Severe Acute Respiratory Syndrome Coronavirus-2 Lambda Variant Collected from a Child from Arkansas and Sequenced.

An eleven-year-old tested positive for SARS-CoV-2 Lambda variant. Sequencing was performed on the Oxford Nanopore and the Illumina NextSeq 500. Both platforms identified all 7 of the synonymous mutations in the sample, while all 28 nonsynonymous mutations were identified from Oxford Nanopore and 20 nonsynonymous mutations were identified from Illumina.

Visualizing ion transport in polymers via ion-chromic indicators.

There is growing interest in polymers with high ionic conductivity for applications including batteries, fuel cells, and separation membranes. However, measuring ion diffusion in polymers can be challenging, requiring complex procedures and instrumentation. Here, a simple strategy to study ion diffusion in polymers is presented that utilizes ion-chromic spiropyan as an indicator to measure the diffusion of LiTFSI, KTFSI, and NaTFSI within poly(ethylene oxide)-based polymer networks. These systems are selected, as these are common ions and polymers used in energy storage applications, however, the approach described is not specific to materials for energy storage. Specifically, to enabling the study of ion diffusion, these salts cause the spiropyran to undergo an isomerization reaction, which results in a significant color change. This colorimetric response enables the determination of the diffusion coefficients of these ions within films of these polymers simply by optically tracking the spatial-temporal evolution of the isomerization product within the film and fitting the data to the relevant diffusion equations. The simplicity of the method makes it amenable to the study of ion diffusion in polymers under a range of conditions, including various temperatures and under macroscopic deformation.

Alternate Graft Options for Staged Flexor Tendon Reconstruction: A Cadaveric Study of Hamstring Autografts Compared to Conventional Autografts.

PURPOSE: To compare the semitendinosus and gracilis tendon lengths and diameters to the palmaris longus, plantaris, flexor digitorum profundus, and flexor pollicis longus (FPL) tendons in a cadaveric model to evaluate the feasibility of hamstring autograft use for staged flexor tendon reconstruction. METHODS: Fifteen fresh cadavers were evaluated for surgical incisions about the knee, forearm, and hand. All flexor digitorum profundus (FDP), FPL, palmaris longus, plantaris, semitendinosus, and gracilis tendons were harvested from each specimen. Diameter and length were recorded and means with SDs were calculated. The mean diameters of the gracilis and semitendinosus were compared to the mean diameters of the FDP and FPL tendons. The hamstring tendon lengths were then compared in terms of percentage of the palmaris longus and plantaris tendon lengths. RESULTS: The gracilis (18.0 cm) and semitendinosus (19.9 cm) means were notably longer than the palmaris longus (16.0 cm) and shorter than the plantaris (30.0 cm). The average gracilis tendon diameter (3.8 mm) was smaller than the flexor tendon diameters except for the little finger FDP (3.8 mm). The semitendinosus tendon diameter (4.8 mm) was larger than all flexor tendons with the exception of the middle finger FDP (4.6 mm). Average gracilis and semitendinosus tendon diameters were 3.7 mm and 4.5 mm in males, and 3.8 mm and 4.8 mm in females. CONCLUSIONS: This study showed the gracilis tendon to have adequate length and diameter for potential autograft use in staged flexor tendon reconstruction in all digits but the little finger. The semitendinosus is larger in diameter than the native flexor tendons, making it a poor autograft option in cases with an intact pulley system. CLINICAL RELEVANCE: Common tendon autograft options for flexor tendon reconstruction are variably present, and the use of gracilis and semitendinosus autograft present potential graft options.

Biomimetic and Biologically Compliant Soft Architectures via 3D and 4D Assembly Methods: A Perspective.

Recent progress in soft material chemistry and enabling methods of 3D and 4D fabrication-emerging programmable material designs and associated assembly methods for the construction of complex functional structures-is highlighted. The underlying advances in this science allow the creation of soft material architectures with properties and shapes that programmably vary with time. The ability to control composition from the molecular to the macroscale is highlighted-most notably through examples that focus on biomimetic and biologically compliant soft materials. Such advances, when coupled with the ability to program material structure and properties across multiple scales via microfabrication, 3D printing, or other assembly techniques, give rise to responsive (4D) architectures. The challenges and prospects for progress in this emerging field in terms of its capacities for integrating chemistry, form, and function are described in the context of exemplary soft material systems demonstrating important but heretofore difficult-to-realize biomimetic and biologically compliant behaviors.

