Evaluation of the performance of a lateral flow device for quantitative detection of anti-SARS-CoV-2 IgG.

INTRODUCTION: The AbC-19™ lateral flow immunoassay (LFIA) performance was evaluated on plasma samples from a SARS-CoV-2 vaccination cohort, WHO international standards for anti-SARS-CoV-2 IgG (human), individuals ≥2 weeks from infection of RT-PCR confirmed SARS-CoV-2 genetic variants, as well as microorganism serology. METHODS: Pre-vaccination to three weeks post-booster samples were collected from a cohort of 111 patients (including clinically extremely vulnerable patients) from Northern Ireland. All patients received Oxford-AstraZeneca COVID-19 vaccination for the first and second dose, and Pfizer-BioNTech for the third (first booster). WHO international standards, 15 samples from 2 variants of concern (Delta and Omicron) and cross-reactivity with plasma samples from other microorganism infections were also assessed on AbC-19™. RESULTS: All 80 (100%) participants sampled post-booster had high positive IgG responses, compared to 38/95 (40%) participants at 6 months post-first vaccination. WHO standard results correlated with information from corresponding biological data sheets, and antibodies to all genetic variants were detected by LFIA. No cross-reactivity was found with exception of one (of five) Dengue virus samples. CONCLUSION: These findings suggest BNT162b2 booster vaccination enhanced humoral immunity to SARS-CoV-2 from pre-booster levels, and that this antibody response was detectable by the LFIA. In combination with cross-reactivity, standards and genetic variant results would suggest LFIA may be a cost-effective measure to assess SARS-CoV-2 antibody status.

Graphene supported graphone/graphane bilayer nanostructure material for spintronics.

We report an investigation into the magnetic and electronic properties of partially hydrogenated vertically aligned few layers graphene (FLG) synthesized by microwave plasma enhanced chemical vapor deposition. The FLG samples are hydrogenated at different substrate temperatures to alter the degree of hydrogenation and their depth profile. The unique morphology of the structure gives rise to a unique geometry in which graphane/graphone is supported by graphene layers in the bulk, which is very different from other widely studied structures such as one-dimensional nanoribbons. Synchrotron based x-ray absorption fine structure spectroscopy measurements have been used to investigate the electronic structure and the underlying hydrogenation mechanism responsible for the magnetic properties. While ferromagnetic interactions seem to be predominant, the presence of antiferromagnetic interaction was also observed. Free spins available via the conversion of sp(2) to sp(3) hybridized structures, and the possibility of unpaired electrons from defects induced upon hydrogenation are thought to be likely mechanisms for the observed ferromagnetic orders.

Comparative in vitro cytotoxicity study of carbon nanotubes and titania nanostructures on human lung epithelial cells.

The aim of this study is to assess in vitro cytotoxic effects of titania nanostructures and carbon nanotubes (CNTs) by exposing A549 lung epithelial cell line to these materials. Titania nanotubes (TiNTs) were grown by hydrothermal treatment of TiO(2) nanoparticles, followed by annealing them at 400°C. The titania nanostructures obtained on annealing (mixture of nanotubes and nanorods) were hollow and open ended, containing 3-5 layers of titania sheets, with an internal diameter ∼3-5 nm and external diameter ∼8-10 nm, and a specific surface area of 265 m(2)/g. As-supplied single walled (SWCNTs) and microwave plasma enhanced chemical vapour deposition (MPCVD) grown multi walled carbon nanotubes (MWCNTs) were used in this study. The lengths and diameters of the SWCNTs were 5-10nm and 0.5-3 nm respectively. The lengths and diameters of the MWCNTs were 25-30 µm and 10-30 nm respectively. The cell viability was evaluated using the MTT (3-(4,-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium) assay. No significant cytotoxic effects of titania nanostructures were observed over a period of a week of testing time, while the presence of CNTs in some cases demonstrated significant cytotoxic effects. Finally, possible reason of cytotoxicity is discussed in the light of microstructures of materials.

