Xist ribonucleoproteins promote female sex-biased autoimmunity.

Autoimmune diseases disproportionately affect females more than males. The XX sex chromosome complement is strongly associated with susceptibility to autoimmunity. Xist long non-coding RNA (lncRNA) is expressed only in females to randomly inactivate one of the two X chromosomes to achieve gene dosage compensation. Here, we show that the Xist ribonucleoprotein (RNP) complex comprising numerous autoantigenic components is an important driver of sex-biased autoimmunity. Inducible transgenic expression of a non-silencing form of Xist in male mice introduced Xist RNP complexes and sufficed to produce autoantibodies. Male SJL/J mice expressing transgenic Xist developed more severe multi-organ pathology in a pristane-induced lupus model than wild-type males. Xist expression in males reprogrammed T and B cell populations and chromatin states to more resemble wild-type females. Human patients with autoimmune diseases displayed significant autoantibodies to multiple components of XIST RNP. Thus, a sex-specific lncRNA scaffolds ubiquitous RNP components to drive sex-biased immunity.

Organization of the human intestine at single-cell resolution.

The intestine is a complex organ that promotes digestion, extracts nutrients, participates in immune surveillance, maintains critical symbiotic relationships with microbiota and affects overall health1. The intesting has a length of over nine metres, along which there are differences in structure and function2. The localization of individual cell types, cell type development trajectories and detailed cell transcriptional programs probably drive these differences in function. Here, to better understand these differences, we evaluated the organization of single cells using multiplexed imaging and single-nucleus RNA and open chromatin assays across eight different intestinal sites from nine donors. Through systematic analyses, we find cell compositions that differ substantially across regions of the intestine and demonstrate the complexity of epithelial subtypes, and find that the same cell types are organized into distinct neighbourhoods and communities, highlighting distinct immunological niches that are present in the intestine. We also map gene regulatory differences in these cells that are suggestive of a regulatory differentiation cascade, and associate intestinal disease heritability with specific cell types. These results describe the complexity of the cell composition, regulation and organization for this organ, and serve as an important reference map for understanding human biology and disease.

Systematically Assessing Natural Compounds' Wound Healing Potential with Spheroid and Scratch Assays.

Understanding cellular processes involved in wound healing is very important given that there are diseases, such as diabetes, in which wounds do not heal. To model tissue regeneration, we focus on two cellular processes: cellular proliferation, to replace cells lost to the wound, and cell motility, activated at the wound edges. We address these two processes in separate, drug responsive, in vitro models. The first model is a scaffold-free three-dimensional (3D) spheroid model, in which spheroids grow larger - to a certain extent - with increased time in culture. The second model, the scratch wound assay, is focused on cell motility. In conjunction with collagen staining, it analyzes changes to the coverage of the wound edge and wound bed. Our workflow gives insights into candidate compounds for wound healing as we show using manuka honey (MH) as an example. Spheroids are responsive to oxidative damage by hydrogen peroxide (H2O2) which affects viability but mostly produces disaggregation. Conversely, MH supports spheroid health, shown by size measurements and viability. In two-dimensional scratch wound assays, MH helps close wounds with relative less collagen production and increases the loose cellular coverage adjacent to and within the wound. We use these methods in the undergraduate research laboratory as teaching and standardization tools, and we hope these will be useful in similar settings.

Single-cell analyses define a continuum of cell state and composition changes in the malignant transformation of polyps to colorectal cancer.

To chart cell composition and cell state changes that occur during the transformation of healthy colon to precancerous adenomas to colorectal cancer (CRC), we generated single-cell chromatin accessibility profiles and single-cell transcriptomes from 1,000 to 10,000 cells per sample for 48 polyps, 27 normal tissues and 6 CRCs collected from patients with or without germline APC mutations. A large fraction of polyp and CRC cells exhibit a stem-like phenotype, and we define a continuum of epigenetic and transcriptional changes occurring in these stem-like cells as they progress from homeostasis to CRC. Advanced polyps contain increasing numbers of stem-like cells, regulatory T cells and a subtype of pre-cancer-associated fibroblasts. In the cancerous state, we observe T cell exhaustion, RUNX1-regulated cancer-associated fibroblasts and increasing accessibility associated with HNF4A motifs in epithelia. DNA methylation changes in sporadic CRC are strongly anti-correlated with accessibility changes along this continuum, further identifying regulatory markers for molecular staging of polyps.

Imaging Sub-Cellular Methionine and Insulin Interplay in Triple Negative Breast Cancer Lipid Droplet Metabolism.

Triple negative breast cancer (TNBC) is a particularly aggressive cancer subtype that is difficult to diagnose due to its discriminating epidemiology and obscure metabolome. For the first time, 3D spatial and chemometric analyses uncover the unique lipid metabolome of TNBC under the tandem modulation of two key metabolites - insulin and methionine - using non-invasive optical techniques. By conjugating heavy water (D2O) probed Raman scattering with label-free two-photon fluorescence (TPF) microscopy, we observed altered de novo lipogenesis, 3D lipid droplet morphology, and lipid peroxidation under various methionine and insulin concentrations. Quantitative interrogation of both spatial and chemometric lipid metabolism under tandem metabolite modulation confirms significant interaction of insulin and methionine, which may prove to be critical therapeutic targets, and proposes a powerful optical imaging platform with subcellular resolution for metabolic and cancer research.

