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At least 343 records · Page 19

Evaluation of a laboratory-developed multiplex real-time PCR assay for diagnosis of syphilis, herpes and chancroid genital ulcers in four public health laboratories in the USA

Objective To evaluate the field performance of a multiplex PCR (M-PCR) assay for detection of herpes simplex virus (HSV)-1 and HSV-2, Treponema pallidum ( T. pallidum ) and Haemophilus ducreyi ( H. ducreyi ) in genital ulcer disease (GUD) specimens. Methods GUD M-PCR was performed on 186 remnant specimens, previously collected for HSV testing, by four public health laboratories (PHLs) and the Laboratory Reference and Research Branch (LRRB) at the Centers for Disease Control and Prevention. The results from the PHLs were compared with those of LRRB, which served as the reference testing method, and percentage agreement was calculated. Results HSV was detected in 31 of 52 (59.6%), 20 of 40 (50%), 43 of 44 (97.7%) and 19 of 50 (38.0%) specimens from PHL1, PHL2, PHL3 and PHL4, respectively. There were seven discrepant results for HSV, and the overall percent agreement between the PHLs and the LRRB was 94%–100%, with a kappa value of 0.922, which demonstrates high agreement. T. pallidum was identified in 7 of 51 (13.7%) specimens from PHL1 with 94.1% agreement and in 2 of 40 (5.0%) specimens from PHL2 with 100% agreement. The LRRB identified three additional T. pallidum -positive specimens from PHL1. The kappa value (0.849) for T. pallidum testing suggests good agreement. Consistent with the LRRB results, no T. pallidum was detected in specimens from PHL3 and PHL4, and H. ducreyi was not detected at any of the study sites. Conclusions The GUD M-PCR assay performed well in four independent PHLs and 12 suspected syphilis cases were identified in this study. The M-PCR assay could provide improved diagnostic options for GUD infections in state and local PHLs.

Infectious Diseases↗

A high-density, high-channel count, multiplexed μECoG array for auditory-cortex recordings

Our understanding of the large-scale population dynamics of neural activity is limited, in part, by our inability to record simultaneously from large regions of the cortex. Here, we validated the use of a large-scale active microelectrode array that simultaneously records 196 multiplexed micro-electrocortigraphical (μECoG) signals from the cortical surface at a very high density (1,600 electrodes/cm 2 ). We compared μECoG measurements in auditory cortex using a custom “active” electrode array to those recorded using a conventional “passive” μECoG array. Both of these array responses were also compared with data recorded via intrinsic optical imaging, which is a standard methodology for recording sound-evoked cortical activity. Custom active μECoG arrays generated more veridical representations of the tonotopic organization of the auditory cortex than current commercially available passive μECoG arrays. Furthermore, the cortical representation could be measured efficiently with the active arrays, requiring as little as 13.5 s of neural data acquisition. Next, we generated spectrotemporal receptive fields from the recorded neural activity on the active μECoG array and identified functional organizational principles comparable to those observed using intrinsic metabolic imaging and single-neuron recordings. Furthermore, this new electrode array technology has the potential for large-scale, temporally precise monitoring and mapping of the cortex, without the use of invasive penetrating electrodes.

60 APPLIED LIFE SCIENCES↗

Data from Development of a CRISPR/Cas9 System for High-Efficiency Multiplexed Gene Deletion in Rhodosporidium toruloides

The oleaginous yeast Rhodosporidium toruloides is considered a promising candidate for production of chemicals and biofuels thanks to its ability to grow on lignocellulosic biomass, and its high production of lipids and carotenoids. However, efforts to engineer this organism are hindered by a lack of suitable genetic tools. Here we report the development of a CRISPR/Cas9 system for genome editing in R. toruloides based on a fusion 5S rRNA–tRNA promoter for guide RNA (gRNA) expression, capable of greater than 95% gene knockout for various genetic targets. Additionally, multiplexed double‐gene knockout mutants were obtained using this method with an efficiency of 78%. This tool can be used to accelerate future metabolic engineering work in this yeast.

