Abstract: The present disclosure relates, in some aspects, to the field of process analytical technology with respect to biotherapeutic protein production processes and products.

Abstract: Hypoxia plays a central role in cancer progression and resistance to therapy. A microdevice platform is engineered to recapitulate the intratumor oxygen gradients that drive the heterogeneous hypoxic landscapes in solid tumors. The microdevice design features a “tumor section”-like culture by incorporating a cell layer between two diffusion barriers, where an oxygen gradient is established by cellular metabolism and physical constraints. The oxygen gradient is confirmed by numerical simulation and imaging-based oxygen sensor measurement. Spatially-resolved hypoxic signaling in cancer cells is also demonstrated through immunostaining, gene expression assay, and hypoxia-targeted drug treatment. The microdevice platform can accurately generate and control oxygen gradients, eliminates complex microfluidic handling, allows for incorporation of additional tumor components, and is compatible with high-content imaging and high-throughput applications.

Abstract: The present invention provides an apparatus and a method for a lysis procedure, in particular for an automated and/or controlled lysis procedure of a sample, in particular a biological sample.

Abstract: The present invention relates to a recombinant cyanobacterium with enhanced halotolerance and compositions thereof, methods of producing the recombinant cyanobacterium, and methods of using the same for biofuel production. The invention also relates to transformed F. diplosiphon strains with enhanced salt tolerance.

Abstract: The present invention provides constructs including a tubular biodegradable polyglycolic acid scaffold, wherein the scaffold may be coated with extracellular matrix proteins and substantially acellular. The constructs can be utilized as an arteriovenous graft, a coronary graft, a peripheral artery bypass conduit, or a urinary conduit. The present invention also provides methods of producing such constructs.

Abstract: The present invention relates to a Apium graveolens L. dulce seed designated 49-08 RZ. The present invention also relates to a Apium graveolens L. dulce plant produced by growing the 49-08 RZ seed. The invention further relates to methods for producing the celery cultivar, represented by celery variety 49-08 RZ.

Abstract: The present invention relates to compositions, methods and kits using cell phenotype altering polynucleotides, cell phenotype altering primary transcripts and cell phenotype altering mmRNA molecules.

Abstract: The present invention discloses a high volume manufacturing process for the production and purification of exosomes. In one embodiment, a hollow fiber perfusion reactor is used to expand stem cells. The growth media used from the stem cell expansion is captured, filtered, centrifuged and exosomes isolated from the waste effluent. In one embodiment, stem cells are derived from bone marrow, fat, blood, cord blood and induced pluripotent stem cells are expanded and the exosomes captured from the growth waste effluent. The exosomes can be used as therapeutics and diagnostics for a number of regenerative and chronic diseases.

Abstract: This invention provides a system for efficiently producing differentiated cells from pluripotent cells, such as human embryonic stem cells. Rather than permitting the cells to form embryoid bodies according to established techniques, differentiation is effected directly in monolayer culture on a suitable solid surface. The cells are either plated directly onto a differentiation-promoting surface, or grown initially on the solid surface in the absence of feeder cells and then exchanged into a medium that assists in the differentiation process. The solid surface and the culture medium can be chosen to direct differentiation down a particular pathway, generating a cell population that is remarkably uniform. The methodology is well adapted to bulk production of committed precursor and terminally differentiated cells for use in drug screening or regenerative medicine.

Abstract: The invention relates to a method for polar body injection, which comprises removing a polar body from a first egg cell and injecting the polar body into a second egg cell that is in a fertilizable state.

Abstract: The presently disclosed subject matter provides for in vitro methods of inducing differentiation of stem cells into enteric neural crest lineage cells, and enteric neural crest lineage cells by such methods. The presently disclosed subject matter also provides for uses of such enteric neural crest lineage cells for preventing and/or treating enteric nervous system disorders (e.g, Hirschsprung's disease), and for screening compounds suitable for preventing and/or treating enteric nervous system disorders (e.g., Hirschsprung's disease).

Abstract: Provided herein are methods of expanding B cells, and in particularly B10 cells capable of producing IL-10, ex vivo. The methods include incubation of harvested B cells in the presence of IL-21. Compositions comprising the ex vivo expanded B cells and methods of using the expanded B cell-containing compositions to treat diseases or conditions are also provided. Methods of assessing B10 cell function in a subject are also provided.