Proportion of U.S. Clinics Offering LGBT-Tailored Mental Health Services Decreased Over Time: A Panel Study of the National Mental Health Services Survey.

Our objective was to characterize the proportion of U.S. mental health clinics that offered LGBT-tailored mental health services between 2014 and 2018. We used data from the National Mental Health Services Survey (NMHSS) to construct a mixed logistic model of availability of LGBT-tailored mental health services over time, by region (Northeast, South, Midwest and West), and by facility type (Veterans Administration, inpatient/residential, outpatient, community mental health centers and mixed). Our results show that the overall proportion of mental health clinics that offered LGBT-tailored services decreased from 2014 to 2018. Our results also indicate that Veteran Affairs clinics and facilities in the West and Northeast were most likely to offer LGBT-tailored mental health services. Given the temporal, regional, and facility gaps in LGBT-tailored mental health services availability, more effort should be dedicated to addressing this disparity.

The Role of Virtual Reality in Screening, Diagnosing, and Rehabilitating Spatial Memory Deficits.

Impairment of spatial memory, including an inability to recall previous locations and navigate the world, is often one of the first signs of functional disability on the road to cognitive impairment. While there are many screening and diagnostic tools which attempt to measure spatial memory ability, they are often not representative of real-life situations and can therefore lack applicability. One potential solution to this problem involves the use of virtual reality (VR), which immerses individuals in a virtually-simulated environment, allowing for scenarios more representative of real-life without any of the associated risks. Here, we review the evidence surrounding the use of VR for the screening and diagnosis of spatial memory impairments, including potential limitations and how it compares to standard neuropsychological tests. We will also discuss the evidence regarding the potential use of VR in the rehabilitation of spatial memory deficits, which has not been well studied, but which could be game-changing if proven successful.

Challenges in the development and establishment of exosome-based drug delivery systems.

Exosomes are extracellular vesicles released from cells and are characterized by a lipid bilayer membrane encapsulating a variety of biological molecules such as nucleic acids or proteins within the lumen or the lipid-bilayer. Under physiological environments, exosomes mediate cell-to-cell communication and cargo transport. Therefore, exosomes have been explored as drug delivery vehicles for improving therapeutic outcomes. Although recent studies have demonstrated promising advances with exosome-based drug delivery systems, several challenges severely hinder further development of exosomes for clinical applications. This review summarizes and emphasizes some of the technical challenges related to the isolation, characterization, and stability testing of exosomes. More importantly, challenges related specifically to the application of exosomes for drug delivery such as cell-uptake, drug loading, drug release, and in vivo distribution will be examined in this article.

Association of State Policies Allowing Medical Cannabis for Opioid Use Disorder With Dispensary Marketing for This Indication.

Importance: Misinformation about cannabis and opioid use disorder (OUD) may increase morbidity and mortality if it leads individuals with OUD to forego evidence-based treatment. It has not been systematically evaluated whether officially designating OUD as a qualifying condition for medical cannabis is associated with cannabis dispensaries suggesting cannabis as a treatment for OUD. Objective: To examine whether state-level policies designating OUD a qualifying condition for medical cannabis are associated with more dispensaries claiming cannabis can treat OUD. Design, Setting, and Participants: This cross-sectional, mixed-methods study of 208 medical dispensary brands was conducted in 2019 using the brands' online content. The study included dispensaries operating in New Jersey, New York, and Pennsylvania, where OUD is a qualifying condition for medical cannabis, and in Connecticut, Delaware, Maryland, Ohio, and West Virginia, where this policy does not exist. Exposures: Presence of OUD on the list of qualifying conditions for a state's medical cannabis program. Main Outcomes and Measures: Binary indicators of whether online content from the brand said cannabis can treat OUD, can replace US Food and Drug Administration-approved medications for OUD, can be an adjunctive therapy to Food and Drug Administration-approved medications for OUD, or can be used as a substitute for opioids to treat other conditions (eg, chronic pain). Results: After excluding duplicates, listings for nonexistent dispensaries, and those without online content, 167 brands across 7 states were included in the analysis (44 [26.3%] in states where OUD was a qualifying condition and 123 [73.7%] in adjacent states). A dispensary listed in a directory for West Virginia was not operational; therefore, comparison states were Connecticut, Delaware, Maryland, and Ohio. In policy-exposed states, 39% (95% CI, 23%-55%) more dispensaries claimed cannabis could treat OUD compared with unexposed states (P < .001). For replacing medications for OUD and being an adjunctive therapy, the differences were 14% (95% CI, 2%-26%; P = .002) and 28% (95% CI, 14%-42%; P < .001), respectively. The suggestion that cannabis could substitute for opioids (eg, to treat chronic pain) was made by 25% (95% CI, 9%-41%) more brands in policy-exposed states than adjacent states (P = .002). Conclusions and Relevance: In this study, state-level policies designating OUD as a qualifying condition for medical cannabis were associated with more dispensaries claiming cannabis can treat OUD. In the current policy environment, in which medical claims by cannabis dispensaries are largely unregulated, these advertisements could harm patients. Future research linking these policies to patient outcomes is warranted.

