Comparison of mesophyll protoplast isolation and transformation between \u3ci\u3ePanicum virgatum\u3c/i\u3e and \u3ci\u3ePanicum hallii\u3c/i\u3e

Protoplasts are appropriate targets for genome editing, DNA functional screens and transgenesis. This project focuses on the comparison of inexpensive mesophyll protoplast isolation via the use of food-grade enzymes and transformation between diploid Panicum hallii Vasey (PAH) and polyploid cellulosic feedstock Panicum virgatum L (switchgrass), a relative of PAH. PAH has great potential as a C4 model species for crop and bioenergy research. Here an inexpensive switchgrass and PAH mesophyll protoplast isolation and transformation system was developed; the first protoplast system for PAH. Using low-cost commercial food-grade enzymes, a cost reduction of ~1000-fold was achieved compared to traditional protoplast isolating enzymes with a cost of $0.003 (USD) per reaction for switchgrass mesophyll protoplasts and$0.0018 (USD) per reaction for switchgrass cell-suspension culture-derived protoplasts. Additionally, PEG-mediated switchgrass mesophyll protoplast transformation was improved to a maximum 30.4 % over the previous transformation efficiency of 9.1 %, achieving higher transformation efficiency with a reduction in DNA quantity. In the first protoplast isolation system for PAH, an average fivefold increase in protoplast yield from PAH leaf tissue over the optimum switchgrass tissue protoplast isolation was shown. PAH yielded an average 7340 ± 1816 viable protoplasts per mg mesophyll tissue and switchgrass yielded 1468 ± 431 viable protoplasts per mg mesophyll tissue with both species having greater than 95 % viable protoplasts. With additional food-grade enzyme concentration optimization, an additional cost decrease to $0.001 (USD) per reaction was shown. PAH mesophyll protoplasts have a diameter from 3.9- 28.1 µm [micrometer], with a mean of 13.5 µm, which are significantly smaller than switchgrass mesophyll protoplasts which range from 6.5- 39.4 µm with a mean of 17.4 µm. Polyethylene glycol (PEG)-mediated transformation of PAH protoplasts revealed an optimum transformation efficiency of 46.7 ± 5.5 % with switchgrass protoplast transformation efficiency of 9.3 ± 1.9 %. The methods in this project provide an essential step toward using P. hallii as a C4 panicoid model species

Resolving the morphological and mechanical properties of palm petioles: shape analysis methods for symmetric sections of natural form

This thesis was submitted for the degree of Doctor of Philosophy and awarded by Brunel University London.Palms support the largest leaves in the world and have evolved on Earth for over 120 million years. They are often reported to be the only structure left standing post-hurricane. Cross-sectional shapes of cantilevered structures are important design factors affecting torsional and bending performance. Understanding the shape contribution of natural sections such as palm petioles (modified leaf stalks) is more difficult than those for simple 2D shapes because conventional methods of calculating section properties are not well suited to these irregular shapes. The role of internal structure, material properties and external shape of palm petioles in cantilever performance has been investigated and three main contributions to knowledge result from this research. Firstly, 3D mapping, i.e., the size, orientation and position, of vascular bundles in the Trachycarpus fortunei palm petiole reveals the distributions of stress and Young’s modulus values, providing a more detailed understanding of petioles than previous work. Secondly, bulk elastic material properties along the longitudinal axis of the same petiole are then input to a bi-layered model of the same petiole establishing the Young’s modulus of the two layers without mechanically testing them individually and for determining that the outer layer is not lignified. Thirdly, the largest contribution is the introduction of modified shape transformers employing the use of circular envelopes, eliminating error caused by approximating second moment of area with the torsional constant. This leads to a novel Shape Edge Mapping (SEM) technique which deconstructs petiole cross section shape elements and enables the structural contribution of these elements to be calculated, improving the understanding of the petiole section and how it relates to its mechanical function. This thesis makes a valuable addition to the knowledge of palm function and presents novel techniques for non-destructive extraction of natural shape data for abstraction and use in preliminary engineering design

Columbus State University Honors College: Senior Theses, Spring 2020

This is a collection of senior theses written by honors students at Columbus State University in Spring 2020.https://csuepress.columbusstate.edu/honors_theses/1001/thumbnail.jp

The Astropy Problem

The Astropy Project (http://astropy.org) is, in its own words, "a community effort to develop a single core package for Astronomy in Python and foster interoperability between Python astronomy packages." For five years this project has been managed, written, and operated as a grassroots, self-organized, almost entirely volunteer effort while the software is used by the majority of the astronomical community. Despite this, the project has always been and remains to this day effectively unfunded. Further, contributors receive little or no formal recognition for creating and supporting what is now critical software. This paper explores the problem in detail, outlines possible solutions to correct this, and presents a few suggestions on how to address the sustainability of general purpose astronomical software

Effects of antiplatelet therapy on stroke risk by brain imaging features of intracerebral haemorrhage and cerebral small vessel diseases: subgroup analyses of the RESTART randomised, open-label trial