Evaluation of a CE approved ambulatory patient monitoring device in a general medical ward.

An evaluation of a newly CE approved bedside monitoring device used in a general hospital ward is presented. This evaluation has shown that it is feasible to use the system within this environment to provide medical staff with supplementary information on patient health, at more frequent intervals than traditional monitoring methods. The physiological data recorded by the body worn device is wirelessly transmitted to a patient management system for storage and display. Good correlation between heart rate values recorded by hospital staff and those recorded by the automated Vitalsens VS100 system was observed. The system has highlighted clinical information that routine observations alone did not readily identify. This can provide clinicians with a better view of the overall health status of the patient. Such medical issues include those witnessed in this study, namely paroxysmal AF, ectopic beats, increasing heart rates recorded prior to a hypoglycaemic event, general high and low heart rate trends and various instances where clinically relevant ECG data has been captured.

Transferring vertically aligned carbon nanotubes onto a polymeric substrate using a hot embossing technique for microfluidic applications.

We explored the hot embossing method for transferring vertically aligned carbon nanotubes (CNTs) into microfluidic channels, fabricated on poly-methyl-methacrylate (PMMA). Patterned and unpatterned CNTs were synthesized by microwave plasma-enhanced chemical vapour deposition on silicon to work as a stamp. For hot embossing, 115 degrees C and 1 kN force for 2 min were found to be the most suitable parameters for the complete transfer of aligned CNTs on the PMMA microchannel. Raman and SEM studies were used to analyse the microstructure of CNTs before and after hot embossing. The PMMA microparticles with dimensions (approx. 10 microm in diameter) similar to red blood cells were successfully filtered using laminar flow through these microfluidic channels. Finally, a microfluidic-based point-of-care device for blood filtration and detection of bio-molecules is drawn schematically.

Endothelial cell growth on silicon modified hydrogenated amorphous carbon thin films.

The biological response of human microvascular endothelial cells (HMEC-1) seeded on Si-DLC films and on control surfaces was evaluated in terms of initial cell enhancement, growth, and cytotoxicity. The microstructure of the films was characterised by Raman spectroscopy and X-ray photoelectron spectroscopy. The effect of changes in microstructure, surface energy, surface electronic state, and electronic conduction, on the biological response of the films to endothelial cells was investigated. Endothelial cell adhesion and growth was found to be affected by changes in the microstructure of the films induced by silicon doping and thermal annealing. We observed a significant statistical difference in endothelial cell count between the as-deposited DLC and Si-DLC films using the one sample t-test at a p-value of 0.05. We also found a statistically significant difference between the adhesion of HMEC films on DLC and Si-DLC films at various annealing temperatures using the one-way ANOVA F statistic test at p < 0.05 and the post-hoc Tukey test. One sample t-test at p < 0.05 of MTT-assay results showed the endothelial cells to be viable when seeded on DLC/Si-DLC films. We suspect that the increased adhesion of endothelial cells induced by increasing the amount of silicon in the Si-DLC films is associated with the development of a suitable surface energy due to silicon addition, which neither favored cell denaturing nor preferential water spreading before cellular attachment on the film surface. The presence of an external positively charged dipole on the Si-DLC films confirmed by our Kelvin probe measurements is also expected to enhance the adhesion of endothelial cells that are well known to carry a negative charge. The Si-DLC films investigated hold potential promise as coatings for haemocompatible artificial implants.

Fabrication of impedimetric sensors for label-free point-of-care immunoassay cardiac marker systems, with passive microfluidic delivery.

Miniaturised point-of-care cardiac marker sensors are being developed, based on impedimetric sensing of cardiac enzyme capture by an antibody layer immobilised on a planar gold electrode sensor. Gold/Ti-on-glass substrates have been used, in a 2 electrode configuration, with antibodies immobilised on the working electrode. Microfluidic structures have been fabricated by a CO2 laser, in 25 mum thick pressure sensitive adhesive (PSA), on a PMMA lid, and the structure bonded on top of the planar sensor. Microfluidic blood/serum delivery has been investigated using a visualisation dye. Some flow problems are observed if the sensor is exposed to air for several days, suggesting that flow channel nanopillars and hermetic encapsulation may be required to guarantee flow properties in commercially produced modules. Work is ongoing to characterise the impedimetric signal changes for myoglobin capture by antimyoglobin, using these sensors. Fifty micron thick PSA, incorporating a robust spacer layer, will be used to give better definition of channel walls.