Dynamic manipulation of droplets using mechanically tunable microtextured chemical gradients.

Materials and strategies applicable to the dynamic transport of microdroplets are relevant to surface fluidics, self-cleaning materials, thermal management systems, and analytical devices. Techniques based on electrowetting, topographic micropatterns, and thermal/chemical gradients have advanced considerably, but dynamic microdroplet transport remains a challenge. This manuscript reports the fabrication of mechano-tunable, microtextured chemical gradients on elastomer films and their use in controlled microdroplet transport. Specifically, discreet mechanical deformations of these films enabled dynamic tuning of the microtextures and thus transport along surface-chemical gradients. The interplay between the driving force of the chemical gradient and the microtopography was characterized, facilitating accurate prediction of the conditions (droplet radius and roughness) which supported transport. In this work, the use of microtextured surface chemical gradients in mechano-adaptive materials with microdroplet manipulation functionality was highlighted.

Detection of Respiratory Pathogens Does Not Predict Risks After Outpatient Adenotonsillectomy.

OBJECTIVES/HYPOTHESIS: To determine whether the presence of detectable upper respiratory infections (URIs) at the time of adenoidectomy/adenotonsillectomy is associated with increased morbidity, complications, and unexpected admissions. STUDY DESIGN: Prospective double-blinded cohort. METHODS: In this prospective cohort study, nasopharyngeal swabs were obtained intraoperatively from 164 pediatric patients undergoing outpatient adenoidectomy/tonsillectomy with or without pressure equalization tubes (PETs) and were analyzed with PCR for the presence of 22 known URIs, including SARS-CoV-2. Surgeons and families were blinded to the results. At the conclusion of the study, rates of detectable infection were determined and intraoperative and postoperative events (unexpected admissions, length of PACU stay, rates of laryngospasm/bronchospasm, oxygen desaturation, bradycardia, and postoperative presentation to an emergency department) were compared between infected and uninfected patients. RESULTS: Of the 164 patients (50% male, 50% female, ages 8 mo-18 y), 136 patients (82.9%) tested positive for one or more URI at the time of surgery. Forty one patients (25.0%) tested positive for three or more URIs concurrently, and 11 (6.7%) tested positive for five or more URIs concurrently. There were no significant differences in admission rates, length of PACU stay, rates of laryngospasm/bronchospasm, oxygen desaturation, bradycardia, or postoperative presentation to an emergency department between positive and negative patients. No patients tested positive for SARS-CoV-2. CONCLUSIONS: A recent positive URI test does not confer any additional intraoperative or postoperative risk in the setting of outpatient adenoidectomy/tonsillectomy in healthy patients. There is no utility in preoperative URI testing, and delaying surgery due to a recent positive URI test is not warranted in this population. LEVEL OF EVIDENCE: 3 Laryngoscope, 131:E2074-E2079, 2021.

Spatiotemporal control of calcium carbonate nucleation using mechanical deformations of elastic surfaces.

Biological systems generate crystalline materials with properties and morphologies that cannot be duplicated using synthetic procedures. Developing strategies that mimic the control mechanisms found in nature would enhance the range of functional materials available for numerous technological applications. Herein, a biomimetic approach based on the mechano-dynamic chemistry of silicone surfaces was used to control the rate of heterogeneous CaCO3 nucleation. Specifically, stretching the silicone surface redistributed functional groups, tuning interfacial energy and thus the rate of CaCO3 crystal formation, as predicted by classical nucleation rate laws. We extended this procedure using microrelief patterns to program surface strain fields to spatially control the location of nucleation. The strategies presented herein represent a fundamental departure from traditional bottom-up crystal engineering, where surfaces are chemically static, to them being active participants in the nucleation process controlling the outcome both spatially and temporally.

Mechanically Induced Hydrophobic Recovery of Poly(dimethylsiloxane) (PDMS) for the Generation of Surfaces with Patterned Wettability.

