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Changes in the global economic model for the use of Natural Resources will increase mainly due to reductions on a point scale of about 90 billion USD native resources. It is expected to reach 190 billion USD by 2060. This will lead to a significant increase in carbon dioxide emissions and a reduction in raw materials needed for high technologies. The only way to counter these challenges is to move to a circular economy. Circular economics calls into question the linear system and requires a rethink of the production process, considering the process of recycling goods. It offers significant potential for innovation, employment opportunities, quality of work, and ultimately a more inclusive economy that serves the needs of all people. The role of the educational system in the process of forming a circular economy. The importance of education is emphasized in the transition to a circular economy. The education system is a key player in the transition from a linear to a circular economy, which should begin to guide this transformation. To do this, the education system must first adapt to new requirements by adopting circular economics teaching in classrooms. Secondly, to ensure an effective large-scale transition by developing students’ proper skills and systems thinking. However, circular economics cannot be tied to a single profession or discipline because its success requires dialogue between different actors. This is an inter-social economic model that is needed in the fields of engineering, design, military operations, and business. Therefore, the teaching of circular economics should also be interdisciplinary and cover different areas of Education.

The authors draw attention to the issue of gender differences in the features of the subjective experience of happiness and unhappiness. The article describes the results of a study conducted with clients of psychologists who report problems related to the experience of destructive interpersonal relationships and consider themselves deeply unhappy in life. The purpose of the study is to reveal the gender specificity of the frustration of the subjects’ life needs underlying the deep experience of unhappiness in their lives. A peculiarity of the approach used by the authors is the consideration of issues of psychological nature in the context of philosophical and religious understanding of eternal values, such as happiness and love for one’s neighbor. A special focus of the authors’ attention is the problems related to the philosophical and religious basis of consideration of the feminine as Sophia the Wisdom of God with the complex of her rejection, the dominant sense of guilt, and the desire to return to the Father. Such methodological symbiosis is facilitated by interdisciplinarity as one of the main principles of the authors’ interaction with the materials of this work. The Thematic Apperception Test (TAT) aimed at the amplification of subjects’ unconscious intentions is used as a diagnostic instrument. The conducted psychological study allows to reveal and describe some of the causes for the manifestation and experience of happiness and unhappiness in the lives of modern men and women and show specific differences between their subjective experiences of happiness and unhappiness in their lives. Unlike previous studies of this kind, the deployed philosophical and religious studies approach provides an opportunity to take a new look at the outlined issues considering them from the point of the philosophical context of values.

The state financial system covers area of public relations which is connected with process of formation, distributions and uses of the centralized and decentralized monetary funds state and municipal unions. The mechanism of accumulation of financial resources along with the organization of the given process can't do without effective and effective institute of financial control. In the light of changes occurring in our country, both administrative, and legal, it is possible to assert with certain confidence that preconditions of successful realization of all problems facing the state, root, first of all, in effective system of financial control. In the given work all aspects of legal maintenance of the state financial control in modern Russia are collected together and analysed. Necessity of acceptance of new decisions concerning legal registration of financial control in Russia is revealed; also existing laws and standard documents are investigated, corresponding conclusions are drawn. Necessary, timely and expedient ways of the decision of an investigated question are offered. The project of possible results owing to acceptance of the offered decisions is prepared.

Cell-free supernatant of Lactobacillus plantarum exhibit a strong antimicrobial effect against a number of pathogenic enterobacteria (E. coli, Shigella flexneri, Salmonella typhimurium, Proteus mirabilis, and Campylobacter jejuni). The degree of growth inhibition in broth culture reached a high level for all tested bacteria. The highest rates were noted for P. mirabilis (by 13 times) and the lowest for S. flexneri (by 5 times) and C. jejuni (by 4.5 times). Significant antiproliferative effect of the supernatant on cells of tumor-derived epithelial cell lines was shown. The highest degree of inhibition (by 22 times) was observed for HT-29 cells (colon carcinoma). Thus, inclusion of probiotics in traditional treatment schemes can increase the effectiveness of antibacterial and antitumor drug therapy., (© 2023. Springer Science+Business Media, LLC, part of Springer Nature.)

In 82 clinical strains of Streptococcus pyogenes (group A streptococci) isolated from patients with various manifestations of streptococcal infection, emm-typing revealed 27 emm-types (n=77) with a predominance of emm-89 (n=15; 18%), emm-75 (n=9; 11%), and emm-1 (n=6; 7%); types emm-3, emm-12, and emm-58 (n=4; 5% each) were found with almost equal frequency; other types were less common. The superantigen genes speC, speG, speH, speI, speJ, speK, speL, speM, smeZ, and SSA were identified in S. pyogenes strains using multiprimer PCR; the genes of the superantigen SpeA and cysteine proteinase SpeB were detected using real-time PCR. All the studied S. pyogenes strains contained superantigen genes, and 98% of the strains had several (from 2 to 7) genes. The number of variants of these sets reached 37; 2% of the strains contained only one superantigen gene. The distribution frequencies of superantigen genes in the studied strains were: speA - 43%; speC - 38%; speG - 93%; speH - 13%; speI - 6%; speJ - 24%; speK - 13%; speL and speM - 11% each; smeZ - 98%; SSA - 15%. All studied S. pyogenes strains contained the speB gene. Our studies have demonstrated that the sets of superantigen genes of group A streptococci are characterized by pronounced diversity to some extent associated with emm-type., (© 2023. Springer Science+Business Media, LLC, part of Springer Nature.)

