Structure-based vaccine design by electron microscopy

Modern vaccine design relies on multiscale, interdisciplinary efforts that take advantage of innovative technologies such as in silico identification of antigens, high throughput screening of antigen immunogenicity, and gene expression profiling to predict host immune responses. In recent years, structural analysis has played an increasingly important role in vaccine development as a means to improve antigen stability, immunogenicity and large scale production. Transmission electron microscopy (TEM), and in particular cryo-TEM, is an established and powerful imaging technique applicable to many specimens, including the three-dimensional (3D) reconstruction of macromolecules and their associated complexes to high resolution. The technique is parsimonious in its material requirements and captures the specimens in their fully hydrated state, close to their native environment. The resolution of cryo-TEM reconstructions was limited to the subnanometer range until the recent development of direct electron detectors and improvements in image processing software, which has led to a so-called “resolution revolution” in the cryo-TEM field. Several protein structures have now been solved at near atomic resolution, establishing the technique as a viable alternative to X-ray analysis for high resolution structure determination. We have determined several structures with and without bound compounds at 2.9 Å – 3.6 Å resolution, which are being integrated into drug discovery and development workflows by our clients. Here we present the 2.4Å resolution structure of apoferritin determined with our Titan Krios electron microscope as an example of the cryo-TEM services available at NIS. These services are significantly enhanced with unique access by NIS to a new instrument, Spotiton, a robotic device that dispenses picoliter-volumes of sample onto a self-blotting nanowire grid as it flies past en route to vitrification. This provides several advantages over standard vitrification methods, including more automated and reproducible preparation of specimens and reducing the deleterious effects of particles interacting with the air-water interface. While high resolution 3D structure determination by cryo-TEM is at the forefront of structural biology, averages of 2D projection images at moderate resolution in negative stain or vitreous ice can also provide a wealth of information that may be difficult to obtain using other methods. This is illustrated in a number of case studies, including (1) mapping of neutralizing epitopes on the CMV pentameric glycoprotein complex; (2) mapping of neutralizing epitopes on the HIV-1 envelope glycoprotein trimer; (3) assessment of structure and conformational stability of pre- and post-fusion RSV-F protein; (4) characterization of novel adjuvants and protein delivery systems. In summary, both the moderate resolution TEM and high resolution cryo-TEM methods are well suited to extensively characterize antigen structure-function relationships, some of which may be refractory to other experimental methods

A revised speech spectrum for STI calculations

The ability of the Speech Transmission Index (STI) to predict speech intelligibility under noisy conditions is highly dependent on the assumed spectrum of the speech signal. Examination of the literature showed that the long-term average speech spectrum of male talkers differs substantially from the speech spectrum recommended for STI calculations (IEC 60268-16). To explore these issues, the long-term average speech spectrum of forty male British English people was first measured, compared with the available literature and proposed for STI calculations. Then, using several voice alarm systems, the influence of the measured spectrum on STI calculations was assessed and comparisons made with the standard speech spectrum. The results showed significant STI differences under noisy conditions and considerable reductions in the required electrical power with the use of the new proposed male spectrum. This indicated that the current STI method could benefit from a revised speech spectrum

Volume 12

Introduction, Dr. Roger A. Byrne, Dean From the Editor, Dr. Larissa Kat Tracy From the Designers, Rachel English, Rachel Hanson Immortality in the Mortal World: Otherworldly Intervention in Lanval and The Wife of Bath\u27s Tale by Haleigh James Analysis of Phenolic Compounds in Moroccan Olive Oils by HPLC by Hannah Meyls Art by Hope Irvin The Effects of Cell Phone Use on Gameplay Enjoyment and Frustration by Megan E. Hlavaty, Samara L. Gall, and Austin J. Funk Care, No Matter What: Planned Parenthood\u27s Use of Organizational Rhetoric to Expand its Reputation by Karyn Keane Analysis of Petroleum Products for Forensic and Environmental Applications by Sarah Ghali, Antonio Harvey, and Katelynn McCrillis Art by Andrew Jones The Triangle Shirtwaist Factory Fire by Rachel Hazelwood Art by Madison Schmitz Ercilla y la imitacion: Araucanos al estilo europeo by Marija Venta Design by Haley Tebo Design by Jeremiah Gilmer White Supremacist\u27s Appropriation of the Persuasion of Passivity in Marvel\u27s Captain America by Bridget Dunn Design by Benjamin Sullivan Art by McKenzie Johnso

