Abstract: Aspects of the disclosure relate to compositions, systems, and methods for evolving nucleic acids and proteins utilizing continuous directed evolution in the periplasm of a host cell. In some embodiments, the methods comprise passing a nucleic acid from cell-to-cell in a desired, function dependent manner. The linkage of the desired function and passage of the nucleic acid from cell-to-cell allows for continuous selection and mutation of the nucleic acid.

Abstract: Provided herein are systems, compositions, kits, and methods for the suppression of pain (e.g., chronic pain). Genes encoding ion channels (e.g., SCN9A) responsible for the propagation pain signals in neurons (e.g., DRG neurons) may be edited using a genome editing agent (e.g., a nucleobase editor). In some embodiments, loss-of-function ion channel mutants are generated, leading to pain suppression. In some embodiments, the genome editing agent is administered locally to the site of pain or to the nerves responsible for propagation of the pain signal.

Abstract: The disclosure provides amino acid sequence variants of Botulinum neurotoxin (BoNT) proteases that cleave (VAMP1, VAMP2, VAMP7, VAMP8, SNAP25, SNAP23, PTEN, etc.) and methods of evolving the same. In some embodiments, proteases described by the disclosure are useful for cleaving proteins found in a cell, that is in an intracellular environment. In some embodiments, proteases described by the disclosure are useful for treating diseases associated with increased or aberrant VAMP7, VAMP8, SNAP23 or PTEN expression or activity, for example, cancer and neurological disorders. Some aspects of this disclosure provide methods for generating BoNT protease variants by continuous directed evolution.

Abstract: An imaging system includes an illumination device for illuminating a target. A surgical microscope receives light from the target, the surgical microscope comprising at least one optical output port at which at least a portion of the received light is provided as an output from the surgical microscope. A tunable filter receives the portion of the received light provided as the output from the surgical microscope, the tunable filter being tunable to pass a filtered portion of the received light, the filtered portion of the received light having a plurality of wavelengths selected by the tunable filter and provided as output from the tunable filter. A high-resolution, broad-bandwidth electronic camera receives the light of a plurality of wavelengths selected by the tunable filter, the electronic camera converting the light of a plurality of wavelengths selected by the tunable filter to a plurality of electrical signals. A processor processes the plurality of electrical signals to form an image of the target.

Abstract: Cellulose nanofibers (CNF) act as a dispersing agent to directly exfoliate graphite in an aqueous solution using sonication. The resulting suspension has graphite flakes, each having 2-20 monolayers, a relatively large lateral dimension, and a plurality of CNF decorating its surfaces and edges. The dispersing effect of the CNF allows the graphite-CNF suspension to be stored without degradation until desired use. The graphite-CNF suspension can be used to form various composite structures, such as by spraying, coating, pouring, extruding, or printing the suspension, and then drying the suspension. The resulting composite structures have improved tensile strength and toughness due to hydrogen bond interactions between the CNF and graphite.

Abstract: A transdermal formulation for the delivery of one or more active agents to a subject is provided. The formulation can be applied to the treatment of various diseases or conditions associated with an elevated CD38 level.

Abstract: The present invention provides a robotic locomotive device (1) that is capable of driving itself forwards and backwards, anchoring and steering itself whilst inside a tubular structure (200), for example, the human colon, or any structure comprising two opposing walls (202, 204). In this respect, the device is made up of two or three segments (102, 104, 106) covered in an elastic material and driven by an internal actuating mechanism. All of the segments (102, 104, 106) have a concertina configuration that enable a shortening and lengthening motion. As well as contracting and extending in length, at least one of the end segments (102, 106) is capable of bending at an angle away from the longitudinal axis such that it becomes wedged or jammed between the walls (202, 204) of the tubular structure (200). That is, the end segments (102, 106) are capable of both a bending action and a contracting and extending action.