Abstract: The present invention relates to cell culture in bioreactors, such as flexible cellbag bioreactors. More specifically the present invention relates to methods for simplifying the production of clinically relevant cell products for use in cell therapy.

Abstract: Embodiments described herein generally provide for expanding cells in a cell expansion system. The cells may be grown in a bioreactor, and the cells may be activated by an activator (e.g., a soluble activator complex). Nutrient and gas exchange capabilities of a closed, automated cell expansion system may allow cells to be seeded at reduced cell seeding densities, for example. Parameters of the cell growth environment may be manipulated to load the cells into a particular position in the bioreactor for the efficient exchange of nutrients and gases. System parameters may be adjusted to shear any cell colonies that may form during the expansion phase. Metabolic concentrations may be controlled to improve cell growth and viability. Cell residence in the bioreactor may be controlled. In embodiments, the cells may include T cells. In further embodiments, the cells may include T cell subpopulations, including regulatory T cells (Tregs), for example.

Abstract: The present invention relates to methods for developing engineered immune cells such as T-cells for immunotherapy that have a higher potential of persistence and/or engraftment in host organism. IN particular, this method involves an inactivation of at least one gene involved in self/non self recognition, combined with a step of contact with at least one non-endogenous immunosuppressive polypeptide. The invention allows the possibility for a standard and affordable adoptive immunotherapy, whereby the risk of GvH is reduced.

Abstract: This disclosure provides methods of making a megakaryocyte-erythroid progenitor cell (MEP), comprising differentiating a MEP precursor cell into a MEP in culture in the presence of an aryl hydrocarbon receptor (AhR) modulator. In some embodiments the AhR modulator is an AhR antagonist. In some embodiments the AhR modulator is an AhR agonist. In some embodiments the methods comprise culturing MEP precursor cells in the presence of an AHR antagonist and then culturing MEP precursor cells in the presence of an AHR agonist. In some embodiments the stem cell is a pluripotent stem cell. In some embodiments the MEP co-expresses CD41 and CD235. In some embodiments the number of MEPs produced in the culture increases exponentially. Methods of making a red blood cell (RBC) by culturing a MEP in the presence of an AhR modulator are also provided. Methods of making a megakaryocyte and/or a platelet, comprising culturing a MEP in the presence of an AhR modulator are also provided.

Abstract: The invention relates to purified, tissue-specific progenitors, methods of making and using such tissue-specific progenitors.

Abstract: A stem cell culturing method includes a second bone marrow aspirate cultured along with a culture medium with first stem cells being fixed on a first culturing vessel bottom surface. When a total surface area of the first stem cells reaches a first target ratio with respect to the bottom surface area of the first culturing vessel, the first stem cells are extracted from the first culturing vessel. Top layer second stem cells are extracted from the first stem cells that are separated into layers and the second stem cells are cultured along with a culture medium. The second stem cells are fixed on a second culturing vessel bottom surface and, when a total surface area of the second stem cells reaches a second target ratio with respect to the bottom surface area of the second culturing vessel, the second stem cells are extracted from the second culturing vessel.

Abstract: The present invention provides preparations of MSCs with important therapeutic potential. The MSC cells are non-primary cells with an antigen profile comprising less than about 1.25% CD45+ cells (or less than about 0.75% CD45+), at least about 95% CD105+ cells, and at least 95% CD166+ cells. Optionally, MSCs of the present preparations are isogenic and can be expanded ex vivo and cryo-preserved and thawed, yet maintain a stable and uniform phenotype. Methods are taught here of expanding these MSCs to produce a clinical scale therapeutic preparations and medical uses thereof.

Abstract: The present invention provides methods, cell cultures and differentiation media to promote differentiation of pluripotent stem cells to pancreatic endocrine cells of a mature phenotype. The resulting pancreatic endocrine cells express single hormonal insulin, PDX1, NKX6.1, and MAFA. In one or more differentiation stages, culturing may be carried out in a culture vessel at the air-liquid interface.

Abstract: This invention discloses a method for the induction of arterial-type of hemogenic endothelium.

