phosphorus-radioisotopes and Muscular-Dystrophies

Recent developments in recombinant DNA technology have made possible the production of gene probes consisting of cloned gene segments, cloned segments of DNA linked to genes, and synthetic gene fragments. Several methods have been developed by which these probes may be used for the diagnosis of human disease. This technology has been outstandingly successful for prenatal diagnosis and carrier detection in many genetic diseases. These methods have also been successfully applied to the analysis of human malignancies, by providing for the determination of cell lineage and clonality in lymphoid neoplasms. Finally, these methods have shown potential for rapid and sensitive diagnosis of some infectious diseases.

Topics: B-Lymphocytes; Bacterial Infections; Biotin; Cloning, Molecular; Deoxyribonucleotides; DNA; DNA Restriction Enzymes; DNA, Recombinant; Female; Genetic Carrier Screening; Genetic Diseases, Inborn; Genotype; Hemophilia A; Humans; Immunoglobulins; Infections; Leukemia, Lymphoid; Leukocytes; Male; Muscular Dystrophies; Nucleic Acid Hybridization; Phenylketonurias; Phosphorus Radioisotopes; Polymorphism, Genetic; Pregnancy; Prenatal Diagnosis; Receptors, Antigen, T-Cell; RNA; T-Lymphocytes; Virus Diseases

This report describes a method for simultaneous quantitative determination of antigens from protein blots based on the highly sensitive "contact-copy" procedure, applying the phosphorylation reaction as a general detection principle. Using ultrasensitive bio-imaging analyzer systems permits us to quickly and easily quantify single bands directly from the picture screen and to achieve a high-resolution printing of the image (pictography). By applying a new kanamycin loading procedure it is possible to use phosphocellulose paper P81 as a substrate matrix. Replacing 32P with 33P as a detection isotope leads to an improvement of the sensitivity, resolution, and safety. The method is applied to analyze the proteins dystrophin, myosin, vinculin, and desmin from human tissue lysates. The high sensitivity of the procedure (detection limit approximately 1 pg dystrophin) permits determination of the quantity of dystrophin in very small tissue biopsy samples, which is of special interest for Duchenne/Becker muscular dystrophy diagnosis.

Topics: Adenosine Triphosphate; Adult; Animals; Blotting, Western; Calibration; Child; Diagnosis, Differential; Dystrophin; Humans; Image Processing, Computer-Assisted; Male; Microchemistry; Muscle Proteins; Muscular Dystrophies; Phosphorus Radioisotopes; Sensitivity and Specificity

The uptake of exogenous 32Pi into the membrane proteins of intact erythrocytes was measured in 8 patients with Duchenne muscular dystrophy. No abnormalities were noted after autoradiographic analysis. This contrasts with earlier results obtained when isolated membranes were phosphorylated with gamma-[32P]ATP, and suggests a possible reinterpretation of those experiments.

Topics: Adolescent; Adult; Autoradiography; Child; Erythrocytes; Humans; Muscular Dystrophies; Myotonic Dystrophy; Phosphorus Radioisotopes; Phosphorylation

Phosphorus 31 nuclear magnetic resonance (31P NMR) signals were recorded from intact human erythrocytes for 16 hours. Total phosphate concentration, which was estimated as the sum of the individual 31P signals, was 25% lower in erythrocytes from men with myotonic dystrophy than in control erythrocytes. The inorganic-phosphate fraction contained the highest average phosphate concentration over the 16-hour period, and made the major contribution to the difference in total phosphate between the two groups. This result was not observed in erythrocytes from either women with myotonic dystrophy or patients with Duchenne's dystrophy and may be due to a change in cell membrane permeability to inorganic phosphate, which leads to lower steady-state concentrations of the intracellular phosphates.

Topics: Adult; Cell Membrane Permeability; Child; Erythrocytes; Female; Humans; Magnetic Resonance Spectroscopy; Male; Muscular Dystrophies; Phosphates; Phosphorus Radioisotopes

Topics: Acetylcholine; Cells, Cultured; Concanavalin A; Epinephrine; Fibroblasts; Humans; Kinetics; Muscular Dystrophies; Phosphatidic Acids; Phosphatidylinositols; Phosphorus Radioisotopes; Skin

Topics: Animals; Axonal Transport; Carbon Radioisotopes; Cell Membrane Permeability; Cholesterol; Mice; Mice, Inbred Strains; Muscles; Muscular Dystrophies; Phosphates; Phospholipids; Phosphorus Radioisotopes; Sciatic Nerve; Spinal Cord

Topics: Adenosine Triphosphatases; Adenosine Triphosphate; Adenylyl Cyclases; Electrophoresis, Polyacrylamide Gel; Humans; Muscles; Muscular Dystrophies; Myotonia; Phosphates; Phosphorus Radioisotopes; Protein Kinases; Sarcolemma; Sarcoplasmic Reticulum; Sodium Dodecyl Sulfate