A method of making nicotinamide riboside, nicotinamide riboside derivatives, or mixtures thereof is disclosed. The method involves contacting at least the following materials to form a solution: i) -D-ribose-1-phosphate, -D-ribose-1-phosphate derivatives, or mixtures thereof; ii) nicotinamide, nicotinamide derivatives, or mixtures thereof; iii) one or more pentosyl transferases (E.C. 2.4.2); iv) and one or more solvents. The resulting solution comprises nicotinamide riboside, nicotinamide riboside derivatives, or mixtures thereof and one or more inorganic orthophosphate anions. The inorganic orthophosphate anions are removed from the solution, leaving a solution of nicotinamide riboside, nicotinamide riboside derivatives, or mixtures thereof.

Provided is aptamers screening method based on graphene without target immobilization and the aptamers obtained from the method, and more particularly, a new GO-SELEX method without target immobilization in which a single-stranded nucleic acid pool may react with a non-bound target material or a counter-target material, after which a single-stranded nucleic acid which has not been bound to the target or counter-target may be separated by using the graphene. Also, the specific aptamer obtained through the above-described method may be used for diagnosing target related diseases.

The present invention relates to productive tissue repair and regeneration. and in particular poylpeptides. compositions including said polypeptides. and methods of using said polypeptides or compositions for productive tissue repair and regeneration. In one aspect. the invention provides a polypeptide comprising. consisting essentially of or consisting of a C-terminal portion of NAMPT comprising a truncated cytokine finger motif (cif) motif. In another aspect. the present invention provides a fusion protein comprising. consisting essentially of or consisting of a polypeptide of a full length NAMPTcif or truncated variants and a tissue delivery or retention enhancing moiety.

A protein of the present invention is a protein having nicotinamide phosphoribosyl transferase activity, described in any one of the following [1] to [3]: [1] a protein consisting of an amino acid sequence set forth in SEQ ID NO: 3 or 5; [2] a mutant protein consisting of an amino acid sequence obtained by deleting, substituting, inserting, or adding 1 to 20 amino acids in the amino acid sequence set forth in SEQ ID NO: 3 or 5; and [3] a homologous protein consisting of an amino acid sequence having 90% or more identity to the amino acid sequence set forth in SEQ ID NO: 3 or 5.

It was discovered that ICIs elevated extracellular NAD levels and conferred cancer cells resistance to ICIs. It was also discovered that NAMPT was induced by IFN-y and ICIs resulting in an increase of extracellular NAD level, which could be reduced by NAMPT knockdown. NAMPT deficiency in HCC sensitized tumors to anti-PD-1 treatment. It was further discovered that extracellular NAD promoted T cell apoptosis, exhaustion, and T cell differentiation to regulatory T cells. The blockade of P2X7R reversed the immunosuppressive effect of NAD and worked effectively in combination with immunotherapies, indicating that it can be a combination therapeutic approach for ICI-resistant tumors. It was shown that serum NAD level can be a predictive biomarker for ICI responses in HCC. Overall, extracellular NAD was shown to act as an immunosuppressive metabolite that could serve as a biomarker for ICI responses and the P2X7R can be a therapeutic target to improve ICI efficacy.

The present invention relates to a method for producing synthetic extracellular vesicles comprising a lipid bilayer including at least two lipids, one or more extracellular vesicle associated proteins, and optionally one or more nucleic acid molecules. The inventive synthetic extracellular vesicles are formed by emulsification using a mechanic emulsifier in the form of polymer shell stabilized synthetic extracellular vesicles. The inventive method allows producing synthetic extracellular vesicles miming the composition and function of natural extracellular vesicles. Therefore, synthetic extracellular vesicles with specific protein and nucleic acids compositions are also disclosed herein, as well as their therapeutic uses.