In vivo CRISPR base editing of PCSK9 durably lowers cholesterol in primates.

Gene-editing technologies, which include the CRISPR-Cas nucleases1-3 and CRISPR base editors4,5, have the potential to permanently modify disease-causing genes in patients6. The demonstration of durable editing in target organs of nonhuman primates is a key step before in vivo administration of gene editors to patients in clinical trials. Here we demonstrate that CRISPR base editors that are delivered in vivo using lipid nanoparticles can efficiently and precisely modify disease-related genes in living cynomolgus monkeys (Macaca fascicularis). We observed a near-complete knockdown of PCSK9 in the liver after a single infusion of lipid nanoparticles, with concomitant reductions in blood levels of PCSK9 and low-density lipoprotein cholesterol of approximately 90% and about 60%, respectively; all of these changes remained stable for at least 8 months after a single-dose treatment. In addition to supporting a 'once-and-done' approach to the reduction of low-density lipoprotein cholesterol and the treatment of atherosclerotic cardiovascular disease (the leading cause of death worldwide7), our results provide a proof-of-concept for how CRISPR base editors can be productively applied to make precise single-nucleotide changes in therapeutic target genes in the liver, and potentially in other organs.

Excitonic Beam Steering in an Active van der Waals Metasurface.

Two-dimensional transition metal dichalcogenides (2D TMDCs) are promising candidates for ultrathin active nanophotonic elements due to the strong tunable excitonic resonances that dominate their optical response. Here, we demonstrate dynamic beam steering by an active van der Waals metasurface that leverages large complex refractive index tunability near excitonic resonances in monolayer molybdenum diselenide (MoSe2). Through varying the radiative and nonradiative rates of the excitons, we can dynamically control both the reflection amplitude and phase profiles, resulting in an excitonic phased array metasurface. Our experiments show reflected light steering to angles between -30° and 30° at different resonant wavelengths corresponding to the A exciton and B exciton. This active van der Waals metasurface relies solely on the excitonic resonances of the monolayer MoSe2 material rather than geometric resonances of patterned nanostructures, suggesting the potential to harness the tunability of excitonic resonances for wavefront shaping in emerging photonic applications.

Quasiparticle and Optical Properties of Carrier-Doped Monolayer MoTe2 from First Principles.

The intrinsic weak and highly nonlocal dielectric screening of two-dimensional materials is well-known to lead to high sensitivity of their optoelectronic properties to environment. Less studied theoretically is the role of free carriers in those properties. Here, we use ab initio GW and Bethe-Salpeter equation calculations, with a rigorous treatment of dynamical screening and local-field effects, to study the doping dependence of the quasiparticle and optical properties of a monolayer transition-metal dichalcogenide, 2H MoTe2. We predict a quasiparticle band gap renormalization of several hundreds of meV for experimentally attainable carrier densities and a similarly sizable decrease in the exciton binding energy. This results in an almost constant excitation energy for the lowest-energy exciton resonance with an increasing doping density. Using a newly developed and generally applicable plasmon-pole model and a self-consistent solution of the Bethe-Salpeter equation, we reveal the importance of accurately capturing both dynamical and local-field effects to understand detailed photoluminescence measurements.

Lifetime-Limited and Tunable Quantum Light Emission in h-BN via Electric Field Modulation.

Color-center-based single-photon emitters in hexagonal boron nitride (h-BN) have shown promising photophysical properties as sources for quantum light emission. Despite significant advances toward such a goal, achieving lifetime-limited quantum light emission in h-BN has proven to be challenging, primarily due to various broadening mechanisms, including spectral diffusion. Here, we propose and experimentally demonstrate suppression of spectral diffusion by applying an electrostatic field. We observe both Stark shift tuning of the resonant emission wavelength and emission line width reduction (down to 89 MHz) nearly to the homogeneously broadened lifetime limit. Finally, we find a cubic dependence of the line width with respect to temperature at the homogeneous broadening regime. Our results suggest that field tuning in electrostatically gated heterostructures is promising as an approach to control the emission characteristics of h-BN color centers, removing spectral diffusion and providing the energy tunability necessary for integrate of quantum light emission in nanophotonic architectures.

