Genetic diversity in Cypripedium calceolus (Orchidaceae) with a focus on north-western Europe, as revealed by plastid DNA length polymorphisms

Background and Aims Cypripedium calceolus, although widespread in Eurasia, is rare in many countries in which it occurs. Population genetics studies with nuclear DNA markers on this species have been hampered by its large nuclear genome size. Plastid DNA markers are used here to gain an understanding of variation within and between populations and of biogeographical patterns. Methods Thirteen length-variable regions (microsatellites and insertions/deletions) were identified in non-coding plastid DNA. These and a previously identified complex microsatellite in the trnL-trnF intergenic spacer were used to identify plastid DNA haplotypes for European samples, with sampling focused on England, Denmark and Sweden. Key Results The 13 additional length-variable regions identified were two homopolymer (polyA) repeats in the rps16 intron and a homopolymer (polyA) repeat and ten indels in the accD-psa1 intergenic spacer. In accD-psa1, most of these were in an extremely AT-rich region, and it was not possible to design primers in the flanking regions; therefore, the whole intergenic spacer was sequenced. Together, these new regions and the trnL-trnF complex microsatellite allowed 23 haplotypes to be characterized. Many were found in only one or a few samples (probably due to low sampling density), but some commoner haplotypes were widespread. Most of the genetic variation was found within rather than between populations (83 vs. 18%, respectively). Two haplotypes occurred from the Spanish Pyrenees to Sweden. Conclusions Plastid DNA data can be used to gain an understanding of patterns of genetic variation and seed-mediated gene flow in orchids. Although these data are less information-rich than those for nuclear DNA, they present a useful option for studying species with large genomes. Here they support the hypothesis of long-distance seed dispersal often proposed for orchid

Disulfide-activated protein kinase G Iα regulates cardiac diastolic relaxation and fine-tunes the Frank-Starling response.

The Frank-Starling mechanism allows the amount of blood entering the heart from the veins to be precisely matched with the amount pumped out to the arterial circulation. As the heart fills with blood during diastole, the myocardium is stretched and oxidants are produced. Here we show that protein kinase G Iα (PKGIα) is oxidant-activated during stretch and this form of the kinase selectively phosphorylates cardiac phospholamban Ser16-a site important for diastolic relaxation. We find that hearts of Cys42Ser PKGIα knock-in (KI) mice, which are resistant to PKGIα oxidation, have diastolic dysfunction and a diminished ability to couple ventricular filling with cardiac output on a beat-to-beat basis. Intracellular calcium dynamics of ventricular myocytes isolated from KI hearts are altered in a manner consistent with impaired relaxation and contractile function. We conclude that oxidation of PKGIα during myocardial stretch is crucial for diastolic relaxation and fine-tunes the Frank-Starling response

Modification of Superoxide Dismutase 1 (SOD1) Properties by a GFP Tag - Implications for Research into Amyotrophic Lateral Sclerosis (ALS)

Since the discovery that mutations in the enzyme SOD1 are causative in human amyotrophic lateral sclerosis (ALS), many strategies have been employed to elucidate the toxic properties of this ubiquitously expressed mutant protein, including the generation of GFP-SOD1 chimaeric proteins for studies in protein localization by direct visualization using fluorescence microscopy. However, little is known about the biochemical and physical properties of these chimaeric proteins, and whether they behave similarly to their untagged SOD1 counterparts. Methodology/Principal Findings Here we compare the physicochemical properties of SOD1 and the effects of GFP-tagging on its intracellular behaviour. Immunostaining demonstrated that SOD1 alone and GFP-SOD1 have an indistinguishable intracellular distribution in PC12 cells. Cultured primary motor neurons expressing GFP or GFP-SOD1 showed identical patterns of cytoplasmic expression and of movement within the axon. However, GFP tagging of SOD1 was found to alter some of the intrinsic properties of SOD1, including stability and specific activity. Evaluation of wildtype and mutant SOD1, tagged at either the N- or C-terminus with GFP, in PC12 cells demonstrated that some chimaeric proteins were degraded to the individual proteins, SOD1 and GFP. Conclusions/Significance Our findings indicate that most, but not all, properties of SOD1 remain the same with a GFP tag.m The Ipsen Fund, the UK Motor Neurone Disease Association, the Research Advisory Board of St Bartholomew’s and the Royal London Charitable Foundation (RAB04/F7), The Brain Research Trust, the Biotechnology and Biological Sciences Research Council, Cancer Research UK and EU FP6 NEST grant 12702

