = 2.35, SD = 0.77), quality of treatment information (mean = 1.92, SD = 0.86), and overall video quality (mean = 1.97, SD = 1.12). In multivariable linear regression, operative videos were associated with lower video quality relative to nonoperative videos (ß = -1.63, 95% confidence interval [CI] = [-2.03 to -1.24], P < .001); the opposite was true for videos from accounts with higher subscriber counts (ß = 0.02, 95% CI = [0.01-0.03], P = .005). Conclusion: The quality and quantity of YouTube's laryngectomy educational content is limited. There is an acute need to increase the quantity and quality of online laryngectomy-related content to better support patients and caregivers as they cope with their diagnosis, prepare for, and recover from surgery.

Mechanisms of SARS-CoV-2 Inactivation Using UVC Laser Radiation.

Severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2) has had a tremendous impact on humanity. Prevention of transmission by disinfection of surfaces and aerosols through a chemical-free method is highly desirable. Ultraviolet C (UVC) light is uniquely positioned to achieve inactivation of pathogens. We report the inactivation of SARS-CoV-2 virus by UVC radiation and explore its mechanisms. A dose of 50 mJ/cm2 using a UVC laser at 266 nm achieved an inactivation efficiency of 99.89%, while infectious virions were undetectable at 75 mJ/cm2 indicating >99.99% inactivation. Infection by SARS-CoV-2 involves viral entry mediated by the spike glycoprotein (S), and viral reproduction, reliant on translation of its genome. We demonstrate that UVC radiation damages ribonucleic acid (RNA) and provide in-depth characterization of UVC-induced damage of the S protein. We find that UVC severely impacts SARS-CoV- 2 spike protein's ability to bind human angiotensin-converting enzyme 2 (hACE2) and this correlates with loss of native protein conformation and aromatic amino acid integrity. This report has important implications for the design and development of rapid and effective disinfection systems against the SARS-CoV-2 virus and other pathogens.

COVID-19 Personal Protective Behaviors during Large Social Events: The Value of Behavioral Observations.

Within the context of reopening society in the summer of 2021, as the UK moved away from 'lockdowns', the Government of Wales piloted the return of organized 'mass gatherings' of people at a number of test events. The current study reports behavioral observations that were made at two of the test events to inform this process. The researchers were particularly interested in four key factors: how (1) context within a venue, (2) environmental design, (3) staffing and social norms, and (4) time across an event, affected the personal protective behaviors of social distancing and face-covering use. Data collection was undertaken by trained observers. Adherence to protective behaviors was generally high, but there is clear evidence that these behaviors were shaped in a systematic way by the environment, situational cues, and the passage of time during the events. Some instances of large-scale non-adherence to personal protective behaviors were documented. An analysis within a dual-process framework suggests ways to understand and respond to supporting target health behaviors in groups of people where intervention is deemed valuable, such as in complex or ambiguous contexts. This is one of the first studies to include a 'true' behavioral measure in understanding human responses to COVID-19. It demonstrates that behavioral observations can add precision and granularity to understanding human behavior in complex real-world contexts. Given the significant physical and mental health burden created acutely and chronically by COVID-19, this work has implications for how governments and organizations support target populations in other complex challenges facing us today, such as in sustainability, and healthy lifestyle behaviors. An individual's intentions are not always matched by their actions, and so the findings support a balanced liberal paternalistic approach where system-level changes support appropriate individual-level decisions to engender collective responsibility and action.

Population Genetic Structure of the Rubber Tree Powdery Mildew Pathogen (Erysiphe quercicola) from China.

In order to manage agricultural pathogens, it is crucial to understand the population structure underlying epidemics. Rubber tree powdery mildew, caused by Erysiphe quercicola, is a serious threat to rubber plantations worldwide, especially in subtropical environments including all rubber tree-growing regions in China. However, the population structure of the pathogen is uncertain. In this study, 16 polymorphic microsatellite markers were used to genotype powdery mildew samples from the main rubber tree-growing regions including Yunnan (YN), Hainan (HN), western Guangdong (WG), and eastern Guangdong (EG). YN had higher genotypic diversity (Simpson's indices), genotypic evenness, Nei's gene diversity, allelic richness, and private allelic richness than the other regions. Cluster analysis, discriminant analysis of principal components, pairwise divergence, and shared multilocus genotype analyses all showed that YN differed significantly from the other regions. The genetic differentiation was small among the other three regions (HN, WG, and EG). Analysis of molecular variance indicated that the variability among regions accounted for 22.37% of the total variability. Genetic differentiation was significantly positively correlated (Rxy = 0.772, P = 0.001) with geographic distance. Linkage equilibrium analysis suggested possible occurrence of sexual recombination although asexual reproduction predominates in E. quercicola. The results suggested that although significant genetic differentiation of E. quercicola occurred between YN and the other regions, pathogen populations from the other three regions lacked genetic differentiation.

