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In this work, new data on the peculiarities of actin immunofluorescent detection in 2-cell mouse embryos using antibodies against C- and N-terminal domains are presented. We studied the distribution of nuclear actin after artificial suppression of transcription and after enzymatic digestion of DNA. The visual observations were supplemented with morphometric analyses of confocal images. In both experimental groups (treated by transcription inhibitors or DNase) the reliable increasing of fluorescence intensity is revealed when antibody against C-, but not against N-terminal domain of actin was used. These finding allows to suppose that antibody against C-terminal domain of actin is suitable with a high efficiency for immunofluorescent studies on the nuclei of cleavage mouse embryo.

The authors present the results of comparison of clinical, instrumental and histological, immunohistochemical and electron microscopic data on two pigment-free ciliochoroidal tumors. Differential diagnosis was made at each of the above stages of diagnosis. There is evidence that despite the similarity of the clinical syndrome, the spindle-cell type of the histological structure of both tumors, the latter were of various histogenesis. This was confirmed by the results of immunohistochemical and electron microscopic studies. Retrospective analysis of the data of immunohistochemical and echographic studies revealed the distinctive signs that could differentiate these two types of tumors at the stage of clinical and instrumental diagnosis.

The investigation of antigenic diversion of tumor cells resulting from the expression of heteroorganic antigens has been continued. Tumor-associated heteroorganic antigens with mol. weight 200-210 kDa (identified before as laminin), 105-130, 75-80 and 43 kDa were detected by anti-kidney serum in fractions of plasmatic membranes of cells of rat ascitic Zajdela hepatoma and cultured HTC hepatoma; the antigen 43 kDa was isolated on immunosorbent and identified by mass spectrometry as beta-actin. Anti-kidney serum revealed laminin in fractions of plasmatic membranes of cultured L8 and L6J1 myoblasts, and L6J1 myotubes; apparently, synthesis of laminin by hepatoma and myogenic cells is not connected with their proliferative activity. Besides, anti-kidney serum detected components 38, 42, 44, 48, 62, 78 and 120 kDa, expression of which on myogenic cells surface might be consequence of active cell proliferation and (or) differentiation.

We studied the capacity of colloidal gold for enhancing specific and nonspecific immune response in laboratory animals (rabbits, rats, and mice) immunized with antigens of various nature. The antibody titers obtained with colloidal gold as a carrier were higher as compared to the standard immunization techniques (free antigen or Freund's adjuvant). Application of colloidal gold increased nonspecific immune responses as well: lysozyme concentration in the blood, activity of the complement system proteins, as well as phagocytic and bactericidal activities. The obtained antibodies were tested by immunodot assay using gold markers. Immunization of the animals with colloidal gold conjugates with haptens as well as complete antigens was shown to induce formation of highly active antibodies without using other antigens such as complete Freund's adjuvant. In addition, antigen quantities for animal immunization with colloidal gold was by one order of magnitude lower as compared to the complete Freund's adjuvant immunization. This fact can point to direct adjuvant activity of colloidal gold.

Some optimized methods of purifying actin from animal, plant, and bacterial cells were developed. Variants of the preparation of antiactin antibodies are described which use both traditional methods of immunization and phase display technology and antigen adsorption on colloidal gold particles. The conjugates of colloidal gold with phalloidin and heavy meromyosin as well as with antibodies were proposed. It was shown that these markers make it possible to reliably identify actins of different origin by the methods of light, and electron microscopy and dot-analysis.

One of the mechanisms of muscular involvement in RA patients was elucidated by identification of antibodies to skeletal muscle components (actin and tropomyosin). Clinical symptoms of skeletal muscles involvement in RA proved to be associated with elevated count of tropomyosin antibodies against an insignificant informative value of actin. Enzyme immunoassay with tropomyosin has established a direct correlation between the number of the antibodies and RA activity.

G-actin was isolated from human myocardium, purified by means of repeated polymerization-depolymerization and immobilized in polyacrylamide gel by the procedure of "emulsion granulation". The granules acquired magnetic properties after addition of iron protoxide hydrophilic particles into the polymerizating mixture. Immobilized G-actin in granules with magnetic properties was used for production of specific antisera, for isolation of pure antibodies by means of an immunosorption procedure, for estimation of specific antibodies in blood sera of patients with rheumatism, of healthy persons and of experimental animals using immunofluorescent, immunoenzymatic and radioimmune procedures.

The capacity of proteins to induce the synthesis of specific immunoglobulins was shown to be correlated with their evolution rate. This correlation can be understood in terms of the clonal-selectional theory. The characteristic correlation parameter is the number of differences in the amino acid sequences between the immunogenic protein and the homological protein of the immunized animal. The above correlation was traced most clearly for the evolutionary conservative proteins.