Characterisation of human bronchial epithelial response to injury and identification of potential resident progenitor cells in vitro

As the first line of defence against external stimuli, the airway epithelium undergoes frequent injury during an adult life. This is countered by repair mechanisms that ensure the integrity of the epithelium. It has been established that there are resident stem/progenitor cells localized within specific niches throughout the respiratory tract. Stem/progenitor cells are activated according to the severity of the insult and are thought to be responsible for repairing the airway epithelium. It has been difficult to isolate those stem cells. In this study, an in vitro human bronchial epithelial model was adapted and characterised. In culture, a pseudostratified epithelium was observed, with basal, ciliated, and secretory cells. Mucus production was also seen in this model. A scrape-wound of the model was employed to study the responses of the airway epithelium to injury. It was observed that migration and then proliferation, of CD44 expressing basal cells, are the first events that take place after scrape-wounding. Up-regulation of CD44 was also observed at the edge of the wounds early post-wounding. This suggested a key role for CD44-expressing basal cells in migration and proliferation after wounding, also suggesting this population may contain a progenitor cell population. Investigation of the secretory profile of the airway epithelium post-wounding revealed an increase in a number of cytokines and growth factors. In particular, IL-6, IL-8, ENA-78, and RANTES were all elevated compared with unwounded cultures. A side population (SP) was identified in differentiated and undifferentiated human bronchial epithelial cells in at least some cultures accounting for 0.1-1.15% of cells present. In summary, the epithelium is important in airway wound repair with basal cells appearing to contain the progenitor population in this human bronchial epithelial cell (HBEC) model. Moreover, SP studies suggested the presence of SP in at least some cultures, which might contribute to airway regeneration. The secretory profile of the airway epithelium post-wounding indicates up-regulation of specific cytokines, which may be important in the pathogenesis of lung diseases such as asthma and COPD. This model should prove useful to assess wound repair pathways and may be of use in the future for proof of concept studies on novel therapeutic agents

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

P2Y Purinergic Receptors, Endothelial Dysfunction, and Cardiovascular Diseases

Purinergic G-protein-coupled receptors are ancient and the most abundant group of G-protein-coupled receptors (GPCRs). The wide distribution of purinergic receptors in the cardiovascular system, together with the expression of multiple receptor subtypes in endothelial cells (ECs) and other vascular cells demonstrates the physiological importance of the purinergic signaling system in the regulation of the cardiovascular system. This review discusses the contribution of purinergic P2Y receptors to endothelial dysfunction (ED) in numerous cardiovascular diseases (CVDs). Endothelial dysfunction can be defined as a shift from a &ldquo;calm&rdquo; or non-activated state, characterized by low permeability, anti-thrombotic, and anti-inflammatory properties, to a &ldquo;activated&rdquo; state, characterized by vasoconstriction and increased permeability, pro-thrombotic, and pro-inflammatory properties. This state of ED is observed in many diseases, including atherosclerosis, diabetes, hypertension, metabolic syndrome, sepsis, and pulmonary hypertension. Herein, we review the recent advances in P2Y receptor physiology and emphasize some of their unique signaling features in pulmonary endothelial cells