Differences in presentation, treatment, and outcomes among minority head and neck cancer patient groups in Los Angeles County.

IMPORTANCE: While minorities represent around 20 % of all HNC patients, these demographics are largely understudied. Furthermore, trends in national studies may not always be fully replicated in locoregional populations, indicating a need for more nuanced study. OBJECTIVE: To better understand our patient population, we sought to understand differences in presentation, management, and outcome between Caucasians and minority groups with HNC. DESIGN: Retrospective cohort analysis of the Los Angeles County Surveillance Epidemiology and End Results (SEER) database. SETTING: Los Angeles County. PARTICIPANTS: All patients in Los Angeles County diagnosed with cancer of the head and neck from January 1, 1988 to December 31, 2018. MAIN OUTCOMES AND MEASURES: The primary outcome in our study was significant differences between racial and ethnic groups in age of diagnosis, sex, socioeconomic quintile, insurance status, stage at diagnosis, treatment modality, time to first treatment, and cancer-specific cause of death. RESULTS: Our 18,510-patient cohort was largely male (64.35 %), white (69.57 %), and were on average 62.84 years old (SD = 20.07). When stratifying patients by race and ethnicity, significant differences were found in average age at diagnosis, sex, socioeconomic quintile, insurance status, and stage at diagnosis, treatment modalities utilized, and time to first treatment (all p < 0.001). Relative to all other head and neck patients, minority groups were significantly younger, had lower proportions of male patients, were less likely to pursue surgery, were more likely to pursue chemotherapy or radiation, and endorsed longer time to first treatment (all p < 0.001). The distribution of socioeconomic quintile (all p < 0.001), insurance status (all p < 0.001), and stage at diagnosis (all p < 0.05) also significantly varied between minority and reference groups. Only African Americans exhibited significantly higher rates of cancer-specific cause of death relative to non-African Americans (p < 0.001). CONCLUSIONS AND RELEVANCE: Pervasive socioeconomic disparities between Caucasian HNC patients and those of other minority racial and ethnic groups in Los Angeles County that likely and significantly impact the diagnosis and management of HNC and its resultant outcomes. We encourage others to similarly examine their local populations to tailor the quality of care provided to patients.

Characterization of Colletotrichum Species Infecting Litchi in Hainan, China.

Litchi (Litchi chinensis) is an evergreen fruit tree grown in subtropical and tropical countries. China accounts for 71.5% of the total litchi cultivated area in the world. Anthracnose disease caused by Colletotrichum species is one of the most important diseases of litchi in China. In this study, the causal pathogens of litchi anthracnose in Hainan, China, were determined using phylogenetic and morphological analyses. The results identified eight Colletotrichum species from four species complexes, including a proposed new species. These were C. karsti from the C. boninense species complex; C. gigasporum and the proposed new species C. danzhouense from the C. gigasporum species complex; C. arecicola, C. fructicola species complex; C. arecicola, C. fructicola and C. siamense from the C. gloeosporioides species complex; and C. musicola and C. plurivorum from the C. orchidearum species complex. Pathogenicity tests showed that all eight species could infect litchi leaves using a wound inoculation method, although the pathogenicity was different in different species. To the best of our knowledge, the present study is the first report that identifies C. arecicola, C. danzhouense, C. gigasporum and C. musicola as etiological agents of litchi anthracnose.

Efficient qPCR estimation and discrimination of airborne inoculum of Leptosphaeria maculans and L. biglobosa, the causal organisms of phoma leaf spotting and stem canker of oilseed rape.