Endopeptidases IdeS and IdeZ (streptococcal virulence factors that specifically cleave IgG heavy chains) are of particular interest because of their potential use in biotechnology, medicine, and veterinary. Genes encoding these enzymes were cloned and expressed in Escherichia coli heterologous expression system (ideS was cloned from a Streptococcus pyogenes collection strain; ideZ from Streptococcus zooepidemicus was synthesized). The 6His-tag was introduced into the amino acid sequence of each endopeptidase, and IdeS and IdeZ were purified by metal affinity chromatography to an apparent homogeneity (according to polyacrylamide gel electrophoresis). Purified enzymes were active against human and animal IgGs; their specificity toward human IgGs was confirmed by polyacrylamide gel electrophoresis. Recombinant IdeZ was used for immunological analysis of equine strangles infection (diagnostics and determination of the titer of specific antibodies in blood). Hence, IdeZ can be used in veterinary and sanitary microbiology to diagnose infections caused by Streptococcus equi and S. zooepidemicus in addition to its application in medicine and biotechnology.

We studied the effect of the L. plantarum strain supernatant on the growth of culture and biofilm of non-fermenting bacteria of the genera Pseudomonas, Achromobacter, and Burkholderia. To obtain a supernatant, the culture of L. plantarum was grown for 48 h at 37°C on a Lactic broth nutrient medium with casein peptone, then centrifuged and filtered through a 0.22-μm Millipore filter. Antimicrobial activity was determined by broth microdilution assay. The inhibitory effect of the supernatant on the growth of bacteria of all three genera was demonstrated. The maximum inhibition was observed for P. aeruginosa (by 13 times compared to the control). For bacteria of the Achromobacter and Burkholderia genera, the inhibition was less pronounced: by 7 and 6 times, respectively. The supernatant also inhibited biofilm formation by P. aeruginosa and A. ruhlandii, but did not affect formed biofilm. Thus, the L. plantarum supernatant obtained by us exhibited pronounced antimicrobial activity against non-fermenting bacteria, the causative agents of nosocomial infections, especially in immunocompromised individuals, very often in cystic fibrosis patients., (© 2022. Springer Science+Business Media, LLC, part of Springer Nature.)

Analysis of the effect of copper and zinc ions on Streptococcus pyogenes and Escherichia coli biofilms revealed significant differences in the effect of these metals in the form of sulfates or chlorides on biofilm formation. Zinc ions in low doses (salt concentration 0.005 M) inhibited the growth of S. pyogenes biofilms by 1.5 times. After increasing salt concentration to 0.05-0.5 M, the growth of biofilm was reduced by 2.5 times in comparison with the positive control. In case of E. coli biofilms, the inhibition was more pronounced: zinc sulfate in a concentration of 0.005 M reduced its growth by 4.6 times in comparison with the positive control. After increasing salt concentration, the growth of E. coli biofilm decreased by 6.8 times. In case of zinc chloride, zinc ions produced weaker effect and reduced biofilm growth by 2.2 and 5 times, respectively. Copper salts in a concentration of 0.005 M had practically no effect on the growth of S. pyogenes biofilm; with increasing salt concentration, the degree of inhibition was close to the effect of zinc. In case of E. coli biofilm, we observed a slight inhibition of the growth by low doses of copper ions (by 1.4-1.3 times); with increasing salt concentration the effect increased by 5.6 and 2.2 times for copper sulfate and chloride, respectively. Copper and zinc cations had no effect on mature biofilm.

Light and electron microscopy enables researchers to study the ultrastructure of GAS biofilms formed on abiotic surfaces. Chains of streptococci surrounded by a bluish film are seen under a light microscope after alcian blue staining of preparations grown on coverslips. The extracellular matrix (indicator of biofilm maturity) becomes visible on ultrathin sections in transmission electron microscopy after additional staining with alcian blue; filamentous structures, characteristic of biofilm, are observed in intercellular spaces. The data obtained by scanning electron microscopy also demonstrate the presence of biofilm.

Our study confirmed the capacity of S. pyogenes strains to form biofilms on abiotic surfaces. Chains of streptococci surrounded by bluish film were seen under a microscope after alcian blue staining of the preparations grown on slides. On ultrathin sections in transmission electron microscope, the extracellular matrix (indicator of biofilm maturity) became visible after staining with alcian blue. Microscopy of the sections shows structures characteristic of a biofilm in spaces between the cells. Scanning electron microscopy also demonstrates the presence of a biomembrane. Importantly that type 1M strain forming in fact no membranes when cultured on plastic plates (Costar) formed biofilms on the glass. It seems that the conditions for the biofilm formation on the plastic and on the glass differ, due to which the exopolymeric matrices formed on different surfaces vary by biochemical composition.

