ABSTRACT
We proposed a new indicator for evaluation of functional activity of Ca(2+) in human blood serum based on lysis of sheep erythrocytes with 10% human blood serum in the presence of 0.55 mM ethylene glycol tetraacetic acid at 37°C for 10 min. After incubation, the degree of sheep erythrocytes lysis inhibition is estimated: inhibition of complement hemolytic activity <30% is considered as high functional Са(2+) activity, inhibition by 31-70% corresponds to normal activity, and >71% indicates low activity. The comparative studies of complement activating function of heterophilic antibodies, complement system reactivity in the presence of 0.29 M NaCl, and functional activity of ionized Ca(2+) make possible estimation of the individual's immune status.
Subject(s)
Calcium/pharmacology , Animals , Calcium/chemistry , Complement Activation/drug effects , Edetic Acid/chemistry , Egtazic Acid/chemistry , Erythrocytes/drug effects , Hemolysis/drug effects , Humans , SheepABSTRACT
It is shown that human serum albumin, previously treated with HOCl (HSA-Cl), enhances luminol-dependent chemiluminescence of neutrophils activated by phorbol-12-myristate-13-acetate (PMA). The enzyme-linked immunosorbent assay revealed that addition of HSA-Cl to neutrophils promotes exocytosis of myeloperoxidase. Inhibitor of myeloperoxidase--4-aminobenzoic acid hydrazide, without any effect on lucigenin-dependent chemiluminescence of neutrophils stimulated with PMA, effectively suppressed luminol-dependent chemiluminescence (IC50 = 20 microM) under the same conditions. The transfer of the cells from medium with HSA-Cl and myeloperoxidase to fresh medium abolished an increase in PMA-induced luminol-dependent chemiluminescence, but not the ability of neutrophils to respond to re-addition of HSA-Cl. A direct and significant (r = 0.75, p) correlation was observed between the intensity of PMA stimulated neutrophil chemiluminescence response and myeloperoxidase activity in the cell-free media after chemiluminescence measurements. These results suggest the involvement of myeloperoxidase in the increase of neutrophil PMA-stimulated chemiluminescence response in the presence of HSA-Cl. A significant positive correlation was found between myeloperoxidase activity in blood plasma of children with severe burns and the enhancing effects of albumin fraction of the same plasma on luminol-dependent chemiluminescence of PMA-stimulated donor neutrophils. These results support a hypothesis that proteins modified in reactions involving myeloperoxidase under oxidative/halogenative stress, stimulate neutrophils, leading to exocytosis of myeloperoxidase, a key element of halogenative stress, and to closing a "vicious circle" of neutrophil activation at the inflammatory site.
Subject(s)
Burns/enzymology , Luminol/chemistry , Neutrophils/drug effects , Peroxidase/metabolism , Serum Albumin/pharmacology , Aniline Compounds/pharmacology , Burns/pathology , Child , Enzyme Inhibitors/pharmacology , Enzyme-Linked Immunosorbent Assay , Humans , Hypochlorous Acid/chemistry , Luminescence , Luminescent Measurements , Neutrophil Activation/drug effects , Neutrophils/enzymology , Neutrophils/metabolism , Neutrophils/pathology , Oxidative Stress , Peroxidase/antagonists & inhibitors , Phorbol Esters/pharmacology , Serum Albumin/chemistryABSTRACT
Soft tissue diseases are pathologic conditions underlain by degenerative, dystrophic, and inflammatory processes developing in muscles, ligaments, synovial bursae and sheaths, fascias, and aponeuroses. Their prevention and medical rehabilitation are achieved with the help of various methods targeted toward different pathogenetic mechanisms and factors facilitating recovery of the affected structures. These methods include instrumental physiotherapy, balneotherapy, peloidotherapy, therapeutic exercises, massage, and pharmacotherapy as appropriate. The rehabilitative strategy implies systematic step-by-step application of these tools in various combinations. Balneotherapy needs to be scheduled to the most favourable climatic seasons. Massage and remedial gymnastics are indispensable procedures in all rehabilitative strategies for the patients with soft tissue diseases. Mechanotherapy, traction, elements of manual therapy, and reflexotherapy should be provided if indicated. They are of special importance whenever the motor function of muscles and circumarticular soft tissues needs to be improved. Regular remedial gymnastics and swimming in a pool are highly efficacious methods of secondary prophylaxys preventing exacerbation of soft tissue diseases. It should be emphasized that early and systematic application of rehabilitative treatment by the above methods helps to restore functional capacity of the affected soft tissues and substantially improve the patients' quality of life.