Refractive Index Modulation in Monolayer Molybdenum Diselenide.

Two-dimensional transition metal dichalcogenides are promising candidates for ultrathin light modulators due to their highly tunable excitonic resonances at visible and near-infrared wavelengths. At cryogenic temperatures, large excitonic reflectivity in monolayer molybdenum diselenide (MoSe2) has been shown, but the permittivity and index modulation have not been studied. Here, we demonstrate large gate-tunability of complex refractive index in monolayer MoSe2 by Fermi level modulation and study the doping dependence of the A and B excitonic resonances for temperatures between 4 and 150 K. By tuning the charge density, we observe both temperature- and carrier-dependent epsilon-near-zero response in the permittivity and transition from metallic to dielectric near the A exciton energy. We attribute the dynamic control of the refractive index to the interplay between radiative and non-radiative decay channels that are tuned upon gating. Our results suggest the potential of monolayer MoSe2 as an active material for emerging photonics applications.

Highly Strain-Tunable Interlayer Excitons in MoS2/WSe2 Heterobilayers.

Interlayer excitons in heterobilayers of transition-metal dichalcogenides (TMDCs) have generated enormous interest due to their permanent vertical dipole moments and long lifetimes. However, the effects of mechanical strain on the optoelectronic properties of interlayer excitons in heterobilayers remain relatively uncharacterized. Here, we experimentally demonstrate strain tuning of Γ-K interlayer excitons in molybdenum disulfide and tungsten diselenide (MoS2/WSe2) wrinkled heterobilayers and obtain a deformation potential constant of ∼107 meV/% uniaxial strain, which is approximately twice that of the intralayer excitons in the constituent monolayers. We further observe a nonmonotonic dependence of the interlayer exciton photoluminescence intensity with strain, which we interpret as being due to the sensitivity of the Γ point to band hybridization arising from the competition between in-plane strain and out-of-plane interlayer coupling. Strain engineering with interlayer excitons in TMDC heterobilayers offers higher strain tunability and new degrees of freedom compared to their monolayer counterparts.

Nanoscale axial position and orientation measurement of hexagonal boron nitride quantum emitters using a tunable nanophotonic environment.

Color centers in hexagonal boron nitride (hBN) have emerged as promising candidates for single-photon emitters (SPEs) due to their bright emission characteristics at room temperature. In contrast to mono- and few-layeredhBN, color centers in multi-layered flakes show superior emission characteristics such as higher saturation counts and spectral stability. Here, we report a method for determining both the axial position and three-dimensional dipole orientation of SPEs in thickhBN flakes by tuning the photonic local density of states using vanadium dioxide (VO2), a phase change material. Quantum emitters under study exhibit a strong surface-normal dipole orientation, providing some insight on the atomic structure ofhBN SPEs, deeply embedded in thick crystals. Next, we optimized a hot pickup technique to reproducibly transfer thehBN flake from VO2/sapphire substrate onto SiO2/Si substrate and relocated the same emitters. Our approach serves as a practical method to systematically characterize SPEs inhBN prior to integration in quantum photonics systems.

Anisotropic Quantum Well Electro-Optics in Few-Layer Black Phosphorus.

The incorporation of electrically tunable materials into photonic structures such as waveguides and metasurfaces enables dynamic, electrical control of light propagation at the nanoscale. Few-layer black phosphorus is a promising material for these applications due to its in-plane anisotropic, quantum well band structure, with a direct band gap that can be tuned from 0.3 to 2 eV with a number of layers and subbands that manifest as additional optical transitions across a wide range of energies. In this Letter, we report an experimental investigation of three different, anisotropic electro-optic mechanisms that allow electrical control of the complex refractive index in few-layer black phosphorus from the mid-infrared to the visible: Pauli-blocking of intersubband optical transitions (the Burstein-Moss effect); the quantum-confined Stark effect; and the modification of quantum well selection rules by a symmetry-breaking, applied electric field. These effects generate near-unity tuning of the BP oscillator strength for some material thicknesses and photon energies, along a single in-plane crystal axis, transforming absorption from highly anisotropic to nearly isotropic. Lastly, the anisotropy of these electro-optical phenomena results in dynamic control of linear dichroism and birefringence, a promising concept for active control of the complex polarization state of light, or propagation direction of surface waves.

