[Cell apoptosis in the culture: specific features of manifestation and influence on the effectiveness of biotechnological production].

The development of large-scale biotechnological production with the use continuous cell-line cultures as producers revealed the role of apoptosis in the culture as a factor producing essential influence on the effectiveness of the process, the level of the accumulation of the target product and its quality. The limiting influence of apoptosis as the programmed death of cells in the culture was determined by a number of factors: the chemical composition, pH and chemical properties of the medium; the levels of glucose, amino-acid and vitamin depletion; the uniformity of oxygenation; temperature conditions; the mode of agitation and the intensity of hydrodynamic forces; the content of mitogenetic factors and toxins in the medium; the presence and activity of the expression of apoptosis stimulators and inhibitors. The presence of correlation between the intensity of apoptosis, cultural properties and the characteristics of the mitotic cycle of the cell line was shown. The latter made it possible to regard the biological properties of the producer culture as one of the leading factors modifying the process of the programmed death of cells.

[Monoclonal antibodies to insulin influencing its biological activity]. / Monoklonal'nye antitela k insulinu, vliiaiushchie na ego biologicheskie aktivnost'.

Somatic hybridization was employed for obtaining 335 hybridomas producing monoclonal antibodies to insulin. Twelve hybridomas were cloned by a method of maximum dilutions, and specific immunoglobulins, secreted by them, were characterized by solid phase immunoenzyme assay. Monoclonal antibodies possessed different activity with relation to human, porcine and cattle insulins, indicating their specificity to different epitopes on the insulin molecule. Antibodies of IN-2 and IN-3 clones were capable of inhibiting insulin biological activity in vivo. In experiments on rabbits antibodies of IN-2 clone decreased a 3-fold insulin hypoglycemic action, and antibodies of IN-3 clone caused an increase in the life span of mice after administration of insulin at high doses to them.

[A receptor for the Fc fragment of human IgG in the causative agent of tularemia]. / Retseptor k Fc-uchastku IgG cheloveka u vozbuditelia tuliaremii.

In 4 Francisella tularensis strains varying in virulence a receptor to Fc site of human IgG has been detected. This receptor consists of two active components with molecular weights of 67,000 and 40,000, competing for binding on Fc site of human IgG with Staphylococcus aureus protein A.

[The dynamics of antibody formation to a vaccinal strain of Francisella tularensis in different immunization regimens]. / Dinamika obrazovaniia antitel k vaktsinnomu shtammu Francisella tularensis pri razlichnykh skhemakh immunizatsii.

The use of different schemes of albino mice immunization either by living or by killed preparations of the vaccine strain of Francisella tularensis when obtaining monoclonal antibodies to the tularemia microbe made it possible to reveal definite regularities in the dynamics of antibody formation. The highest titres of antibodies in sera of animals-donors of splenocytes were obtained during the daily (for 3 days) intraperitoneal immunization of mice with living vaccine or with its thrice administration to the spleen thrice with the interval of 10 days. Revaccination against a background of high titres of antibodies decreased their quantity in blood serum of mice, while that against a background of low titres increased them.

[Additions to the theory of the dual nature of photoreception]. / Dopolneniia k teorii i dvoistvennoi prirode fotoretseptsii.

On the grounds of clinico-physiologic investigations the author revises the existing dual theory of photoreception. The paper describes complex clinical investigations of visual functions in patients with alimentary and essential hemeralopias and shows that rods and cones do not function in conditions of scotopic illumination without the presence of rhodopsin. On the grounds of investigations of the phenomenon of inversion and depression in the short-wave part of the visible spectrum in normal persons after dark adaptation in conditions of scotopic illumination, the author comes to a conclusion that there is a so-called "rhodopsin filter" just the presence of which in the process of dark adaptation creates conditions for appearance of this phenomenon. The author believes that there exist two kinds of photoreception: a direct disc two-component and a mediated rhodopsin one-component, as well as the third, transitory, that corresponds to those conditions of illumination in which the Purkinje's phenomenon appears.

[Monoclonal antibodies to antigens of cholera vibrios of the Ogava serovar]. / Monoklonal'nye antitela k antigenam kholernykh vibrionov serovara Ogava.

A set of hybrids is obtained synthesizing monoclonal antibodies to the surface antigenic determinants of the choleric vibrio of the Ogava serovar. The antigenic structure of vibrios of the typical and atypical strains isolated from a man and from environmental objects is studied using a collection of highly specific immunoglobulins. The high-specific diagnostic preparations for identification of the 01-group cholera agent serovar may be created on their base.

[Characteristics of the monoclonal antibodies induced by the Crimean hemorrhagic fever virus]. / Kharakteristika monoklonal'nykh antitel, indutsirovannykh virusom krymskoi gemorragicheskoi likhoradki.

Twelve lines of hybridomas secreting monoclonal antibodies (MCA) have been generated by fusion of spleen lymphocytes of BALB/c mice immunized with crude Crimean hemorrhagic fever virus (CHF). The hybridomas multiply actively in vitro (over 50 passages) and as ascitic tumors in the abdominal cavity of BALB/c mice. MCA were characterized by indirect immunofluorescence (IF), complement fixation (CF), biological neutralization test (NT), agar gel diffuse precipitation tests, and by the type of immunoglobulins. In the indirect IF, all kinds of MCA reacted with CHF virus, in CF only GEMA 12 and GEMA 31 which also precipitated the virus antigen in AGDPT. The CF-positive MCA belonged to the IgG2a and IgG2b subclasses, but 2 more MCA species of the same immunoglobulin type did not react in CF. No neutralizing properties have been found with MCA. All MCA reacted similarly in indirect IF and CF both with CHF and Congo viruses. Among other members of the CHF-Congo antigenic group, only GEMA 4 MCA interacted with Hazara virus. MCA generated in this study are intended for use in identification of CHF and Congo viruses in diagnostic tests.

