Publications by authors named "Novokhatskiĭ A"

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.

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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.

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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.

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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.

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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.

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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.

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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.

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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.

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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.

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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.

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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.

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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.

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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%.

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Monoclonal MAK-14-7 antibodies to the surface antigen of Venezuelan equine encephalomyelitis virus and OKA series to vaccinia virus antigens were studied by enzyme-immunoassay (EIA) on the solid phase of the infected Vero, BHK-21, and HeLa cells. The immunoglobulins under study from the culture and ascitic fluids of hybridomas were shown to bind specifically with the appropriate antigens of the infected cells. The activity of monoclonal antibodies to viral antigens in EIA on the solid phase of the infected cells was 10-fold higher than in indirect immunofluorescence test.

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The dynamics of accumulation of infectious Pichinde virus in the culture medium and virus-specific antigens in cells was studied in relation to multiplicity of infection in a multicycle experiment. Differences in the fluorescence pattern of Pichinde virus antigens in IFAT were found to depend on the use of acetone or formaldehyde for fixation of the infected cells.

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Six hybridomas of the EJ series producing monoclonal antibodies to Japanese encephalitis virus antigens were generated by hybridization of immune splenocytes with the parental line of mouse myeloma cells NS-0, and one hybridoma (EJ-10) with the X63-Ag8/653 line. Among 7 species of monoclonal antibodies examined by Ouchterlony method, 3 were identified as IgM and 4 as IgG. The highest clone-producing efficacy was shown by hybridoma EJ-10 generated on the basis of X-653 cells and the least by hybridoma EJ-20.

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The conditions of immunoenzyme assay have been studied on the solid state phase of infected cells using the model of monoclonal antibodies MAK-14-7 to the virus of Venezuelan equine encephalomyelitis (VVEE) and monoclonal antibodies OKA-1 to vaccine virus in the systems of VNK-21 cells or 4647 cells infected by VVEE, or HeLa cells infected by vaccine virus. The titer of monoclonal antibodies detected grows with the dose of infected cells fixed in the holes of micropanel used for reaction and with the multiplicity of infection. The most intensive and contrasting dyeing of conjugate has been registered when the cells have been fixed with 0.

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A panel of 6 hybridomas "XEJIMA" producing monoclonal antibodies specific to HeLa cells is prepared. Monoclonal antibodies do not bind to antigens of human diploid fibroblasts, human continuous B- and T-lymphocytes and animal cell lines. The specificity of monoclonal antibodies to cellular antigens of 5 HeLa-like cell lines and 6 human tumour cells lines, not contaminated with HeLa cells, is determined.

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