Pan-Family Assays for Rapid Viral Screening: Reducing Delays in Public Health Responses During Pandemics.

BACKGROUND: Coronavirus disease 2019 has highlighted deficiencies in the testing capacity of many developed countries during the early stages of pandemics. Here we describe a strategy using pan-family viral assays to improve early accessibility of large-scale nucleic acid testing. METHODS: Coronaviruses and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) were used as a case study for assessing utility of pan-family viral assays during the early stages of a novel pandemic. Specificity of a pan-coronavirus (Pan-CoV) assay for a novel pathogen was assessed using the frequency of common human coronavirus (HCoV) species in key populations. A reported Pan-CoV assay was assessed to determine sensitivity to 60 reference coronaviruses, including SARS-CoV-2. The resilience of the primer target regions of this assay to mutation was assessed in 8893 high-quality SARS-CoV-2 genomes to predict ongoing utility during pandemic progression. RESULTS: Because of common HCoV species, a Pan-CoV assay would return false positives for as few as 1% of asymptomatic adults, but up to 30% of immunocompromised patients with respiratory disease. One-half of reported Pan-CoV assays identify SARS-CoV-2 and with small adjustments can accommodate diverse variation observed in animal coronaviruses. The target region of 1 well-established Pan-CoV assay is highly resistant to mutation compared to species-specific SARS-CoV-2 reverse transcriptase-polymerase chain reaction assays. CONCLUSIONS: Despite cross-reactivity with common pathogens, pan-family assays may greatly assist management of emerging pandemics through prioritization of high-resolution testing or isolation measures. Targeting highly conserved genomic regions make pan-family assays robust and resilient to mutation. A strategic stockpile of pan-family assays may improve containment of novel diseases before the availability of species-specific assays.

Detection of voluntary dehydration in paediatric populations using non-invasive point-of-care saliva and urine testing.

AIM: Voluntary dehydration, or lack of fluid intake despite water availability, is common in otherwise healthy children, and can lead to adverse effects. Most dehydration biomarkers are impractical for routine assessment in paediatric populations. This study aimed to assess two non-invasive hydration assessment tools, urine specific gravity (USG ) and a novel point-of-care (POC) salivary osmolarity (SOSM) sensor, in healthy children. METHODS: Volunteers were tested by colorimetric USG and a handheld SOSM system. Observed values were compared against previous studies to determine hydration status, as was the concordance between parameters. RESULTS: At the common USG threshold of 1.020, 42.4% of the 139 healthy children were dehydrated. The same prevalence was found using the 70-mOSM cut-off value. Comparative analysis of SOSM at varying USG thresholds demonstrated significantly higher SOSM in dehydrated children with a USG  ≥ 1.030 (P = 0.002). CONCLUSION: At the USG threshold of 1.020 and SOSM threshold of 70 mOSM, 42.4% of healthy children were found to be voluntarily dehydrated. Significantly higher SOSM was observed in dehydrated children (USG ≥ 1.030). As the first study on the utility of POC SOSM measurements for detecting dehydration, these results provide a foundation for future POC characterisation of SOSM in other populations and clinical contexts.

Meander Thin-Film Biosensor Fabrication to Investigate the Influence of Structural Parameters on the Magneto-Impedance Effect.

Thin-film magneto-impedance (MI) biosensors have attracted significant attention due to their high sensitivity and easy miniaturization. However, further improvement is required to detect weak biomagnetic signals. Here, we report a meander thin-film biosensor preparation to investigate the fabrication parameters influencing the MI effect. Specifically, we hypothesized that an optimal film thickness and sensing area size ratio could be achieved to obtain a maximum MI ratio. A meander multilayer MI biosensor based on a NiFe/Cu/NiFe thin-film was designed and fabricated into 3-, 6-, and 9-turn models with film thicknesses of 3 µm and 6 µm. The 9-turn biosensor resembled the largest sensing area, while the 3- and 6-turn biosensors were designed with identical sensing areas. The results indicated that the NiFe film thickness of 6 µm with a sensing area size of 14.4 mm2 resembling a 9-turn MI biosensor is the optimal ratio yielding the maximum MI ratio of 238%, which is 70% larger than the 3- and 6-turn structures. The 3- and 6-turn MI biosensors exhibited similar characteristics where the MI ratio peaked at a similar value. Our results suggest that the MI ratio can be increased by increasing the sensing area size and film thickness rather than the number of turns. We showed that an optimal film thickness to sensing area size ratio is required to obtain a high MI ratio. Our findings will be useful for designing highly sensitive MI biosensors capable of detecting low biomagnetic signals.

