Purification and quantification of recombinant Epstein-Barr viral glycoproteins gp350/220 from Chinese hamster ovary cells

Truncated Epstein-Barr virus (EBV) membrane antigen gp350/220 (EBV-MA) lacking the membrane anchor was expressed and secreted into the medium of recombinant Chinese hamster ovary cells that had been cultured in Plasmapur hollow-fibre modules using defined serum-free medium. The EBV-MA in the medium was concentrated by 70% (w/v) ammonium sulphate precipitation and subsequently purified by immunoaffinity chromatography using an anti-EBV-MA (EBV.0T6) monoclonal antibody (mAb) column. Adsorbed antigen was eluted with 3 M MgCl2 in phosphate-buffered saline, concentrated by Mono Q anion-exchange chromatography and analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, silver staining and Western blotting using EBV-positive serum and anti-EBV-MA specific mAbs. Monospecific polyclonal rabbit antibodies against the purified EBV-MA were raised and purified by protein G affinity chromatography. For the measurement of EBV-MA antigen levels a sandwich enzyme-linked immunosorbent assay using rabbit polyclonal antibodies and a horseradish peroxidase-conjugated anti-MA mAb was developed having a detection level of 10 ng/ml

Morphological effects of porous poly-D,L-lactic acid/hydroxyapatite scaffolds produced by supercritical CO2 foaming on their mechanical performance

A novel supercritical CO2 foaming technique was used to fabricate scaffolds of controllable morphology and mechanical properties, with the potential to tailor the scaffolds to specific tissue engineering applications. Biodegradable scaffolds are widely used as temporary supportive structures for bone regeneration. The scaffolds must provide a sufficient mechanical support while allowing cell attachment and growth as well as metabolic activities. In this study, supercritical CO2 foaming was used to prepare fully interconnected porous scaffolds of poly-D,L-lactic acid and poly-D,L-lactic acid/hydroxyapatite. The morphological, mechanical and cell behaviours of the scaffolds were measured to examine the effect of hydroxyapatite on these properties. These scaffolds showed an average porosity in the range of 86%–95%, an average pore diameter of 229–347 µm and an average pore interconnection of 103–207 µm. The measured porosity, pore diameter, and interconnection size are suitable for cancellous bone regeneration. Compressive strength and modulus of up to 36.03 ± 5.90 and 37.97 ± 6.84 MPa were measured for the produced porous scaffolds of various compositions. The mechanical properties presented an improvement with the addition of hydroxyapatite to the structure. The relationship between morphological and mechanical properties was investigated. The matrices with different compositions were seeded with bone cells, and all the matrices showed a high cell viability and biocompatibility. The number of cells attached on the matrices slightly increased with the addition of hydroxyapatite indicating that hydroxyapatite improves the biocompatibility and proliferation of the scaffolds. The produced poly-D,L-lactic acid/hydroxyapatite scaffolds in this study showed a potential to be used as bone graft substitutes

Artificial light at night causes diapause inhibition and sex-specific life history changes in a moth

Rapidly increasing levels of light pollution subject nocturnal organisms to major alterations of their habitat, the ecological consequences of which are largely unknown. Moths are well-known to be attracted to light at night, but effects of light on other aspects of moth ecology, such as larval development and life-history, remain unknown. Such effects may have important consequences for fitness and thus for moth population sizes. To study the effects of artificial night lighting on development and life-history of moths, we experimentally subjected Mamestra brassicae (Noctuidae) caterpillars to low intensity green, white, red or no artificial light at night and determined their growth rate, maximum caterpillar mass, age at pupation, pupal mass and pupation duration. We found sex-specific effects of artificial light on caterpillar life-history, with male caterpillars subjected to green and white light reaching a lower maximum mass, pupating earlier and obtaining a lower pupal mass than male caterpillars under red light or in darkness. These effects can have major implications for fitness, but were absent in female caterpillars. Moreover, by the time that the first adult moth from the dark control treatment emerged from its pupa (after 110 days), about 85% of the moths that were under green light and 83% of the moths that were under white light had already emerged. These differences in pupation duration occurred in both sexes and were highly significant, and likely result from diapause inhibition by artificial night lighting. We conclude that low levels of nocturnal illumination can disrupt life-histories in moths and inhibit the initiation of pupal diapause. This may result in reduced fitness and increased mortality. The application of red light, instead of white or green light, might be an appropriate measure to mitigate negative artificial light effects on moth life history

Dynamical confirmation of a stellar-mass black hole in the transient X-ray dipping binary MAXI J1305-704