Effects of Miramistin and Phosprenil on biofilms of S. pyogenes, S. aureus, E. coli, L. acidophilus, and L. plantarum were studied. Significant differences in the effects of these substances on mature biofilms of microorganisms and the process of their formation were observed. Miramistin had significant inhibiting effects on the forming of biofilms and on the formed biofilms of all studied microorganisms. Treatment with Miramistin inhibited biofilm formation by 2-3 times compared to the control. This effect was found already after using of Miramistin in the low doses (3.12 μg/ml). Inhibition of the growth of a formed biofilm was observed only after treatment with Miramistin in the high doses (25-50 μg/ml). Phosprenil in the high doses (15-30 mg/ml) inhibited the forming of biofilms, especially the biofilms of S. pyogenes and L. plantarum (by 3-4.5 times). Treatment of formed biofilms with the agent in doses of 6.0 and 0.6 mg/ml was associated with pronounced stimulation of its growth in S. pyogenes, S. aureus, and L. acidophilus.

Dynamics of IL-6 level was studied in blood serum of CBA mice receiving intraperitoneal injections of killed and live culture of group A Streptococcus and its supernatants. It was found that administration of killed culture was followed by a significant rise in IL-6 level (by 4.7 times in 1 h and by 9.3 times in 5 h in comparison with the control). By 24 h, cytokine content was below the control. The highest levels of IL-6 were found after treatment with supernatants of Streptococcus cultures (by 10.5 times in 1 h and by 14.9 times in 5 h, in comparison with the control). Administration of live culture was accompanied by an increase in IL-6 concentration by 3.2 times in 3 h. In this experimental series, the maximum level of IL-6 was found in 48 h (by 5.2 times), and then it gradually decreased below the control. Different dynamics of changes in IL-6 level after administration of killed and live cultures of group A Streptococcus may suggests that they activate different signal pathways.

Administration of bacterial antigens to CBA mice induced an increase in serum concentration of virtually all cytokines with a peak in 4 h after administration of S. typhimurium antigens and in 7 h after administration of streptococcus antigens. In 20 h, cytokine concentrations returned to the control level or were slightly below it. In 4 h after administration of S. typhimurium antigens preceded 3 h before by administration of streptococcus antigens, we observed a significant decrease in serum concentrations of IFN-γ, IL-10, GM-CSF, IL-12, and TNF-α, in comparison with injection S. typhimurium antigens alone and IL-5, IL-10, GM-CSF, and TNF-α in comparison with injection of streptococcus antigens alone; the concentrations of IL-2 and IFN-γ, in contrast, increased by 1.5 times in this case. In 20 h after administration of S. typhimurium antigens, the number of multipotential stromal cells (MSC) in the bone marrow and their cloning efficiency (ECF-MSC) increased by 4.8 and 4.4 times, respectively, in comparison with the control, while after administration of streptococcus antigens by 2.6 and 2.4 times, respectively. In 20 h after administration of S. typhimurium antigens preceded 3 h before by administration of streptococcus antigens, these parameters increased by 3.2 and 2.9 times, respectively, in comparison with the control, i.e. the observed increase in the level of MSC count and ECF-MSC is more consistent with the response of the stromal tissue to streptococcus antigens. Thus, successive administration of two bacterial antigens corrected both serum cytokine profiles and MSC response to administration of each antigen separately, which indicates changeability of the stromal tissue in response to changes in the immune response.

Comparative analysis of serum cytokine profiles of CBA mice was carried out 1, 5, 24, and 48 h after intraperitoneal injection of killed culture of different streptococcus A types. The production of cytokines in response to different streptococcus types varied. The highest level was recorded in response to types 1M and 3T+M, more often detected in invasive streptococcal infection. The highest levels of IL-2 were recorded in response to 1M (47-fold increase in comparison with the control) and 3T+M streptococcus types (more than 10-fold increase). Injections of these types also led to an increase of IFN-γ level (15.6 and 11.3 times, respectively). The level of TNF-α increased less (3.6 times in response to 3T+M and 2.6 times in response to 1M type). The levels of IL-5, IL-10, and IL-12 increased 2-3-fold. Injections of 1T and 5M types led to just a 2-fold increase of cytokine levels. These data indicated induction of the immune response trend by mainly Th1 or mixed Th1/Th2 pattern in response to group A streptococcus antigens.

Aim: Evaluation of the ability to form biofilms by various M, T and MT-types of group A streptococci (GAS), as well as study of the effect of various antibiotics on biofilm formation., Materials and Method: 43 strains of various M and T type GAS were studied. The cultures were grown in Todd-Hewitt broth with the addition of 0.5% yeast extract. Comparative evaluation of the ability to form biofilm was carried out using photometry. Benzylpenicillin, oxacillin, cepha- losporin, cefuroxime and ceftriaxone antibiotics were used at various concentrations., Results: GAS differ significantly by their ability to form biofilms. The highest ability was noted in 8 strains--2M, 9M, 12M, 13M, 19M, 30M, 36M-types and 6MT type. Simultaneous introduction of GAS cultures and antibiotics into the culture well, except for ceftriaxone, resulted in growth inhibition of both plankton cells and biofilms., Conclusion: The ability of GAS to form biofilm depends on streptococci serotype. During simultaneous introduction of GAS with antibiotics into the well, the biofilm does not form.