Tranexamic acid for spontaneous intracerebral hemorrhage: a randomized controlled pilot trial (ISRCTN50867461)

Background: Spontaneous intracerebral hemorrhage (ICH) can be devastating, particularly if hematoma expansion (HE) occurs. Tranexamic acid (TA), an antifibrinolytic drug, significantly reduced mortality in bleeding patients after trauma in the large CRASH-2 trial. The CRASH-2 ICH substudy found that TA nonsignificantly reduced mortality and dependency in traumatic ICH. The aim of this study was to assess the feasibility of performing a randomized controlled trial of tranexamic acid in spontaneous ICH, ahead of a definitive study. Methods: We performed a single-center, prospective, randomized (2:1), double-blind, placebo-controlled blinded endpoint trial of TA (intravenous 1 g bolus, 1 g infusion/8 h) in acute (<24 hours) spontaneous ICH. The primary objective was to test the feasibility of recruiting to the trial. Other objectives included tolerability (adverse events) and the effect of TA on HE and death and dependency. Results: The trial was feasible, with 24 patients enrolled (TA, n 5 16; placebo, n 5 8) between March 2011 and March 2012, and acceptable—only 3 patients declined to participate. All patients received the correct randomized treatment; 1 patient in the TA group did not complete the infusion because of neurologic deterioration. There were no significant differences in secondary outcomes including adverse events, HE, death, and dependency. One patient in the TA group had a deep vein thrombosis. Conclusions: This, the first randomized controlled trial of TAin ICH, found that the protocol could be delivered on schedule (2 patients/mo) and was feasible. Larger studies are needed to assess safety and efficacy of TA in ICH

ABA Criminal Justice Section Task Force on College Due Process Rights and Victim Protections: Recommendations for Colleges and Universities in Resolving Allegations of Campus Sexual Misconduct

The Executive Committee of the ABA Criminal Justice Section commissioned the Task Force on College Due Process Rights and Victim Protections in November 2016. Immediately after, extensive efforts were made to find members that represented all interested parties: victims, the accused, universities, other stakeholders, and national experts. The Task Force was fully constituted in the winter of 2017, and it ended up including two voting members who were originally liaisons from the ABA Commission on Domestic and Sexual Violence and the ABA Section of Civil Rights and Social Justice. This elevation was made in recognition of their significant contributions

Basic science232. Certolizumab pegol prevents pro-inflammatory alterations in endothelial cell function

Background: Cardiovascular disease is a major comorbidity of rheumatoid arthritis (RA) and a leading cause of death. Chronic systemic inflammation involving tumour necrosis factor alpha (TNF) could contribute to endothelial activation and atherogenesis. A number of anti-TNF therapies are in current use for the treatment of RA, including certolizumab pegol (CZP), (Cimzia ®; UCB, Belgium). Anti-TNF therapy has been associated with reduced clinical cardiovascular disease risk and ameliorated vascular function in RA patients. However, the specific effects of TNF inhibitors on endothelial cell function are largely unknown. Our aim was to investigate the mechanisms underpinning CZP effects on TNF-activated human endothelial cells. Methods: Human aortic endothelial cells (HAoECs) were cultured in vitro and exposed to a) TNF alone, b) TNF plus CZP, or c) neither agent. Microarray analysis was used to examine the transcriptional profile of cells treated for 6 hrs and quantitative polymerase chain reaction (qPCR) analysed gene expression at 1, 3, 6 and 24 hrs. NF-κB localization and IκB degradation were investigated using immunocytochemistry, high content analysis and western blotting. Flow cytometry was conducted to detect microparticle release from HAoECs. Results: Transcriptional profiling revealed that while TNF alone had strong effects on endothelial gene expression, TNF and CZP in combination produced a global gene expression pattern similar to untreated control. The two most highly up-regulated genes in response to TNF treatment were adhesion molecules E-selectin and VCAM-1 (q 0.2 compared to control; p > 0.05 compared to TNF alone). The NF-κB pathway was confirmed as a downstream target of TNF-induced HAoEC activation, via nuclear translocation of NF-κB and degradation of IκB, effects which were abolished by treatment with CZP. In addition, flow cytometry detected an increased production of endothelial microparticles in TNF-activated HAoECs, which was prevented by treatment with CZP. Conclusions: We have found at a cellular level that a clinically available TNF inhibitor, CZP reduces the expression of adhesion molecule expression, and prevents TNF-induced activation of the NF-κB pathway. Furthermore, CZP prevents the production of microparticles by activated endothelial cells. This could be central to the prevention of inflammatory environments underlying these conditions and measurement of microparticles has potential as a novel prognostic marker for future cardiovascular events in this patient group. Disclosure statement: Y.A. received a research grant from UCB. I.B. received a research grant from UCB. S.H. received a research grant from UCB. All other authors have declared no conflicts of interes