MAXI J1305-704 has been proposed as a high-inclination candidate black hole X-ray binary in view of its X-ray properties and dipping behaviour during outburst. We present photometric and spectroscopic observations of the source in quiescence that allow us to reveal the ellipsoidal modulation of the companion star and absorption features consistent with those of an early K-type star (Teff = 4610 +130 -160 K). The central wavelengths of the absorption lines vary periodically at Porb = 0.394 +- 0.004 d with an amplitude of K2 = 554 +- 8 km/s . They imply a mass function for the compact object of f(M1) = 6.9 +- 0.3 Msun, confirming its black hole nature. The simultaneous absence of X-ray eclipses and the presence of dips set a conservative range of allowed inclinations 60 deg < i < 82 deg, while modelling of optical light curves further constrain it to i = 72 +5 -8 deg. The above parameters together set a black hole mass of M1 = 8.9 +1.6 -1.0 Msun and a companion mass of M2=0.43 +- 0.16 Msun, much lower than that of a dwarf star of the observed spectral type, implying it is evolved. Estimates of the distance to the system (d = 7.5 +1.8 -1.4 kpc) and space velocity (vspace = 270 +- 60 km/s ) place it in the Galactic thick disc and favour a significant natal kick during the formation of the BH if the supernova occurred in the Galactic Plane.Comment: 14 pages, 6 figures. Submitted to MNRA

The Chandra Galactic Bulge Survey: optical catalogue and point-source counterparts to X-ray sources

As part of the Chandra Galactic Bulge Survey (GBS), we present a catalogue of optical sources in the GBS footprint. This consists of two regions centered at Galactic latitude b = 1.5 degrees above and below the Galactic Centre, spanning (l x b) = (6x1) degrees. The catalogue consists of 2 or more epochs of observations for each line of sight in r', i' and H{\alpha} filters. It is complete down to r' = 20.2 and i' = 19.2 mag; the mean 5{\sigma} depth is r' = 22.5 and i' = 21.1 mag. The mean root-mean-square residuals of the astrometric solutions is 0.04 arcsec. We cross-correlate this optical catalogue with the 1640 unique X-ray sources detected in Chandra observations of the GBS area, and find candidate optical counterparts to 1480 X-ray sources. We use a false alarm probability analysis to estimate the contamination by interlopers, and expect ~ 10 per cent of optical counterparts to be chance alignments. To determine the most likely counterpart for each X-ray source, we compute the likelihood ratio for all optical sources within the 4{\sigma} X-ray error circle. This analysis yields 1480 potential counterparts (~ 90 per cent of the sample). 584 counterparts have saturated photometry (r'<17, i'<16), indicating these objects are likely foreground sources and the real counterparts. 171 candidate counterparts are detected only in the i'-band. These sources are good qLMXB and CV candidates as they are X-ray bright and likely located in the Bulge.Comment: 18 pages, 18 figures. Published in MNRAS. 2016MNRAS.458.4530

Bivalirudin started during emergency transport for primary PCI.

BACKGROUND: Bivalirudin, as compared with heparin and glycoprotein IIb/IIIa inhibitors, has been shown to reduce rates of bleeding and death in patients undergoing primary percutaneous coronary intervention (PCI). Whether these benefits persist in contemporary practice characterized by prehospital initiation of treatment, optional use of glycoprotein IIb/IIIa inhibitors and novel P2Y12 inhibitors, and radial-artery PCI access use is unknown. METHODS: We randomly assigned 2218 patients with ST-segment elevation myocardial infarction (STEMI) who were being transported for primary PCI to receive either bivalirudin or unfractionated or low-molecular-weight heparin with optional glycoprotein IIb/IIIa inhibitors (control group). The primary outcome at 30 days was a composite of death or major bleeding not associated with coronary-artery bypass grafting (CABG), and the principal secondary outcome was a composite of death, reinfarction, or non-CABG major bleeding. RESULTS: Bivalirudin, as compared with the control intervention, reduced the risk of the primary outcome (5.1% vs. 8.5%; relative risk, 0.60; 95% confidence interval [CI], 0.43 to 0.82; P=0.001) and the principal secondary outcome (6.6% vs. 9.2%; relative risk, 0.72; 95% CI, 0.54 to 0.96; P=0.02). Bivalirudin also reduced the risk of major bleeding (2.6% vs. 6.0%; relative risk, 0.43; 95% CI, 0.28 to 0.66; P<0.001). The risk of acute stent thrombosis was higher with bivalirudin (1.1% vs. 0.2%; relative risk, 6.11; 95% CI, 1.37 to 27.24; P=0.007). There was no significant difference in rates of death (2.9% vs. 3.1%) or reinfarction (1.7% vs. 0.9%). Results were consistent across subgroups of patients. CONCLUSIONS: Bivalirudin, started during transport for primary PCI, improved 30-day clinical outcomes with a reduction in major bleeding but with an increase in acute stent thrombosis. (Funded by the Medicines Company; EUROMAX ClinicalTrials.gov number, NCT01087723.)