Heater Integrated Lab-on-a-Chip Device for Rapid HLA Alleles Amplification towards Prevention of Drug Hypersensitivity.

HLA-B*15:02 screening before administering carbamazepine is recommended to prevent life-threatening hypersensitivity. However, the unavailability of a point-of-care device impedes this screening process. Our research group previously developed a two-step HLA-B*15:02 detection technique utilizing loop-mediated isothermal amplification (LAMP) on the tube, which requires two-stage device development to translate into a portable platform. Here, we report a heater-integrated lab-on-a-chip device for the LAMP amplification, which can rapidly detect HLA-B alleles colorimetrically. A gold-patterned micro-sized heater was integrated into a 3D-printed chip, allowing microfluidic pumping, valving, and incubation. The performance of the chip was tested with color dye. Then LAMP assay was conducted with human genomic DNA samples of known HLA-B genotypes in the LAMP-chip parallel with the tube assay. The LAMP-on-chip results showed a complete match with the LAMP-on-tube assay, demonstrating the detection system's concurrence.

A meta-analysis of in vitro exposures to weak radiofrequency radiation exposure from mobile phones (1990-2015).

To function, mobile phone systems require transmitters that emit and receive radiofrequency signals over an extended geographical area exposing humans in all stages of development ranging from in-utero, early childhood, adolescents and adults. This study evaluates the question of the impact of radiofrequency radiation on living organisms in vitro studies. In this study, we abstract data from 300 peer-reviewed scientific publications (1990-2015) describing 1127 experimental observations in cell-based in vitro models. Our first analysis of these data found that out of 746 human cell experiments, 45.3% indicated cell changes, whereas 54.7% indicated no changes (p = 0.001). Realizing that there are profound distinctions between cell types in terms of age, rate of proliferation and apoptosis, and other characteristics and that RF signals can be characterized in terms of polarity, information content, frequency, Specific Absorption Rate (SAR) and power, we further refined our analysis to determine if there were some distinct properties of negative and positive findings associated with these specific characteristics. We further analyzed the data taking into account the cumulative effect (SAR × exposure time) to acquire the cumulative energy absorption of experiments due to radiofrequency exposure, which we believe, has not been fully considered previously. When the frequency of signals, length and type of exposure, and maturity, rate of growth (doubling time), apoptosis and other properties of individual cell types are considered, our results identify a number of potential non-thermal effects of radiofrequency fields that are restricted to a subset of specific faster-growing less differentiated cell types such as human spermatozoa (based on 19 reported experiments, p-value = 0.002) and human epithelial cells (based on 89 reported experiments, p-value < 0.0001). In contrast, for mature, differentiated adult cells of Glia (p = 0.001) and Glioblastoma (p < 0.0001) and adult human blood lymphocytes (p < 0.0001) there are no statistically significant differences for these more slowly reproducing cell lines. Thus, we show that RF induces significant changes in human cells (45.3%), and in faster-growing rat/mouse cell dataset (47.3%). In parallel with this finding, further analysis of faster-growing cells from other species (chicken, rabbit, pig, frog, snail) indicates that most undergo significant changes (74.4%) when exposed to RF. This study confirms observations from the REFLEX project, Belyaev and others that cellular response varies with signal properties. We concur that differentiation of cell type thus constitutes a critical piece of information and should be useful as a reference for many researchers planning additional studies. Sponsorship bias is also a factor that we did not take into account in this analysis.

