Hair follicle-resident progenitor cells are a major cellular contributor to heterotopic subcutaneous ossifications in a mouse model of Albright hereditary osteodystrophy.

Heterotopic ossifications (HOs) are the pathologic process by which bone inappropriately forms outside of the skeletal system. Despite HOs being a persistent clinical problem in the general population, there are no definitive strategies for their prevention and treatment due to a limited understanding of the cellular and molecular mechanisms contributing to lesion development. One disease in which the development of heterotopic subcutaneous ossifications (SCOs) leads to morbidity is Albright hereditary osteodystrophy (AHO). AHO is caused by heterozygous inactivation of GNAS, the gene that encodes the α-stimulatory subunit (Gαs) of G proteins. Previously, we had shown using our laboratory's AHO mouse model that SCOs develop around hair follicles (HFs). Here we show that SCO formation occurs due to inappropriate expansion and differentiation of HF-resident stem cells into osteoblasts. We also show in AHO patients and mice that Secreted Frizzled Related Protein 2 (SFRP2) expression is upregulated in regions of SCO formation and that elimination of Sfrp2 in male AHO mice exacerbates SCO development. These studies provide key insights into the cellular and molecular mechanisms contributing to SCO development and have implications for potential therapeutic modalities not only for AHO patients but also for patients suffering from HOs with other etiologies.

Optimising Patient Outcomes in Tongue Cancer: A Multidisciplinary Approach.

A multidisciplinary approach to the management of tongue cancer is vital for achieving optimal patient outcomes. Nursing and allied health professionals play essential roles within the team. We developed symposia comprising a series of online lectures offering a detailed perspective on the role each discipline and consumer perspective has in the management of patients with tongue cancer. The topics, including epidemiology and prevention, diagnosis, treatment planning, surgery, adjuvant care, and the management of recurrent or metastatic disease, were thoroughly examined. The symposia highlighted the significance of fostering collaboration and continuous learning through a multidisciplinary approach. This initiative should be relevant to healthcare professionals, researchers, and policymakers striving to enhance patient outcomes in tongue cancer care through innovative collaboration.

vSPACE: Exploring Virtual Spatial Representation of Articular Chondrocytes at the Single-Cell Level.

Single cell RNA sequencing technology has been dramatically changing how gene expression studies are performed. However, its use has been limited to identifying subtypes of cells by comparing cells' gene expression levels in an unbiased manner to produce a 2D plot (e.g., UMAP/tSNE). We developed a new method of placing cells in 2D space. This system, called vSPACE, shows a virtual spatial representation of scRNAseq data obtained from human articular cartilage by emulating the concept of spatial transcriptomics technology, but virtually. This virtual 2D plot presentation of human articular cartage cells generates several zonal distribution patterns, in one or multiple genes at a time, reveling patterns that scientists can appreciate as imputed spatial distribution patterns along the zonal axis. The discovered patterns are explainable and remarkably consistent across all six healthy doners despite their respectively different clinical variables (age and sex), suggesting the confidence of the discovered patterns.

CGCom: a framework for inferring Cell-cell Communication based on Graph Neural Network.

Cell-cell communication is crucial in maintaining cellular homeostasis, cell survival and various regulatory relationships among interacting cells. Thanks to recent advances of spatial transcriptomics technologies, we can now explore if and how cells' proximal information available from spatial transcriptomics datasets can be used to infer cell-cell communication. Here we present a cell-cell communication inference framework, called CGCom, which uses a graph neural network (GNN) to learn communication patterns among interacting cells by combining single-cell spatial transcriptomic datasets with publicly available ligand-receptor information and the molecular regulatory information down-stream of the ligand-receptor signaling. To evaluate the performance of CGCom, we applied it to mouse embryo seqFISH datasets. Our results demonstrate that CGCom can not only accurately infer cell communication between individual cell pairs but also generalize its learning to predict communication between different cell types. We compared the performance of CGCom with two existing methods, CellChat and CellPhoneDB, and our comparative study revealed both common and unique communication patterns from the three approaches. Commonly found communication patterns include three sets of ligand-receptor communication relationships, one between surface ectoderm cells and spinal cord cells, one between gut tube cells and endothelium, and one between neural crest and endothelium, all of which have already been reported in the literature thus offering credibility of all three methods. However, we hypothesize that CGCom is superior in reducing false positives thanks to its use of cell proximal information and its learning between specific cell pairs rather than between cell types. CGCom is a GNN-based solution that can take advantage of spatially resolved single-cell transcriptomic data in predicting cell-cell communication with a higher accuracy.