Effects of hospital facilities on patient outcomes after cancer surgery: an international, prospective, observational study

Background Early death after cancer surgery is higher in low-income and middle-income countries (LMICs) compared with in high-income countries, yet the impact of facility characteristics on early postoperative outcomes is unknown. The aim of this study was to examine the association between hospital infrastructure, resource availability, and processes on early outcomes after cancer surgery worldwide.Methods A multimethods analysis was performed as part of the GlobalSurg 3 study-a multicentre, international, prospective cohort study of patients who had surgery for breast, colorectal, or gastric cancer. The primary outcomes were 30-day mortality and 30-day major complication rates. Potentially beneficial hospital facilities were identified by variable selection to select those associated with 30-day mortality. Adjusted outcomes were determined using generalised estimating equations to account for patient characteristics and country-income group, with population stratification by hospital.Findings Between April 1, 2018, and April 23, 2019, facility-level data were collected for 9685 patients across 238 hospitals in 66 countries (91 hospitals in 20 high-income countries; 57 hospitals in 19 upper-middle-income countries; and 90 hospitals in 27 low-income to lower-middle-income countries). The availability of five hospital facilities was inversely associated with mortality: ultrasound, CT scanner, critical care unit, opioid analgesia, and oncologist. After adjustment for case-mix and country income group, hospitals with three or fewer of these facilities (62 hospitals, 1294 patients) had higher mortality compared with those with four or five (adjusted odds ratio [OR] 3.85 [95% CI 2.58-5.75]; p<0.0001), with excess mortality predominantly explained by a limited capacity to rescue following the development of major complications (63.0% vs 82.7%; OR 0.35 [0.23-0.53]; p<0.0001). Across LMICs, improvements in hospital facilities would prevent one to three deaths for every 100 patients undergoing surgery for cancer.Interpretation Hospitals with higher levels of infrastructure and resources have better outcomes after cancer surgery, independent of country income. Without urgent strengthening of hospital infrastructure and resources, the reductions in cancer-associated mortality associated with improved access will not be realised

SPARC 2022 book of abstracts

Welcome to the Book of Abstracts for the 2022 SPARC conference. Our conference is called “Moving Forwards” reflecting our re-emergence from the pandemic and our desire to reconnect our PGR community, in celebration of their research. PGRs have continued with their research endeavours despite many challenges, and their ongoing successes are underpinned by the support and guidance of dedicated supervisors and the Doctoral School Team. To recognise supervision excellence we will be awarding our annual Supervisor of the Year prizes, based on the wonderful nominations received from their PGR students.Once again, we have received a tremendous contribution from our postgraduate research community; with over 60 presenters, 12 Three-Minute Thesis finalists, and 20 poster presentations, the conference showcases our extraordinarily vibrant, inclusive, and resilient PGR community at Salford. This year there will be prizes to be won for ‘best in conference’ presentations, in addition to the winners from each parallel session. Audience members too could be in for a treat, with judges handing out spot prizes for the best questions asked, so don’t miss the opportunity to put your hand up. These abstracts provide a taster of the diverse and impactful research in progress and provide delegates with a reference point for networking and initiating critical debate. Take advantage of the hybrid format: in online sessions by posting a comment or by messaging an author to say “Hello”, or by initiating break time discussions about the amazing research you’ve seen if you are with us in person. Who knows what might result from your conversation? With such wide-ranging topics being showcased, we encourage you to take up this great opportunity to engage with researchers working in different subject areas from your own. As recent events have shown, researchers need to collaborate to meet global challenges. Interdisciplinary and international working is increasingly recognised and rewarded by all major research funders. We do hope, therefore, that you will take this opportunity to initiate interdisciplinary conversations with other researchers. A question or comment from a different perspective can shed new light on a project and could lead to exciting collaborations, and that is what SPARC is all about. SPARC is part of a programme of personal and professional development opportunities offered to all postgraduate researchers at Salford. More information about this programme is available on our website: Doctoral School | University of Salford. Registered Salford students can access full details on the Doctoral School hub: Doctoral School Hub - Home (sharepoint.com) You can follow us on Twitter @SalfordPGRs and please use the #SPARC2022 to share your conference experience.We particularly welcome taught students from our undergraduate and master’s programmes as audience members. We hope you enjoy the presentations on offer and that they inspire you to pursue your own research career. If you would like more information about studying for a PhD here at the University of Salford, your lecturers can advise, or you can contact the relevant PGR Support Officer; their details can be found at Doctoral School | University of Salford. We wish you a rich and rewarding conference experience