Silicone elastomers are used in a variety of "stretchable" technologies (e.g., wearable electronics and soft robotics) that require the elastomeric components to accommodate varying magnitudes of mechanical stress during operation; however, there is limited understanding of how mechanical stress influences the surface chemistry of these elastomeric components despite the potential importance of this property with regards to overall function. In this study, plasma-oxidized silicone (poly(dimethylsiloxane)) films were systematically subjected to various amounts of tensile stress and the resulting surface chemical changes were monitored using contact angle measurements, X-ray photoelectron spectroscopy, and gas chromatography-mass spectrometry. Understanding the influence of mechanical stress on these materials made possible the development of a facile method for the rapid, on-demand switching of surface wettability and the generation of surface wettability patterns and gradients. The use of mechanical stress to control surface wettability is broadly applicable to the fields of microfluidics, soft robotics, printing, and to the design of adaptable materials and sensors.

Covalent Bonding of Thermoplastics to Rubbers for Printable, Reel-to-Reel Processing in Soft Robotics and Microfluidics.

The lamination of mechanically stiff structures to elastic materials is prevalent in biological systems and popular in many emerging synthetic systems, such as soft robotics, microfluidics, stretchable electronics, and pop-up assemblies. The disparate mechanical and chemical properties of these materials have made it challenging to develop universal synthetic procedures capable of reliably adhering to these classes of materials together. Herein, a simple and scalable procedure is described that is capable of covalently laminating a variety of commodity ("off-the-shelf") thermoplastic sheets to silicone rubber films. When combined with laser printing, the nonbonding sites can be "printed" onto the thermoplastic sheets, enabling the direct fabrication of microfluidic systems for actuation and liquid handling applications. The versatility of this approach in generating thin, multifunctional laminates is demonstrated through the fabrication of milliscale soft actuators and grippers with hinged articulation and microfluidic channels with built-in optical filtering and pressure-dependent geometries. This method of fabrication offers several advantages, including technical simplicity, process scalability, design versatility, and material diversity. The concepts and strategies presented herein are broadly applicable to the soft robotics, microfluidics, and advanced and additive manufacturing communities where hybrid rubber/plastic structures are prevalent.

Impact of thrombophilia screening on venous thromboembolism management practices.

BACKGROUND: It is unclear whether thrombophilia testing provides any further information on risk of recurrence or guidance in management of patients with a first episode of idiopathic venous thromboembolism (VTE). Furthermore, after the introduction to clinical practice of clinical prediction rules, thrombophilia screening could be less relevant in anticoagulation decision making. We assessed the potential impact of thrombophilia screening on the decision of maintaining anticoagulation beyond the initially planned anticoagulation period in patients with an unprovoked VTE, before and after the introduction of a clinical prediction rule into practice. PATIENTS/METHODS: We conducted a single center, retrospective cohort study of consecutive patients with a diagnosis of unprovoked VTE, including a study period of 12years. Two groups were compared, before and after 2008. RESULTS: We included 1033 patients of which 85.2% were tested for thrombophilia and 26.2% were identified with any thrombophilia. A similar proportion of patients continued on anticoagulation after 6months (54.1% vs 57.1%, respectively). The proportion of patients continuing anticoagulation based on the thrombophilia screen remained small (13.9% vs 12.7%, respectively). Continuing anticoagulation beyond the initial period planned resulted in a 75% risk reduction in VTE recurrence, independent of the presence of thrombophilia (HR 0.25, 95% CI 0.12-0.55; P<0.001). CONCLUSIONS: Thrombophilia screening continues to have little relevance in clinical decision making for anticoagulation. Prolonging anticoagulation beyond 6months in an at-risk population decreased the risk of VTE recurrence regardless of their thrombophilia status.

Stretchable Substrates for the Assembly of Polymeric Microstructures.

