Use of Self-Collected Saliva Samples for the Detection of SARS-CoV-2.

OBJECTIVE: Using a US Food and Drug Administration (FDA) emergency use authorization (EUA) reverse transcription polymerase chain reaction (RT-PCR) method, we examined the analytic performance accuracy of saliva specimens as compared to nasopharyngeal (NP) specimens in symptomatic patients. Correlation between test results and symptoms was also evaluated. METHODS: Over a 5-week period in 2020, 89 matched saliva and nasopharyngeal swabs were collected from individuals exhibiting symptoms consistent with SARS-CoV-2. Specimens were tested with an FDA EUA-approved RT-PCR method, and performance characteristics were compared. RESULTS: The concordance rate between saliva and nasopharyngeal testing was 93.26%. The mean cycle threshold value of saliva when compared to the NP specimen was 3.56 cycles higher. As compared to NP swab, saliva testing demonstrates acceptable agreement but lower sensitivity. CONCLUSION: When compared to a reference method using NP swabs, the use of saliva testing proved to be a reliable method. Self-collected saliva testing for SARS-CoV-2 allows for a viable option when trained staff or collection materials are in short supply.

Performance Accuracy of a Laboratory-Developed Real-Time RT-PCR Method for Detection of SARS-CoV-2 in Self-Collected Saliva Specimens.

OBJECTIVE: The ongoing COVID-19 pandemic caused by SARS-CoV-2 has challenged diagnostic laboratories to re-examine traditional methods for collecting specimens and sample types used in molecular testing. Our goal was to demonstrate that saliva can be used for detecting SARS-CoV-2 and correlates well with established molecular methods using nasopharyngeal (NP) swabs. METHODS: We examined use of a saliva collection device in conjunction with a laboratory-developed real-time reverse transcription-polymerase chain reaction (LDPCR) method for detecting SARS-CoV-2 in a symptomatic population and compared results with 2 US Food and Drug Administration (FDA)-approved methods (emergency use authorization [EUA]) that use specimens from NP swabs. RESULTS: The sensitivity of LDPCR compared with the reference methods was 75.0% (21/28); specificity, 98.1% (104/106). When cycle threshold values were compared between paired specimens using the LDPCR and a EUA reverse transcription PCR method, both targeting the open-reading frame gene, the mean value for saliva was 4.66 cycles higher than for NP specimens. CONCLUSION: Use of self-collected saliva in conjunction with an LDPCR for SARS-CoV-2 compared favorably with 2 FDA EUA methods using NP swabs. The use of an alternative sample type and assay method will aid in expanding the availability of testing during the ongoing COVID-19 pandemic.

Laser Sintering of Liquid Metal Nanoparticles for Scalable Manufacturing of Soft and Flexible Electronics.

Soft, flexible, and stretchable electronics are needed to transmit power and information, and track dynamic poses in next-generation wearables, soft robots, and biocompatible devices. Liquid metal has emerged as a promising material for these applications due to its high conductivity and liquid phase state at room temperature; however, surface oxidation of liquid metal gives it unique behaviors that are often incompatible with scalable manufacturing techniques. This paper reports a rapid and scalable approach to fabricate soft and flexible electronics composed of liquid metal. Compared to other liquid metal patterning approaches, this approach has the advantages of compatibility with a variety of substrates, ease of scalability, and efficiency through automated processes. Nonconductive liquid metal nanoparticle films are sintered into electrically conductive patterns by use of a focused laser beam to rupture and ablate particle oxide shells, and allow their liquid metal cores to escape and coalesce. The laser sintering phenomenon is investigated through comparison with focused ion beam sintering and by studying the effects of thermal propagation in sintered films. The effects of laser fluence, nanoparticle size, film thickness, and substrate material on resistance of the sintered films are evaluated. Several devices are fabricated to demonstrate the electrical stability of laser-patterned liquid metal traces under flexing, multilayer circuits, and intricately patterned circuits. This work merges the precision, consistency, and speed of laser manufacturing with the material benefits of liquid conductors on elastic substrates to demonstrate decisive progress toward commercial-scale manufacturing of soft electronics.

Reducing Actuator Requirements in Continuum Robots Through Optimized Cable Routing.

Continuum manipulators offer many advantages compared to their rigid-linked counterparts, such as increased degrees of freedom and workspace volume. Inspired by biological systems, such as elephant trunks and octopus tentacles, many continuum manipulators are made of multiple segments that allow large-scale deformations to be distributed throughout the body. Most continuum manipulators currently control each segment individually. For example, a planar cable-driven system is typically controlled by a pair of cables for each segment, which implies two actuators per segment. In this article, we demonstrate how highly coupled crossing cable configurations can reduce both actuator count and actuator torque requirements in a planar continuum manipulator, while maintaining workspace reachability and manipulability. We achieve highly coupled actuation by allowing cables to cross through the manipulator to create new cable configurations. We further derive an analytical model to predict the underactuated manipulator workspace and experimentally verify the model accuracy with a physical system. We use this model to compare crossing cable configurations to the traditional cable configuration using workspace performance metrics. Our work here focuses on a simplified planar robot, both in simulation and in hardware, with the goal of extending this to spiraling-cable configurations on full 3D continuum robots in future work.

