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Methods of diagnosing muscle damageMethods of diagnosing muscle damage description/claimsThe Patent Description & Claims data below is from USPTO Patent Application 20090148860, Methods of diagnosing muscle damage. Brief Patent Description - Full Patent Description - Patent Application Claims
This application claims the benefit of priority of U.S. Provisional Patent Application No. 60/052,697, filed Jul. 16, 1997, the contents of which is incorporated herein by reference. This invention relates to methods for assessing the extent of cellular damage of muscle tissue, particularly skeletal and cardiac muscle. When circulation of blood, and therefore oxygen, to muscle is interrupted (ischemia), the ability of the muscle to contract is impaired. Even if circulation is restored (e.g., by reperfusion), muscle function can remain depressed. In certain types of muscle, such as cardiac muscle, the effects of ischemia can have severe consequences, and inadequate circulation of blood to the heart is one of the most important causes of morbidity in developed countries. Clinically, ischemia and reperfusion injury manifest as a spectrum. In its mildest form, ischemia is transient, reperfusion is established quickly and the reduced contractility of the muscle tissue is temporary and reversible. However, longer and more severe ischemia produces irreversible damage and cellular necrosis. The functions of myofilament specific proteins, such as troponin I and troponin T (members of the regulatory complex), myosin light chain 1 (MLC1), and α-actinin are affected during hypoxemia (i.e., reduced delivery of oxygen due to a reduced partial pressure and/or arterial content of oxygen), ischemia, and/or ischemia/reperfusion injury. These changes affect contraction of muscle by apparently altering the interaction of troponin I, troponin T, myosin light chain 1, and α-actinin with other proteins critical for normal muscle contraction. Troponin I is a key component of the troponin regulatory complex which directly controls striated (cardiac and skeletal) muscle contraction and relaxation. Troponin T is also part of the troponin complex and is involved in assembly of troponin-tropomyosin on the actin filament. α-actinin is a cytoskeletal protein, the main component of the Z lines. Traditionally α-actinin was believed to keep the actin filaments aligned. However, the large changes in conformation of Z lines during the cross bridge cycle suggest that α-actinin has a dynamic role during muscle contraction. Myosin light chain 1 is an integral part of the myosin myofibril. Myosin light chain 1 is found in slow and fast skeletal and atrial and ventricular cardiac muscles. To date, the underlying molecular changes responsible for the reduced contractility of injured muscles resulting from hypoxemia, hypoxemia, ischemia, and reperfusion are not known. As a result, early diagnosis of the above-mentioned states and assessment of the extent of muscle damage in a subject, particularly the difference between reversible and irreversible muscle damage, has not been possible. The current serum diagnostic indicators used in diagnosing myocardial infarction (e.g., anti-troponin I, anti-troponin T and anti-creatine kinase MB antibodies by Spectral Diagnostics Inc., Toronto, Canada; anti-troponin T, anti-MLC1 antibodies by Baxter Inc., Chicago, Ill.; and anti-troponin I antibody by Baxter Inc.) are indicators of myocardial necrosis (irreversible damage) since they detect proteins which are released from the heart myocyte following the loss of cellular membrane integrity. These indicators provide no information as to the extent or type of myocardial damage, or the molecular nature of which would clearly benefit the art. To date there are no commercially-available serum or urine markers for skeletal muscle damage. The present invention provides methods for assessing cardiac and skeletal muscle damage in a subject. The method includes obtaining a biological sample from a subject being assessed for muscle damage and evaluating the sample for presence or absence of one or more myofilament protein modification products, including, for example, individual protein fragments, or covalent or non-covalent complexes formed from two or more myofilament proteins, which may be intact proteins or protein fragments in the biological sample. According to one aspect of the invention, the amount of myofilament protein modification product present in a biological sample can be assessed as an indication of the extent of muscle damage in the subject. In accordance with the invention, a method of assessing muscle damage in a subject comprises obtaining a biological sample from a subject, incubating the biological sample with at least one compound which specifically binds to one or more different myofilament proteins or myofilament protein modification products present in the sample, under conditions which allow the compound to form one or more complexes with the myofilament proteins or myofilament protein modification products, detecting said one or more complexes, and characterizing the profile of said one or more myofilament proteins or myofilament protein modification products contained in said one or more complexes, as an indication of the extent or type of muscle damage in the subject. The compound can for example be an antibody, a protein, a peptide or a peptidomimetic that forms a complex with the myofilament protein modification product. In certain embodiments, the myofilament protein is troponin I, troponin T, troponin C, myosin light chain 1, α-actinin or a fragment(s) or combination(s) thereof. The invention also provides a kit for assessing myocardial damage in a biological sample obtained from a subject. In one embodiment, the kit includes a compound which specifically binds to a myofilament protein modification product and instructions explaining how to use the kit to assess muscle damage in a biological sample obtained from a subject. In other embodiments the compound may bind to one or more myofilament protein modification products. The kit may also include a label or labelled compound used to identify the myofilament protein modification product(s) thereof. The kit may further include a reagent(s) appropriate for detecting the label. The invention further provides assays, e.g., screening tests, for identifying an agent which modulates the level of one or more myofilament protein modification products in a biological sample. The assay involves obtaining a biological sample containing a myofilament protein modification product from a subject, testing the biological sample with an agent (e.g., contacting the sample with the agent), and determining the effect of the agent on the level of the myofilament protein modification product in the biological sample, wherein an agent(s) which modulate the level of the myofilament protein modification product in a biological sample are identified. In accordance with the invention, the presence and level of myofilament modification products in a biological sample are detected. The biological sample can be obtained from any subject exhibiting, exposed to, suspected of having, or being treated for, a condition or conditions which could cause hypoxemic/ischemic damage to muscle tissue. The invention therefore also provides for the assessment of efficacy of, for example, treatments such as cardioplegia (preservation) and preconditioning of the myocardium, and rehabilitation following heart disease-related injury such as infarction. The invention is also applicable to rehabilitation of patients with skeletal muscle damage, disease such as rhabdomyolysis, respiratory diseases such as, but not restricted to, chronic obstructive pulmonary disease, emphysema, asthma and bronchitis, bullectomy (lung reduction surgery), and following insult due to surgery or other trauma. The invention further provides for the assessment of the appropriateness of the level of training in athletes and animals such as race horses, where myofilament modification products as indicators of skeletal muscle breakdown can be detected. Yet other applications of the invention include the diagnosis of respiratory muscle dysfunction, wherein myofilament modification products can also exist. Other features and advantages of the invention will be apparent from the following detailed description, and from the claims. Embodiments of the invention will now be described, by way of example, with reference to the accompanying drawings, wherein: Continue reading about Methods of diagnosing muscle damage... Full patent description for Methods of diagnosing muscle damage Brief Patent Description - Full Patent Description - Patent Application Claims Click on the above for other options relating to this Methods of diagnosing muscle damage patent application. 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