Total synthesis of naturally occurring abietane diterpenoids via a late-stage Fe(iii)-bTAML catalysed Csp3-H functionalization.

The synthesis of diverse trans-fused decalins, including the abietane diterpenoids scaffold, using an efficient selective oxidation strategy is described. The abietane core was demonstrated to be a versatile scaffold that can be site-selectively functionalized. The utility of this novel oxidation strategy was showcased in a concise total synthesis of six abietane congeners.

Paper Sensor Modified with MoS2 for Detection of Dopamine Using a Machine-Intelligent Web App Interface.

The sensing behavior of a MoS2-functionalized paper sensor towards dopamine was explored through a combinatorial approach of theoretical analysis, subsequent experimental validation, and machine-learning-driven predictive modeling of the measured electrochemical outputs. The suitability of the chosen 2D material for efficient detection of dopamine was confirmed using density functional theory. The physisorption behavior along with electrostatic interaction due to the incorporation of dopamine on MoS2 was unraveled under the purview of theoretically estimated noncovalent interaction and charge density difference plot. The theoretical Löwdin population analysis elucidates the alteration in oxidation potential of dopamine, as observed in electrochemical experiments. The electrochemical responses of the developed sensor with the spiked serum samples showed an average accuracy of more than 96% with a limit of detection of 10 nM. Furthermore, implementation of a machine-intelligent interactive web app interface improved the resolution of the sensing platform significantly with an enhanced accuracy of nearly 99%.

Edge-Confined Excitons in Monolayer Black Phosphorus.

Quantum confinement of two-dimensional excitons in van der Waals materials via electrostatic trapping, lithographic patterning, Moiré potentials, and chemical implantation has enabled significant advances in tailoring light emission from nanostructures. While such approaches rely on complex preparation of materials, natural edges are a ubiquitous feature in layered materials and provide a different approach for investigating quantum-confined excitons. Here, we observe that certain edge sites of monolayer black phosphorus (BP) strongly localize the intrinsic quasi-one-dimensional excitons, yielding sharp spectral lines in photoluminescence, with nearly an order of magnitude line width reduction. Through structural characterization of BP edges using transmission electron microscopy and first-principles GW plus Bethe-Salpeter equation (GW-BSE) calculations of exemplary BP nanoribbons, we find that certain atomic reconstructions can strongly quantum-confine excitons resulting in distinct emission features, mediated by local strain and screening. We observe linearly polarized luminescence emission from edge reconstructions that preserve the mirror symmetry of the parent BP lattice, in agreement with calculations. Furthermore, we demonstrate efficient electrical switching of localized edge excitonic luminescence, whose sites act as excitonic transistors for emission. Localized emission from BP edges motivates exploration of nanoribbons and quantum dots as hosts for tunable narrowband light generation, with future potential to create atomic-like structures for quantum information processing applications as well as exploration of exotic phases that may reside in atomic edge structures.

Rydberg Excitons and Trions in Monolayer MoTe2.

Monolayer transition metal dichalcogenide (TMDC) semiconductors exhibit strong excitonic optical resonances, which serve as a microscopic, noninvasive probe into their fundamental properties. Like the hydrogen atom, such excitons can exhibit an entire Rydberg series of resonances. Excitons have been extensively studied in most TMDCs (MoS2, MoSe2, WS2, and WSe2), but detailed exploration of excitonic phenomena has been lacking in the important TMDC material molybdenum ditelluride (MoTe2). Here, we report an experimental investigation of excitonic luminescence properties of monolayer MoTe2 to understand the excitonic Rydberg series, up to 3s. We report a significant modification of emission energies with temperature (4 to 300 K), thereby quantifying the exciton-phonon coupling. Furthermore, we observe a strongly gate-tunable exciton-trion interplay for all the Rydberg states governed mainly by free-carrier screening, Pauli blocking, and band gap renormalization in agreement with the results of first-principles GW plus Bethe-Salpeter equation approach calculations. Our results help bring monolayer MoTe2 closer to its potential applications in near-infrared optoelectronics and photonic devices.