Background Findings from the RESTART trial suggest that starting antiplatelet therapy might reduce the risk of recurrent symptomatic intracerebral haemorrhage compared with avoiding antiplatelet therapy. Brain imaging features of intracerebral haemorrhage and cerebral small vessel diseases (such as cerebral microbleeds) are associated with greater risks of recurrent intracerebral haemorrhage. We did subgroup analyses of the RESTART trial to explore whether these brain imaging features modify the effects of antiplatelet therapy

Promoting active learning in universities

Thematic peer group repor

Mortality and pulmonary complications in patients undergoing surgery with perioperative SARS-CoV-2 infection: an international cohort study

Background: The impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on postoperative recovery needs to be understood to inform clinical decision making during and after the COVID-19 pandemic. This study reports 30-day mortality and pulmonary complication rates in patients with perioperative SARS-CoV-2 infection. Methods: This international, multicentre, cohort study at 235 hospitals in 24 countries included all patients undergoing surgery who had SARS-CoV-2 infection confirmed within 7 days before or 30 days after surgery. The primary outcome measure was 30-day postoperative mortality and was assessed in all enrolled patients. The main secondary outcome measure was pulmonary complications, defined as pneumonia, acute respiratory distress syndrome, or unexpected postoperative ventilation. Findings: This analysis includes 1128 patients who had surgery between Jan 1 and March 31, 2020, of whom 835 (74·0%) had emergency surgery and 280 (24·8%) had elective surgery. SARS-CoV-2 infection was confirmed preoperatively in 294 (26·1%) patients. 30-day mortality was 23·8% (268 of 1128). Pulmonary complications occurred in 577 (51·2%) of 1128 patients; 30-day mortality in these patients was 38·0% (219 of 577), accounting for 81·7% (219 of 268) of all deaths. In adjusted analyses, 30-day mortality was associated with male sex (odds ratio 1·75 [95% CI 1·28–2·40], p\textless0·0001), age 70 years or older versus younger than 70 years (2·30 [1·65–3·22], p\textless0·0001), American Society of Anesthesiologists grades 3–5 versus grades 1–2 (2·35 [1·57–3·53], p\textless0·0001), malignant versus benign or obstetric diagnosis (1·55 [1·01–2·39], p=0·046), emergency versus elective surgery (1·67 [1·06–2·63], p=0·026), and major versus minor surgery (1·52 [1·01–2·31], p=0·047). Interpretation: Postoperative pulmonary complications occur in half of patients with perioperative SARS-CoV-2 infection and are associated with high mortality. Thresholds for surgery during the COVID-19 pandemic should be higher than during normal practice, particularly in men aged 70 years and older. Consideration should be given for postponing non-urgent procedures and promoting non-operative treatment to delay or avoid the need for surgery. Funding: National Institute for Health Research (NIHR), Association of Coloproctology of Great Britain and Ireland, Bowel and Cancer Research, Bowel Disease Research Foundation, Association of Upper Gastrointestinal Surgeons, British Association of Surgical Oncology, British Gynaecological Cancer Society, European Society of Coloproctology, NIHR Academy, Sarcoma UK, Vascular Society for Great Britain and Ireland, and Yorkshire Cancer Research

Impact of opioid-free analgesia on pain severity and patient satisfaction after discharge from surgery: multispecialty, prospective cohort study in 25 countries

Background: Balancing opioid stewardship and the need for adequate analgesia following discharge after surgery is challenging. This study aimed to compare the outcomes for patients discharged with opioid versus opioid-free analgesia after common surgical procedures.Methods: This international, multicentre, prospective cohort study collected data from patients undergoing common acute and elective general surgical, urological, gynaecological, and orthopaedic procedures. The primary outcomes were patient-reported time in severe pain measured on a numerical analogue scale from 0 to 100% and patient-reported satisfaction with pain relief during the first week following discharge. Data were collected by in-hospital chart review and patient telephone interview 1 week after discharge.Results: The study recruited 4273 patients from 144 centres in 25 countries; 1311 patients (30.7%) were prescribed opioid analgesia at discharge. Patients reported being in severe pain for 10 (i.q.r. 1-30)% of the first week after discharge and rated satisfaction with analgesia as 90 (i.q.r. 80-100) of 100. After adjustment for confounders, opioid analgesia on discharge was independently associated with increased pain severity (risk ratio 1.52, 95% c.i. 1.31 to 1.76; P < 0.001) and re-presentation to healthcare providers owing to side-effects of medication (OR 2.38, 95% c.i. 1.36 to 4.17; P = 0.004), but not with satisfaction with analgesia (beta coefficient 0.92, 95% c.i. -1.52 to 3.36; P = 0.468) compared with opioid-free analgesia. Although opioid prescribing varied greatly between high-income and low- and middle-income countries, patient-reported outcomes did not.Conclusion: Opioid analgesia prescription on surgical discharge is associated with a higher risk of re-presentation owing to side-effects of medication and increased patient-reported pain, but not with changes in patient-reported satisfaction. Opioid-free discharge analgesia should be adopted routinely