The directed assembly of micro-/nanoscale objects relies on physical or chemical processes to generate structures that are not possible via self-assembly alone. A relatively unexplored strategy in directed assembly is the "active" manipulation of building blocks through deformations of elastomeric substrates. This manuscript reports a method which uses macroscopic mechanical deformations of chemically modified silicone films to realize the rational assembly of microscopic polymer structures. Specifically, polystyrene microparticles are deposited onto polydimethylsiloxane substrates using microcontact-printing where, through a process that involved stretching/relaxing the substrates and bonding of the particles, they are elaborated into microstructures of various sizes, shapes, symmetries, periodicities, and functionalities. The resulting polymeric microstructures can be released and redeposited onto planar/nonplanar surfaces. When building blocks with different properties (e.g., those with fluorescent and catalytic properties) are used, it is possible to fabricate structures with heterogeneous functionality. This method can be extended to the assembly of numerous micro-/nanoscale building blocks (e.g., colloidal organic/inorganic materials) with rational control over the size, shape, and functionality of the product. As a strategy, the use of substrate deformations to enable the micromanipulation and fabrication of a potentially diverse set of assemblies represents a powerful tool useful to, for example, nanotechnology and micromanufacturing.

Modeling the role of healthcare access inequalities in epidemic outcomes.

Urban areas, with large and dense populations, offer conditions that favor the emergence and spread of certain infectious diseases. One common feature of urban populations is the existence of large socioeconomic inequalities which are often mirrored by disparities in access to healthcare. Recent empirical evidence suggests that higher levels of socioeconomic inequalities are associated with worsened public health outcomes, including higher rates of sexually transmitted diseases (STD's) and lower life expectancy. However, the reasons for these associations are still speculative. Here we formulate a mathematical model to study the effect of healthcare disparities on the spread of an infectious disease that does not confer lasting immunity, such as is true of certain STD's. Using a simple epidemic model of a population divided into two groups that differ in their recovery rates due to different levels of access to healthcare, we find that both the basic reproductive number (R0) of the disease and its endemic prevalence are increasing functions of the disparity between the two groups, in agreement with empirical evidence. Unexpectedly, this can be true even when the fraction of the population with better access to healthcare is increased if this is offset by reduced access within the disadvantaged group. Extending our model to more than two groups with different levels of access to healthcare, we find that increasing the variance of recovery rates among groups, while keeping the mean recovery rate constant, also increases R0 and disease prevalence. In addition, we show that these conclusions are sensitive to how we quantify the inequalities in our model, underscoring the importance of basing analyses on appropriate measures of inequalities. These insights shed light on the possible impact that increasing levels of inequalities in healthcare access can have on epidemic outcomes, while offering plausible explanations for the observed empirical patterns.

Soft Surfaces for the Reversible Control of Thin-Film Microstructure and Optical Reflectance.

A micromechano-optical material is rapidly and reversibly switched between distinct states of reflectance by simply stretching and relaxing the hybrid structure. The material is fabricated and controlled by leveraging the ability of soft elastic substrates to regulate the growth and morphological evolution of a chemically deposited polycrystalline thin film.

Reconfigurable microfluidic systems with reversible seals compatible with 2D and 3D surfaces of arbitrary chemical composition.

Microfluidic channels are typically fabricated in polydimethylsiloxane (PDMS) using soft lithography and sealed against a support substrate using various irreversible/reversible techniques-the most widely used method is the irreversible bonding of PDMS to glass using oxygen plasma. These techniques are limited in their ability to seal channels against rough, uneven, and/or three-dimensional substrates. This manuscript describes the design and fabrication of soft microfluidic systems from combinations of silicone elastomers that can be reversibly sealed against an array of materials of various topographies/geometries using compression. These soft systems have channels with cross-sectional dimensions that can be decreased, reversibly, by hundreds of microns using compressive stress, and the ability to interface with virtually any support substrate. These capabilities go beyond that achievable with devices fabricated in PDMS alone and enable the integration of microfluidic functionality directly with rough and/or 3D surfaces, providing new opportunities in solution processing useful to, for example, materials science and the analytical/forensic sciences.

Bilayer surface association of the pHLIP peptide promotes extensive backbone desolvation and helically-constrained structures.