Magneto-Impedance Biosensor Sensitivity: Effect and Enhancement.

Biosensors based on magneto-impedance (MI) effect are powerful tools for biomedical applications as they are highly sensitive, stable, exhibit fast response, small in size, and have low hysteresis and power consumption. However, the performance of these biosensors is influenced by a variety of factors, including the design, geometry, materials and fabrication procedures. Other less appreciated factors influencing the MI effect include measuring circuit implementation, the material used for construction, geometry of the thin film sensing element, and patterning shapes compatible with the interface microelectronic circuitry. The type magnetic (ferrofluid, Dynabeads, and nanoparticles) and size of the particles, the magnetic particle concentration, magnetic field strength and stray magnetic fields can also affect the sensor sensitivity. Based on these considerations it is proposed that ideal MI biosensor sensitivity could be achieved when the sensor is constructed in sandwich thick magnetic layers with large sensing area in a meander shape, measured with circuitry that provides the lowest possible external inductance at high frequencies, enclosed by a protective layer between magnetic particles and sensing element, and perpendicularly magnetized when detecting high-concentration of magnetic particles.

Cross-ethnicity tagging SNPs for HLA alleles associated with adverse drug reaction.

Reduction of adverse drug reaction (ADR) incidence through screening of predisposing human leucocyte antigen (HLA) alleles is a promising approach for many widely used drugs. However, application of these associations has been limited by the cost burden of HLA genotyping. Use of single nucleotide polymorphisms (SNPs) that can approximate ('tag') HLA alleles of interest has been proposed as a cost-effective and simple alternative to conventional genotyping. However, most reported SNP tags have not been validated and there is concern regarding clinical utility of this approach due to tagging inconsistency across different populations. We assess the ability of 67 previously reported and 378 novel tagging SNPs, identified here in 5 HLA reference panels, to tag 15 ADR-associated HLA alleles in a panel of 955 ethnically diverse samples. Tags for 8 HLA alleles of interest were identified with 100% sensitivity and >95% specificity. These SNPs may act as a reliable genotyping approach for the routine screening of patients, without the need to account for patient ethnicity.

Rapid Detection of HLA-B*57:01-Expressing Cells Using a Label-Free Interdigitated Electrode Biosensor Platform for Prevention of Abacavir Hypersensitivity in HIV Treatment.

Pre-treatment screening of individuals for human leukocyte antigens (HLA) HLA-B*57:01 is recommended for the prevention of life-threatening hypersensitivity reactions to abacavir, a drug widely prescribed for HIV treatment. However, the implementation of screening in clinical practice is hindered by the slow turnaround time and high cost of conventional HLA genotyping methods. We have developed a biosensor platform using interdigitated electrode (IDE) functionalized with a monoclonal antibody to detect cells expressing HLA-B*57:01. This platform was evaluated using cell lines and peripheral blood mononuclear cells expressing different HLA-B alleles. The functionalized IDE sensor was able to specifically capture HLA-B*57:01 cells, resulting in a significant change in the impedance magnitude in 20 min. This IDE platform has the potential to be further developed to enable point-of-care HLA-B*57:01 screening.

Development of an Ultrasensitive Impedimetric Immunosensor Platform for Detection of Plasmodium Lactate Dehydrogenase.

Elimination of malaria is a global health priority. Detecting an asymptomatic carrier of Plasmodium parasites to receive treatment is an important step in achieving this goal. Current available tools for detection of malaria parasites are either expensive, lacking in sensitivity for asymptomatic carriers, or low in throughput. We investigated the sensitivity of an impedimetric biosensor targeting the malaria biomarker Plasmodium lactate dehydrogenase (pLDH). Following optimization of the detection protocol, sensor performance was tested using phosphate-buffered saline (PBS), and then saliva samples spiked with pLDH at various concentrations. The presence of pLDH was determined by analyzing the sensor electrical properties before and after sample application. Through comparing percentage changes in impedance magnitude, the sensors distinguished pLDH-spiked PBS from non-spiked PBS at concentrations as low as 250 pg/mL (p = 0.0008). Percentage changes in impedance magnitude from saliva spiked with 2.5 ng/mL pLDH trended higher than those from non-spiked saliva. These results suggest that these biosensors have the potential to detect concentrations of pLDH up to two logs lower than currently available best-practice diagnostic tools. Successful optimization of this sensor platform would enable more efficient diagnosis of asymptomatic carriers, who can be targeted for treatment, contributing to the elimination of malaria.

