[Comprehensive rehabilitation of the subjects presenting with occupational muscular-skeletal abnormalities].

The methodological approach to the rehabilitative treatment of the subjects presenting with occupational muscular-skeletal abnormalities in the upper limb girdle associated with their occupational activities implies the combined application of a pulsed magnetic field, therapeutic peloids and ultrasound therapy to the neuromuscular apparatus and tendinous-capsular structures of the rotator cuff undergoing dystrophic degeneration. This therapeutic modality makes it possible to improve the biomechanical conditions of the patients by broadening the range of active painless movements in the affected shoulder joint by 42% (p < 0.05), normalizing tonal and load-bearing characteristics of the muscles (increase of the initially reduced muscular tone at rest by 27% (p < 0.05) at a maximum voluntary tension (16%, p > 0.05), changing trophicity of periarticular tissues (elevation of the pain sensitivity threshold of tendons and painful indurations in the functionally active muscles of the thoracic girdle of the upper extremity by 76% (p < 0.05). It is concluded that these changes contribute to the improvement of professional activities of the patients.

[Administration of ehf-therapy for rehabilitation of locomotor diseases].

Noise EHF radiation under low-frequency (10 Hz) modulation both in monovariant and in combination with spinal tractions promotes regression of neuro-orthopedic disorders. Good effects were achieved due to improvement of zonal hemodynamics, conditions of functioning of the neuromuscular and locomotor system of the spine and limbs, favourable shifts in biochemical and autonomic status of patients with lumbar osteochondrosis and osteoarthrosis.

Production and purification of melanoma gp100 antigen and polyclonal antibodies.

gp100 is a melanoma-associated antigen found to carry immunogenic epitopes that can induce a CTL response against tumor cells. Production and purification of large quantities of this polypeptide may be important in the context of diagnosis and vaccinating against melanoma. To overcome the hydrophobic nature of gp100, we cloned and expressed only a part of the protein, and obtained a hydrophilic recombinant polypeptide (HR-gp100) that contained most of the immunogenic peptides. High yield was achieved in an Escherichia coli expression system. The protein was purified by AKTA Prime using anionic-columns. Polyclonal antibodies developed in chicken against HR-gp100 were efficient at detecting gp100 in melanoma cells, as determined by Western blot analysis and by immunohistochemistry. HR-gp100 can be used to develop a vaccine against melanoma. Antibodies to HR-gp100 may be used to detect tumors of melanocytic origin or to determine the level of gp100 expression in tumors prior to immunotherapy with the protein or one of its peptides.

Purification and characterization of a novel tripeptidyl aminopeptidase from Streptomyces lividans 66.

An extracellular tripeptidyl aminopeptidase has been purified from Streptomyces lividans 66 cell-free cultures. The enzyme is a major component of the secreted proteolytic activity. The protease removes only the N-terminal tripeptide from recombinant human GM-CSF and IL-3 but does not cleave recombinant human IL-6. The enzyme cleaves the synthetic tripeptide substrates APA-pNA and APM-pNA but does not cleave substrates with blocked amino terminals. Smaller substrates are not cleaved. The enzyme appears to be a serine protease of 55 kDa molecular weight. The pH optimum is between 7.5 and 8.5 but varies slightly with the substrate. The N-terminal sequence and amino acid composition have been determined.

High-level expression of enzymatically active human Cu/Zn superoxide dismutase in Escherichia coli.