Abstract: Provided herein are artificial lung organoids. The artificial lung organoids may include an epithelial cell layer comprising mammalian lung epithelial cells, a stromal cell layer comprising mammalian lung fibroblast cells and an endothelial cell layer comprising mammalian endothelial cells. The artificial lung organoids may optionally include a porous membrane between said epithelial cell layer and said stromal cell layer and/or between said stromal cell layer and said endothelial lung cell layer.

Abstract: The invention provides compositions and methods useful to prepare segmented, negative strand RNA viruses, e.g., orthomyxoviruses such as influenza A viruses, entirely from cloned cDNAs and in the absence of helper virus.

Abstract: Transgenic plants having enhanced yield and having enhanced seed yield are disclosed. The transgenic plants are transformed with a transgenic polynucleotide encoding one or more metabolic enzymes. The metabolic enzymes can be from any biological source. The transgenic polynucleotide(s) comprises a nucleic acid sequences encoding the metabolic enzymes under the control of functional plant promoters, the one or more metabolic enzymes are targeted to the plastids by the addition of plastid targeting signals. Optionally the functional plant promoters are seed specific promoters and the metabolic enzymes are targeted to the plastids by the addition of plastid targeting peptide heterologous to the metabolic enzymes. Methods of making the transgenic plants and transgenic polynucleotides are disclosed. The magnitude of the increases in seed yield achieved with these transgenic plants are simply unprecedented.

Abstract: The present disclosure relates generally to the production of alcohols, and more specifically to biological platforms for the production of alcohols using monofunctional aldehyde dehydrogenases and monofunctional alcohol dehydrogenases.

Abstract: The present disclosure relates to a mutant 3-hydroxybutyrate dehydrogenase (3-HBDH) with improved performance relative to the wild-type 3-HBDH, a nucleic acid encoding the mutant 3-HBDH, a cell having the mutant 3-HBDH or the nucleic acid, and/or a method of determining the amount or concentration of 3-hydroxybutyrate in a sample. Also disclosed is the use of the mutant 3-HBDH for determining the amount or concentration of 3-hydroxybutyrate in a sample, and a device for determining the amount or concentration of 3-hydroxybutyrate in a sample.

Abstract: A rubber polymerase protein consisting of the amino acid sequence of SEQ ID NO: 2 for biosynthesis of natural rubber originating from a para rubber tree (Hevea brasiliensis). A gene encoding the rubber polymerase protein can be advantageously used for a technique of increasing a production amount of natural rubber from a rubber tree, a technique of producing natural rubber in other plants, microalgae, or microorganisms, or a technique for in vitro production of biorubber.

Abstract: The present invention provides lipolytic enzyme variants. Specifically, the present invention provides lipolytic enzyme variants having one or more modifications as compared to a parent lipolytic enzyme having at least one improved property. In addition, the present invention provides compositions comprising a lipolytic enzyme variant of the invention. The present invention also provides methods of cleaning using compositions comprising a lipolytic enzyme variant of the invention.

Abstract: The present invention discloses a lysin that is capable of killing Staphylococcus and the use thereof, belonging to the field of biological agents. The present invention discloses the amino acid sequence and the encoding gene sequence of the lysin. This lysin keeps active in a wide range of pH. It has lytic activity against Staphylococcus in pH 4-11. The recombinant protease constructed by the encoding gene can be solubly expressed in E. coli strain BL21 (DE3). The lysin can be used to effectively kill multiple species Staphylococcus in vitro, including methicillin sensitive Staphylococcus aureus (MSSA) and methicillin resistant Staphylococcus aureus (MRSA) isolated in clinics. This lysin can be used as an antibiotic for the treatment of staphylococcal infections in vivo. This lysin is also able to rapidly lyse staphylococcal cell wall; as a result, intracellular substances such as ATP and DNA are released. Those released substances can be used to detect the type of Staphylococcus.

Abstract: The present invention relates to isolated polypeptides having xylanase activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods for producing and using the polypeptides.

Abstract: The present invention provides cell-targeting molecules comprising binding regions for cell-type specific targeting and Shiga toxin A Subunit effector regions for Shiga toxin effector functions, wherein the Shiga toxin effector regions are at and/or proximal to an amino-terminus of a polypeptide component of the cell targeted molecule, and optionally comprising a disrupted, furin-cleavage motif between the Shiga toxin effector region and the binding region. The cell-targeting molecules of the invention exhibit a more optimized cytotoxicity and/or improved, in vivo tolerability as compared to related molecules comprising less amino-terminus proximal, Shiga toxin effector regions and/or furin-cleavage sensitive, wild-type, Shiga toxin effector regions. The cell targeting molecules of the invention have uses, such as, e.g.

