Dynamic Gene Regulatory Networks Drive Hematopoietic Specification and Differentiation.

Metazoan development involves the successive activation and silencing of specific gene expression programs and is driven by tissue-specific transcription factors programming the chromatin landscape. To understand how this process executes an entire developmental pathway, we generated global gene expression, chromatin accessibility, histone modification, and transcription factor binding data from purified embryonic stem cell-derived cells representing six sequential stages of hematopoietic specification and differentiation. Our data reveal the nature of regulatory elements driving differential gene expression and inform how transcription factor binding impacts on promoter activity. We present a dynamic core regulatory network model for hematopoietic specification and demonstrate its utility for the design of reprogramming experiments. Functional studies motivated by our genome-wide data uncovered a stage-specific role for TEAD/YAP factors in mammalian hematopoietic specification. Our study presents a powerful resource for studying hematopoiesis and demonstrates how such data advance our understanding of mammalian development.This work was funded by a Longer Larger (LoLa) consortium grant from the Biotechnology and Biological Sciences Research Council, UK, to the senior authors and the corresponding author, computing infrastructure grants from the Wellcome Trust and National Institute for Health Research to B.G., grants from Cancer Research UK to G.L. and V.K., and funding from the Bloodwise charity to C.B.This is the final version of the article. It first appeared from Cell Press via http://dx.doi.org/10.1016/j.devcel.2016.01.02

Effect of remote ischaemic conditioning on clinical outcomes in patients with acute myocardial infarction (CONDI-2/ERIC-PPCI): a single-blind randomised controlled trial.

BACKGROUND: Remote ischaemic conditioning with transient ischaemia and reperfusion applied to the arm has been shown to reduce myocardial infarct size in patients with ST-elevation myocardial infarction (STEMI) undergoing primary percutaneous coronary intervention (PPCI). We investigated whether remote ischaemic conditioning could reduce the incidence of cardiac death and hospitalisation for heart failure at 12 months. METHODS: We did an international investigator-initiated, prospective, single-blind, randomised controlled trial (CONDI-2/ERIC-PPCI) at 33 centres across the UK, Denmark, Spain, and Serbia. Patients (age >18 years) with suspected STEMI and who were eligible for PPCI were randomly allocated (1:1, stratified by centre with a permuted block method) to receive standard treatment (including a sham simulated remote ischaemic conditioning intervention at UK sites only) or remote ischaemic conditioning treatment (intermittent ischaemia and reperfusion applied to the arm through four cycles of 5-min inflation and 5-min deflation of an automated cuff device) before PPCI. Investigators responsible for data collection and outcome assessment were masked to treatment allocation. The primary combined endpoint was cardiac death or hospitalisation for heart failure at 12 months in the intention-to-treat population. This trial is registered with ClinicalTrials.gov (NCT02342522) and is completed. FINDINGS: Between Nov 6, 2013, and March 31, 2018, 5401 patients were randomly allocated to either the control group (n=2701) or the remote ischaemic conditioning group (n=2700). After exclusion of patients upon hospital arrival or loss to follow-up, 2569 patients in the control group and 2546 in the intervention group were included in the intention-to-treat analysis. At 12 months post-PPCI, the Kaplan-Meier-estimated frequencies of cardiac death or hospitalisation for heart failure (the primary endpoint) were 220 (8·6%) patients in the control group and 239 (9·4%) in the remote ischaemic conditioning group (hazard ratio 1·10 [95% CI 0·91-1·32], p=0·32 for intervention versus control). No important unexpected adverse events or side effects of remote ischaemic conditioning were observed. INTERPRETATION: Remote ischaemic conditioning does not improve clinical outcomes (cardiac death or hospitalisation for heart failure) at 12 months in patients with STEMI undergoing PPCI. FUNDING: British Heart Foundation, University College London Hospitals/University College London Biomedical Research Centre, Danish Innovation Foundation, Novo Nordisk Foundation, TrygFonden

Should Recipients Choose Their Donor? Experience of a New Consent Process in Lung Transplantation