Modeling of intramembranous ossification using human pluripotent stem cell-derived paraxial mesoderm derivatives.

Vertebrates form their skeletal tissues from three distinct origins (the neural crest, paraxial mesoderm, and lateral plate mesoderm) through two distinct modes of ossification (intramembranous and endochondral ossification). Since the paraxial mesoderm generates both intramembranous and endochondral bones, it is thought to give rise to both osteoprogenitors and osteo-chondroprogenitors. However, it remains unclear what directs the paraxial mesoderm-derived cells toward these different fates in distinct skeletal elements during human skeletal development. To answer this question, we need experimental systems that recapitulate paraxial mesoderm-mediated intramembranous and endochondral ossification processes. In this study, we aimed to develop a human pluripotent stem cell (hPSC)-based system that models the human intramembranous ossification process. We found that spheroid culture of the hPSC-derived paraxial mesoderm derivatives generates osteoprogenitors or osteo-chondroprogenitors depending on stimuli. The former induced intramembranous ossification, and the latter endochondral ossification, in mouse renal capsules. Transcriptional profiling supported the notion that bone signatures were enriched in the intramembranous bone-like tissues. Thus, we developed a system that recapitulates intramembranous ossification, and that enables the induction of two distinct modes of ossification by controlling the cell fate of the hPSC-derived paraxial mesoderm derivatives.

Death Due to Drug Reaction with Eosinophilia and Systemic Symptoms (DRESS) Syndrome: A Case Report.

Background: Drug reaction with eosinophilia and systemic symptoms (DRESS) syndrome is a life-threatening, multi-organ adverse drug reaction with an incidence of 1 in 1000 to 1 in 10 000 high-risk drug exposures. Case Presentation: An elderly female presented to the hospital with progressive weakness and a diffuse erythematous macular rash covering most of her body that started 3 days prior. Over the next 3 days, the patient quickly deteriorated, developing disorientation with acute onset left-sided weakness, leukocytosis, thrombocytopenia, eosinophilia, liver and kidney failure, and hypoxia. Clinical and histological changes supported the diagnosis of DRESS syndrome caused by intravenous (IV) ampicillin administered during a prior hospitalization for a urinary tract infection. Systemic corticosteroids were initiated quickly thereafter, but the patient ultimately succumbed to complications caused by DRESS syndrome. Conclusion: There are currently no randomized trials evaluating treatments for DRESS, and evidenced-based guidelines are lacking. Viral reactivation has also been suggested as a possible complication of DRESS syndrome, though the true incidence and association remain unclear. Although we started our patient on high-dose IV corticosteroids early in her course, she still succumbed to complications of DRESS syndrome. Further research into the treatment of DRESS syndrome and its association with viral reactivation is essential.

A Comparison of Objectively Measured Free-Living Physical Behaviour in Adults with and without Lower Limb Amputation.

Objectively monitored free-living physical behaviours of adults with and without lower limb amputation (LLA) were compared. METHODS: 57 adults with LLA wore an activPAL3™ for 8 days. A comparison data set (n = 57) matched on gender, age and employment status was used. Variables included: time sitting; standing; stepping; sit-to-stand transitions; step count and cadence. Comparisons were made between adults with and without LLA and between gender, level and cause of amputation. RESULTS: Participants with LLA due to trauma versus circulatory causes were less sedentary and more active; however, no difference in physical behaviour was recorded across gender or level of amputation. Participants with LLA spent more time sitting (p < 0.001), less time standing and stepping (p < 0.001) and had a lower step count (p < 0.001). Participants with LLA took more steps in cadence bands less than 100 steps·min-1 and fewer steps in cadence bands greater than 100 steps·min-1 compared to participants without LLA. CONCLUSIONS: People with LLA were less active and more sedentary than people without LLA and participated in less activity at a moderate or higher intensity when matched on age, gender and employment. Interventions are needed to promote active lifestyles in this population.