GalNAc-Lipid nanoparticles enable non-LDLR dependent hepatic delivery of a CRISPR base editing therapy.

Lipid nanoparticles have demonstrated utility in hepatic delivery of a range of therapeutic modalities and typically deliver their cargo via low-density lipoprotein receptor-mediated endocytosis. For patients lacking sufficient low-density lipoprotein receptor activity, such as those with homozygous familial hypercholesterolemia, an alternate strategy is needed. Here we show the use of structure-guided rational design in a series of mouse and non-human primate studies to optimize a GalNAc-Lipid nanoparticle that allows for low-density lipoprotein receptor independent delivery. In low-density lipoprotein receptor-deficient non-human primates administered a CRISPR base editing therapy targeting the ANGPTL3 gene, the introduction of an optimized GalNAc-based asialoglycoprotein receptor ligand to the nanoparticle surface increased liver editing from 5% to 61% with minimal editing in nontargeted tissues. Similar editing was noted in wild-type monkeys, with durable blood ANGPTL3 protein reduction up to 89% six months post dosing. These results suggest that GalNAc-Lipid nanoparticles may effectively deliver to both patients with intact low-density lipoprotein receptor activity as well as those afflicted by homozygous familial hypercholesterolemia.

Preconcentration and analysis of trace volatile carbonyl compounds.

We describe a preconcentration device that may be suitable for quantitative analysis of trace volatile ketones and aldehydes in ambient air as well as in human breath. The approach is based on microreactor chips fabricated from silicon wafers. The microreactors have thousands of micropillars in microfluidic channels for uniformly distributing a gaseous sample flowing through the chips. The surfaces of the micropillars are functionalized with a quaternary ammonium aminooxy salt, [2-(aminooxy)ethyl]-N,N,N-trimethylammonium iodide (ATM), for trapping trace ketones and aldehydes by means of oximation reactions. ATM adducts and unreacted ATM are eluted from the microreactor with less than 40 µL of methanol and directly analyzed by nanospray Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS). Ketones and aldehydes at levels of 1 ppbv have been detected using this microreactor and FTICR-MS system.

Machine learning based urinary pH sensing using polyaniline deposited paper device and integration of smart web app interface: Theory to application.

The present study employs density functional theory-based first principle calculation to investigate the electron transport properties of polyaniline following exposure to acidic and alkaline pH. In-situ deposited polyaniline-based paper device maintains emeraldine salt form while it is exposed to acidic pH and converts to emeraldine base when it is subjected to alkaline pH solutions. These structural changes at acidic and alkaline pH are validated experimentally by Raman spectra. Furthermore, the Raman spectra computed from density functional theory are validated with the experimental spectra. The changes in the theoretical energy band gap of polyaniline obtained from first principle calculations were correlated with the changes in the experimental impedimetric response of the sensor after exposure to acidic and alkaline solutions. Finally, the impedimetric responses were used to predict urine pH through a machine learning based smart and interactive web application. Different machine learning based regression models were implemented to acquire the best possible outcome. Gradient Boosting Regressor with least square loss model was selected as it showed lowest mean square, mean absolute, and root mean square error than other models. The smart sensing platform successfully predicts the unknown pH of urine samples with an average accuracy of more than 98%. The locally deployed smart web app can be accessed within a local area network by the end-user, which holds promise towards effective detection of urinary pH.

American college of radiology thyroid imaging reporting and data system score has high diagnostic value in the diagnosis of malignant thyroid nodules: A prospective single-center cross-sectional study.

