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

Investigating the genetic control of postharvest shelf life and vitamin C content in broccoli (Brassica oleracea var. italica)

Broccoli (Brassica oleracea var. italica) is a popular vegetable, known for its nutritional benefits. However, the marketability of broccoli is limited by a short shelf life. Broccoli is susceptible to rapid postharvest senescence, which causes visible head yellowing and wilting from dehydration. Visible quality loss is also accompanied by a decline in nutrients, resulting in a product with reduced postharvest nutritional value. These factors combined cause broccoli to become unmarketable, leading to severe wastage in the retail chain. Postharvest yellowing in broccoli has been shown to be controlled by genotype, as a doubled haploid population (MGDH) created from an F1 cross between GD33, a poor performing DH line (yellow in 2 days) and Mar34, a good performing DH line (staying green > 4 days) exhibited natural variation for shelf life. Therefore, to investigate the genetic control of quality in broccoli, the fixed mapping population was assessed for shelf life, morphological traits and vitamin C content and stability in replicated field trials. Visual inspections identified head yellowing, stem turgor and bud compactness as the main traits affecting the marketability of broccoli. Two methods to quantify head yellowing were also evaluated. Spectrophotometer readings were found to be more sensitive than Image J in detecting colour change, but Image J data was more reproducible. Vitamin C quantification using HPLC, confirmed that natural variation was present in the MGDH population at harvest. Vitamin C content during postharvest storage, detected by plate assays, found vitamin C to be unstable, degrading quickly after harvest. A unique broccoli x broccoli linkage map, covering ~72.9% of the B.oleracea genome, was also constructed by genotyping the MGDH population with SSR and AFLP markers. QTL analysis of the trait data positioned 48 significant QTL in the linkage map for head yellowing (4), colour co-ordinates (17), morphological traits (17), bud quality (2) and postharvest vitamin C content (3) and stability (5). The identification of QTLs associated with the above traits has provided useful information for breeders to breed for improved nutritional and quality in broccoli using marker-assisted selection (MAS). The location of QTLs has also provided targets for fine-scale mapping and for the identification of candidate genes underlying traits

Mapping and analysis of genetic loci controlling quality traits in broccoli - a new broccoli linkage map and mapped quality trait QTLs

Aims of the project 1. Identify and map additional QTL linked to a range of quality traits a. Shelf life (turgor); QTL for yellowing already identified b. Nutrient content (different components) c. Stability of nutrients during storage d. Head morphology 2. Analyse nutrients and potential flavour attributes a. Global metabalome analysis b. Key nutrients (antioxidants, glucosinolates, flavanoids, vitamins) 3. Fine scale mapping of QTL, potential gene identification a. Backcross selected lines for fine scale mapping b. Microarray analysis c. Candidate gene nominatio

Mapping and analysis of genetic loci controlling quality traits in broccoli

The consumption of Brassica vegetables in the UK has increased over the past 10 years. In particular, the demand for fresh broccoli has increased in line with increased public awareness of the nutritional profile and health benefits associated with this crop. Broccoli has an unpredictable shelf life attributed primarily to genetic and environmental variation. Differing supply-chain components exacerbate the situation. Most leafy vegetables are supplied on a three-day delivery cycle; broccoli requires a 2-day delivery cycle. Improving broccoli shelf life by 1 day would benefit the supply chain considerably. The rapid post-harvest senescence of broccoli heads is characterised by visible phenotypic changes such as head yellowing, browning and reduced turgor. However, these changes in organoleptic quality are preceded by a decline in the nutritional status of the heads. Since post-harvest yellowing is not visible until the later stages of head senescence, consumers may be eating broccoli of inferior nutritional status. At WHRI we have developed a unique ‘broccoli x broccoli’ doubled haploid mapping population derived from the F1 of the cross between a poor performing line (yellow in 2 days) with a line that stayed green for more than 4 days at 16°C. The fixed mapping population captures the complete range of diversity in days to yellowing and has been scored for quality traits in field trails at WHRI over 4 growing seasons. Data for shelf life traits including bud yellowing are being used to identify putative QTL positions on our framework linkage map. We are extending this work to investigate the nutritional status of the broccoli head at harvest and how the metabolite profile changes through storage. This project will deliver valuable breeding resources that will benefit the supply chain and ultimately the consumer