Media Framing of Women's Football During the COVID-19 Pandemic.

This article examines British media coverage of women's association football during the coronavirus (COVID-19) pandemic, to identify how the media framed the women's game and how these frames could shape the public perceptions of it. Through a database search of British-based news coverage of women's football, 100 news articles were identified in the first 6 months after the start of the pandemic. A thematic analysis was conducted, and five dominant frames were detected in the context of COVID-19: 1) financial precariousness of women's football; 2) the commercial prioritisation of men's football; 3) practical consideration of the sport (e.g., alterations to national and international competitions); 4) debating the future of women's football and 5) concern for players (e.g., welfare, uncertain working conditions). These frames depart from the past trivialisation and sexualisation of women's sport, demonstrate the increased visibility of women's football, and shift the narrative towards the elite stratum of the game. Most of this reporting was by women journalists, while men were shown to write less than women about women's football. This research advocates continued diversification of the sports journalism workforce to dissolve the hegemonic masculine culture that still largely dominates the industry.

Stem cell-based modeling and single-cell multiomics reveal gene-regulatory mechanisms underlying human skeletal development.

Although the skeleton is essential for locomotion, endocrine functions, and hematopoiesis, the molecular mechanisms of human skeletal development remain to be elucidated. Here, we introduce an integrative method to model human skeletal development by combining in vitro sclerotome induction from human pluripotent stem cells and in vivo endochondral bone formation by implanting the sclerotome beneath the renal capsules of immunodeficient mice. Histological and scRNA-seq analyses reveal that the induced bones recapitulate endochondral ossification and are composed of human skeletal cells and mouse circulatory cells. The skeletal cell types and their trajectories are similar to those of human embryos. Single-cell multiome analysis reveals dynamic changes in chromatin accessibility associated with multiple transcription factors constituting cell-type-specific gene-regulatory networks (GRNs). We further identify ZEB2, which may regulate the GRNs in human osteogenesis. Collectively, these results identify components of GRNs in human skeletal development and provide a valuable model for its investigation.

Ultrasound-derived mechanical stimulation of cell-laden collagen hydrogels for bone repair.

Cell therapy is emerging as an effective treatment strategy for many diseases. Here we describe a novel approach to bone tissue repair that combines hydrogel-based cell therapy with low intensity pulsed ultrasound (LIPUS), an FDA approved treatment for fracture repair. Bone marrow-derived stromal cells (BMSCs) have been encapsulated in type I collagen hydrogels and mechanically stimulated using LIPUS-derived acoustic radiation force (ARF). We observed the expression and upward trend of load-sensitive, osteoblast-specific markers and determined that the extent of cell response is dependent on an optimal combination of both hydrogel stiffness and ARF intensity. Specifically, cells encapsulated in hydrogels of optimal stiffness respond at the onset of ultrasound by upregulating early bone-sensitive markers such as calcium, cyclooxygenase-2, and prostaglandin E2 , and later by supporting mineralized tissue formation after 21 days of culture. In vivo evaluation of a critical size calvarial defect in NOD scid gamma (NSG) mice indicated that the implantation of BMSC-laden hydrogels of optimal stiffness improved healing of calvarial defects after daily administration of ARF over 4 weeks. Collectively, these findings validate the efficacy of our system of localized cell delivery for treating bone defects where undifferentiated BMSCs are induced to the osteoblastic lineage. Further, in vivo healing may be enhanced via non-invasive transdermal mechanical stimulation of implanted cells using ARF.

Comparison of virtual reality and physical simulation training in first-year radiography students in South America.