The content of multipotent stromal cells (MSC) in the bone marrow and efficiency of their cloning (ECF-MSC) increased by 3 times 1 day after administration of complex S. typhimurium antigens to CBA mice, while the relative content of alkaline phosphatase-positive MSC colonies (marker of osteogenesis; P(+) colonies) decreased from 14% (control) to 3%. After administration of the complex S. typhimurium antigens to CBA mice 3 h after (or 3 h before) curettage or treatment with morphogenetic protein (BMP-2), the content of MSC and ECF-MSC decreased on the next day by ~3 times in comparison with animals receiving antigens alone and approached the control level. The relative content of P(+) colonies increased to 20 and 35%, respectively, in comparison with animals receiving antigens (3%), but was significantly lower than after curettage (34%) or BMP-2 (42%) administration. Expression of IL-1β, IL-6, IL-12, TNF-α, and IFN-γ genes in the primary cultures of stromal bone marrow cells induced by antigen administration was suppressed, while the concentrations of IL-12 and TNF-α in the culture medium sharply decreased after antigen treatment in combination with curettage or BMP-2 administration. Administration of complex S. typhimurium antigens after pretreatment with BMP-2 (3 h before) was associated with a decrease in serum levels of IL-2, IFN-γ, IL-12, and TNF-α in mice receiving BMP-2+S. typhimurium group 4 h after treatment in comparison with the animals receiving only S. typhimurium antigens alone by 1.9, 4.4, 1.5, and 6 times, respectively, i.e. to normal level or below it, while the concentration of IL-10 increased by almost 2 times, which probably reflected anti-inflammatory properties of BMP-2. These data probably attest to competitive relations between osteogenesis and immune response at the level of MSC.

We studied the effect of BMP-2 added to the culture medium on osteogenic and proliferative properties of multipotent stromal cells (MSC) and on the expression of cytokine genes induced by immunization of experimental animals with bacterial antigens. It is shown that the presence of BMP-2 in the culture medium stimulates proliferation of bone marrow MSC and especially spleen MSC (which was seen from enlargement of MSC colonies); improves the efficiency of MSC cloning; increases osteogenic activity of mouse bone marrow MSC; induces osteogenic differentiation of splenic MSC (osteogenesis is normally not observed in the spleen); reduces the number of macrophages in cultures; inhibits synthesis of mRNA for proinflammatory cytokines (IL-1β, IL-6, IL-8, TNF-α) that typically occurs in cultures of the bone marrow and spleen from animals immunized with S. typhimurium or group A streptococcus antigens. Bearing in mind that proinflammatory cytokines negatively affect osteogenic activity of the bone marrow, we can hypothesize that BMP-2 not only stimulates osteogenesis, but also provides optimal conditions for its realization by suppressing the expression of genes encoding these cytokines.

Injection of polyvinylpyrrolidone (synthetic type 2 T-independent antigen) stimulated the efficiency of clone-forming efficiency and the content of stromal precursor cells in CBA mice in the femoral bone marrow (almost 3-fold) and in the spleen (by 1.7 times) with the peak within 24 h and normalization by day 3 after immunization. The expression of IL-6, IL-8, and TNF-α genes in bone marrow and spleen cultures from immunized animals appeared on day 1 and disappeared on day 3. Hence, stimulation of stromal tissue in response to polyvinylpyrrolidone immunization was significantly less pronounced in comparison with immunization with S. typhimurium antigens. The counts of stromal precursor cells in these organs did not increase in CBA/N mice not responding to polyvinylpyrrolidone because they had no xid-mutation in Brutton's tyrosine kinase (Btk) gene, and the proinflammatory cytokine genes expression in primary cultures derived from these animals did not increase either. These data indicated that the degree of stromal tissue stimulation in immunized mice correlated with the immune response intensity. This indicated a close relationship between the stromal tissue and immune system. Stromal tissue seemed to be stimulated not only and not so much through the stromal cell Toll-like receptors, but mainly through interactions of immunocompetent and stromal cells, the former presumably playing the leading role in this process.

