Purpose Single cell (sc) analyses of key embryonic, fetal and adult stages were performed to generate a comprehensive single cell atlas of all the corneal and adjacent conjunctival cell types from development to adulthood. Methods Four human adult and seventeen embryonic and fetal corneas from 10 to 21 post conception week (PCW) specimens were dissociated to single cells and subjected to scRNA- and/or ATAC-Seq using the 10x Genomics platform. These were embedded using Uniform Manifold Approximation and Projection (UMAP) and clustered using Seurat graph-based clustering. Cluster identification was performed based on marker gene expression, bioinformatic data mining and immunofluorescence (IF) analysis. RNA interference, IF, colony forming efficiency and clonal assays were performed on cultured limbal epithelial cells (LECs). Results scRNA-Seq analysis of 21,343 cells from four adult human corneas and adjacent conjunctivas revealed the presence of 21 cell clusters, representing the progenitor and differentiated cells in all layers of cornea and conjunctiva as well as immune cells, melanocytes, fibroblasts, and blood/lymphatic vessels. A small cell cluster with high expression of limbal progenitor cell (LPC) markers was identified and shown via pseudotime analysis to give rise to five other cell types representing all the subtypes of differentiated limbal and corneal epithelial cells. A novel putative LPCs surface marker, GPHA2, expressed on the surface of 0.41% ± 0.21 of the cultured LECs, was identified, based on predominant expression in the limbal crypts of adult and developing cornea and RNAi validation in cultured LECs. Combining scRNA- and ATAC-Seq analyses, we identified multiple upstream regulators for LPCs and demonstrated a close interaction between the immune cells and limbal progenitor cells. RNA-Seq analysis indicated the loss of GPHA2 expression and acquisition of proliferative limbal basal epithelial cell markers during ex vivo LEC expansion, independently of the culture method used. Extending the single cell analyses to keratoconus, we were able to reveal activation of collagenase in the corneal stroma and a reduced pool of limbal suprabasal cells as two key changes underlying the disease phenotype. Single cell RNA-Seq of 89,897 cells obtained from embryonic and fetal cornea indicated that during development, the conjunctival epithelium is the first to be specified from the ocular surface epithelium, followed by the corneal epithelium and the establishment of LPCs, which predate the formation of limbal niche by a few weeks. Conclusions Our scRNA-and ATAC-Seq data of developing and adult cornea in steady state and disease conditions provide a unique resource for defining genes/pathways that can lead to improvement in ex vivo LPCs expansion, stem cell differentiation methods and better understanding and treatment of oc...

Fibroblasts have different roles during physiology and disease in regulating myocardial function via soluble mediators. A crosstalk between fibroblasts and cardiomyocytes, controlled by TGF-β, is crucial in this interaction.

These findings suggest that the clinical manifestations of patients with GO may be changing over time in Europe.

The aim of this study was to compare the incidence, nature, and cause of injuries sustained in rugby union played on artificial turf and grass. The study comprised a two-season investigation of match injuries sustained by six teams competing in Hong Kong's Division 1 and training injuries sustained by two teams in the English Premiership. Injury definitions and recording procedures were compliant with the international consensus statement on epidemiological studies of injuries in rugby union. There were no significant differences in the overall incidence (rate ratio = 1.42; P = 0.134) or severity (P = 0.620) of match injuries sustained on the two surfaces. The lower limb and joint (non-bone)/ligament injuries were the most common location and type of match injury on both surfaces; the incidence of anterior cruciate ligament injuries was nearly four times higher on artificial turf than grass but the difference was not statistically significant (rate ratio = 3.82; P = 0.222). There were no significant differences in the overall incidence (rate ratio = 1.36; P = 0.204) or severity (P = 0.302) of training injuries sustained on artificial turf and grass. The lower limb and muscle/tendon injuries were the most common location and type of training injury on both surfaces. The results indicate that the overall risks of injury on artificial turf are not significantly different from those experienced on grass; however, the difference in the incidence of anterior cruciate ligament injuries on the two surfaces is worthy of further study.