INTRODUCTION: The aim of this study was to comparatively evaluate the learning outcomes achieved by first-year radiography students educated with either virtual reality (VR) simulation training or physical simulation training. The implementation of VR has been proposed to enhance learning in radiography students and provide a more effective and efficient approach to simulation. However, the learning outcomes achieved with this approach have not been widely investigated. METHODS: Through stratified randomisation, 188 radiography students were allocated to one of two matched groups: a VR group (using Virtual Medical Coaching's Radiography simulation) and a physical simulation group (using Philips' X-ray equipment). Both groups were taught 31 radiography views over one 25-week semester. Both groups were assessed in an Objective Structured Clinical Examination (OSCE), using actors as patients in a physical X-ray environment. Assessment was conducted by assigning objective count scores for five assessment criteria. RESULTS: The VR group achieved shorter OSCE duration and fewer errors in moving equipment and patient positioning: these results were statistically significant (P < 0.00). There was no significant difference in the frequency of errors in radiographic exposure setting between the VR and the physical simulation group. The current findings concur with the limited number of published studies concerning VR simulation in radiography. CONCLUSIONS: The results of this study demonstrated superior effectiveness and efficiency in the VR group. This provides preliminary evidence to introduce VR simulation in the host institution and provide evidence that it may be possible to replace the use of physical simulation across other years of the degree. Further research investigating these possibilities is warranted.

CRISPR-Cas12a induced DNA double-strand breaks are repaired by multiple pathways with different mutation profiles in Magnaporthe oryzae.

CRISPR-Cas mediated genome engineering has revolutionized functional genomics. However, understanding of DNA repair following Cas-mediated DNA cleavage remains incomplete. Using Cas12a ribonucleoprotein genome editing in the fungal pathogen, Magnaporthe oryzae, we detail non-canonical DNA repair outcomes from hundreds of transformants. Sanger and nanopore sequencing analysis reveals significant variation in DNA repair profiles, ranging from small INDELs to kilobase size deletions and insertions. Furthermore, we find the frequency of DNA repair outcomes varies between loci. The results are not specific to the Cas-nuclease or selection procedure. Through Ku80 deletion analysis, a key protein required for canonical non-homologous end joining, we demonstrate activity of an alternative end joining mechanism that creates larger DNA deletions, and uses longer microhomology compared to C-NHEJ. Together, our results suggest preferential DNA repair pathway activity in the genome that can create different mutation profiles following repair, which could create biased genome variation and impact genome engineering and genome evolution.

Prediction of Neonatal Respiratory Distress Biomarker Concentration by Application of Machine Learning to Mid-Infrared Spectra.

The authors of this study developed the use of attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) combined with machine learning as a point-of-care (POC) diagnostic platform, considering neonatal respiratory distress syndrome (nRDS), for which no POC currently exists, as an example. nRDS can be diagnosed by a ratio of less than 2.2 of two nRDS biomarkers, lecithin and sphingomyelin (L/S ratio), and in this study, ATR-FTIR spectra were recorded from L/S ratios of between 1.0 and 3.4, which were generated using purified reagents. The calibration of principal component (PCR) and partial least squares (PLSR) regression models was performed using 155 raw baselined and second derivative spectra prior to predicting the concentration of a further 104 spectra. A three-factor PLSR model of second derivative spectra best predicted L/S ratios across the full range (R2: 0.967; MSE: 0.014). The L/S ratios from 1.0 to 3.4 were predicted with a prediction interval of +0.29, -0.37 when using a second derivative spectra PLSR model and had a mean prediction interval of +0.26, -0.34 around the L/S 2.2 region. These results support the validity of combining ATR-FTIR with machine learning to develop a point-of-care device for detecting and quantifying any biomarker with an interpretable mid-infrared spectrum.

Predicting the targets of IRF8 and NFATc1 during osteoclast differentiation using the machine learning method framework cTAP.

BACKGROUND: Interferon regulatory factor-8 (IRF8) and nuclear factor-activated T cells c1 (NFATc1) are two transcription factors that have an important role in osteoclast differentiation. Thanks to ChIP-seq technology, scientists can now estimate potential genome-wide target genes of IRF8 and NFATc1. However, finding target genes that are consistently up-regulated or down-regulated across different studies is hard because it requires analysis of a large number of high-throughput expression studies from a comparable context. METHOD: We have developed a machine learning based method, called, Cohort-based TF target prediction system (cTAP) to overcome this problem. This method assumes that the pathway involving the transcription factors of interest is featured with multiple "functional groups" of marker genes pertaining to the concerned biological process. It uses two notions, Gene-Present Sufficiently (GP) and Gene-Absent Insufficiently (GA), in addition to log2 fold changes of differentially expressed genes for the prediction. Target prediction is made by applying multiple machine-learning models, which learn the patterns of GP and GA from log2 fold changes and four types of Z scores from the normalized cohort's gene expression data. The learned patterns are then associated with the putative transcription factor targets to identify genes that consistently exhibit Up/Down gene regulation patterns within the cohort. We applied this method to 11 publicly available GEO data sets related to osteoclastgenesis. RESULT: Our experiment identified a small number of Up/Down IRF8 and NFATc1 target genes as relevant to osteoclast differentiation. The machine learning models using GP and GA produced NFATc1 and IRF8 target genes different than simply using a log2 fold change alone. Our literature survey revealed that all predicted target genes have known roles in bone remodeling, specifically related to the immune system and osteoclast formation and functions, suggesting confidence and validity in our method. CONCLUSION: cTAP was motivated by recognizing that biologists tend to use Z score values present in data sets for the analysis. However, using cTAP effectively presupposes assembling a sizable cohort of gene expression data sets within a comparable context. As public gene expression data repositories grow, the need to use cohort-based analysis method like cTAP will become increasingly important.