Generation of amphidiploids from hybrids of wheat and related species from the genera Aegilops, Secale, Thinopyrum and Triticum as a source of genetic variation for wheat improvement

We aim to improve diversity of domesticated wheat by transferring genetic variation for important target traits from related wild and cultivated grass species. The present study describes the development of F1 hybrids between wheat and related species from the genera Aegilops, Secale, Thinopyrum, and Triticum and production of new amphidiploids. Amphidiploid lines were produced from 20 different distant relatives. Both colchicine and caffeine were successfully used to double the chromosome numbers. The genomic constitution of the newly formed amphidiploids derived from seven distant relatives was determined using genomic in situ hybridization (GISH). Altogether, 42 different plants were analysed, 19 using multicolour GISH separating the chromosomes from the A, B, and D genomes of wheat, as well as the distant relative, and 23 using single colour GISH. Restructuring of the allopolyploid genome, both chromosome losses and aneuploidy, was detected in all the genomes contained by the amphidiploids. From the observed chromosome numbers there is an indication that in amphidiploids the B genome of wheat suffers chromosome losses less frequently than the other wheat genomes. Phenotyping to realize the full potential of the wheat – related grass germplasm is underway, linking the analyzed genotypes to agronomically important target traits

A new broccoli × broccoli immortal mapping population and framework genetic map: tools for breeders and complex trait analysis

A unique broccoli x broccoli doubled haploid (DH) population has been created from the F-1 of a cross between two DH broccoli lines derived from cultivars Green Duke and Marathon. We genotyped 154 individuals from this population with simple sequence repeat and amplified fragment length polymorphism markers to create a B. oleracea L. var. italica 'intra-crop' specific framework linkage map. The map is composed of nine linkage groups with a total length of 946.7 cM. Previous published B. oleracea maps have been constructed using diverse crosses between morphotypes of B. oleracea; this map therefore represents a useful breeding resource for the dissection of broccoli specific traits. Phenotype data have been collected from the population over five growing seasons; the framework linkage map has been used to locate quantitative trait loci for agronomically important broccoli traits including head weight (saleable yield), head diameter, stalk diameter, weight loss and relative weight loss during storage, as well as traits for broccoli leaf architecture. This population and associated linkage map will aid breeders to directly map agronomically important traits for the improvement of elite broccoli cultivars

Empagliflozin in Patients with Chronic Kidney Disease

Background The effects of empagliflozin in patients with chronic kidney disease who are at risk for disease progression are not well understood. The EMPA-KIDNEY trial was designed to assess the effects of treatment with empagliflozin in a broad range of such patients. Methods We enrolled patients with chronic kidney disease who had an estimated glomerular filtration rate (eGFR) of at least 20 but less than 45 ml per minute per 1.73 m(2) of body-surface area, or who had an eGFR of at least 45 but less than 90 ml per minute per 1.73 m(2) with a urinary albumin-to-creatinine ratio (with albumin measured in milligrams and creatinine measured in grams) of at least 200. Patients were randomly assigned to receive empagliflozin (10 mg once daily) or matching placebo. The primary outcome was a composite of progression of kidney disease (defined as end-stage kidney disease, a sustained decrease in eGFR to < 10 ml per minute per 1.73 m(2), a sustained decrease in eGFR of & GE;40% from baseline, or death from renal causes) or death from cardiovascular causes. Results A total of 6609 patients underwent randomization. During a median of 2.0 years of follow-up, progression of kidney disease or death from cardiovascular causes occurred in 432 of 3304 patients (13.1%) in the empagliflozin group and in 558 of 3305 patients (16.9%) in the placebo group (hazard ratio, 0.72; 95% confidence interval [CI], 0.64 to 0.82; P < 0.001). Results were consistent among patients with or without diabetes and across subgroups defined according to eGFR ranges. The rate of hospitalization from any cause was lower in the empagliflozin group than in the placebo group (hazard ratio, 0.86; 95% CI, 0.78 to 0.95; P=0.003), but there were no significant between-group differences with respect to the composite outcome of hospitalization for heart failure or death from cardiovascular causes (which occurred in 4.0% in the empagliflozin group and 4.6% in the placebo group) or death from any cause (in 4.5% and 5.1%, respectively). The rates of serious adverse events were similar in the two groups. Conclusions Among a wide range of patients with chronic kidney disease who were at risk for disease progression, empagliflozin therapy led to a lower risk of progression of kidney disease or death from cardiovascular causes than placebo