Purpose Consent for lung transplantation in the UK was recently reviewed after public concern when a patient died after receiving lungs from a donor who was a smoker without the specific consent of the recipient. The family insisted they would have refused the organs given the choice. The British Transplantation Society subsequently recommended that all patients should be offered a choice about potential donors. Since February 2012 we implemented a form allowing lung transplant recipients to make choices on donor decisions. Guidance notes were provided to inform patients of the risks associated with organs from donors who: •had a history of smoking •had engaged in high risk sexual or recreational activities •had a malignancy •were over the age of 60 years or •had a significant viral or bacterial infection. Alternatively, patients could choose to accept organs from any donor the transplant team deemed suitable for transplantation. Methods and Materials Donor choices were discussed with patients at the time of transplant assessment. Consent form and guidance notes were provided with a request to complete and return the form. A follow up call was conducted if no response was received. Responses were analysed to obtain a view of the donor preferences of our patients. Results 52 patients were invited to complete a donor choices form and 43 (83%) responded. 84% (n=36) of responders chose to accept any organ deemed suitable by the team, irrespective of specific donor characteristics. Of the 7 (16%) patients who made a defined decision: 3 declined to consider donors with high risk sexual or recreational behaviour; 2 declined to consider donors aged over 60; 1 declined to consider donors with a smoking history and 1 declined to consider donors with a history of malignancy. Conclusions Whilst public demand has warranted a change in practice, and it is imperative to offer recipients a choice about potential donors, the majority trust the judgement of the transplant team to make the right decisions on their behalf

Blood transfusion after lung transplantation: Impact on early function and survival

Objectives: Blood transfusion is associated with higher morbidity and mortality after general cardiothoracic surgery but little is known of the impact on the transplant population. We investigated the profile of blood product transfusion in the bilateral lung transplant (BLT) population and the impact on function and survival outcomes. Methods: A total of 311 adult patients who underwent BLT between 2003 and 2013 were retrospectively reviewed. Patients were stratified according to pretransplant diagnosis and amount of blood products transfused within 24 h of surgery. Results: Patients, 174 male, 137 female (mean age 41.4 ± 14.0 years) underwent BLT, using cardiopulmonary bypass for cystic fibrosis (48.87%), fibrotic lung disease (12.21%), emphysema (27.01%), bronchiectasis (5.79%), pulmonary hypertension (1.29%) and others (4.50%). Median number of red blood cells (RBC) in the first 24 h was 3 (0–40) units, fresh frozen plasma (FFP) was 2 (0–26) units, platelets = 1 (0–7) units. There were no differences in transfusion rates according to pretransplant diagnosis. Patients were divided according to the number of units transfused in the first 24 h. Survival was not influenced by whether patients were transfused with more or less than the median number of units of RBC (P = 0.162) or FFP (P = 0.298) (Fig. 1). However, survival was adversely affected by platelet transfusion (P = 0.032). Mean FEV1 at 6 months was significantly better for patients transfused with more than the median number of units of RBC (2.66 vs 2.83, P < 0.0001), FFP (2.61 vs 2.89, P < 0.0001) and platelets (2.73 vs 2.82, P < 0.0001). Conclusion: Unlike general cardiothoracic surgery, blood transfusion has no effect on survival, but administration of platelets has an adverse effect. Blood product administration does not differ significantly with pretransplant diagnosis. Interestingly, lung function at 6 months is significantly better for patients with transfusion with more blood products

Feasibility and Acceptability of a Physical Activity Behavioural Modification Tele-Coaching Intervention in Lung Transplant Recipients

BACKGROUND: Despite improvements in pulmonary function following lung transplantation (LTx), physical activity levels remain significantly lower than the general population. To date, there is little research investigating interventions to improve daily physical activity in LTx recipients. This study assessed the feasibility and acceptability of a novel, 12-weeks physical activity tele-coaching (TC) intervention in LTx recipients. METHODS: Lung transplant recipients within 2 months of hospital discharge were recruited and randomised (1:1) to TC or usual care (UC). TC consists of a pedometer and smartphone app, allowing transmission of activity data to a platform that provides feedback, activity goals, education, and contact with the researcher as required. Recruitment and retention, occurrence of adverse events, intervention acceptability and usage were used to assess feasibility. RESULTS: Key criteria for progressing to a larger study were met. Of the 15 patients eligible, 14 were recruited and randomised to TC or UC and 12 completed (67% male; mean ± SD age; 58 ± 7 years; COPD n = 4, ILD n = 6, CF n = 1, PH n = 1): TC (n = 7) and UC (n = 5). TC was well accepted by patients, with 86% indicating that they enjoyed taking part. Usage of the pedometer was excellent, with all patients wearing it for over 90% of days and rating the pedometer and telephone contact as the most vital aspects. There were no adverse events related to the intervention. After 12 weeks, only TC displayed improvements in accelerometry steps/day (by 3475 ± 3422; p = .036) and movement intensity (by 153 ± 166 VMU; p = .019), whereas both TC and UC groups exhibited clinically important changes in physical SF-36 scores (by 11 ± 14 and 7 ± 9 points, respectively). CONCLUSION: TC appears to be a feasible, safe, and well-accepted intervention in LTx