High-speed indoor optical wireless communication system employing a silicon integrated photonic circuit.

Beam-steering-based optical wireless technologies are being widely investigated due to the capability of providing high-speed wireless connectivity in indoor applications. However, high-speed indoor optical wireless systems are traditionally realized with discrete bulky components, significantly limiting their practical applications. In this Letter, we demonstrate an infrared optical wireless communication system employing a miniaturized silicon integrated photonic circuit for beam steering for the first time. Experimental results show that up to 12.5 Gb/s optical wireless communication can be achieved with error-free performance over a free-space range of 140 cm, and limited mobility of users can be realized. The experimental results of this Letter open the way for realizing integrated high-speed optical wireless communications.

Mind the prevalence rate: overestimating the clinical utility of psychiatric diagnostic classifiers.

Currently, there is an intense pursuit of pathognomonic markers and diagnostic ('risk-based') classifiers of psychiatric conditions. Commonly, the epidemiological prevalence of the condition is not factored into the development of these classifiers. By not adjusting for prevalence, classifiers overestimate the potential of their clinical utility. As valid predictive values have critical implications in public health and allocation of resources, development of clinical classifiers should account for the prevalence of psychiatric conditions in both general and high-risk populations. We suggest that classifiers are most likely to be useful when targeting enriched populations.

Attentional set-shifting and social abilities in children with schizotypal and comorbid autism spectrum disorders.

OBJECTIVE: While diagnostically independent, autism and schizotypal disorders can co-occur. Their concurrent impact on outcomes and phenotypes has not been investigated. We investigated the impact of comorbid autism and schizotypal disorders in children on executive functioning and socio-pragmatic skills - core features of both disorders. METHOD: Executive functioning (assessed with the Cambridge Neuropsychological Test Automated Battery) and socio-pragmatic skills (assessed using the Melbourne Assessment of Schizotypy in Kids) were investigated in a total of 67 (6-12 year old) children with autism ( n = 15; M/F = 10/5), schizotypal disorder ( n = 8; M/F = 5/3) and comorbid autism and schizotypal disorder ( n = 12; M/F = 5/7) and typically developing children ( n = 32; M/F = 17/15). RESULTS: Both the autism and schizotypal disorder groups performed more poorly than the typically developing group on socio-pragmatic skills and overall performance (i.e. number of stages completed) of the intra-/extra-dimensional set-shifting task (all ps < 0.001). Clear distinctions between the autism and schizotypal groups were present in the intra-/extra-dimensional task relative to the typically developing group - the autism group had difficulties with extra-dimensional shifts ( p < 0.001), and the schizotypal disorder group with intra-dimensional shifts ( p = 0.08). Interestingly, the overall performance of the comorbid group on the intra-/extra-dimensional task was not significantly different from the typically developing group, and they were superior to both the autism ( p = 0.019) and schizotypal disorder ( p = 0.042) groups on socio-pragmatic skills. CONCLUSION: The phenotypical overlap between autism and schizotypal disorders may be precipitated by different cognitive styles and/or mechanisms associated with attention and information processing. We propose that sustaining and switching attention represent two poles of irregularities across the autism and schizotypal spectra, which appear to converge in a compensatory manner in the comorbid group. Our findings highlight the importance of investigating children with a dual diagnosis of autism and schizotypal disorders, and raise intriguing questions about possible mechanisms to explain the attenuated impairment observed in the group of children with comorbid autism and schizotpyal disorders.