Introduction: A palpable thyroid swelling is a very common finding and is seen in almost 12% of Asian Indian population. Thyroid imaging reporting and data system (TI-RADS) can be used as a risk stratification system to determine malignant or benign thyroid nodules and necessity of further intervention. Objective: The objective of this study was to determine the positive predictive value (PPV) of TI-RADS category/ultrasound using TI-RADS categories in the diagnosis of malignancy in clinically suspected thyroid nodule and necessity for further intervention in the case of malignant thyroid nodules. Materials and Methods: We conducted a prospective study evaluating 110 patients (140 thyroid nodules) from March 2018 to April 2020 including patients with thyroid swelling. Ultrasound was performed by a radiologist on the patients, and targeted fine-needle aspiration cytology from thyroid nodules was interpreted by an experienced pathologist. Ultrasound features and TI-RADS category were compared with cytology and surgical histopathology. Sensitivity, specificity, PPV, and positive likelihood ratio in different categories of TI-RADS nodules were calculated. Results: A total of 113 thyroid nodules were assessed. Out of 113 nodules (right lobe - 64, isthmus - 6, and left lobe - 43), 84 nodules were benign and 29 nodules were malignant. Eleven (10%) patients were found to have metastatic cervical lymphadenopathy. There was no significant difference in the occurrence of malignant nodules according to gender, location of the nodule, or size of the nodule. The mean anteroposterior and transverse diameter of benign nodule was 14.1 ± 6.9 mm and 20.9 ± 9.9 mm, respectively, whereas in the case of malignant nodules, it was 15.6 ± 7.1 mm and 19.5 ± 9.0 mm, respectively. A TI-RADS score of ≥4 had 84% PPV for malignancy. The PPV for malignancy was 32.2%, 49.1%, and 100% for TI-RADS 2, 3, and 5 categories. Conclusion: TI-RADS is a simple, practical, and cost-effective tool for assessing the malignancy rates of thyroid nodules. TI-RADS categories 4 and 5 have high PPV for malignancy in thyroid nodules.

Résumé Introduction: Un gonflement palpable de la thyroïde est une constatation très courante et est observé chez près de 12 % de la population indienne d'Asie. Le système de rapport et de données d'imagerie thyroïdienne (TI-RADS) peut être utilisé comme système de stratification des risques pour déterminer les nodules thyroïdiens malins ou bénins et la nécessité d'une intervention supplémentaire. Objectif: L'objectif de cette étude était de déterminer la valeur prédictive positive (VPP) de la catégorie/échographie TI-RADS à l'aide des catégories TI-RADS dans le diagnostic de malignité dans un nodule thyroïdien cliniquement suspecté et la nécessité d'une intervention supplémentaire dans le cas d'une tumeur maligne de la thyroïde. nodules. Matériels et méthodes: Nous avons mené une étude prospective évaluant 110 patients (140 nodules thyroïdiens) de mars 2018 à avril 2020 incluant des patients présentant un gonflement de la thyroïde. L'échographie a été réalisée par un radiologue sur les patients, et la cytologie par aspiration à l'aiguille fine ciblée des nodules thyroïdiens a été interprétée par un pathologiste expérimenté. Les caractéristiques échographiques et la catégorie TI-RADS ont été comparées à la cytologie et à l'histopathologie chirurgicale. La sensibilité, la spécificité, la VPP et le rapport de vraisemblance positif dans différentes catégories de nodules TI-RADS ont été calculés. Résultats: Au total, 113 nodules thyroïdiens ont été évalués. Sur 113 nodules (lobe droit - 64, isthme - 6 et lobe gauche - 43), 84 nodules étaient bénins et 29 nodules malins. Onze (10%) patients présentaient une lymphadénopathie cervicale métastatique. Il n'y avait pas de différence significative dans la survenue de nodules malins selon le sexe, la localisation du nodule, ou la taille du nodule. Le diamètre moyen antéropostérieur et transversal du nodule bénin était de 14,1 ± 6,9 mm et 20,9 ± 9,9 mm, respectivement, alors que dans le cas des nodules malins, il était de 15,6 ± 7,1 mm et 19,5 ± 9,0 mm, respectivement. Un score TI-RADS ≥ 4 avait une VPP de 84 % pour la malignité. La VPP pour la malignité était de 32,2 %, 49,1 % et 100 % pour les catégories TI-RADS 2, 3 et 5. Conclusion: TI-RADS est un outil simple, pratique et rentable pour évaluer les taux de malignité des nodules thyroïdiens. Les catégories TI-RADS 4 et 5 ont une VPP élevée pour la malignité des nodules thyroïdiens. Mots-clés : Nodule, système de rapport et de données d'imagerie thyroïdienne, thyroïde, échographie.

Immunological profiling and development of a sensing device for detection of IL-13 in COPD and asthma.