BACKGROUND: Detection of the inoculum of phytopathogens greatly assists in the management of diseases, but is difficult for pathogens with airborne fungal propagules. Here, we present experiments to determine the abundance and distribution frequencies of the ascospores of Leptosphaeria (Plenodomus) species that were collected on the tapes of volumetric Hirst-type traps near oilseed rape fields in Poznan, Poland and Harpenden, UK. Fungal detection and species discrimination were achieved using a SYBR-Green quantitative polymerase chain reaction (qPCR) with two different pairs of primers previously reported to differentiate Leptosphaeria maculans (Plenodomus lingam) or L. biglobosa (P. biglobosus). RESULTS: Detection was successful even at fewer than five spores per m3 of air. The primer pairs differed in the correlation coefficients obtained between DNA yields and the daily abundance of ascospores that were quantified by microscopy on duplicate halves of the spore trap tapes. Important differences in the specificity and sensitivity of the published SYBR-Green assays were also found, indicating that the Liu primers did not detect L. biglobosa subclade 'canadensis', whereas the Mahuku primers detected L. biglobosa subclade 'canadensis' and also the closely related Plenodomus dezfulensis. CONCLUSIONS: Comparisons confirmed that application of qPCR assays to spore trap samples can be used for the early detection, discrimination and quantification of aerially dispersed L. maculans and L. biglobosa propagules before leaf spot symptoms are visible in winter oilseed rape fields. The specificity of the primers must be taken into consideration because the final result will greatly depend on the local population of the pathogen. © 2023 Society of Chemical Industry.

Opioid-limiting legislation and prescribing habits of otolaryngologists among Medicare beneficiaries.

Objectives: To identify changes in otolaryngologists' opioid prescribing trends for Medicare beneficiaries associated with the enactment of state laws that limit the duration of prescriptions to 3-7 days in the years 2016 and 2017 in the United States. Methods: Through the Centers for Medicare and Medicaid Services (CMS) database, we retrieved data on Medicare enrollment and on the total days prescribed and total number of beneficiaries for the drugs codeine/acetaminophen, hydrocodone/acetaminophen, oxycodone HCl, oxycodone/acetaminophen, and tramadol HCl, by each otolaryngologist prescriber in 13 states from January 2013 to December 2019. We modeled trends using linear spline regression models that controlled for Medicare beneficiaries' state-level socio-demographic characteristics' fixed effects. Results: Across the 13 states, the number of days of all five opioids prescribed per beneficiary declined by 8.35 (SD = 12.61). The most commonly prescribed opioid type by otolaryngologists during the 5-year study period was tramadol HCl (28.72 days/beneficiary) followed by oxycodone HCl (19.99 days/beneficiary). All opioids had declines in prescription days over this time window and higher rates of decline in the years following law passage. Four states experienced statistically significant declines in the prescriptions of all opioids after the year of legislation passage (p < .05). Some states that had the greatest inclines in opioid prescriptions in the years prior to law enactment also experienced the greatest reductions in the time after legislation enactment. Conclusions: Opioid prescribing practices of otolaryngologists may have been affected by opioid prescription duration limiting laws passed in 13 states in 2016 and 2017. Level of Evidence: Level 4.

Flow rate accuracy of infusion devices within healthcare settings: a systematic review.

Background: One in five patients admitted to the hospital treated with intravenous (IV) fluid therapy suffer complications due to inappropriate administration. Errors have been reported in 13-84% of the preparation and administration of IV medications. The safe delivery of IV fluids requires precise rate administration. Objectives: This systematic review aims to determine the accuracy of infusion sets and devices and examine the factors that affect the flow rate accuracy of devices. Data Sources and Methods: Six databases (CINAHL, MEDLINE PubMed, EMBASE, Web of Science and Cochrane Database of systematic reviews) were systematically searched. Search terms included infusion pumps, infusion devices, flow rate accuracy, fluid administration rate, gravity-led infusion set and fluid balance. Studies were included if they examined infusion devices' flow rate accuracy and drop rates for fluids or non-oncological drugs. Findings were tabulated and synthesised qualitatively. The quality of the studies was examined based on the design of the studies due to their heterogeneity. Results: Eight studies were included: Four studies were conducted on human subjects in the hospital environment; studies recruited 182 participants between the ages of 18 and 94 years. Two studies examined flow rate accuracy in recruited patients across 509 observations and 2387 drip hours. No trials prospectively assessed the accuracy of infusion pumps in the clinical domain, and no studies were reported on patient safety outcomes. Four studies examined the impact of mechanical and physiological factors on the flow rate accuracies of infusion devices. Height and back pressure simulated vibrating conditions, the viscosity of IV fluid and the positions of patients were reported to have a significant impact on infusion volume and flow rates of infusion devices. Additionally, giving sets that vary from the manufacturer's specifications are reported to increase error percent by 10-20%. Conclusion: Infusion devices are an important source of error in administering IV fluids. Yet, there needs to be more prospective trial data to support their clinical accuracy and the impact on patient outcomes. Future flow variability and accuracy studies should capture their impact on patient safety and clinical outcomes.