Expression of human Cu/Zn superoxide dismutase (SOD) with activity comparable to the human erythrocyte enzyme was achieved in Escherichia coli by using a vector containing a thermoinducible lambda PL promoter and a beta-lactamase-derived ribosomal binding site. The recombinant human SOD was found in the cytosol of disrupted bacteria and represented greater than 10% of the total bacterial protein. The enzyme was purified to homogeneity by salt precipitation, gel filtration chromatography, and ion exchange chromatography. The active enzyme was obtained in high yield only when 1 mol of copper and 1 mol of zinc were incorporated into each mol of subunit during bacterial growth or by reconstitution of the apoenzyme. Human Cu/Zn SOD produced in bacteria has an apparent subunit molecular mass of 19 kDa on NaDodSO4/polyacrylamide gels. The native enzyme behaves as a dimer of 32 kDa as determined by gel filtration. Sequence analysis of the NH2 terminus revealed that the first 14 amino acids corresponded to authentic human SOD except that the NH2-terminal alanine was not acetylated. Thus, the bacterial processing system readily removes the NH2-terminal methionine residue from recombinant human SOD.

Molecular aspects of experimental autoimmune myasthenia gravis.

Structure and function studies on acetylcholine receptor (AChR) were attained by studying various derivatives of the receptor molecule, by analysis of monoclonal antibodies, and by investigating the possible structural relationship between AChR and the thymus. Pharmacologically inactive denatured AChR preparation does not induce myasthenia in rabbits, although it elicits antibodies that cross-react with the intact receptor. Denatured AChR has both preventive and therapeutic effects on experimental autoimmune myasthenia gravis. Mild tryptic digestion of AChR does not abolish the pharmacological specificity and myasthenic activity of the receptor. Trypsinated AChR, which on SDS gel electrophoresis shows one major band with a molecular weight of 27,000, represents an active receptor derivative with a relatively low structural complexity. Monoclonal antibodies against defined determinants of AChR were elicited and characterized. One monoclonal antibody line is directed against the cholinergic binding site. Experiments demonstrating that thymic lymphocytes bear a surface antigen that cross-reacts with AChR are described.

Immune regulation of experimental myasthenia.

Experimental autoimmune myasthenia gravis (EAMG) is an appropriate model for studying the molecular origin, immunological mechanism and regulation of myasthenia gravis. Several approaches are being utilised for the regulation of the immune response to AChR and for immunosuppression of EAMG: Corticosteriods and azathioprine can suppress EAMG concomitantly with suppression of immune responses to AChR. High dose cyclophosphamide treatment in mice facilitates the onset of EAMG and results in a selective suppression of the humoral response to AChR whereas the cellular response is enhanced. Specific immunosuppression of EAMG is achieved by using a nonmyasthenic, denatured AChR preparation which cross reacts with the intact receptor. Various degradations and modifications of AChR are being performed in order to identify the smallest molecular entity responsible for the myasthenic activity of AChR. Studies on specific monoclonal antibodies, anti-idiotypes, and on the effect of measles virus on EAMG are being described and their possible significance in regulating myasthenia are being discussed.

Specific immunosuppression of experimental autoimmune myasthenia gravis by denatured acetylcholine receptor.

Specific immunosuppression of experimental autoimmune myasthenia gravis (EAMG) was achieved by the use of a denatured preparation of the acetylcholine receptor (AcChoR) that did not in itself induce the disease. Torpedo californica AcChoR was irreversibly denatured by complete reduction and carboxymethylation in 6 M guanidine hydrochloride. Rabbits immunized with reduced carboxymethylated receptor (RCM-AcChoR) produced antibodies that reacted with both RCM-AcChoR and intact AcChoR. The specificity of anti-RCM-AcChoR antibodies is different from that of anti-AcChoR antibodies because the former are directed to only part of the antigenic determinants present in the intact receptor. RCM-AcChoR, which by itself is completely nonmyasthenic, was shown to be capable of both preventing the onset of EAMG and of reversing the clinical symptoms in myasthenic rabbits. In all cases the therapeutic effect of RCM-AcChoR administration on EAMG was accompanied by a change in the immunological specificity of the antibodies. The crossreactivity between AcChoR and RCM-AcChoR and the nonpathogenicity of RCM-AcChoR appear to be crucial in governing the specific immunosuppressive effects of RCM-AcChoR on EAMG.