Chronic obstructive pulmonary disease (COPD) and asthma are the two most common obstructive lung diseases which affects millions worldwide and impose an enormous burden on global healthcare. The overlapping features shared by these two diseases often make differential diagnosis difficult to achieve, leading to misdiagnosis of these patients. Both asthma and COPD are associated with chronic inflammation of the airways which is perpetuated by the interplay between immunological mediators. The crucial role played by these mediators make them attractive targets for disease diagnosis. The present study investigates the immunological mediator profile in these patients as compared with controls. Further, a potential biomarker for the development of a sensing platform is identified. Multiplexed analysis of 8 commonly studied immunological markers (IL-4, IL-5, IL-6, IL-13, TGF-ß, IFN-γ, MCP-1 and NGAL) in serum showed distinct dysregulation pattern, with IL-13 showing the highest potential for differential diagnosis. An impedimetric self-assembled monolayer (SAM) based sensor for detecting IL-13 is developed to distinguish between asthma and COPD. The device shows reliable output with high accuracy and sensitivity towards the detection of IL-13.

A novel microreactor approach for analysis of ketones and aldehydes in breath.

We report a fabricated microreactor with thousands of micropillars in channels. Each micropillar surface is chemically functionalized to selectively preconcentrate gaseous ketones and aldehydes of exhaled breath and to enhance ultra-trace, rapid analysis by direct-infusion Fourier transform-ion cyclotron resonance (FT-ICR) mass spectrometry (MS). The micropillar reactive coating contains the quaternary ammonium aminooxy salt 2-(aminooxy)ethyl-N,N,N-trimethylammonium iodide (ATM) for capturing trace carbonyl VOCs by means of an oximation reaction. We demonstrate the utility of this approach for detection of C(1) to C(12) aldehydes and ketones in exhaled breath, but the approach is applicable to any gaseous sample.

Highly specific, NIR fluorescent contrast agent with emission controlled by gold nanoparticle.

Nanoparticles are currently being intensively studied for in vivo molecular imaging because of their unique and beneficial properties. Among these particles, some metal particles possess strong surface plasmon fields that can effectively alter fluorescence. Using this fluorescence alteration, an NIR fluorophore based, nanosized contrast agent for breast cancer diagnosis is being developed. The fluorophore is conjugated to gold nanoparticles (GNP) via a short spacer whose length was specially adjusted to have the strong plasmon field to quench the fluorescence. The spacer also has a special molecular sequence that can be cleaved by an enzyme secreted by targeted cancer cells. Normally, the entity does not fluoresce. If it is delivered to the cancer site, the short spacer would be cleaved by the enzyme secreted by the cancer cell at which point the fluorescence would be restored. This entity can incorporate a cancer targeting molecule for a cancer specific delivery. The entity specifically targets cancer cells and fluoresce only when the spacer is cleaved by a specific cancer secreting biomolecule, providing dual specificity for cancer diagnosis. In the future, this entity will be combined with cancer drugs for seamless detection and personalized therapy.

Raman spectroscopy assisted biochemical evaluation of L929 fibroblast cells on differentially crosslinked gelatin hydrogels.

Biochemical evaluation of cell-matrix interaction using conventional labelling techniques often possesses limitations due to dye entrapment. In contrast, Raman spectroscopy guided approach offers label-free determination of cell-matrix biochemistry. Herein, gelatin (Gel) matrices modified with 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide/ N-Hydroxysuccinimide (EDC/NHS) and glutaraldehyde (GTA) was used as standards for comparative evaluation. Raman spectroscopy was deployed as a label-free approach to investigate interaction of cells with Gel hydrogels. Raman-based approach assisted in evaluation of cell-matrix interactions by identifying key biomolecular signatures retrospecting the fact that L929 fibroblast cells portrayed excellent growth and proliferation kinetics in crosslinked Gel as compared to its bare counterpart. EDC crosslinked hydrogels exhibited superior cell proliferation than its GTA counterparts. Cell proliferation on differentially crosslinked gel was also confirmed using standard MTT Assay and Rhodamine-DAPI staining thus corroborating the fact that Raman spectroscopy can be deployed as a superior label-free alternative towards real-time determination of cell proliferation and growth.