Are the flow rate of infusion devices accurate in fluid administration? Background/Why was this study done? Nearly all patients in healthcare settings undergo treatment with fluid therapy that is administered through a vein. Inaccurate intravenous fluid administration causes patient harm. However, very little information in the literature explains how precisely intravenous fluid is administered. What did the researchers do? We reviewed the literature on flow rate accuracies of specific infusion devices and examined the factors that affect the flow rate accuracies of intravenous fluid administered to patients. What did the researchers find? We found that the flow rate accuracies of infusion devices vary greatly, and they are often affected by physiological and mechanical factors. However, the precise impact of this on patients' clinical outcomes is not often reported, representing a significant knowledge gap. What do the findings mean? We conclude that there is an urgent need to improve the reporting and precision of intravenous fluid rate administration and to understand how this impacts patient safety and clinical outcomes.

Single-cell RNA-sequence analysis of human bone marrow reveals new targets for isolation of skeletal stem cells using spherical nucleic acids.

There is a wealth of data indicating human bone marrow contains skeletal stem cells (SSC) with the capacity for osteogenic, chondrogenic and adipogenic differentiation. However, current methods to isolate SSCs are restricted by the lack of a defined marker, limiting understanding of SSC fate, immunophenotype, function and clinical application. The current study applied single-cell RNA-sequencing to profile human adult bone marrow populations from 11 donors and identified novel targets for SSC enrichment. Spherical nucleic acids were used to detect these mRNA targets in SSCs. This methodology was able to rapidly isolate potential SSCs found at a frequency of <1 in 1,000,000 in human bone marrow, with the capacity for tri-lineage differentiation in vitro and ectopic bone formation in vivo. The current studies detail the development of a platform to advance SSC enrichment from human bone marrow, offering an invaluable resource for further SSC characterisation, with significant therapeutic impact therein.

Local Reaction Causing Airway Compromise Following Vocal Fold Injection Augmentation with Carboxymethylcellulose.

OBJECTIVES: This case report aims to identify local allergic reaction as a rare complication of vocal fold injection augmentation with carboxymethylcellulose and discuss the management of airway swelling following this reaction. INTRODUCTION: Glottis insufficiency due to true vocal fold immobility is important to manage to reduce risk of aspiration and improve voice function. Vocal fold injection augmentation with carboxymethylcellulose is considered a safe and effective treatment for glottis insufficiency, commonly due to vocal fold immobility. METHODS: Retrospective medical records review and case report. RESULTS: We report a unique case of an adult female with vocal fold immobility that was treated with injection laryngoplasty using carboxymethylcellulose and subsequently developed a local reaction requiring intubation and tracheostomy placement. CONCLUSION: Otolaryngologists should be aware of this rare, but life-threatening complication and counsel patients accordingly when obtaining consent. If there are signs and symptoms of airway edema, the patient should be transferred to the ICU for airway watch, intravenous steroids, and possible intubation.

Single-cell analysis reveals prognostic fibroblast subpopulations linked to molecular and immunological subtypes of lung cancer.

Fibroblasts are poorly characterised cells that variably impact tumour progression. Here, we use single cell RNA-sequencing, multiplexed immunohistochemistry and digital cytometry (CIBERSORTx) to identify and characterise three major fibroblast subpopulations in human non-small cell lung cancer: adventitial, alveolar and myofibroblasts. Alveolar and adventitial fibroblasts (enriched in control tissue samples) localise to discrete spatial niches in histologically normal lung tissue and indicate improved overall survival rates when present in lung adenocarcinomas (LUAD). Trajectory inference identifies three phases of control tissue fibroblast activation, leading to myofibroblast enrichment in tumour samples: initial upregulation of inflammatory cytokines, followed by stress-response signalling and ultimately increased expression of fibrillar collagens. Myofibroblasts correlate with poor overall survival rates in LUAD, associated with loss of epithelial differentiation, TP53 mutations, proximal molecular subtypes and myeloid cell recruitment. In squamous carcinomas myofibroblasts were not prognostic despite being transcriptomically equivalent. These findings have important implications for developing fibroblast-targeting strategies for cancer therapy.