Abstract: A method comprising applying an object recognition pipeline to frames of video data. The object recognition pipeline provides a mask output of objects detected in the frames. The method includes fusing the mask output of the object recognition pipeline with depth data associated with the frames of video data to generate a map of object instances, including projecting the mask output to a model space for the map of object instances using a camera pose estimate and the depth data. An object instance in the map of object instances is defined using surface-distance metric values within a three-dimensional object volume, and has an object pose estimate indicating a transformation of the object instance to the model space. The object pose estimate and the camera pose estimate form nodes of a pose graph for the map of model instances.

Abstract: The present application provides methods for preparing soluble lipidated ligand agents comprising a ligand entity and a lipid entity, and in some embodiments, provides relevant parameters of each of these components, thereby enabling appropriate selection of components to assemble active agents for any given target of interest.

Abstract: An imaging apparatus for imaging a sample (7) comprises an array of electronically addressable pixels (6) wherein each pixel is arranged to support a surface plasmon resonance therein to generate an evanescent electromagnetic field. This field extends transversely from the pixel so as to be salient from the array at a first side of the array for illuminating the sample at said first side. A light source (15) is arranged to illuminate the array with excitation light therewith to generate said surface plasmon resonance. An optical detector (12A, 12B, 12C) is arranged at a second side of the array which is opposite to said first side of the array for detecting optical radiation returned from the array in response to illumination of the array by said excitation light. A processing unit (4) is arranged to associate the detected optical radiation with the address of the pixel or pixels within the array at which the surface plasmon resonance was generated.

Abstract: Transcriptomes of individual neurons provide rich information about cell types and dynamic states. However, it is difficult to capture rare dynamic processes, such as adult neurogenesis, because isolation from dense adult tissue is challenging, and markers for each phase are limited. Here, Applicants developed Nuc-seq, Div-Seq, and Dronc-Seq. Div-seq combines Nuc-Seq, a scalable single nucleus RNA-Seq method, with EdU-mediated labeling of proliferating cells. Nuc-Seq can sensitively identify closely related cell types within the adult hippocampus. Div-Seq can track transcriptional dynamics of newborn neurons in an adult neurogenic region in the hippocampus. Dronc-Seq uses a microfluidic device to co-encapsulate individual nuclei in reverse emulsion aqueous droplets in an oil medium together with one uniquely barcoded mRNA-capture bead. Finally, Applicants found rare adult newborn GABAergic neurons in the spinal cord, a non-canonical neurogenic region.

Abstract: Nucleic acid constructs, vectors, and recombinant cells harboring the nucleic acid constructs or vectors are disclosed. The nucleic acid constructs include genes encoding a chimeric antigen receptor (CAR) and/or one or more transcription factors, optionally mutated. The transcription factors include those that mediate proinflammatory cytokine expression, e.g., T-bet, STAT1, or STAT4. Methods are disclosed of co-expression of the CAR and the transcription factor in a human or non-human immune cell, preferably human T cells. Also disclosed are methods for using these cells for immunotherapy, e.g., in treating cancer, infection, autoimmunity, allergy or inflammation diseases by the administration of a prophylactically or therapeutically effective amount of one or more of the nucleic acid constructs, vectors, and/or immune cells, e.g., human CAR-T cells, described herein.

Abstract: A system and a method for predicting susceptibility of genus Klebsiella to amikacin are provided. The system includes a calculating unit containing a computer-readable storage medium stored with a computer program; the computer program is used to, when is executed by a processor, implement a calculation method for an Exp(?k) power value, and the calculation method for the Exp(?k) power value includes: step 1, calculating a k value according to formula I; and step 2, calculating the Exp(?k) power value with an Euler's constant e as a base and a ?k value as an exponential; and C1 to C5 are respectively copy numbers of an arr-2 gene, an acrB gene, an armA gene, an oqxB gene, and a rmtB gene in to-be-predicted Klebsiella strains. When the method and the system are adopted to predict the susceptibility of the genus Klebsiella to amikacin, an accuracy rate is about 98.8%.