Parental Origin of Gsα Inactivation Differentially Affects Bone Remodeling in a Mouse Model of Albright Hereditary Osteodystrophy.

Albright hereditary osteodystrophy (AHO) is caused by heterozygous inactivation of GNAS, a complex locus that encodes the alpha-stimulatory subunit of heterotrimeric G proteins (Gsα) in addition to NESP55 and XLαs due to alternative first exons. AHO skeletal manifestations include brachydactyly, brachymetacarpia, compromised adult stature, and subcutaneous ossifications. AHO patients with maternally-inherited GNAS mutations develop pseudohypoparathyroidism type 1A (PHP1A) with resistance to multiple hormones that mediate their actions through G protein-coupled receptors (GPCRs) requiring Gsα (eg, parathyroid hormone [PTH], thyroid-stimulating hormone [TSH], growth hormone-releasing hormone [GHRH], calcitonin) and severe obesity. Paternally-inherited GNAS mutations cause pseudopseudohypoparathyroidism (PPHP), in which patients have AHO skeletal features but do not develop hormonal resistance or marked obesity. These differences between PHP1A and PPHP are caused by tissue-specific reduction of paternal Gsα expression. Previous reports in mice have shown loss of Gsα causes osteopenia due to impaired osteoblast number and function and suggest that AHO patients could display evidence of reduced bone mineral density (BMD). However, we previously demonstrated PHP1A patients display normal-increased BMD measurements without any correlation to body mass index or serum PTH. Due to these observed differences between PHP1A and PPHP, we utilized our laboratory's AHO mouse model to address whether Gsα heterozygous inactivation differentially affects bone remodeling based on the parental inheritance of the mutation. We identified fundamental distinctions in bone remodeling between mice with paternally-inherited (GnasE1+/-p) versus maternally-inherited (GnasE1+/-m) mutations, and these findings were observed predominantly in female mice. Specifically, GnasE1+/-p mice exhibited reduced bone parameters due to impaired bone formation and enhanced bone resorption. GnasE1+/-m mice, however, displayed enhanced bone parameters due to both increased osteoblast activity and normal bone resorption. These in vivo distinctions in bone remodeling between GnasE1+/-p and GnasE1+/-m mice could potentially be related to changes in the bone microenvironment driven by calcitonin-resistance within GnasE1+/-m osteoclasts. Further studies are warranted to assess how Gsα influences osteoblast-osteoclast coupling. © 2021 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

The Effect of Haematocrit on Measurement of the Mid-Infrared Refractive Index of Plasma in Whole Blood.

Recent advances suggest that miniaturised mid-infrared (MIR) devices could replace more time-consuming, laboratory-based techniques for clinical diagnostics. This work uses Fourier transform infrared spectroscopy to show that the MIR complex refractive index of whole blood varies across a range of haematocrit. This indicates that the use of an evanescent measurement is not sufficient to optically exclude the cellular content of blood in the MIR, as previously assumed. Here, spectral refractive index data is presented in two ways. First, it is given as whole blood with varying haematocrit. Second, it is given as the percentage error that haematocrit introduces to plasma. The maximum error in the effective plasma refractive index due to the haematocrit of healthy adults was 0.25% for the real part n and 11% for the imaginary part k. This implies that calibration measurements of haematocrit can be used to account for errors introduced by the cellular content, enabling plasma spectra and analyte concentrations to be indirectly calculated from a whole blood sample. This methodological advance is of clinical importance as plasma concentration of analytes such as drugs can be determined using MIR without the preprocessing of whole blood.