Open Bedside Tracheostomy: Safe and Cost Saving but Underutilized Nationally.

OBJECTIVE: To evaluate open bedside tracheostomy (OBT) and compare it with open operating room (OR) tracheostomy and bedside percutaneous dilatational tracheostomy (PDT) in complications and cost. To determine the tracheostomy practice patterns of academic otolaryngology programs. STUDY DESIGN: Retrospective cohort study and cross-sectional study. SETTING: Public hospital and tertiary care hospital. METHODS: Otolaryngology program directors were surveyed to determine their institutions' tracheostomy practice patterns and the factors preventing the implementation of open bedside tracheostomies. A retrospective chart review was done of tracheostomies performed at our institutions from 2009 to 2019 for prolonged mechanical ventilation. Complications, length of intubation, comorbidities, body mass index, demographics, mortality rates, and decannulation rates were recorded. A cost analysis between OBT and PDT was conducted. RESULTS: Data from 802 patients were analyzed for 449 OBTs, 206 PDTs, and 147 open OR tracheostomies. Complication rates were low. PDTs were more likely to have perioperative tracheal bleeding (P = .028) and mucus plugging (P = .006). OBTs were performed on sicker patients with a higher Charlson Comorbidity Index than PDT and OR tracheostomies. The cost of OBT was less than that of PDT. The survey response rate of tracheostomy practice patterns was 46%. The otolaryngologists at the responding programs all conducted OR tracheostomies, while 52.7% did OBTs and 30.9% PDTs. CONCLUSION: OBT can be done safely in patients with multiple comorbidities and has a cost that can be less than PDT. Despite these benefits, only 50% of academic institutions routinely performed OBT.

Genetic Analysis of Colletotrichum siamense Populations from Different Hosts and Counties in Hainan, China, Using Microsatellite Markers.

Colletotrichum siamense was demonstrated as the dominant species among Colletotrichum spp. that infected rubber tree, areca palm, and coffee in Hainan, China. However, the extent of genetic differentiation within the species C. siamense in relation to geographical regions and host species is not known. In this study, 112 C. siamense isolates were genotyped with 12 microsatellite markers. In total, there were 99 multilocus genotypes. Results from permutational multivariate analysis of variance and analysis of molecular variance indicated that there was no significant genetic differentiation between fungal populations with respect to host, location (county), and year. Discriminant analysis of principal components and STRUCTURE analysis showed that C. siamense isolates grouped into three clusters; further analysis confirmed that there were significant (P < 0.001) genetic differences among the three clusters. However, each cluster had isolates from different hosts, counties, or years, supporting the lack of genetic differentiation with respect to host, county, and year. Statistical analyses of allelic associations indicated some evidence for recombination within the populations defined on the basis of host or county. The present findings provide insights into the genetic structure of C. siamense on the three perennial host species in Hainan and suggest that the disease on these three crops can be effectively considered as one disease and, hence, needs to be controlled simultaneously in mixed plantations.

Virtual Versus In-Person Head and Neck Physical Examination Training in Medical Students: A Pilot Assessment.

OBJECTIVE: To compare the effect of virtual and in-person head and neck physical examination training events on medical student confidence in performing examination maneuvers and seeking mentorship from otolaryngology faculty and residents. METHODS: Training events were held with first-year medical student volunteers in 2020 (in-person) and 2021 (virtual). Participants in both cohorts were given didactics on head and neck cancer, trained to perform a head and neck physical examination, and demonstrated their clinical skills to otolaryngology faculty and residents. Pre- and post-training surveys were utilized to assess the following outcomes: participant head and neck cancer knowledge, confidence in performing examination maneuvers, and confidence in seeking mentorship in otolaryngology. Differences in outcomes between training settings were assessed by comparing participant survey responses pre- and post- training. RESULTS: Both in-person and virtual training modalities improved participant confidence in performing the physical examination. There was no significant difference in the degree of improvement between training types. In-person training significantly increased participant confidence in seeking mentorship from otolaryngology faculty and residents (P = .003), while virtual training did not (P = .194). CONCLUSION: Virtual training modalities are feasible methods of teaching the head and neck physical examination. Instruction through a video conferencing platform has the potential to be incorporated into traditional in-person medical education in a permanent fashion. This pilot study can inform future studies directly comparing in-person and virtual physical examination training modalities.