Ultra-broadband, compact, and high-reflectivity circular Bragg grating mirror based on 220 nm silicon-on-insulator platform.

A compact (4.49 µm × 4.54 µm) and ultra-broadband circular Bragg grating mirror with relaxed fabrication requirements is proposed and demonstrated based on the 220 nm silicon-on-insulator (SOI) platform. Based on FDTD-simulations, the proposed grating mirror can achieve a reflectivity of >90% over a ultrabroad bandwidth of 500 nm (1263 - 1763 nm), and a high reflectivity of >95% over a broad bandwidth of 397 nm (1340 - 1737 nm), which covers the entire E- to U-bands. The circular grating is fabricated, and the experimental measurement results exhibit a high reflectivity of 93% - 98% within the measured band of 1530 to 1610 nm, which agrees well with simulations. Based on the proposed broadband and high-efficiency circular Bragg mirror, a compact notch filter with high rejection ratio (>10 dB) and low transmission loss (<0.5 dB) is also fabricated and presented, and the proposed filter could find various potential applications in optical communications and sensing applications. With its ultrabroad bandwidth, high reflectivity and compact size, the proposed circular Bragg mirror is expected to be a promising element for large-scale photonic integrated circuits and applications which require ultra-broadband and high-efficiency on-chip reflections.

Indoor infrared optical wireless localization system with background light power estimation capability.

The indoor user localization function is in high demand for high-speed wireless communications, navigations and smart-home applications. The optical wireless technology has been used to localize end users in indoor environments. However, its accuracy is typically very limited, due to the ambient light, which is relatively strong. In this paper, a novel high-localization-accuracy optical wireless based indoor localization system, based on the use of the mechanism that estimates background light intensity, is proposed. Both theoretical studies and demonstration experiments are carried out. Experimental results show that the accuracy of the proposed optical wireless indoor localization system is independent on the localization light strength, and that an average localization error as small as 2.5 cm is attained, which is 80% better than the accuracy of previously reported optical wireless indoor localization systems.

High contrast circular grating reflector on silicon-on-insulator platform.

A compact circular high contrast grating (HCG) reflector with a footprint of only 4.03 µm×4.32 µm on 220 nm silicon-on-insulator (SOI) platform is proposed and experimentally demonstrated. The proposed device breaks the high wavelength selectivity limitation for the conventional grating reflectors on a thin SOI platform by using the circular structure in a compact region. In addition, the device provides a high polarization selectivity over a wide wavelength range which is useful for applications such as tunable laser cavities and resonators to provide wide tuning range and high polarization stability. The circular structure based HCG reflector has an ultra-wide operational bandwidth (Δλ) of over 385 nm with the center wavelength (λ) set at 1550 nm, providing a Δλ/λ=24.83%. An average reflectance high of 94.15% is observed from 1525 to 1610 nm in the experimental measurement. The polarization extinction ratio is greater than 13 dB over the entire measured wavelength range.

Ultra-broadband and low-loss 3 dB optical power splitter based on adiabatic tapered silicon waveguides.

A broadband, low-loss and polarization-insensitive 3 dB optical power splitter based on adiabatic tapered silicon waveguides is proposed and investigated. 3D-FDTD simulation results show that the splitter achieves an output transmission efficiency of nearly 50% over an ultra-broad wavelength range from 1200 to 1700 nm. The device is fabricated, and experimental results show that the splitter exhibits a low excess loss of <0.19 dB for the TE polarization and <0.14 dB for the TM polarization over the entire measured wavelength range from 1530 to 1600 nm, while having an adiabatic taper length of only 5 µm. In addition, the measured power uniformity of the cascaded 1×8 splitter is only 0.47 dB, and 0.17 dB for the TE and TM polarizations, respectively. With the advantages of low loss, broad bandwidth, and compact size, the proposed splitter is a promising element for large-scale silicon integrated photonic circuits.

Lab on a chip sensor for rapid detection and antibiotic resistance determination of Staphylococcus aureus.