Intensive Dialectical Behavior Treatment for Individuals With Borderline Personality Disorder With and Without Substance Use Disorders.

Treatment of borderline personality disorder (BPD) with comorbid substance use disorder can be challenging due to symptom overlap and limited assessment methods. Preliminary evidence has shown promising effectiveness of dialectical behavioral therapy (DBT) for BPD with comorbid substance use disorders. The current study compared the benefits of a 28-day transitional DBT treatment program for individuals with BPD with and without substance use disorders through evaluating the changes in coping skills, generalized anxiety, and depression symptom scales at admission and discharge. A total of 76 patients were split into two groups: Group 1 consisted of individuals with BPD without substance use disorders (n = 41), and Group 2 involved individuals with BPD and a substance use disorder (SUD) (n = 35). A univariate general linear model showed significant differences between the two groups in improvement of coping skills and depressive symptoms. After a 28-day transitional DBT treatment program there were significant decreases from severe to moderate depression scores in both groups. Our findings support the effectiveness of DBT treatment in patients with comorbid BPD and SUD.

Music Tempo: A Tool for Regulating Walking Cadence and Physical Activity Intensity in Overweight Adults?

This study investigated if music tempo can prompt a desired walking cadence, and if music can provide a stimulus to regulate physical activity intensity in a longitudinal physical activity intervention with free-living adults. Overweight adults (n = 37; 94.26 ± 17.11 kg; 49.63 ± 12.37 years) were randomly assigned to an intervention (IG, n = 17) or usual care group (UC, n = 20) as part of a novel nine-month walking intervention. IG participants walked to self-selected music with a predetermined tempo and received a behavioural change support programme. At baseline, four-, six- and nine-months participants were asked to walk around an elliptical track at their habitual pace (0-2 min) and then in time to a predetermined tempo (2-8 min) designed to elicit moderate intensity. Cadence response (steps/min) was assessed and intensity (heart rate (bpm) recorded using wireless telemetry. A repeated measures general linear model (GLM) examined differences between groups over time (p < 0.05). All data is presented as means ± SD. At each assessment point both groups displayed an immediate cadence adjustment in response to music tempo (p < 0.01) i.e., habitual cadence vs. 3 METs target cadence (p < 0.05) and 3 METs target cadence vs. 5 METs target cadence (p < 0.05). Additionally, IG participants displayed an increased habitual cadence (0-2 min) at each assessment point (p < 0.05; 110 ± 9, 121.80 ± 7.5, 121.46 ± 10, 121.93 ± 7 steps/min respectively). UC participant's habitual cadence was unchanged from 0-9 months (p > 0.05; 120 ± 10, 116 ± 13, 119 ± 12 and 119 ± 9 steps/min respectively). Music tempo may be a useful regulatory tool to prompt the free-living individual to reach an appropriate stride rate to achieve a walking pace that is at least moderate intensity. It also appears that results may be trainable as throughout the study an increased habitual walking cadence was observed, in the absence of music.

Reconstruction of Chronic Wounds Secondary to Injectable Drug Use with a Biodegradable Temporizing Matrix.

Injectable drug use in the upper extremity often leads to chronic wounds complicated by osteomyelitis. Conventional reconstructive options are often not feasible and/or are contraindicated in this patient population. We have started using a synthetic, biodegradable temporizing matrix (BTM) for the treatment of these patients. We hypothesize that BTM is a safe, low-risk, and low-morbidity alternative reconstructive option. We report outcomes after staged debridement and BTM application followed by split-thickness skin grafting for two patients with large, chronic bilateral forearm wounds with concomitant osteomyelitis confirmed by MRI and biopsy. No acute surgical complications were encountered and at a mean follow-up of 13 months, both patients had maintained stable soft-tissue coverage. Reconstruction using BTM is a novel treatment option that can simplify the reconstruction, reduce donor-site morbidity, and optimize success for patients with chronic wounds resulting from injectable drug use. Initial outcomes are promising; however, further comparative studies are needed to better evaluate long-term outcomes of this technique.