Despite their presence in many aspects of biology, the study of membrane proteins lags behind that of their soluble counterparts. Improving structural analysis of membrane proteins is essential. Deep-UV resonance Raman (DUVRR) spectroscopy is an emerging technique in this area and has demonstrated sensitivity to subtle structural transitions and changes in protein environment. The pH low insertion peptide (pHLIP) has three distinct structural states: disordered in an aqueous environment, partially folded and associated with a lipid membrane, and inserted into a lipid bilayer as a transmembrane helix. While the soluble and membrane-inserted forms are well characterized, the partially folded membrane-associated state has not yet been clearly described. The amide I mode, known to be sensitive to protein environment, is the same in spectra of membrane-associated and membrane-inserted pHLIP, indicating comparable levels of backbone dehydration. The amide S mode, sensitive to helical structure, indicates less helical character in the membrane-associated form compared to the membrane-inserted state, consistent with previous findings. However, the structurally sensitive amide III region is very similar in both membrane-associated and membrane-inserted pHLIP, suggesting that the membrane-associated form has a large amount of ordered structure. Where before the membrane-associated state was thought to contain mostly unordered structure and reside in a predominantly aqueous environment, we have shown that it contains a significant amount of ordered structure and rests deeper within the lipid membrane.

Kisspeptin/KISS1R signaling potentiates extravillous trophoblast adhesion to type-I collagen in a PKC- and ERK1/2-dependent manner.

During the first trimester of human pregnancy, cytotrophoblasts proliferate within the tips of the chorionic villi to form cell columns that anchor the placenta to the uterus. This migration coincides with a widespread change in the adhesion molecule repertoire of these trophoblasts. Kisspeptin and its receptor, KISS1R, are best known as potent triggers of gonadotropin-releasing hormone secretion. The kisspeptin/KISS1R signaling system is also highly expressed in the human placenta, where it was demonstrated to inhibit extra-villous trophoblast (EVT) migration and invasion in vitro. Here we show that kisspeptin, in a dose- and time-dependent manner, induces increased adhesion of human EVTs to type-I collagen, a major component of the human placenta. This increased adhesion was both rapid and transient, suggesting that it likely occurred through the activation of KISS1R secondary effectors such as PKC and ERK, which underwent rapid and transient kisspeptin-dependent activation in EVTs. We then showed that inhibition of both PKC and ERK1/2 attenuated the kisspeptin-dependent increase in EVT adhesion, suggesting that these molecules are key positive regulators of trophoblast adhesion. We therefore propose that kisspeptin/KISS1R signaling potentiates EVT adhesion to type-I collagen via "inside-out signaling." Furthermore, kisspeptin treatment increased mouse blastocyst adhesion to collagen I, suggesting that kisspeptin signaling is a key regulator of trophoblast function during implantation as well as early placentation.

Regulation of GPR54 signaling by GRK2 and {beta}-arrestin.

Kisspeptin and its receptor, GPR54, are major regulators of the hypothalamic-pituitary-gonadal axis as well as regulators of human placentation and tumor metastases. GPR54 is a G(q/11)-coupled G protein-coupled receptor (GPCR), and activation by kisspeptin stimulates phosphatidy linositol 4, 5-biphosphate hydrolysis, Ca(2+) mobilization, arachidonic acid release, and ERK1/2 MAPK phosphorylation. Physiological evidence suggests that GPR54 undergoes agonist-dependent desensitization, but underlying molecular mechanisms are unknown. Furthermore, very little has been reported on the early events that regulate GPR54 signaling. The lack of information in these important areas led to this study. Here we report for the first time on the role of GPCR serine/threonine kinase (GRK)2 and beta-arrestin in regulating GPR54 signaling in human embryonic kidney (HEK) 293 cells, a model cell system for studying the molecular regulation of GPCRs, and genetically modified MDA MB-231 cells, an invasive breast cancer cell line expressing about 75% less beta-arrestin-2 than the control cell line. Our study reveals that in HEK 293 cells, GPR54 is expressed both at the plasma membrane and intracellularly and also that plasma membrane expression is regulated by cytoplasmic tail sequences. We also demonstrate that GPR54 exhibits constitutive activity, internalization, and association with GRK2 and beta- arrestins-1 and 2 through sequences in the second intracellular loop and cytoplasmic tail of the receptor. We also show that GRK2 stimulates the desensitization of GPR54 in HEK 293 cells and that beta-arrestin-2 mediates GPR54 activation of ERK1/2 in MDA-MB-231 cells. The significance of these findings in developing molecular-based therapies for treating certain endocrine-related disorders is discussed.