Transcriptional programming of immunoregulatory responses in human Langerhans cells.

Human epidermal Langerhans cells (LCs) maintain immune homeostasis in the skin. To examine transcriptional programming of human primary LCs during homeostasis, we performed scRNA-seq analysis of LCs before and after migration from the epidermis, coupled with functional assessment of their regulatory T cell priming capabilities. The analysis revealed that steady-state LCs exist in a continuum of maturation states and upregulate antigen presentation genes along with an immunoregulatory module including the genes IDO1, LGALS1, LAMTOR1, IL4I, upon their migration. The migration-induced transition in genomic state is accompanied by the ability of LCs to more efficiently prime regulatory T cell responses in co-culture assays. Computational analyses of the scRNAseq datasets using SCENIC and Partial Information Decomposition in Context identified a set of migration-induced transcription factors including IRF4, KLF6 and RelB as key nodes within a immunoregulatory gene regulatory network. These findings support a model in which efficient priming of immunoregulatory responses by LCs is dependent on coordinated upregulation of a migration-coupled maturation program with a immunoregulation-promoting genomic module.

Effective control of Leptosphaeria maculans increases importance of L. biglobosa as a cause of phoma stem canker epidemics on oilseed rape.

BACKGROUND: Phoma stem canker is a damaging disease of oilseed rape caused by two related fungal species, Leptosphaeria maculans and L. biglobosa. However, previous work has mainly focused on L. maculans and there has been little work on L. biglobosa. This work provides evidence of the importance of L. biglobosa to stem canker epidemics in the UK. RESULTS: Quantification of L. maculans and L. biglobosa DNA using species-specific quantitative PCR showed that L. biglobosa caused both upper stem lesions and stem base cankers on nine oilseed rape cultivars in the UK. Upper stem lesions were mainly caused by L. biglobosa. For stem base cankers, there was more L. maculans DNA than L. biglobosa DNA in the susceptible cultivar Drakkar, while there was more L. biglobosa DNA than L. maculans DNA in cultivars with the resistance gene Rlm7 against L. maculans. The frequency of L. biglobosa detected in stem base cankers increased from 14% in 2000 to 95% in 2013. Ascospores of L. biglobosa and L. maculans were mostly released on the same days and the number of L. biglobosa ascospores in air samples increased from the 2010/2011 to 2012/2013 growing seasons. CONCLUSION: Effective control of L. maculans increased infection by L. biglobosa, causing severe upper stem lesions and stem base cankers, leading to yield losses. The importance of L. biglobosa to phoma stem canker epidemics can no longer be ignored. Effective control of phoma stem canker epidemics needs to target both L. maculans and L. biglobosa. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Osteoclast-derived extracellular vesicles are implicated in sensory neurons sprouting through the activation of epidermal growth factor signaling.

BACKGROUND: Different pathologies, affecting the skeletal system, were reported to display altered bone and/or cartilage innervation profiles leading to the deregulation of the tissue homeostasis. The patterning of peripheral innervation is achieved through the tissue-specific expression of attractive or repulsive axonal guidance cues in specific space and time frames. During the last decade, emerging findings attributed to the extracellular vesicles (EV) trading a central role in peripheral tissue innervation. However, to date, the contribution of EV in controlling bone innervation is totally unknown. RESULTS: Here we show that sensory neurons outgrowth induced by the bone resorbing cells-osteoclasts-is promoted by osteoclast-derived EV. The EV induced axonal growth is achieved by targeting epidermal growth factor receptor (EGFR)/ErbB2 signaling/protein kinase C phosphorylation in sensory neurons. In addition, our data also indicate that osteoclasts promote sensory neurons electrophysiological activity reflecting a possible pathway in nerve sensitization in the bone microenvironment, however this effect is EV independent. CONCLUSIONS: Overall, these results identify a new mechanism of sensory bone innervation regulation and shed the light on the role of osteoclast-derived EV in shaping/guiding bone sensory innervation. These findings provide opportunities for exploitation of osteoclast-derived EV based strategies to prevent and/or mitigate pathological uncontrolled bone innervation.