Conversion↗

Signal Processing of Multiplexed Optical PWM Signals for Sensor Arrays in Nuclear Environments

Safe and effective generation of terrestrial nuclear power greatly benefits from the actionable data provided by the array of sensors located throughout a plant to provide a holistic online indication of reactor operation. This array includes sensors for monitoring coolant flow and pressure, temperature and heat transfer, radiation levels, structure health monitoring, and other critical parameters for reactor operation. While sensors capable of measuring these parameters have been developed, the electronics used in the pre-amplification and analog-to-digital conversion of the small signals they produce are extremely sensitive and susceptible to damage by high-temperatures and radiation environments nuclear reactors encounter while generating power. The small signals from sensors in nuclear power plants (NPPs) are transmitted over long cable runs which introduce dispersion artifacts into the signals of interest as well as electromagnetic interference (EMI) from lighting fixtures, pumps, mains electricity, and other equipment. To overcome these challenges, a front-end digitization (FREND) platform has been developed to use radiation-tolerant electronics to multiplex, amplify, and optically encode signals from an array of sensors for transmission over an optical fiber to mitigate dispersion and EMI artifacts from long runs of electrical cabling. To recover the optically transmitted data, a signal processing scheme based on 1-dimensional template matching to an indexing channel is described herein and demonstrated to have an effective bit-depth of 9.2 bits (1%). This scheme has been validated in proof-of-concept, non-nuclear testing and preliminary experimental results show good agreement between the measured optical output and and sensor input signals. The FREND system represents a low-loss data link between sensors in nuclear environments and data acquisition hardware which is aimed at improving the signal-to-noise ratio of the data acquired from these sensors to provide better information to operators and researchers.

Sweeney, Dan↗

Practical Multiplexing For Third Generation Quantum Repeaters

We restrict quantum error correcting code sizes in third-generation quantum repeaters to explore qubit channel multiplexing. We find that the single link optimization is not necessarily the optimal configuration for improving the practical encoded-qubit throughput.

97 MATHEMATICS AND COMPUTING↗

Silicon photonic chip for 16-channel wavelength division (de-)multiplexing in the O-band

We experimentally demonstrate a silicon photonic chip-scale 16-channel wavelength division multiplexer (WDM) operating in the O-band. The silicon photonic chip consists of a common-input bus waveguide integrated with a sequence of 16 spectral add-drop filters implemented by 4-port contra-directional Bragg couplers and resonant cladding modulated perturbations. The combination of these features reduces the spectral bandwidth of the filters and improves the crosstalk. An apodization of the cladding modulated perturbations between the bus and the add/drop waveguides is used to optimize the strength of the coupling coefficient in the propagation direction to reduce the intra-channel crosstalk on adjacent channels. The fabricated chip was validated experimentally with a measured intra-channel crosstalk of ∼−18.9 dB for a channel spacing of 2.6 nm. The multiplexer/demultiplexer chip was also experimentally tested with a 10 Gbps data waveform. The resulting eye-pattern indicates that this approach is suitable for datacenter WDM-based interconnects in the O-band with large aggregate bandwidths.

Davis, Jordan A. (ORCID:0000000343483295)↗

Cryogenic C-band wavelength division multiplexing system using an AIM Photonics Foundry process design kit

Cryogenic environments make superconducting computing possible by reducing thermal noise, electrical resistance and heat dissipation. Heat generated by the electronics and thermal conductivity of electrical transmission lines to the outside world constitute two main sources of thermal load in such systems. As a result, higher data rates require additional transmission lines which come at an increasingly higher cooling power cost. Hybrid or monolithic integration of silicon photonics with the electronics can be the key to higher data rates and lower power costs in these systems. We present a 4-channel wavelength division multiplexing photonic integrated circuit (PIC) built from modulators in the AIM Photonics process development kit (PDK) that operate at 25 Gbps at room temperature and 10 Gbps at 40 K. We further demonstrate 2-channel operation for 20 Gbps aggregate data rate at 40 K using two different modulators/wavelengths, with the potential for higher aggregate bit rates by utilizing additional channels.

42 ENGINEERING↗

Parametric amplification of angularly multiplexed waves for application to beam smoothing

The parametric amplification of angularly multiplexed waves is studied via modeling and experiments. The phase-matching properties of the amplification process with a single pump, including parasitic interactions of pairs of signal/idler, are investigated. An experimental demonstration with up to six signal waves is presented, with a quantification of the output properties, including stability relative to the input pump and seeds. A proof-of-concept study of beam smoothing induced by combining up to 12 speckle fields generated from six signal waves and six idler waves is shown.