Immunization of CBA mice with killed group A streptococcus (type 5) vaccine changed the counts of stromal precursor cells (CFC-F) in bone marrow transplants at different donor-recipient combinations (normal, N, or immune, I). CFC-F counts in bone marrow transplants from normal mice transplanted to immunized animals decreased 4-6-fold depending on the transplant age in comparison with similar transplants in normal recipients. The percentage of CFC-F colonies with alkaline phosphatase (osteogenesis marker) activity decreased more than 2-fold. Similarly, the count of CFC-F in the transplants was 2-fold lower during delayed (7 months) period after bone marrow transplantation from immunized donors (8-12 days after the end of immunization) to intact recipients, while 2 months after transplantation it was 3-fold lower. The mean optical density of the bone capsule in preparations stained for glycogen and alkaline phosphatase was 1.5-3 times lower in the N-->I and I-->N experiments in comparison with the control (N-->N). On the other hand, CFC-F count in the femoral bone marrow of immunized animals was significantly (3.5-2.5 times) higher during the period from 8 days to 8 months after the end of immunization compared to CFC-F count in the femoral bone marrow of intact mice. These results attest to a significant prolonged effect of streptococcal antigens on the bone marrow stromal tissue. These data also indicate that not all CFC-F, the counts of which increased in response to antigens, are responsible for transplantability of the stromal tissue in heterotopic transplantation. Immunization by streptococcal antigens seemed to suppress transplantability and osteogenic activity of stromal stem cells. The efficiency of CFC-F cloning in mouse bone marrow cultures increased significantly (2-3-fold) in the presence of sera from immune mice. The levels of TNF-α and IFN-γ were low in this serum (2.7 and 6 times lower, respectively) in comparison with normal serum. Presumably, the effects of streptococcal antigens on stromal tissue were mediated through serum cytokines.

Injection of S. typhimurium antigens significantly (9-fold) increased cloning efficiency and, hence, the content of stromal precursor cells in the spleen as soon as after 24 h. These parameters returned to normal by days 6-15 after immunization. Cultured splenocytes collected from immune (but not intact) animals expressed the genes of proinflammatory cytokines IL-1β (on days 1, 6, 15) and IL-6 (on days 1 and 6), TNF-α (on days 6 and 15), and of IFN-α and IL-18 (on days 6 and 15). The expression of IL-4 gene was suppressed on day 6 after immunization, of IL-10 gene on days 1 and 6, of IL-6 gene on day 15. Hence, no signs of immune response suppression by stromal cells were found in this system. The spectrum and dynamics of the expression of pro- and anti-inflammatory cytokine genes in stromal cell cultures from the spleen of immunized mice seemed to correspond to those needed for support of the immune response to S. typhimurium antigens, observed in immunized animals. The results indicate possible involvement of stromal cells in the realization of immune response in vivo. The increase of stromal precursor cells cloning efficiency in response to antigen injection could not be reproduced in vitro: the presence of S. typhimurium antigens in primary cultures of intact mouse bone marrow and spleen throughout the entire period of culturing ≈ 20-fold reduced cloning efficiency in cultures.

The study was carried out on CBA mice using the method of heterotopic transplantation. A fragment of the femoral bone marrow (1/2) or spleen (1/5 of the organ) was transplanted under the renal capsule of a recipient. The following donor-recipient cross-transplantation variants were studied: young-young (Y-Y), young-old (Y-O), old-old (O-O), and old-young (O-Y). Cell suspensions were prepared from 2-month transplants inoculated in monolayer cultures and the cloning efficiency (ECF-F) of stromal precursor cells (CFC-F) was evaluated. The bone marrow transplant ECF-F and the count of CFC-F in the O-O group were 8-fold lower than in the Y-Y group. In the O-Y group, ECF-F was 3-fold higher than in the O-O group, but by 2.5 times lower than in the Y-Y group. ECF-F in Y-O group was 2-fold lower than in Y-Y group. The ECF-F and CFC-F count in spleen transplants in the O-O group were 4- and 6-fold lower, respectively, than in Y-Y group. However, in O-Y group ECF-F was 7-fold higher than in O-O group and higher than even in Y-Y group. The weight of induced ectopic bone tissue after transplantation of the osteoinductor (fragments of the allogenic urinary bladder mucosa) was 2-fold lower in the O-O vs. Y-Y group. However, comparison of the ectopic bone tissue weights in different experimental groups showed that osteoinductor activity of the bladder epithelium did not decrease, but increased 3-fold with age (O-Y:Y-Y). A 5-fold reduction of this proportion in groups where the osteoinductor was transplanted from old donors to old and young recipients (O-Y:O-O) could be attributed to age-specific reduction of the count of inducible osteogenic precursor cells (IOPC). The data in general suggest that age-specific reduction of the stromal precursor count and functional activity could be caused by the true reduction (exhaustion) of cell pool (bone marrow CFC-F; presumably, IOPC) and by the regulatory effects of the organism (bone marrow and splenic CFC-F, IOPC). These data seem to be significant for understanding of the role of osteogenic stromal precursor cells in the development of age-associated bone tissue defects, for example, senile osteoporosis.

