The present disclosure relates to an acetylcholine receptor-binding peptide and, more particularly, to novel peptides which exhibit a wrinkle amelioration effect by binding the peptides to an acetylcholine receptor on which acetylcholine acts, thereby blocking secretion of acetylcholine. Peptides according to the present disclosure suppress secretion of acetylcholine by having a high binding strength with the acetylcholine receptor, thereby strongly binding the peptides to acetylcholine. Therefore, a cosmetic composition and a pharmaceutical composition comprising the peptides according to the present disclosure as an active ingredient exhibit an excellent wrinkle ameliorating effect.

The present invention relates to the use of compounds that block the α9α10 subtype of the nicotinic acetylcholine receptor (nAChR) for treating pain, such as neuropathic pain and inflammatory pain, and inflammatory disorders, such as arthritis. The present invention also relates to screening compounds to identify analgesic agents that block the α9α10 subtype of the nAChR.

The present invention pertains to fields of biochemistry and molecular biology, relates to an αO-superfamily conotoxin peptide, pharmaceutical composition thereof, preparation method and use thereof. The present invention further relates to a propeptide of the conotoxin peptide, nucleic acid construct thereof, expression vector and transformed cell thereof, and fusion protein thereof. The present invention further relates to a method for blocking acetylcholine receptors as well as a use of the conotoxin peptide in the manufacture of a medicament. The new αO-superfamily conotoxin peptide of the present invention is capable of specifically blocking acetylcholine receptor (nAChRs) (e.g., α9α10 nAChR), and NMDA receptor (e.g., NR2C NMDAR), and has activity for treatment of neuralgia, addiction, and activity for treatment of chemotherapy of cancers, breast cancer, lung cancer, wound healing, epilepsia, ischemia, and thus is promising in the manufacture of analgesic, a medicament for treatment of addiction, and a tool drug for neuroscience.

A method of diagnosing the risk that a subject has or has an increased risk of developing overactive bladder syndrome is described. The method includes detecting the levels of choline and/or acetylcholine in a biological sample from the subject, comparing the detected levels to reference values, and characterizing the subject as having an increased risk of having or developing overactive bladder syndrome if the choline and/or acetylcholine values are higher than the reference values. Kits for determining choline and/or acetylcholine levels in a subject, and methods of treating a subject for overactive bladder syndrome are also described.

Disclosed is a method for simultaneous analysis of neurotransmitters and/or their metabolites. The method includes (a) separating analytes including a plurality of neurotransmitters and/or their metabolites from a sample selected from body tissues, body fluids, secretions, and excretions, (b) derivatizing the analytes with ethyl chloroformate to obtain derivatives of the plurality of neurotransmitters and/or their metabolites, (c) separating the derivatives of the plurality of neurotransmitters and/or their metabolites by liquid chromatography, and (d) subjecting the separated derivatives of the neurotransmitters or their metabolites to multiple reaction monitoring (MRM) using a mass spectrometer. According to the method, a plurality of neurotransmitters in a very small amount of sample can be simultaneously analyzed in an accurate and rapid manner based on derivatization to increase the stability and ionization efficiency of the substances.

Provided herein are methods for determining the level of muscarinic acetylcholine receptor subtype-1 (M1 receptor) anticholinergic activity in a blood serum sample. The methods include radioactive methods and non-radioactive methods. The method comprises the steps of removing protein from the blood serum sample by treatment with perchloric acid (PCA) to produce a PCA-treated serum sample, incubating the PCA-treated serum sample with a membrane preparation from cultured cells expressing the M1 receptor and an M1 receptor ligand; detecting an amount of binding of the M1 receptor ligand to the M1 receptor and comparing the amount of binding to a standard to determine the level of M1 receptor anticholinergic activity in the blood serum sample. Alternatively, the method may comprise loading calcium sensitive dye into the cells and testing serum from a subject in a fluorescence assay to determine the anticholinergic activity relative to a sample known to contain little to no anticholinergic activity. Also provided are kits for performing the method.

