The invention is directed to compositions to screen for small molecule drugs that inhibit proteases, such as viral proteases, e.g., HIV proteases; and methods for making and using these compositions. The invention provides compositions and methods for identifying compositions, e.g., drug molecules, that can inhibit proteases, e.g., HIV proteases. In alternative embodiments, the invention provides cell-based assays to screen for compositions, e.g., small molecules or drugs, that inhibit or modify the activity of enzymes such as calcium-dependent protein convertases involved in HIV envelop protein processing, including cleavage of the HIV gp160 envelope precursor, resulting in gp120 and gp41 envelope products.

Disclosed are methods, compositions and kits for the isolation of exosomes from biological fluids and tissues. Volume-excluding polymers are used to precipitate exosomes from biological samples thereby allowing exosome isolation by low-speed (benchtop) centrifugation or filtration. Further fractionation of exosomes after precipitation is also described.

The disclosure provides compositions and methods relating to aromatic-cationic peptides. The methods comprise administering to the subject an effective amount of an aromatic-cationic peptide to subjects in need thereof. For example, the peptides may be administered to subjects in need of a mitochondrial-targeted antioxidant.

The present invention relates to the preparation and use of therapeutic compounds for the treatment of diseases at specific subcellular target areas such as specific cellular organelles. In particular, the therapeutic compounds of the invention are specific for modifying enzyme activity within targeted organelles or structures of cells and tissues. Subcellular organelles and structures that may be specifically targeted by compounds of the present invention include lysosomes, autophagasomes, the endoplasmic reticulum, the Golgi complex, peroxisomes, the nucleus, membranes and the mitochondria.

Systems and methods for identifying molecules that are biologically active against a disease, where the method can comprise culturing a first mammalian cell population under organoid formation conditions in the presence of a test molecule to obtain a first organoid, wherein the first mammalian cell population, when cultured under the organoid formation conditions in the absence of the test molecule, results in an organoid with a disease phenotype; imaging the first organoid following exposure to the test molecule; analyzing one or more images of the first organoid using a neural network that has been trained to assign a probability score of disease or non-disease ranging between 0% and 100%; assigning the first organoid a probability score ranging between 0% and 100%; wherein the test molecule is biologically active against the disease if the probability score of the first organoid is greater than a cutoff probability score of non-disease or lower than a cutoff probability score of disease.

This document relates to methods and materials involved in treating mammals having a cancer (e.g., a cancer having one or more CD19 positive extracellular vesicles (EVs) and/or one or more small EVs in the tumor microenvironment). For example, methods of treating a mammal having a blood cancer including one or more CD19 positive EVs and/or one or more small EVs in the blood by administering one or more cancer immunotherapies (e.g., one or more chimeric antigen receptor (CAR) T-cell therapies) to the mammal are provided.

In some aspects, the present disclosure provides a method for identifying one or more biomolecules. In some embodiments, the method comprises generating a set of biomolecules by incubating a cell in a supplemented medium under conditions sufficient for the cell to generate the set of biomolecules. In some embodiments, the method comprises contacting at least a portion of the supplemented medium with one or more surfaces to adsorb the set of biomolecules. In some embodiments, the method comprises removing the one or more surfaces and the set of biomolecules from the at least the portion of the supplemented medium to produce a separated sample. In some embodiments, the method comprises releasing, in the separated sample, the set of biomolecules from the one or more surfaces. In some embodiments, the method comprises detecting at least a subset of the set of biomolecules, thereby identifying one or more biomolecules.

Compositions and methods for phase-separation of chromatin in distinct states are provided. In particular, differentially decorated chromatin in different states can be isolated by phase separation according to the methods disclosed herein. In some cases, specific chromatin interacting proteins are added to chromatin to recreate specific, biologically relevant chromatin states in vitro, which can be phase-separated using the disclosed methods. In addition, methods of using phase-separated chromatin for in vitro screening of pharmacological agents that modulate chromatin states, including small molecule chemicals and biomolecules, are also provided.

Techniques for screening. A lipid and gramicidin are mixed and prepared for processing. The mixture is frozen and then thawed in a series of cycles before being extruded. The extruded mixture is frozen and thawed. Once thawed, the extruded mixture may be desalted to provide a liposome solution. The liposome solution may be stored for subsequent use such as, but not limited to, for use in screening tests.

The present document describes an in vitro method for determining a level of mitochondrial function from a subject by determining a change in a labeled compound labeling a viable cell or tissue sample isolated from a subject following introduction of a volume of a solution over said viable cell or tissue sample. The comparison of the change to a normal reference is indicative of mitochondrial function in the subject.