How to Teach Engineering Ethics?: A Retrospective and Prospective Sketch of TU Delft’s Approach to Engineering Ethics Education

This paper provides a retrospective and prospective overview of TU Delft’s approach to engineering ethics education. For over twenty years, the Ethics and Philosophy of Technology Section at TU Delft has been at the forefront of engineering ethics education, offering education to a wide range of engineering and design students. The approach developed at TU Delft is deeply informed by the research of the Section, which is centered around Responsible Research and Innovation, Design for Values, and Risk Ethics. These theoretical approaches are premised on the notion that technologies are inherently value-laden, and as such contain the possibility of fostering or hindering moral values. Each of these approaches encourages students to take a proactive attitude with respect to their projects and profession, thinking creatively about – and taking responsibility for – how to both prevent harm and do good via the technologies they help develop. To explain how this is put into practice, this paper sketches a brief history of ethics teaching at TU Delft, outlines current activities, and presents future plans for Bachelor and Master’s level engineering ethics education at TU Delft.Ethics & Philosophy of TechnologyValues Technology and Innovatio

The characteristics and pre-hospital management of blunt trauma patients with suspected spinal column injuries:a retrospective observational study

Background Pre-hospital spinal immobilisation by emergency medical services (EMS) staff is currently the standard of care in cases of suspected spinal column injuries. There is, however, a lack of data on the characteristics of patients who received spinal immobilisation during the pre-hospital phase and on the adverse effects of immobilisation. The objectives of this study were threefold. First, we determined the pre-hospital characteristics of blunt trauma patients with suspected spinal column injuries who were immobilised by EMS staff. Second, we assessed the choices made by EMS staff regarding spinal immobilisation techniques and reasons for immobilisation. Third, we researched the possible adverse effects of immobilisation. Design A retrospective observational study in a cohort of blunt trauma patients. Study method Data of blunt trauma patients with suspected spinal column injuries were collected from one EMS organisation between January 2008 and January 2013. Coded data and free text notes were analysed. Results A total of 1082 patients were included in this study. Spinal immobilisation was applied in 96.3 % of the patients based on valid pre-hospital criteria. In 2.1 % of the patients immobilisation was not based on valid criteria. Data of 1.6 % patients were missing. Main reasons for spinal immobilisation were posterior midline spinal tenderness (37.2 % of patients) and painful distracting injuries (13.5 % of patients). Spinal cord injury (SCI) was suspected in 5.7 % of the patients with posterior midline spinal tenderness. A total of 15.8 % patients were immobilised using non-standard methods. The reason for departure from the standard method was explained for 3 % of these patients. Reported adverse effects included pain (n = 10, 0.9 %,); shortness of breath (n = 3, 0.3 %); combativeness or anxiety (n = 6, 0.6 %); and worsening of pain when supine (n = 1, 0.1 %). Conclusion/recommendation Spinal immobilisation was applied in 96.3 % of all included patients based on pre-hospital criteria. We found that consensus among EMS staff on how to interpret the criterion 'distracting injury' was lacking. Furthermore, the adverse effects of spinal immobilisation were incompletely documented in pre-hospital care reports. To provide validated information on potential symptoms of SCI, a uniform EMS scoring system for motoric assessment should be developed

Excessive toxicity of cabozantinib in a phase II study in patients with recurrent and/or metastatic salivary gland cancer

AIM: Because the tyrosine kinases c-MET and vascular endothelial growth factor receptors (VEGFR) are often overexpressed in salivary gland cancer (SGC), this study evaluated the efficacy and safety of cabozantinib in patients with recurrent/metastatic (R/M) SGC. PATIENTS AND METHODS: A single-centre phase II study was conducted. Patients with immunohistochemical c-MET-positive R/M SGC were included in three cohorts: adenoid cystic carcinoma (ACC); salivary duct carcinoma (SDC) and other miscellaneous SGCs. No prior systemic treatments were required. Patients started cabozantinib 60 mg once daily. The primary outcome was the objective response rate (ORR). Secondary outcomes included survival, safety and quality of life. Per Simon-two-stage design, depending on efficacy, a maximum of 43 patients would be included. RESULTS: In total, 25 patients were included until premature closure owing to severe toxicity. Six patients (24%) had grade 3-5 wound complications, occurring at a median of 7.1 months on cabozantinib treatment (range 2.1-12.6). Remarkably, four of these six patients developed this complication in the area prior exposed to high-dose radiotherapy. Other grade ≥3 adverse events in >1 patient were hypertension (20%), diarrhoea (8%) and dehydration (8%). Twenty-one patients were evaluable for response; 1/15 ACC (ORR: 7%); 1/4 SDC and 0/2 patients with other miscellaneous SGC responded. Median progression-free survival was 9.4 months (95% confidence interval [CI] 7.4-11.4 months), 7.2 months (95%CI 0.0-15.1) and 6.9 months (95%CI 0.0-15.1), respectively. CONCLUSION: This study showed too many severe cabozantinib-associated wound complications in patients with SGC, especially in prior irradiated areas. Therefore, the study closed prematurely. The efficacy in the limited number of evaluable patients was low to moderate. TRIAL REGISTRATION: This trial was registered on ClinicalTrials.gov: NCT03729297