The present invention includes a method for identifying a patient response to treatment for Alzheimer's Disease with a non-steroidal anti-inflammatory drug (NSAID) or an acetylcholinesterase (AChE) inhibitor drug comprising: obtaining a blood or serum sample from the patient; determining the presence of a proinflammatory endophenotype in the blood or serum sample of the patient; using the proinflammatory endophenotype to detect treatment response (a responder, a stable, a non-responder or an adverse responder); and treating the patient with the NSAID or the AChE inhibitor if the patient is in the responder or the stable treatment response phenotype group; or preventing a treatment with the NSAID or the AChE inhibitor if the patient is a non-responder or an adverse responder.

The invention provides a method of diagnosing overactive bladder disorder (OAB), the method comprising: measuring the concentrations of one or more of adenosine triphosphate (ATP), acetylcholine (ACh), nitric oxide (NO) and interleukin 5 (IL-5) in a sample obtained from a subject; normalising the concentrations to the concentration of creatinine (Cr) in the sample; range standardising the normalised concentrations and subject's age to the following values: Age to 120 years old; ATP/Cr to 0.000001; ACh/Cr to 0.1; NO to 20000; IL-5/Cr to 100; wherein the likelihood of having OAB (p.sub.OAB)=1/1+e.sup.−x, where X=one or more of the following: (a) (−2.688±1.050)+5.472±2.098×subject's age+1.356±0.559×Gender (Female=1, Male=0)+(−7.998±40.273)×[IL-5/Cr]; (b) (−2.141±0.966)+4.506±1.902×subject's age+1.034±0.519×Gender (Female=1, Male=0)+(−5294.063±9075.456)×[ACh/Cr]; (c) (−2.825±1.072)+5.964±2.167×subject's age+1.312±0.562×Gender (Female=1, Male=0)+17.790±58.762×[IL-5/Cr]+(−9180.821±12700.057)×[ACh/Cr]; (d) (−2.993±1.197)+5.580±2.309×subject's age+1.724±0.719×Gender (Female=1, Male=0)+63.571±73.444×[IL-5/Cr]+(−0908.523±13606.752)×[ACh/Cr]+(−566.991±636.589)×[ATP/Cr]; (e) (−3.090±1.200)+5.393±2.256×subject's age+1.797±0.717×Gender (Female=1, Male=0)+34.767±56.331×[IL-5/Cr]+(−562.743±629.316)×[ATP/Cr]; or (f) (−2.650±1.067)+5.516±2.120×subject's age+1.389±0.583×Gender (Female=1, Male=0)+(−4.060±45.238)×[IL-5/Cr]+(−1.456±6.833)×[NO/Cr]; and wherein a pOAB above a threshold indicates that the subject has a high likelihood of having or developing OAB and a pOAB below a threshold indicates that the subject does not have OAB.

The present invention provides methods for treating inflammatory or autoimmune diseases or conditions using activators tin of nicotinic acetylcholine receptors (nAChRs) containing a cholinergic receptor nicotinic alpha 6 subunit (nAChRS), such as activating antibodies that bind to a nAChR containing a nAChP6 subunit and small molecule agonists of nAChRs containing a nAChRS subunit. The invention also features compositions containing 6*nAChR activators, methods of diagnosing patients with an 6*nAChR-associated inflammatory or autoimmune disease or condition, and methods of predicting the response of an inflammatory or autoimmune disease or condition in a subject to treatment with 6*nAChR activators.

Methods for treating nerve damage in a muscle, e.g., denervated muscle tissue (e.g., muscle damaged from traumatic injury), in a patient in need thereof featuring performing a pre-operative muscle biopsy on the denervated muscle tissue; making visible motor end plates (MEPs) in neuromuscular junctions (NMJs) in the biopsy; and performing a nerve transfer (e.g., partial radial nerve to axillary transfer) if (i) the MEPs shown in the biopsy persist and (ii) the MEPs shown in the biopsy retain their structures and exhibit certain morphometric properties. The nerve transfer helps regain neuromuscular function of the denervated muscle tissue. The biopsy may feature the use of two-photon microscopy. In certain embodiments, the method is performed at least 6 months after injury to the patient.