Point of care biomedical sensors.

With the development of critical areas of interdisciplinary research, scientists will lead to an enhanced understanding and control of the key processing stages involved in the design and manufacture of sensor-and electrode-based micro-devices such as micro total analysis systems (muTAS), bio-chips, bio-arrays and DNA sensors. It is anticipated that this fundamental work will lead to tangible technology transfer in the areas of blood analysis, cardiac enzyme detectors, liver and renal function, infectious diseases, gene disorders, etc. These will obviously have major benefits to healthcare delivery. The development of miniaturised, integrated sensor/electrode-based devices are revolutionising both the delivery of health care. In this paper, some of the most exciting research themes in these areas are identified, as well as novel underpinning fabrication technologies and transduction principles, which should make future advances possible.

Macrophage responses to vascular stent coatings.

Diamond-like carbon (DLC) films have been proposed as potential coatings for blood-contacting devices. In this study, tetrahedral amorphous carbon (ta-C) films deposited by filtered cathodic vacuum arc system (FCVA) were compared with commercially deposited polyurethane coatings (PU) and uncoated stainless steel samples. X-ray reflectivity (XRR) measurements were performed to check density and thickness of the ta-C coatings, and contact angles measurements were used to assess surface wettability. J774 macrophages were used to assess the cell responses to the materials. Cell number, metabolic activity, and hydrogen peroxide production were measured by using biochemical assays, and the cell attachment and morphology were determined by using scanning electron microscopy and confocal laser scanning microscopy. Wettability measurements showed that of the materials, the stainless steel was the most hydrophilic, and the ta-C coatings were the most hydrophobic. Although the initial attachment and morphology did not appear to be dependent on the wettability, the cell numbers did increase with increasing wettability. Macrophages on the stainless steel samples were the most active in producing hydrogen peroxide. These data show that ta-C samples performed as well as commercial PU-coated samples in blocking cell reactions to the substrate and may prove to be effective coatings for blood-contacting materials.

Experimental determination of compound action potential direction and propagation velocity from multi-electrode nerve cuffs.

Information extracted from whole-nerve electroneurograms, recorded using electrode cuffs, can provide signals to neuroprostheses. However, the amount of information that can be extracted from a single tripole is limited. This communication demonstrates how previously unavailable information about the direction of action potential propagation and velocity can be obtained using a multi-electrode cuff and that the arrangement acts as a velocity-selective filter. Results from in vitro experiments on frog nerves are presented.

Platelet adhesion on silicon modified hydrogenated amorphous carbon films.

We have investigated the effect of changes in microstructure, surface energy, surface charge condition and electronic conduction on the interaction of human platelets with silicon modified hydrogenated amorphous carbon films (a-C:H:Si or Si-DLC). Results based on Raman spectroscopy, Scanning electron microscopy, X-ray photo-electron spectroscopy, surface energy measurements, electrical resistivity, contact potential difference, and thermal annealing indicates a correlation between some of the measured values and the interaction of the films with human blood platelets. Statistical analysis of platelet aggregation on the films using the Student's t-test indicated differences between platelet aggregation on the modified films compared to the as-deposited film at a p-value of <0.05.

First Report of Armillaria mellea s. str. in Hardwood Forests of Southern Ontario.