Aim: To assess effect of continued immunization of mice from CBA line with inactivated group A streptococcal vaccine on levels of serum cytokines and number of stromal bone marrow progenitor cells in immunized mice and in heterotopic transplants after different variants of transplantation., Materials and Methods: CBA mice were immunized during 3 weeks with heat-killed vaccine prepared from group A streptococci type 5. Levels of pro- and antiinflammatory cytokines were measured with BioPlex device. Number of stromal progenitor cells was determined on quantity of colonies formed by cells explanted to monolayer cultures., Results: Significant 2.3-fold increase of number of stromal progenitor cells in femoral bone marrow of immune mice was demonstrated. Experiments with heterotopic transplants showed that bone marrow in transplants of mice immunized with streptococci--variant Normal-->Immune (N-->1)-- is defective both on efficacy of cloning and number of stromal progenitor cells. Even short-term presence of stromal tissue in immune organism (variant I -->N) significantly changed these parameters, especially at late time after transplantation. In serum of immune mice changes of cytokines levels, especially TNFalpha, were observed. The level of the latter was decreased (mean--2.5-fold) in all immune serum samples compared to normal serum., Conclusion: Immunization of mice with group A streptococci leads to changes in stromal tissue and, possibly, to damage of microenvironment functions including hemo- and lymphopoiesis.

Administration of S. typhimurium microbial mass to mice was followed by a significant increase (by 3-4 times) in the efficiency of cloning and number of stromal precursors in the femoral bone marrow. These parameters were maximum on days 1-3, but returned to normal by the 8th-15th day after immunization. As differentiated from intact animals, the expression of genes for proinflammatory cytokines IL-1β (day 1 after immunization), IL-6 (days 1-3), TNF-α (days 1, 3, and 6), and IFN-α (days 1-3) was detected in bone marrow cultures from immunized mice. The expression of genes for IFN-γ, IL-18, and IFN-α was decreased on days 1, 3, and 6 after immunization of animals, respectively. Gene expression for the anti-inflammatory cytokine IL-4 was observed on day 6 after immunization. Therefore, this system was not characterized by a decrease in the immune response of stromal cells. The stromal component of hemopoietic and lymphoid organs has the vector of influences in response to bacterial antigens. This vector is directed to the stimulation and progression, but not to the suppression of immune reactions. Our results indicate that resident stromal cells play a role in the immune response of the body.

The counts of stromal precursor cells in bone marrow transplants obtained from animals 2 months after their immunization with killed type 5 group A streptococcus vaccine drop almost 3-fold in comparison with transplants from normal donors. Six months after donor immunization, the count of stromal precursor cells in the transplants reaches the normal level. The count of stromal precursor cells in bone marrow transplants from normal mice transplanted to recipients 6 months after their immunization with killed streptococcus vaccine also virtually did not change in comparison with the counts in bone marrow transplants from normal donors transplanted to normal recipients. The weight and size of bone capsules of 6.5-month bone marrow transplants in intact recipients after transplantation from donors immunized 2 months before with killed type 5 group A streptococcus vaccine was 3-fold lower than in bone marrow transplants collected from intact donors. The content of stromal precursor cells in the femoral bone marrow of animals immunized with killed streptococcus vaccine was 2.5 time higher in comparison with the parameter in the femoral bone marrow of normal mice even 8 months after immunization. The results indicate a significant long-acting effect of streptococcal antigens on the bone marrow stromal tissue, specifically, on its osteogenesis potential.

The efficiency of cloning of stromal precursor cell in mouse bone marrow culture increases significantly (2-3-fold) in the presence of serum from mice immunized with type 5 group A streptococcus antigens (5-20 microl serum/ml culture medium) in comparison with intact animal serum. The levels of TNF-alpha and IFN-gamma are significantly reduced (2.7 times and more than 6-fold, respectively) in the sera of immunized mice in comparison with normal serum. Serum levels of IL-2, -4, -5, -10, and -12 were about the same in both groups; no granulocyte-macrophage CSF was detected. These data attest to appreciable effect of immunization with streptococcal antigens on the bone marrow stromal tissue; this effect is presumably mediated through serum cytokines.

The count of stromal precursor cells in bone marrow transplants from CBA mice, transplanted to animals immunized with killed type 5 group A streptococcus vaccine, decreased 4.5-6.5 times (depending on the transplant age) in comparison with the grafts transplanted to normal recipients. The counts of stromal precursor cells in 1.5-3-month bone marrow transplants from animals immunized with killed streptococcal vaccine transplanted to normal mice were virtually the same, while in 7-month transplants they decreased 2-fold in comparison with their counts in bone marrow transplants from normal CBA mice transplanted to normal animals. The content of stromal precursor cells in the femoral bone marrow of animals immunized with killed streptococcal vaccine was appreciably (3.5 times) higher than in the bone marrow of normal mice. The results attest to an appreciable effect of streptococcal antigens on the bone marrow stromal tissue and suggest that not all stromal precursor cells, whose count increases after injection of antigens, are responsible for transplantability of the stromal tissue in case of its heterotopic transplantation.

Normal human and rabbit sera, as well as IgG isolated from them, have proved to be capable of reacting with the cells of the valve endothelium of the human and bovine heart. As shown in this study, these reactions are linked with the presence of Fc receptors on the epithelial cells. This is confirmed by the positive reactions of the endothelial cells with the Fc fragments of IgG, as well as with pure antibodies to egg albumin and to group A streptococcal polysaccharide and their complexes. As revealed in this study, Fc receptors on endothelial cells and staphylococcal Fc receptors bind with the definite fraction of normal human serum IgG with, probably, more pronounced cytophil properties. This fraction is not linked with IgG subclasses. The suggestion may be made that the presence of IgG Fc binding activity in group A streptococci, coinciding with the binding activity of Fc receptors in some cells of the human body, is probably of importance for pathogenic streptococci, facilitating their successful invasion.