Dorrer, C. [University of Rochester, NY (United St↗

Real-time multiplexed digital cavity-enhanced spectroscopy

Cavity-enhanced spectroscopy is a sensitive optical absorption technique but one where the practical applications have been limited to studying small wavelength ranges. In addition, this Letter shows that wideband operation can be achieved by combining techniques usually reserved for the communications community with that of cavity-enhanced spectroscopy, producing a multiplexed real-time cavity-enhanced spectrometer. We use multiple collinear laser sources operating asynchronously and simultaneously while being detected on a single photodetector. This is synonymous with radio frequency (RF) cellular systems in which signals are detected on a single antenna but decoded uniquely. Here, we demonstrate results with spectra of methyl salicylate and show parts-per-billion per root hertz sensitivity measured in real-time.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Point-by-point inscribed sapphire parallel fiber Bragg gratings in a fully multimode system for multiplexed high-temperature sensing

In this work, we study the point-by-point inscription of sapphire parallel fiber Bragg gratings (sapphire pFBGs) in a fully multimode system. A parallel FBG is shown to be critical in enabling detectable and reliable high-order grating signals. The impacts of modal volume, spatial coherence, and grating location on reflectivity are examined. Three cascaded seventh-order pFBGs are fabricated in one sapphire fiber for wavelength multiplexed temperature sensing. Using a low-cost, fully multimode 850-nm interrogator, reliable measurement up to 1500°C is demonstrated.

42 ENGINEERING↗

Multiplexed color centers in a silicon photonic cavity array

Entanglement distribution is central to the modular scaling of quantum processors and establishing quantum networks. Color centers with telecom-band transitions and long spin coherence times are suitable candidates for long-distance entanglement distribution. However, high-bandwidth memory-enhanced quantum communication is limited by high-yield, scalable creation of efficient spin-photon interfaces. Here, we develop a silicon photonics platform consisting of arrays of bus-coupled cavities. The coupling to a common bus waveguide enables simultaneous access to individually addressable cavity-enhanced T center arrays. We demonstrate frequency-multiplexed operation of two T centers in separate photonic crystal cavities. In addition, we investigate the cavity enhancement of a T center through hybridized modes formed between physically distant cavities. Our results show that bus-coupled arrays of cavity-enhanced color centers could enable efficient on-chip and long-distance entanglement distribution.

Komza, Lukasz↗

Multiplexed Digital mRNA Profiling of the Inflammatory Response in the West Nile Swiss Webster Mouse Model

Background and purpose: The ability to track changes in gene expression following viral infection is paramount to understanding viral pathogenesis. This study was undertaken to evaluate the nCounter, a high throughput digital gene expression system, as a means to better understand West Nile virus (WNV) dissemination and the inflammatory response against WNV in the outbred Swiss Webster (SW) mouse model over the course of infection. Methodology: The nCounter Mouse Inflammation gene expression kit containing 179 inflammation related genes was used to analyze gene expression changes in multiple tissues over a nine day course of infection in SW mice following intraperitoneal injection with WNV. Protein expression levels for a subset of these cytokine/chemokine genes were determined using a multiplex protein detection system (BioPlex) and comparisons of protein/RNA expression levels made. Results: Expression analysis of spleen, lung, liver, kidney and brain of SW mice infected with WNV revealed that Cxcl10 and Il12b are differentially expressed in all tissues tested except kidney. Data stratification of positively confirmed infected (WNV (+)) versus non-infected (WNV (2) tissues allowed differentiation of the systemic inflammatory gene response from tissue specific responses arising from WNV infection. Significant (p,0.05) decrease in C3ar1 was found in WNV (2) spleen. Il23a was significantly upregulated, while Il10rb was down-regulated in WNV (2) lung. Il3 and Mbl2 were down-regulated in WNV (2) liver. In WNV (+) livers, Stat1, Tlr2, chemokines Cxcl1, Cxcl3, Cxcl9, Cxcl10, cytokines Il6, Il18, cytokine-related gene Il1r and cytokine agonist Ilrn were significantly upregulated. In WNV (2) brain tissues, Csf2 and Cxcl10 were significantly upregulated. Similar gene and protein expression kinetics were found for Ccl2, Ccl3, Ccl4 and Ccl5 and correlated with the presence of infectious virus. In summary, the utility of the nCounter platform for rapid identification of gene expression changes in SW mice associated with WNV infection was demonstrated.