Armillaria mellea sensu stricto (Vahl:Fr.) Kummer has been found infrequently in studies conducted in northeastern North America (2) and it was not found in an earlier survey of northern Ontario (3). In a 1999 survey of Armillaria spp. conducted in central and southern Ontario, A. mellea s. str. was found at four of 110 locations surveyed, two approximately 100 km west and southwest of Ottawa, one 15 km north of Belleville, and one on the Niagara Peninsula in the Balls Falls Conservation Area. At the Ottawa area sites, A. mellea s. str. was found on red oak (Quercus rubra L.) and beech (Fagus grandifolia Ehrh.) stumps. A. ostoyae (Romagn.) Herink and A. gallica Marxmuller & Romagn. were also found on these sites. At the Belleville site, it was found on a dead co-dominant white birch (Betula papyrifera Marsh.), while at the Niagara Peninsula site it was found on a dead, suppressed beech. A. gallica was also found at these latter sites. Sites ranged from moisture regimes fresh to moist, and from sandy to silty soil from both calcareous and non-calcareous parent material. Four isolates were obtained from infected wood and rhizomorphs and identified by the polymerase chain reaction, yielding restriction fragment length polymorphism (RFLP) products of 500 and 183 base pairs. These fragment sizes are similar to findings reported in an earlier study (4). The identification was confirmed by haploid/diploid pairing. The Balls Falls Conservation Area is in close proximity to extensive fruit orchards and the presence of A. mellea is cause for concern, especially in light of its proven pathogenicity to fruit trees (1). This is the first confirmation of A. mellea s. str. in Ontario. References: (1) J. E. Adaskaveg and J. M. Ogawa. Plant Dis. 74:341-352, 1990. (2) J. T. Blodgett and J. J. Worrall. Plant Dis. 76:166-170, 1992. (3) M. T. Dumas. Can. J. For. Res. 18:872-874, 1988. (4) T. C. Harrington and B. D. Wingfield. Mycologia 87:280-288, 1995.

Glenoid replacement in total shoulder arthroplasty.

The outcome of total shoulder arthroplasty is largely based on the quality of glenoid component fixation, which, in turn, is related to the amount and quality of bone stock, glenohumeral stability, and rotator cuff function. This article discusses the anatomic and pathologic indications for glenoid replacement, surgical techniques, and results.

Axillary artery injury as a complication of proximal humerus fractures.

Proximal humerus fractures are common injuries and represent approximately 5% of all fractures. These fractures are infrequently associated with neurovascular injuries. Brachial plexus injuries are uncommon, whereas axillary artery injuries are rare. A review of 19 previously reported cases of axillary artery injury after proximal humerus fracture revealed that 84% occurred in patients older than 50 years, 53% were associated with brachial plexus injury, and 21% resulted in upper extremity amputation. This study describes a case of axillary artery injury after proximal humerus fracture and, on the basis of a literature review, offers suggestions for the early diagnosis and effective treatment of this uncommon injury.

Calcium buffer injections inhibit cytokinesis in Xenopus eggs.

A slow cortical wave of high calcium accompanies the elongation of cleavage furrows in medaka fish eggs as well as in Xenopus eggs. We explored the role of such waves by injecting calcium buffers into Xenopus eggs at various times before and during first and second cleavage. Injection earlier than about 15 minutes before first cleavage normally starts delays it for hours. Injection between about 15 minutes and a few minutes before cleavage normally starts allows a (short) furrow to form on time but usually yields an eccentric one. This forms away from the injection side, often as far off-center as the egg's equator, and then regresses. Injection soon after it starts quickly arrests elongation of the furrow and eventually induces its regression; while injection a bit later likewise soon arrests elongation but allows delocalized furrow deepening to continue. The dependence of these inhibitory actions upon the dissociation constants and final cytosolic concentrations of the injected buffers indicates that they act as shuttle buffers to suppress needed zones of high calcium in the micromolar range. We conclude that the high calcium that is found within these furrows is needed to induce them, to extend them and even to maintain them. Moreover, while short, eccentric furrows often form as far off center as the equator, they somehow always form along a meridian through the animal pole. This seems difficult to explain by the orthodox, diastral model. Rather, it suggests that the cleavage furrows in Xenopus--and perhaps in animal cells quite generally--are directly induced by a diastema or telophase disc rather than by the asters.