In this review the superantigens (SAG) of group A Streptococcus, mainly pyrogenic toxins, and the coding genes are characterized. Specific sites on the molecules of the main histocompatibility complex, class II, and T-receptors, interacting with SAG, are described. This interaction results in hyperproduction of cytokines posing a systemic influence and leading to severe consequences. The data on the state of protective immunity in SAG-associated diseases and some approaches to their treatment are given.

Modern data on the etiology and pathogenesis of invasive streptococcal infection and the syndrome of streptococcal toxic shock are presented. In the course of the last 10-15 years essential changes in the system of interaction of group A streptococci and the macroorganism have been noted. The growth of morbidity in severe invasive forms of streptococcal infection with different clinical manifestations, including the syndrome of toxic shock, is observed. Most often this disease develops in elderly people, making up a group of risk, but sometimes affects healthy young people. Different pathogenicity factors of streptococci, capable of inducing the development of infection, are analyzed. Special attention is given to superantigens: pyrogenic toxins and M-protein. The suggestion that the development of the disease is seemingly linked with the state of specific protective immunity is substantiated. In spite of achievements in the field of the microbiology and immunology of group A streptococci, the causes of the appearance and development of invasive streptococcal infection have not yet been determined.

A panel of monoclonal antibodies (McAb) to different determinants of group A Streptococcus polysaccharide (A-PS) has been studied. As revealed in this study, A-PS contains at least 4 determinants, common with different epidermal antigens. McAb, cross-reacting (CR) with different mammalian tissue antigens, have not been found to be group-specific. Experiments on the inhibition of the immunoenzyme reaction of McAb with A-PS indicate that CR determinants include rhamnose and beta-N-acetyl-D-glucosamine of rhamnose alone. In addition, we have confirmed our earlier suggestion that group-specific high-affinity precipitating antibodies acted on different sites of A-PS. Contrary to earlier opinions, antibodies to group-specific determinants of A-PS have been found not to react with epidermal antigens.

Normal human and rabbit sera, and the IgG isolated from them, have been shown by immunofluorescence to react with bovine and human heart valve fibroblasts. Analogous results were obtained with sera of children under the age of 2. Positive reactions were observed also with fibroblasts, chondrocytes and osteocytes of human fetal joint tissues. The reactions are mostly due to monomeric immunoglobulins, since soluble immune complexes give much weaker reactions with the fibroblasts. The reactions are apparently dependent on the presence of Fc receptors on these cells. This conclusion is confirmed by positive reactions with IgG Fc fragments, with pure antibodies to ovalbumin and with human monoclonal IgG. The monoclonal IgG1 possesses the strongest ability to bind with fibroblast Fc receptors. No Fc-IgG receptors have been revealed on the fibroblasts and other structures of the interstitial connective tissue of human and bovine myocardium.

The study of serum samples, obtained from 15 cosmonauts before and after space flights, with the use of the indirect fluorescent method showed that in 7 cosmonauts antibodies to different elements of the human heart muscle appeared after flights. Strong and very strong luminescence of the elements of heart muscle tissue was detected in the cosmonauts after the third space flight. When studying the sera on sections of bovine heart muscle tissue, the reactions of the sera taken before and after flights were found to have no essential differences.

The data are confirmed about the presence of Fc-receptors reacting with human and rabbit IgG on the fibroblasts of human and bovine heart valves. It is shown the similar receptors are present on the fibroblasts of joints and L-cells. The receptors react largely with monomeric IgG. Clear-cut distinctions are revealed between the fibroblasts of heart valves and those of the myocardium which like other cells of myocardial interstitial connective tissue do not contain Fc-receptors.

It was demonstrated using the indirect immunofluorescent technique that normal human and rabbit sera, and IgG isolated from them intensively reacted with fibroblasts of human and bovine heart valves. The results obtained with Fab and Fc fragments of IgG sugges that this reaction is due to the Fc region of the IgG molecule and related to the presence of the Fc receptor on fibroblasts of heart valves.

The sera from patients with active rheumatic fever possess unmarked ability to react with antigens of human fibroblasts. More intense reactions were obtained with guinea-pig fibroblasts. In rheumatoid arthritis, positive reactions with human and guinea-pig fibroblasts were disclosed in half of the test sera. No complement-dependent cytotoxic antibodies were found in the patients' sera.

The reaction of heterophile antibodies with interstitial connective tissue cells of bovine myocardium was inhibited with alpha-D-galactose. D-glucose, D-mannose, L-fucose, N-acetylgalactosamine and other monosaccharides demonstrated the lack of inhibitory activity. Disaccharide melibiose and trisaccharide raffinose containing terminal alpha-galactose also had an inhibitory effect on the reaction of sera with bovine myocardium. The results show that heterophile antibodies to heterophile bovine antigen revealed in rheumatic patients' sera differ from antibodies to I and i antigens and T-antibodies, whose specificity is associated with terminal beta-galactose. They also differ from other heterophile antibodies described earlier.