Abstract: The present disclosure provides polypeptides having aminopeptidase activity and isolated nucleic acid sequences encoding the polypeptides. In some embodiments, the disclosure also provides to nucleic acid constructs, vectors, and host cells comprising the nucleic acid sequences as well as methods for producing the polypeptides. In some embodiments, the present disclosure further provides to methods of obtaining protein hydrolysates useful as flavor improving agents.

Abstract: The present invention relates to isolated polypeptides having protease activity, and polynucleotides encoding the polypeptides. The invention further relates to the use of such polypeptides in detergent and/or in cleaning processes. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing the polypeptides.

Abstract: The present invention relates to a method for producing a lysine carboxylase mutant strain, characteristics of the mutant strain, a gene encoding the lysine decarboxylase mutant strain, and a method for producing cadaverine using the same. The present invention provides lysine decarboxylase derived from E. coli improved through a protein engineering variation. In addition, the lysine decarboxylase mutant strain of the present invention increases activity, pH stability, and thermal stability at the time of producing cadaverine, thereby reducing production costs, through increasing a yield and productivity.

Abstract: Provided herein are recombinant microorganisms having two or more copies of a nucleic acid sequence encoding xylose isomerase, wherein the nucleic acid encoding the xylose isomerase is an exogenous nucleic acid. Optionally, the recombinant microorganisms include at least one nucleic acid sequence encoding a xylulose kinase and/or at least one nucleic acid sequence encoding a xylose transporter. The provided recombinant microorganisms are capable of growing on xylose as a carbon source.

Abstract: An apparatus and methods are provided for the magnetic separation of target bioentities. The apparatus include a fluid chamber and a magnetic element for drawing target bioentities toward a collection surface of the fluid chamber. The apparatus may include a positioning assembly operable to variable change the position and orientation of the fluid chamber relative to the magnetic element.

Abstract: The present invention provides a poly(alkylene oxide) polymer based size selective DNA isolation method for isolating DNA molecules having a size above a certain cut-off value from a DNA containing sample, comprising (a) preparing a binding mixture comprising the DNA containing sample, at least one poly(alkylene oxide) polymer and at least one divalent cation, wherein said binding mixture has a pH that lies in the range of 8 to 10 and binding precipitated DNA molecules to a solid phase having an unmodified silicon containing surface, thereby providing a solid phase having bound thereto DNA molecules having a size above the cut-off value, wherein under the used binding conditions DNA molecules having a size which is less than the cut-off value substantially do not bind to the solid phase; (b) separating the bound DNA molecules from the remaining sample; —optionally washing the bound DNA molecules; and —optionally eluting the bound DNA molecules from the solid phase.

Abstract: A method of altering the functional state of any mRNA enabling its selective and specific recognition and subsequent selective manipulation and a universal principle for increasing the specificity and selectivity of molecular target recognition at the level of nucleic acids. The principle of the specific and selective recognition of nucleic acids is based on simultaneous recognition of two or more sequences of the target nucleic acid, whereas these have to be spaced from each other by a certain defined distance. Such method of nucleic acid recognition through specific recognition of well-defined sequences of the nucleic acid that are spaced from each other by a defined distance, minimizes the probability of stable binding of the interfering construct to inadvertent nucleic acids, thereby dramatically increasing the selectivity of recognition of the targeted nucleic acid.

Abstract: The present invention relates to a highly automatable method for isolation and/or purification of nucleic acids from a biological sample, which is particularly suitable for nucleic acids-shorter than 250 bp and can be performed without a proteolytic pre-digestion step in an automated system, preferably a cartridge-based system. In a further aspect, the present invention also provides automated nucleic acid detection methods based on said isolation and/or purification method, as well as buffers and kits to be used in performing said methods.

Abstract: Focused libraries of vectors or genetic packages that display, display and express, or comprise a member of a diverse family of antibody peptides, polypeptides or proteins and collectively display, display and express, or comprise at least a portion of the focused diversity of the family. The libraries have length and sequence diversities that mimic that found in native human antibodies.