Adult Popillia japonica as an Otorhinolaryngologic Invasive Foreign Body in a Rural Area.

Otorhinolaryngologic foreign bodies may be encountered in-office visits, the emergency department, and speciality consultations. These include food, toys, and other small items, are present in pediatric patients. Because patients may be asymptomatic and the insertion of the foreign body not observed, obtaining medical care may be delayed. Conversely, insects as foreign bodies, especially in the external ear canal, can cause a patient significant pain and distress, directing the patient to seek immediate care. Here, we present a case of an adult Japanese beetle (Popillia japonica) as a foreign body in the ear of a 14-year-old female. A review of otorhinolaryngologic foreign bodies is also discussed, with particular attention to the ear and rural location. This case highlights the potential for agricultural insects to act as invasive foreign bodies, especially in areas where they are known to be endemic pests and the consequences of delayed treatment.

Does the Format of Preclass Reading Quizzes Matter? An Evaluation of Traditional and Gamified, Adaptive Preclass Reading Quizzes.

Preclass reading quizzes (RQs) have been shown to enhance student performance. Many instructors implementing evidence-based teaching assign preclass RQs to ensure their students are prepared to engage in class activities. Textbook companies now offer a gamified, adaptive-learning RQ format. In these RQs, students answer point-valued questions until they reach a threshold. If students answer incorrectly, the question decreases in point value on the next attempt. These RQs also give students who answer questions incorrectly more questions on that topic and direct students to sections of a textbook they need to review. We assessed the impact of gamified, adaptive preclass RQs compared with more traditional preclass RQs on in-class RQs and course exam performance as well as students' perceptions of RQs. Students in the gamified, adaptive treatment performed equally compared with students in the traditional, static treatment on in-class RQs and course exams. While students in the gamified, adaptive treatment did have a more positive perception of preclass RQs, this factor explained less than 3% of the variation in RQ perception. Our findings suggest that instructors should verify that gamified, adaptive technologies impact student learning in their course before integrating them into their course and asking students to pay for them.

Principal component analysis of binding energies for single-point mutants of hT2R16 bound to an agonist correlate with experimental mutant cell response.

Directed evolution is a technique that enables the identification of mutants of a particular protein that carry a desired property by successive rounds of random mutagenesis, screening, and selection. This technique has many applications, including the development of G protein-coupled receptor-based biosensors and designer drugs for personalized medicine. Although effective, directed evolution is not without challenges and can greatly benefit from the development of computational techniques to predict the functional outcome of single-point amino acid substitutions. In this article, we describe a molecular dynamics-based approach to predict the effects of single amino acid substitutions on agonist binding (salicin) to a human bitter taste receptor (hT2R16). An experimentally determined functional map of single-point amino acid substitutions was used to validate the whole-protein molecular dynamics-based predictive functions. Molecular docking was used to construct a wild-type agonist-receptor complex, providing a starting structure for single-point substitution simulations. The effects of each single amino acid substitution in the functional response of the receptor to its agonist were estimated using three binding energy schemes with increasing inclusion of solvation effects. We show that molecular docking combined with molecular mechanics simulations of single-point mutants of the agonist-receptor complex accurately predicts the functional outcome of single amino acid substitutions in a human bitter taste receptor.

The phagosomal nutrient transporter (Pht) family.

Phagosomal transporters (Phts), required for intracellular growth of Legionella pneumophila, comprise a novel family of multispanning alpha-helical proteins within the major facilitator superfamily (MFS). The members of this family derive exclusively from bacteria. Multiple paralogues are present in a restricted group of Alpha- and Gammaproteobacteria, but single members were also found in Chlamydia and Cyanobacteria. Their protein sequences were aligned, yielding a phylogenetic tree showing the relations of the proteins to each other. Topological analyses revealed a probable 12 alpha-helical transmembrane segment (TMS) topology. Motif identification and statistical analyses provided convincing evidence that these proteins arose from a six TMS precursor by intragenic duplication. The phylogenetic tree revealed some potential orthologous relationships, suggestive of common function. However, several probable examples of lateral transfer of the encoding genetic material between bacteria were identified and analysed. The Pht family most closely resembles a smaller MFS family (the UMF9 family) with no functionally characterized members. However, the UMF9 family occurs in a broader range of prokaryotic organism types, including Archaea. These two families differ in that organisms bearing members of the Pht family often have numerous paralogues, whereas organisms bearing members of the UMF9 family never have more than two. This work serves to characterize two novel families within the MFS and provides compelling evidence for horizontal transfer of some of the family members.

Bronchoscopic diagnosis of an extensive tracheal laceration.

Tracheal lacerations resulting from endotracheal intubation are extremely rare. We report a case where the initial diagnosis was misled by the radiological findings. Our case uniquely emphasizes several issues pertinent to the management of tracheal lacerations. We review the published works on this topic and emphasize the role of fibre-optic bronchoscopy in the assessment of airway injuries before operative management.