The Gram-positive bacterium, Staphylococcus aureus (S. aureus), is a major pathogen responsible for a variety of infectious diseases ranging from cellulitis to more serious conditions such as septic arthritis and septicaemia. Timely treatment with appropriate antibiotic therapy is essential to ensure clinical defervescence and to prevent further complications such as infective endocarditis or organ impairment due to septic shock. To date, initial antibiotic choice is empirical, using a "best guess" of likely organism and sensitivity- an approach adopted due to the lack of rapid identification methods for bacteria. Current culture based methods take up to 5 days to identify the causative bacterial pathogen and its antibiotic sensitivity. This paper provides proof of concept for a biosensor, based on interdigitated electrodes, to detect the presence of S. aureus and ascertain its sensitivity to flucloxacillin rapidly (within 2 hours) in a cost effective manner. The proposed method is label-free and uses non-faradic measurements. This is the first study to successfully employ interdigitated electrodes for the rapid detection of antibiotic resistance. The method described has important potential outcomes of faster definitive antibiotic treatment and more rapid clinical response to treatment.

Vertical Nanowire Electrode Arrays as Novel Electrochemical Label-Free Immunosensors.

A new method for the fabrication of a label-free electrochemical immunosensor based on vertical nanowires (VNWs) is proposed. The VNWs are functionalized to detect antibodies against a major astrocytic structural protein component, glial fibrillary acidic protein (GFAP). It is revealed that the interaction of GFAP-antibody with functionalized VNWs leads to a clear change in device conductance and the corresponding capacitance.

Experimental demonstration of a novel indoor optical wireless localization system for high-speed personal area networks.

In this Letter, we propose a novel indoor localization system based on optical wireless technology. By using the same architecture as the high-speed full-duplex indoor optical wireless communication system, the "search and scan" process, and the added transmission power and beam footprint information in the "search and scan" message, indoor localization functionality is achieved. Proof-of-concept experiments are carried out, and results show that an average error of <15 cm is achieved with a localization beam size of 1 m. In addition, the major localization-accuracy-limiting factors are analyzed both theoretically and experimentally. When incorporated with the optical wireless communication system, high-speed indoor wireless personal area networks can be achieved.

Decreased expression of mGluR5 within the dorsolateral prefrontal cortex in autism and increased microglial number in mGluR5 knockout mice: Pathophysiological and neurobehavioral implications.

Metabotropic glutamate receptor 5 (mGluR5) and microglial abnormalities have been implicated in autism spectrum disorder (ASD). However, controversy exists as to whether the receptor is down or upregulated in functioning in ASD. In addition, whilst activation of mGluR5 has been shown to attenuate microglial activation, its role in maintaining microglial homeostasis during development has not been investigated. We utilised published microarray data from the dorsolateral prefrontal cortex (DLPFC) of control (n=30) and ASD (n=27) individuals to carry out regression analysis to assess gene expression of mGluR5 downstream signalling elements. We then conducted a post-mortem brain stereological investigation of the DLPFC, to estimate the proportion of mGluR5-positive neurons and glia. Finally, we carried out stereological investigation into numbers of microglia in mGluR5 knockout mice, relative to wildtype littermates, together with assessment of changes in microglial somal size, as an indicator of activation status. We found that gene expression of mGluR5 was significantly decreased in ASD versus controls (p=0.018) as well as downstream elements SHANK3 (p=0.005) and PLCB1 (p=0.009) but that the pro-inflammatory marker NOS2 was increased (p=0.047). Intensity of staining of mGluR5-positive neurons was also significantly decreased in ASD versus controls (p=0.016). Microglial density was significantly increased in mGluR5 knockout animals versus wildtype controls (p=0.011). Our findings provide evidence for decreased expression of mGluR5 and its signalling components representing a key pathophysiological hallmark in ASD with implications for the regulation of microglial number and activation during development. This is important in the context of microglia being considered to play key roles in synaptic pruning during development, with preservation of appropriate connectivity relevant for normal brain functioning.