Abstract: A method and kit for constructing a barcoded single-stranded DNA library are disclosed. The method includes preparing single-stranded DNA molecules each having a dephosphorylated 5? end, ligating a first adaptor to a 3? end of each single-stranded DNA molecule, and synthesizing a complementary strand of each single-stranded DNA molecule ligated to the first strand of the first adaptor. The kit includes the first adaptor having a first strand, which includes, from a 5? end to a 3? end, a phosphate group, a barcode sequence, and a first primer recognition sequence. The kit also includes a DNA ligase for a ligation between the 5? end of the first strand of the first adaptor to each single-stranded DNA molecule, and a first primer for the synthesis of the complementary strand. The method allows for analysis of rare mutations and from nucleic acid samples of low quality and quantity.

Abstract: Disclosed herein are methods of using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated (Cas) 9-based epigenomic editing systems for high-throughput screening of regulatory element function.

Abstract: Provided are a method for constructing a nucleic acid single-stranded cyclic library and the reagents used therein. By the combination of interruption via a transposase with a restricted nick translation reaction, the method realizes a simple and rapid nucleic acid single-stranded cyclic library construction.

Abstract: The present disclosure is directed, in some embodiments, to engineered nucleic acids comprising a promoter operably linked to a nucleotide sequence encoding a guide ribonucleic acid (gRNA) that comprises a specificity determining sequence (SDS) and a protospacer adjacent motif (PAM). The present disclosure is directed, in some embodiments, to cells comprising, vectors comprising, and methods of producing the engineered nucleic acids.

Abstract: The specification relates to the use of Mcl-1 inhibitors to promote apoptosis in vascular endothelial cells undergoing neovascularisation in disease states.

Abstract: Provided herein are methods, kits and compositions to increase ANP levels in a subject for the treatment of hypertension and/or cardiovascular diseases, comprising at least one anti-miR agent that inhibits at least one of, or a combination of miR-103, miR-105, miR-107 and/or miR-155, alone, or in combination with an inhibitor of miR-425. Anti-miR agents can be small molecules or an oligonucleotide complementary to at least part of the sequence of hsa-miR-103, hsa-miR-105, hsa-miR-107 and/or hsa-miR-155, or their respective seed sequences or.their binding site in the 3?UTR of NPPA gene. Also provided herein are methods, kits and compositions to decrease ANP levels in a subject for the treatment of low blood pressure, comprising at least one or a combination of agonists of miR-103, miR-105, miR-107 and/or miR-155, alone or in combination with a miR-425 agonist.

Abstract: Described herein are composition and methods related to targeting the Nf1-Ras-Ets axis, that when perturbed, is identified as playing a role in initiation and maintenance in glioma. A postnatal, mosaic, autochthonous, glioma model that captures the first hours and days of gliomagenesis in more resolution than conventional genetically engineered mouse models of cancer demonstrates that disruption of the Nf1-Ras pathway in the ventricular zone at multiple signaling nodes uniformly results in rapid neural stem cell depletion, progenitor hyperproliferation, and gliogenic lineage restriction. By abolishing Ets subfamily activity, which is upregulated downstream of Ras, there is block of glioma initiation, thereby providing new therapeutic avenues for targeting glioma.

Abstract: Disclosed herein are compounds, compositions and methods for modulating splicing of SMN2 mRNA in a cell, tissue or animal. Also provided are uses of disclosed compounds and compositions in the manufacture of a medicament for treatment of diseases and disorders, including spinal muscular atrophy.

Abstract: Among the various aspects of the present disclosure is the provision of a blood brain barrier (BBB) opening agent and uses thereof. An aspect of the present disclosure provides for methods of opening up the BBB; increasing the permeability of the tricellular junction; and using a BBB opening agent for the treatment of a brain pathology or neurological disease, disorder, or condition.

Abstract: The presently disclosed subject matter relates, in certain embodiments, to compositions and methods for the inhibition of the JAK-STAT pathway in order to induce hair growth. In certain embodiments, the presently disclosed subject matter relates to topical treatments with small molecule inhibitors of the JAK-STAT pathway to induce hair growth.