Abstract: Provided herein are engineered nucleic acids (e.g., expression vectors, including viral vectors, such as lentiviral vectors, adenoviral vectors, AAV vectors, herpes viral vectors, and retroviral vectors) that encode OCT4; KLF4; SOX2; or any combination thereof that are useful, for example, in inducing cellular reprogramming, tissue repair, tissue regeneration, organ regeneration, reversing aging, or any combination thereof. Also provided herein are recombinant viruses (e.g., lentiviruses, alphaviruses, vaccinia viruses, adenoviruses, herpes viruses, retroviruses, or AAVs) comprising the engineered nucleic acids (e.g.

Abstract: A bioink for extrusion-based printing includes an extracellular matrix (ECM) precursor comprising an uncrosslinked polymer, and sacrificial microparticles dispersed in the ECM precursor. The sacrificial microparticles have a melting temperature above a crosslinking temperature of the uncrosslinked polymer. A method of fabricating a tissue/organ model or therapeutic construct comprises extruding a bioink comprising a first ECM precursor and first sacrificial microparticles through a nozzle moving relative to a deposition bath, and depositing an extruded filament comprising the bioink into the deposition bath as the nozzle moves. After deposition, the first ECM precursor is crosslinked to form a first ECM material, and after the crosslinking, the first sacrificial microparticles are melted to form pores in the first ECM material. The pores may have a width or diameter comparable to that of individual cells.

Abstract: The present invention generally relates to systems and methods for imaging or determining nucleic acids or other desired targets, for instance, within cells or tissues. In one aspect, a sample is exposed to a plurality of nucleic acid probes that are determined within the sample. In some cases, however, background fluorescence or off-target binding may make it more difficult to determine properly bound nucleic acid probes. Accordingly, other components of the samples that may be contributing to the background, such as proteins, lipids, and/or other non-targets, may be “cleared” from the sample to improve determination. However, in certain embodiments, nucleic acids or other desired targets may be prevented from also being cleared, e.g., using polymers or gels within the sample. Other aspects are generally directed to compositions or kits involving such systems, methods of using such systems, or the like.

Abstract: A large-span corridor structure composed of arc-shaped cantilever trusses and stay cable supporting structures is provided, comprising cantilever trusses with arc-shaped facades, orthogonal connecting trusses at a corridor, connecting trusses at cantilever portions, corridor truss structures, and stay cable supporting structures; the cantilever trusses with arc-shaped facades are composed of two sets of ultra-long cantilever trusses with arc-shaped facades and triquetrous planes which are supported by ground and symmetrical, and each are set of ultra-long cantilever trusses with arc-shaped facades and triquetrous planes are formed of several ultra-long cantilever trusses with arc-shaped facades and triquetrous planes arranged in parallel at a certain distance.

Abstract: The invention provides ultrasensitive methods for detection and quantification of target analytes in samples. The methods can be multiplexed to allow simultaneous detection and quantification of multiple target analytes. The methods can achieve an attomolar limit of detection. The invention also provides related compositions and kits.

Abstract: The present invention is directed to aqueous and hybrid aqueous electrolytes that comprise a lithium salt. The present invention is also directed to methods of making the electrolytes and methods of using the electrolytes in batteries and other electrochemical technologies.

Abstract: Methods of making safe extracts of Curcuma are provided. The processes provided include methods that use an extraction solvent that is at least substantially non-toxic and useful also as a pharmaceutically acceptable carrier in liquid dosage forms. The processes can produce a significantly higher yield from a single extraction than the state-of-the-art processes. For example, liquid dosage forms can be produced directly from the extraction process without requiring removal of the extraction solvent, reducing complexity and cost of processing over the state-of-the-art. Methods of making microemulsions and nanoemulsions are also provided to enhance the bioavailability and stability of the extracts.