OmniSkins: Robotic skins that turn inanimate objects into multifunctional robots.

Robots generally excel at specific tasks in structured environments but lack the versatility and the adaptability required to interact with and locomote within the natural world. To increase versatility in robot design, we present robotic skins that can wrap around arbitrary soft bodies to induce the desired motions and deformations. Robotic skins integrate actuation and sensing into a single conformable material and may be leveraged to create a multitude of controllable soft robots with different functions or gaits to accommodate the demands of different environments. We show that attaching the same robotic skin to a soft body in different ways, or to different soft bodies, leads to distinct motions. Further, we show that combining multiple robotic skins enables complex motions and functions. We demonstrate the versatility of this soft robot design approach in a wide range of applications-including manipulation tasks, locomotion, and wearables-using the same two-dimensional (2D) robotic skins reconfigured on the surface of various 3D soft, inanimate objects.

A Soft Parallel Kinematic Mechanism.

In this article, we describe a novel holonomic soft robotic structure based on a parallel kinematic mechanism. The design is based on the Stewart platform, which uses six sensors and actuators to achieve full six-degree-of-freedom motion. Our design is much less complex than a traditional platform, since it replaces the 12 spherical and universal joints found in a traditional Stewart platform with a single highly deformable elastomer body and flexible actuators. This reduces the total number of parts in the system and simplifies the assembly process. Actuation is achieved through coiled-shape memory alloy actuators. State observation and feedback is accomplished through the use of capacitive elastomer strain gauges. The main structural element is an elastomer joint that provides antagonistic force. We report the response of the actuators and sensors individually, then report the response of the complete assembly. We show that the completed robotic system is able to achieve full position control, and we discuss the limitations associated with using responsive material actuators. We believe that control demonstrated on a single body in this work could be extended to chains of such bodies to create complex soft robots.

Hybrid Self-Assembly during Evaporation Enables Drop-on-Demand Thin Film Devices.

We propose and demonstrate a hybrid self-assembly process as the mechanism for producing strikingly uniform deposits from evaporating drops composed of cosolvents. This assembly process leverages both particle-fluid interactions to carry the particles to the drop surface and particle-interface interactions to assemble the particles into a uniform film. We anchor our results in a cosolvent evaporation model that agrees with our experimental observations. We further employ the process to produce thin film devices such as flexible broadband neutral density filters and semitransparent mirrors. Our observations suggest that this assembly process is free of particle-substrate interactions, which indicates that the results should be transferable across a multitude of material/substrate systems.

Mechanically sintered gallium-indium nanoparticles.

Liquid metal nanoparticles that are mechanically sintered at and below room temperature are introduced. This material can be sintered globally on large areas of entire deposits or locally to create liquid traces within deposits. The metallic nanoparticles are fabricated by dispersing a liquid metal in a carrier solvent via sonication. The resulting dispersion is compatible with inkjet printing, a process not applicable to the bulk liquid metal in air.

Reflections on the specificity of synaptic connections.

The principal focus of this treatise is the specificity of synaptic connectivity in the mammalian central nervous system. The occurrence of stereotypical patterns of connection at the macro level (e.g., the general consistency with which axonal pathways impinge on and originate within specific cortical areas and layers) implies that the cerebral cortex is a highly ordered structure. Order is seen also at the more micro level of synaptic connectivity, for instance, in the contrasting synaptic patterns of spiny vs. non-spiny neurons. Quantitative electron microscopic studies of synapses between identified neurons and correlative anatomical/electrophysiological investigations indicate that the high degree of order characterizing many aspects of cortical organization is mirrored by an equally ordered arrangement of synaptic connections between specific types of neurons. The recognition of recurring synaptic patterns has generated increased support for the notion of synaptic specificity as opposed to randomness, and we have begun now to understand the role of specificity in cortical function. At the core of cortical processing lie myriad possibilities for computation provided by the wealth of synaptic connections involving each neuron. Specificity, by limiting possibilities for connection, imposes an order on synaptic interactions even as processes of dynamic selection or synaptic remodeling ensure the constant formation and dissolution of cortical circuits. Collectively, these operations make maximal use of the richness of cortical synaptic connections to produce a highly flexible system, irrespective of the degree of hard-wiring, mutability, randomness or specificity that obtains for cortical wiring at any particular time. A brief, historical account of developments leading to our current understanding of cortical synaptic organization will precede the presentation of evidence for synaptic specificity.

Quantitative analysis of synaptic distribution along thalamocortical axons in adult mouse barrels.