Sera of patients suffering from rheumatic diseases and myocarditis were examined on the sections of human and bovine myocardial tissue by indirect immunofluorescence with the use of pure IgG antibodies or monospecific sera against IgG, IgA and IgM. It was shown that antibodies reacting with different myofibers and interstitial connective tissue of the heart belong to the main immunoglobulin classes (IgG, IgA and IgM). There was a significant predominance of IgG antibodies as shown by the frequency of their detection and by the titer height. The predominance of antibodies to certain classes of immunoglobulins did not correlate with a specific disease entity. The frequency of detecting antibodies to a certain immunoglobulin class was in good agreement with the time of the disease onset. Moreover, the frequency of positive reactions due to IgG, IgA, and IgM antibodies correlated with the level of the appropriate immunoglobulins in the test sera.

Antibodies against connective tissue elements of various bovine organs were isolated from the sera of rheumatic fever patients with the aid of immunosorbents (bovine connective tissue extract and erythrocyte stroma). The antibody preparations obtained were not identical and contained antibodies against different antigens of bovine connective tissue. The antibody preparations failed to react with human connective tissue components.

The antibodies reactive in immunofluorescence with the fibroblasts of human and bovine heart valves were isolated from the rheumatic patients' sera with the aid of immunoadsorbent prepared from human heart valve tissue. No reaction between the antibodies and the fibroblasts of the interstitial connective tissue of the myocardium was observed. It seems that the fibroblasts of the heart valves and the myocardium contain different antigens; the presence of common antigenic components in human and bovine valve fibroblasts is shown.

Investigation of antibodies to the specific determinant of streptococcal group A polysaccharide in indirect immunofluorescence experiments has revealed the existence of a cross-reactive antigen in the epithelial cells of the thymus and skin. This CR antigen is contained by the epithelial cells of man and animals of different species. It has been demonstrated in all the individuals studied including animals producing antibodies to the polysaccharide of Group A streptococci. The principal cause of autoimmune thymitis characteristic of rheumatic fever and other autoimmune processes is probably damage done to the thymus by autoantibodies resulting from immunization with microbial cross-reactive antigens shared by the thymus. Reaction of the autoantibodies with thymic antigens may affect the immunosuppressive function of the thymus and the maturation process of suppressor T cells. These events probably constitute the basic stage in the development of an autoimmune process.

The indices of cellular and humoral immunity were studied in 46 patients with heart diseases of rheumatic origin prior to and alter operation for replacement of the heart valves with synthetic prostheses (in 19) and pig aortic valves treated with glutaraldehyde and methaperiodate solutions ater Karpantje (in 27). The indirect fluorescence method and the leukocyte migration inhibition test to the myocardial antigens showed that the operation for valve prosthetics causes an increase in the content of antibodies and activation of specific leukocyte sensitization to the antigens of the myocardium and heart valves. These changes do not depend on the type of the valve prosthesis and are normalized by the 6th month. The activation of immunological processes encountered in some patients in late periods after the operation (1-8 years) corresponded to an aggravation of their clinical condition which in most cases was not associated with impaired functioning of the biological prosthesis. The implantation of xenogenous valve prostheses treated by Karpantje's method did not cause the development of immunological processes which could have induced destruction of the prostheses.

Purified antibodies against a specific determinant of group A streptococcal polysaccharide were obtained. These antibodies were investigated by the immunofluorescent method on tissue sections of the heart and heart valves. No cross-reactions between group A streptococcal polysaccharide and mammalian connective tissue were revealed.

Reactions of heterophilic antibodies with antigens of the interstitial connective tissue (ICT) of the myocardium of different animals were studied and compared. The reactions were identical on bovine, pig, sheep and rabbit myocardium. The heterophilic antigen of the ICT of bovine myocardium was also found on bovine, rabbit and rat red cells and was absent on sheep red blood cells. It is suggested that heterophilic antigen of the ICT of the bovine myocardium is a novel heterophilic antigen, the specificity of which is linked with D-galactose and which differs from antigens of other heterophilic systems.

The work discusses the results of studying by indirect immunofluorescence and the passive hemagglutination test antibodies to sacolemma antigens of myocardial muscle fiber in patients with rheumatic fever as compared to those in patients with affection of myocardium of nonrheumatic origin. The frequency of the detection of antibodies to sarcolemma antigens in sera of patients with rheumatic fever by immunofluorescence correlates with the activity of the process. No differences were revealed in the frequency of detection of antibodies reacting with sarcolemma in rheumatic fever and in other heart diseases in the active phase. Antibodies to sarcolemma antigens contained in HCL-extracts from human myocardium preparations, which are detected in the passive hemagglutination test are revealed significantly more often in sera of patients with rheumatic fever. Control experiments with IgG fractions obtained from whole sera were conducted to remove any possible nonspesific reactions of tissue extracts with the sera tested.