Abstract: The disclosure provides methods and compositions for treating blood diseases/disorders, such as sickle cell disease, hemochromatosis, hemophilia, and beta-thalassemia. For example the disclosure provides therapeutic guide RNAs that target the promotor of HBG1/2 to generate point mutations that increase expression of fetal hemoglobin. As another example, the disclosure provides therapeutic guide RNAs that target mutations in HBB, Factor VIII, and HFE to treat sickle cell disease, beta-thalassemia, hemophilia and hemochromatosis. The disclosure also provides fusion proteins comprising a Cas9 (e.g., a Cas9 nickase) domain and adenosine deaminases that deaminate adenosine in DNA. In some embodiments, the fusion proteins are in complex with nucleic acids, such as guide RNAs (gRNAs), which target the fusion proteins to a DNA sequence (e.g., an HBG1 or HBG2 protmoter sequence, or an HFE, GBB, or F8 gene sequence).

Abstract: Some aspects of this disclosure provide methods for phage-assisted continuous evolution (PACE) of proteases. Some aspects of this invention provide methods for evaluating and selecting protease inhibitors based on the likelihood of the emergence of resistant proteases as determined by the protease PACE methods provided herein. Some aspects of this disclosure provide strategies, methods, and reagents for protease PACE, including fusion proteins for translating a desired protease activity into a selective advantage for phage particles encoding a protease exhibiting such an activity and improved mutagenesis-promoting expression constructs. Evolved proteases that recognize target cleavage sites which differ from their canonical cleavage site are also provided herein.

Abstract: The use of compatible, semi-miscible or miscible polymer compositions as support structures for the 3D printing of objects, including those made from polyether-block-amide copolymers such as PEBAX® block copolymers from Arkema, polyamides such as RILSAN® polyamides from Arkema, polyether ketone ketone such as KEPSTAN® PEKK from Arkema, and fluoropolymers, such a KYNAR® PVDF from Arkema, especially objects of polyvinylidene fluoride and its copolymers. One particularly useful miscible polymer is an acrylic polymer, which is miscible with the fluoropolymer in the melt. The support structure composition provides the needed adhesion to the build plate and to the printed object and support strength during the 3D printing process, yet it is removable after the fluoropolymer object has cooled. The support polymer composition is selected to be stiff and low warping, yet flexible enough to be formed into filaments.

Abstract: The present invention is directed to material extrusion additive manufacturing processes, including fused filament fabrication, used to manufacture improved parts, devices, and prototypes using polyetherketoneketones (“PEKK”) and polyetheretherketones (“PEEK”). Using the improved processes of the invention, PEKK or PEEK polymer readily is 3D printed by FFF such that it crystallizes slowly enough during deposition for the resulting part to remain mostly or substantially amorphous during printing and thus have low percentage and/or more uniform shrinkage per layer and little to no warping from the base during print, and yet fast enough so that the resulting part crystallizes in post-print processing without substantial or any loss of its printed structure.

Abstract: A decorative cap and fastener for fastening a wheel to a hub of a vehicle are provided. The fastener includes a fastener head including an outer surface and an internal surface defining a cavity. The cap includes an end wall having an external surface and an internal surface. The cap further includes a side wall extending from an outer periphery of the end wall. The cap also includes a central member extending longitudinally from the internal surface of the end wall within the side wall.

Abstract: A decorative cap and fastener for fastening a wheel to a hub of a vehicle are provided. The fastener includes a fastener head including an outer surface and an internal surface defining a cavity. The cap includes an end wall having an external surface and an internal surface. The cap further includes a side wall extending from an outer periphery of the end wall. The cap also includes a central member extending longitudinally from the internal surface of the end wall within the side wall.

Abstract: A wheel includes a rim having an outer radius, R1, extending from a center point of the wheel to an outer edge of the rim, and a hub having an outer radius, R2, extending from the center point of the wheel to an outer edge of the hub. The hub is positioned coaxially with the rim and has a plurality of circumferentially positioned and equiangularly spaced bolt holes for attachment of the wheel to the vehicle. The wheel further includes a plurality of equiangularly spaced, Y-shaped spokes having a trunk portion contacting the hub, two limb portions each contacting the rim, and a branch point at which the trunk portion and limb portions converge. The spokes extend radially outward from the hub to the rim. The bolt holes are interspaced between the trunk portions. A distance between the center point of the wheel and the branch point is D1.

Abstract: A wheel assembly for a motor vehicle, which includes a driveshaft member with a shortened stem portion. The wheel assembly also includes a bearing arrangement with an asymmetric configuration. An outboard bearing row may be disposed radially further from a central axis of a hub member than an inboard bearing row, and may have a diameter that is smaller than a diameter of the inboard bearing row. An outboard end of the stem portion may be generally aligned in the axial direction with the outboard bearing row. The stem portion length may approximately correspond to the bearing assembly length. Contact angles of a bearing assembly may be configured to distribute force over the wheel assembly. A fastening member may engage an outboard end of the stem portion and apply force to the hub member at a location inboard of the outboard bearing row.

Abstract: A wheel assembly for a motor vehicle is disclosed. The wheel assembly includes a driveshaft member with a shortened stem portion. The wheel assembly also includes a bearing arrangement with an asymmetric configuration.