Quantitative data on thalamocortical synapses in adult mouse barrels have been obtained largely by using lesion-nduced degeneration to label thalamic afferents. By the time degenerating axons can be identified with the electron microscope, they have broken up into many separate pieces, making it impossible to assess the distribution of synapses along unbroken lengths of afferent. Here, this deficiency is rectified by examining intact lengths of axon labeled by the injection of biotinylated dextran amine into ipsilateral thalamus. Serial thin section reconstructions were analyzed to determine the numbers of synapses per axon length made with dendritic spines vs. shafts and the locations of synapses with respect to axonal varicosities. Results for seven axonal segments from six mice showed an average of 0.2 synapses/microm; 80% were made with spines and 20% with dendritic shafts. Just over two-thirds of axonal varicosities formed one synapse; most of the remainder formed two and rarely three, whereas 8% formed none. Although most synapses occurred at varicosities (88%), more than 12% were made at cylindrically shaped regions of the reconstructed axonal segments. These results serve as a caveat for the use of light microscopy to quantify synapses, wherein the usual approach is to equate one varicosity with one synapse. For thalamocortical afferents to mouse barrels, equating one varicosity with one synapse would prove to be incorrect more than 30% of the time and would exclude the roughly 12% of synaptic connections made at cylindrical regions of thalamocortical afferents.

Changes in mouse barrel synapses consequent to sensory deprivation from birth.

Neonatal sensory deprivation induced by whisker trimming affects significantly the functional organization of receptive fields in adult barrel cortex. In this study, the effects of deprivation on thalamocortical synapses and on asymmetrical and symmetrical synapses not of thalamic origin were examined. Thalamocortical synapses were labeled by lesion-induced degeneration in adult (postnatal day 60) mice subjected to whisker trimming from birth, other synaptic types were unlabeled. Brains were processed for electron microscopy, and numerical densities of synapses were evaluated by using stereologic approaches for whisker trimmed vs. control animals. Results demonstrated no change in nonthalamic, asymmetrical synapses; however, a decrease of 52% in the numerical density of symmetrical synapses (46.3 vs. 88.5 million per mm(3); Z = -2.121; P < 0.05) and a decrease of 43% in the numerical density of thalamocortical synapses (57.5 vs. 102.33 million per mm(3); Z = -2.121; P < 0.05) were observed after deprivation. Thus, experience-dependent plasticity of receptive fields in barrel cortex involves directly axons of both extrinsic and intracortical origin. The proportion of thalamocortical axospinous to axodendritic synapses was the same in control vs. deprived animals: in each instance, 80% of the synapses were axospinous (Z = 0.85; P = 0.2). These results suggest that neither excitatory neurons, whose thalamocortical synapses are primarily axospinous, nor inhibitory neurons, whose thalamocortical synapses are mainly axodendritic (White [1989] Cortical Circuits. Synaptic Organization of the Cerebral Cortex; Structure, Function, and Theory. 1989; Boston: Birkhauser), are affected preferentially by the deprivation-associated decrease in thalamocortical synapses.

Synaptic patterns of thalamocortical afferents in mouse barrels at postnatal day 11.

This study focuses on the synaptic output patterns of thalamocortical axons in mouse barrel cortex at postnatal day (P) 11. Axons were labeled by biotinylated dextran amine transported anterogradely following injection in vivo into the ventrobasal thalamus. Labeled axons in the posteromedial barrel subfield were examined by light and electron microscopy and then reconstructed in three dimensions to assess the spatial distribution of their synapses. Thalamocortical axons form asymmetrical synapses, both at varicosities and along cylindrical portions of the axons; usually, only one synapse occurs per site, contrasting with the case in the adult, in which multiple synapses are typical. At P11, varicosities without synapses are common. As in adult barrels, approximately 80% of synapses formed by thalamocortical axons are with dendritic spines; 20% are with dendritic shafts. The similarity in the distribution of thalamocortical synapses onto spines vs. dendrites in developing and mature barrels indicates that adult synaptic patterns already are specified at a very early stage of thalamocortical synaptogenesis.

Specificity of cortical synaptic connectivity: emphasis on perspectives gained from quantitative electron microscopy.

This report traces the historical development of concepts regarding the specificity of synaptic connectivity in the cerebral cortex as viewed primarily from the perspective of electron microscopy. The occurrence of stereotypical patterns of connection (e.g., contrasting synaptic patterns on the surfaces of spiny vs. non-spiny neurons, the general consistency with which axonal pathways impinge on and originate within specific cortical areas and layers, triadic synaptic relationships) implies that cortical connectivity is highly structured. The high degree of order characterizing many aspects of cortical organization is mirrored by an equally ordered arrangement of synaptic connections between specific types of neurons. This observation is based on quantitative electron microscopic studies of synapses between identified neurons and from the results of correlative anatomical/electrophysiological investigations. The recognition of recurring synaptic patterns and responses between specific neurons has generated increased support for the notion of specificity of synaptic connections at the expense of randomness, but the role of specificity in cortical function is an unresolved question. At the core of cortical processing lie myriad possibilities for computation provided by the wealth of synaptic connections involving each cortical neuron. Specificity, by limiting possibilities for connection, can impose an order on synaptic interactions even as processes of dynamic selection or synaptic remodeling ensure the constant formation and dissolution of cortical circuits. These operations make maximal use of the richness of cortical synaptic connections to produce a highly flexible system, irrespective of the degree of randomness or specificity that obtains for cortical wiring at any particular time.