The James Webb Space Telescope Mission

Twenty-six years ago a small committee report, building on earlier studies, expounded a compelling and poetic vision for the future of astronomy, calling for an infrared-optimized space telescope with an aperture of at least $4m$. With the support of their governments in the US, Europe, and Canada, 20,000 people realized that vision as the $6.5m$ James Webb Space Telescope. A generation of astronomers will celebrate their accomplishments for the life of the mission, potentially as long as 20 years, and beyond. This report and the scientific discoveries that follow are extended thank-you notes to the 20,000 team members. The telescope is working perfectly, with much better image quality than expected. In this and accompanying papers, we give a brief history, describe the observatory, outline its objectives and current observing program, and discuss the inventions and people who made it possible. We cite detailed reports on the design and the measured performance on orbit.Comment: Accepted by PASP for the special issue on The James Webb Space Telescope Overview, 29 pages, 4 figure

Effectiveness of safe and sustainable cleaning practices in childcare centres

The Faculty of Life and Social Science, Swinburne University of Technology, conducted an investigation into the effectiveness of a range of cleaning practices in two childcare centres within inner Melbourne. The investigation was a component of The Safe and Sustainable Indoor Cleaning Project, funded by the Sustainability Fund, managed by Sustainability Victoria, of which Fresh Clean Green were the funding recipients. The investigation took place between June and August 2007. Cleaning is necessary for creating a good indoor environment, maintaining an acceptable level of perceived cleanliness, preventing surface degradation, controlling the potential risk of infection from microorganisms and dust exposure in general, and is economically beneficial. However, the use of cleaning agents can contaminate indoor air and outdoor environments. Over recent years there has been an increased shift to the use of non-toxic 'green' or 'environmentally friendly' products, as a means to reduce exposure to cleaning agents. Anecdotally, childcare centres have also been showing an increased interest in the use of 'green' or 'environmentally friendly' materials, including disinfectants, as part of their cleaning practices. However, literature surrounding the effectiveness of non-toxic or 'green' substances as a means of reducing microbial load is limited. The purpose of this research was to investigate the efficacy of the current cleaning regimes in two centres, involving the use of disinfectants, and compare this with the application of a plant-based, non-hazardous detergent diluted with warm water. The investigation was an experimental design, and involved two sets of sampling for bacteria, including the bacteria Staphylococcus aureus and Escherichia coli which are indicators of hygenic practices, in conjunction with a Total Plate Count, a common test applied to indicate microbial presence. The results indicated that using a plant-based, non-hazardous detergent diluted with warm water was as effective in reducing the amount of bacteria on the surfaces tested as were other materials, such as disinfectants, in these two centres. This is consistent with the guidelines set by the National Health and Medical Research Council (NH&MRC) 'Staying healthy in child care: preventing infectious disease: 4th edition'. These guidelines highlight that the cleaning of these surfaces with detergents followed by rinsing and drying, removes the bulk of germs and that disinfectants are usually unnecessary. An exception to this is an infectious disease breakout situation, where health authorities may stipulate the use of a specific detergent, in addition to the normal cleaning processes. [Executive summary