Allogeneic blood transfusion in bilateral lung transplantation: impact on early function and mortality

Objectives: Blood transfusion is associated with higher morbidity and mortality after general cardiothoracic surgery but its impact within the transplant population is unclear. We investigated the profile of blood product transfusion in the bilateral lung transplant population and its impact on function and mortality. Methods: Three hundred and eleven adult patients who underwent bilateral lung transplant between 2003 and 2013 were retrospectively reviewed. Patients were stratified according to pretransplant diagnoses and amount of blood products transfused within 24 h of transplant. All-cause mortality at the 1-year follow-up was analysed using a Cox proportional hazards regression model. Results: One hundred and seventy-four male patients and 137 female patients (mean age = 41.4 ± 14.0 years) underwent bilateral lung transplant using cardiopulmonary bypass for cystic fibrosis (48.9%), fibrotic lung disease (12.2%), emphysema (27.0%), bronchiectasis (5.8%), pulmonary hypertension (1.3%) and others (4.5%). The median number of red blood cells in the first 24 h was 3 (0–40) units, fresh frozen plasma (FFP) = 2 (0–26) units and platelets = 1 (0–7) units. The unadjusted all-cause mortality at the 1-year follow-up did not appear to be different between patient subgroups stratified by the median number of units of red blood cells (P = 0.827) or FFP transfused (P = 0.456). However, 1-year mortality was adversely affected when more than the median number of units of platelets was transfused (P = 0.010). Upon adjustment for confounding variables, 1-year mortality was noted to be greater among patients transfused more than the median unit of platelets (adjusted hazard ratios: 2.3, 95% confidence interval: 1.15–4.61, P = 0.019) and those with longer bypass times (P = 0.046). No significant difference in the number of units transfused was noted when patients were stratified by pretransplant diagnosis. Predicted lung function at 3 and 6 months was not significantly affected by greater blood product use. Conclusions: Unlike general cardiothoracic surgery, blood transfusion had no effect on all-cause mortality, whereas a greater administration of platelets was observed to be associated with higher adjusted 1-year mortality. Transfusion rates were not significantly influenced by pretransplant diagnoses. Interestingly, lung function at 3 and 6 months was similar for patients who received more blood product transfusion

Mycobacterium ulcerans low infectious dose and mechanical transmission support insect bites and puncturing injuries in the spread of Buruli ulcer

Addressing the transmission enigma of the neglected disease Buruli ulcer (BU) is a World Health Organization priority. In Australia, we have observed an association between mosquitoes harboring the causative agent, Mycobacterium ulcerans, and BU. Here we tested a contaminated skin model of BU transmission by dipping the tails from healthy mice in cultures of the causative agent, Mycobacterium ulcerans. Tails were exposed to mosquito (Aedes notoscriptus and Aedes aegypti) blood feeding or punctured with sterile needles. Two of 12 of mice with M. ulcerans contaminated tails exposed to feeding A. notoscriptus mosquitoes developed BU. There were no mice exposed to A. aegypti that developed BU. Eighty-eight percent of mice (21/24) subjected to contaminated tail needle puncture developed BU. Mouse tails coated only in bacteria did not develop disease. A median incubation time of 12 weeks, consistent with data from human infections, was noted. We then specifically tested the M. ulcerans infectious dose-50 (ID50) in this contaminated skin surface infection model with needle puncture and observed an ID50 of 2.6 colony-forming units. We have uncovered a biologically plausible mechanical transmission mode of BU via natural or anthropogenic skin punctures

<i>M</i>. <i>ulcerans</i> incubation period and infectious dose₅₀.

<p>(A) Incubation period of <i>M</i>. <i>ulcerans</i> based on the time between sterile-needle puncture of an <i>M</i>. <i>ulcerans</i> contaminated mouse-tail and first observation of a lesion. (B) Estimated <i>M</i>. <i>ulcerans</i> ID50 for contaminated skin surface transmission model. Dashed lines indicate 95% confidence intervals. Dotted lines depicts ID50.</p