54 ENVIRONMENTAL SCIENCES↗

Pooled PPIseq: Screening the SARS-CoV-2 and human interface with a scalable multiplexed protein-protein interaction assay platform

Protein-Protein Interactions (PPIs) are a key interface between virus and host, and these interactions are important to both viral reprogramming of the host and to host restriction of viral infection. In particular, viral-host PPI networks can be used to further our understanding of the molecular mechanisms of tissue specificity, host range, and virulence. At higher scales, viral-host PPI screening could also be used to screen for small-molecule antivirals that interfere with essential viral-host interactions, or to explore how the PPI networks between interacting viral and host genomes co-evolve. Current high-throughput PPI assays have screened entire viral-host PPI networks. However, these studies are time consuming, often require specialized equipment, and are difficult to further scale. Here, we develop methods that make larger-scale viral-host PPI screening more accessible. This approach combines the mDHFR split-tag reporter with the iSeq2 interaction-barcoding system to permit massively-multiplexed PPI quantification by simple pooled engineering of barcoded constructs, integration of these constructs into budding yeast, and fitness measurements by pooled cell competitions and barcode-sequencing. We applied this method to screen for PPIs between SARS-CoV-2 proteins and human proteins, screening in triplicate >180,000 ORF-ORF combinations represented by >1,000,000 barcoded lineages. Our results complement previous screens by identifying 74 putative PPIs, including interactions between ORF7A with the taste receptors TAS2R41 and TAS2R7, and between NSP4 with the transmembrane KDELR2 and KDELR3. We show that this PPI screening method is highly scalable, enabling larger studies aimed at generating a broad understanding of how viral effector proteins converge on cellular targets to effect replication.

60 APPLIED LIFE SCIENCES↗

Highly Multiplexed Assays for Detection of Biothreat and Food Safety Agents, CRADA No. TC02156.0

This was a collaborative effort between Lawrence Livermore National Security, LLC as manager and operator of Lawrence Livermore National Laboratory (LLNL) and BioSearch Technologies Inc. (BTI) of Novato, California, to develop Multiplexed Molecular Assays for the Rapid and Sensitive Detection of Microbial Agents of Concern for Bioterrorism and Food Safety for the Food and Drug Administration (FDA) Center for Food Safety and Applied Nutrition (CFSAN). There are no outstanding leans for this project.

45 MILITARY TECHNOLOGY, WEAPONRY, AND NATIONAL DEF↗

Low-Noise Amplifiers and Superconducting Flex Circuits for Frequency Domain Multiplexed Readout of Detector Arrays

The next-generation ground-based cosmic microwave background (CMB) experimental program aims to provide definitive measurements of the early universe using telescopes configured with large focal plane arrays requiring ~100,000 detectors and associated readout amplifiers and interconnecting components that are currently unavailable commercially. We describe research to demonstrate the feasibility of developing low-noise cryogenic amplifiers based on series SQUID arrays that are suitable for frequency domain multiplexed readout of large numbers of CMB detectors, and the feasibility of developing robust superconducting flex cables that will simplify cryogenic systems integration for large CMB instruments. Two iterations of prototype series SQUID array amplifier designs were completed. The designs were optimized using modeling and simulation tools to meet the requirements for CMB detector readouts. Two fabrication runs were completed, and a large number of devices from each design iteration were tested and characterized at 4 K. We also completed the development of processes for the fabrication of prototype superconducting flex cables on 4" wafers. Samples were tested for strength and robustness, and to determine the critical current at 4 K as a function of trace width. The new generation of series SQUID array amplifier designs exhibit excellent performance characteristics – very smooth output characteristics that simplify tuning and operation, low input inductance, low current noise referred to the input, and low power dissipation that for the first time will enable these devices to be operated at the focal plane array stage of CMB instruments. The processes developed in Phase I to produce superconducting flex cables were used to complete the fabrication of a full wafer with several test and sample flex circuits. The 6-micron thick flex material is very strong with a breaking strength of several pounds for a 20-mm wide strip. Critical current density was reasonable and scaled with trace width as expected.

79 ASTRONOMY AND ASTROPHYSICS↗

Highly Multiplexed Assays for Detection of Biothreat and Food Safety Agents Final Report CRADA No. TC02156.0

This was a collaborative effort between Lawrence Livermore National Security, LLC as manager and operator of Lawrence Livermore National Laboratory (LLNL) and BioSearch Technologies Inc. (BTI) of Novato, California, to develop Multiplexed Molecular Assays for the Rapid and Sensitive Detection of Microbial Agents of Concern for Bioterrorism and Food Safety for the Food and Drug Administration (FDA) Center for Food Safety and Applied Nutrition (CFSAN). There are no outstanding leans for this project.

59 BASIC BIOLOGICAL SCIENCES↗