[Central physiologic relative scotoma after dark adaptation]. / Fiziologicheskaia tsentral'naia otnositerl'naia skotoma posle temnovo adaptatsii.

The paper describes data on conditions for appearance of a physiologic central retinal scotoma of dark adaptation. Examinations of 80 eyes with normal visual functions have shown that central scotoma of dark adaptation is revealed in conditions of scotopic illumination 20-25 minutes after dark adaptation. The optimal duration of dark adaptation is 40-45 minutes. More frequently scotoma is displaced upwards from the point of fixation, its sizes varying from 4 degrees to 10 degrees by the horizontal meridian and from 3 degrees to 8 degrees by the vertical meridian. The presence of this scotoma should be considered when examining the state of the visual analyser in conditions of scotopic illumination. This can play a certain role in differential diagnostics of clinical ophthalmopathology.

[Biological activity of human alpha-interferons studied using specific antisera]. / Issledovanie biologicheskoi aktivnosti chelovecheskikh al'fa-interferonov pri pomoshchi spetsificheskikh antisyvorotok.

Preparation of highly active rabbit antisera (AS) to human recombinant alpha 2-interferon and their use for studying biological properties of natural and plasmid alpha-interferons are described. By exhaustion of AS by alpha 3-interferon there were prepared practically monospecific AS not reacting with antigenic determinants of alpha 3-interferon. It was found that alpha 3-interferon represented a significant portion of human lymphoblastoid interferons and was included in PH-labile alpha-interferon from serum of patients with Kaposi carcinoma. AS to alpha 2-interferon completely neutralized antiviral and antiproliferative activity of the homologous subtype alpha-interferon and stimulation of cytotoxicity of human natural killer cells induced by it. It neutralized also the same effects of the heterologous subtypes (alpha 3 and alpha F/D) and leukocytic interferon, but the neutralization level was lower. The results of the study confirmed the polyfunctional nature of the interferon molecule.

[Monoclonal antibodies to the yellow fever virus]. / Monoklonal'nye antitela k virusu zheltoi likhoradki.

A highly technological Yel-2 hybrid cell clone was isolated. The clone produces monoclonal antibodies to protein E of the yellow fever virus (Dakar strain). The Yel-2 hybridoma was cloned by the method of limiting dilutions. By the data of indirect immunofluorescence the titre of the specific antibodies reached 1:128 in the culture fluid and 1:10240 in the ascitic fluid. The Yel-2 antibodies had no cross reactions with forest-spring encephalitis, Dengue 2 and Dengue 4 viruses. In the hemagglutination inhibition test the activity of the Yel-2 ascitic preparations was observed at a dilution of 1:50000 while the complement fixation test did not reveal it. Administration of the Yel-2 cells into the abdominal cavity of mice BALB/c induced development of ascitic tumors in 97 per cent of pristane-sensitized mice and in 90 per cent of nonsensitized mice. The hybridoma was successfully transplanted from mouse to mouse (more than 20 passages during the observation period) and maintained high constant levels of specific antibody secretion. The high ascite forming capacity of the Yel-2 hybridoma provided decreasing of the cell dose administered to the mice from 10(7) to 10(4) cells per mouse.

[Quantitative characteristics of degeneration of the fibers of the optic nerve after section]. / Kolichestvennaia kharakteristika degeneratsii volokon zritel'nogo nerva posle ego peresecheniia.

The optic nerve of the dog has been histologically studied. In the intact nerve 160,000 axons have been revealed. In 3 weeks after the nerve section in the orbit and near the nerve disk 72% retinoencephal fibers, 28% encephaloretinal and 0.23% of centrofugal axons are found.

[Increase in the specificity of immunoenzyme analysis by using a monoclonal antibody-based conjugate]. / Povyshenie spetsifichnosti immunofermentnogo analiza za schet ispol'zovaniia kon"iugata na osnove monoklonal'nykh antitel.

Examinations by EIA of laboratory cultures of various orthopoxviruses and specimens from human-monkey pox cases demonstrated the advantages of a monoclonal preparation over the peroxidase polyclonal conjugate prepared from hyperimmune vaccinia antiserum.

[Conditions for forming ascitic tumors in hybridoma cultivation in vivo]. / Izuchenie uslovii obrazovaniia astsitnykh opukholei pri kul'tivirovanii gibridom in vivo.

The conditions of the formation of ascitic cells in BALB/c mice injected with hybridoma cells were studied. All the hybridomas under study, producing monoclonal antibodies to viral antigens, induced the formation of ascitic tumors when introduced into the abdominal cavity of BALB/c mice pretreated with sensitizing agents. In the mice pretreated with pristane hybridoma cells took at a rate of 43-80% and in the mice pretreated with Freund's complete adjuvant, 31-70%. Angara oil and perfume oil, as well as Bayol F, were less effective. The time of the formation of ascites was inversely proportional to the dose of the injected cells, while the volume of ascitic fluid depended rather on the type of hybridoma and not on the dose of the injected cells. The study showed that the use of physiological saline or culture medium without serum for washing the abdominal cavity of mice after withdrawing ascites permitted the additional collection of 2.6-13.7 million hybrid cells, as well as a considerable amount of immunoglobulins.