[Immunoenzyme test system for detecting antibodies to group-specific antigens of group A Streptococcus on the base of conjugated N-acetylglucosamine and its use in medical practice]. / Immunofermentnaia test-sistema dlia opredeleniia antitel k gruppospetsificheskomu antigenu streptokokka gruppy A na osnove kon'iugirovannogo N-atsetilgliukozamina i ee primenenie v meditsinskoi praktike.

Enzyme immunoassay kit has been created for detecting antibodies to group A Streptococcus, based on N-acetylglucosamine. N-acetylglucosamine was selected as the group-specific determinant due to the structure of group A Streptococcus polysaccharide, in which this monosaccharide residue is lateral to the main polysaccharide chain and hence more available for antibodies. Water-soluble polyacrylamide is the carrier in this kit, for this carrier is stable and not liable to nonspecific reaction with proteins. In addition, the synthesis of polyacrylamide conjugates ensures reproducible results. Use of this kit permits the identification of group A streptococcal etiology of the disease and thus carry out appropriate therapy; moreover, it helps predict the outcome of an acute streptococcal infection and detect the poststreptococcal complications in the early period of the disease.

[The M-protein heterogeneity of Streptococcus pyogenes strains isolated from clinically healthy and ill children in an organized collective]. / Geterogennost' po M-belku shtammov Streptococcus pyogenes, vydelennykh ot klinicheski zdorovykh i bol'nykh detei v organizovannom kollektive.

The study of the kinetic curves of the lysis of group A S. pyogenes cell-walls (cells), serovars 29 and 12 M, variants M+ and M-; with endo-N-acetyl-muramidase revealed that the kinetics of the lysis of virulent and avirulent strains was different: variant M- was lyzed faster than M+. This difference in the cell-wall lysis of both variants made it possible to use this method for the identification of M+ and M- states of the strains. 18 group A S. pyogenes cultures were studied. The cultures isolated from healthy and sick children in an organized group belonged to variants both M+ and M-. The pepsin fragments of M-proteins of group A S. pyogenes, type M 1, isolated in dynamics from a carrier and a patient, were studied. Their amino acid compositions were studied, and differences between them were established. The data thus obtained indicate that heterogeneity is characteristic of both the surface structure of streptococci and the individual components of their surface, in particular, M-protein.

[The production of erythrogenic toxin and the epidemic process in respiratory streptococcal infection]. / Produktsiia éritrogennogo toksina i épidemicheskii protsess respiratornoi streptokokkovoi infektsii.

The capacity of Streptococcus pyogenes, isolated from clinically healthy children in a large organized group, for producing erythrogenic toxin A, B and C was studied. The dynamics of toxin production was compared with changes in the levels of morbidity and carrier state, as well as with some characteristics of the interaction between the populations of the infective agent and the host by such values as virulence and susceptibility. Similarity in the dynamics of the parameters under study was noted. The seasonal dynamics of characteristics which determined the capacity for toxin production significantly influenced the levels and one year morbidity dynamics with respiratory streptococcal infection. The study demonstrated the information value of the characteristics of toxin production for understanding both the interval mechanisms of the epidemic process and the influence of the external ones on subsequent epidemic situation.

[The heterogeneity of Streptococcus pyogenes strains isolated from clinically healthy and sick children in an organized collective]. / Geterogennost' shtammov Streptococcus pyogenes, vydelennykh ot klinicheski zdorovykh i bol'nykh detei v organizovannom kollektive.

Experiments on typed strains used as an example revealed that kinetic curves obtained in the lysis by endo-N-aretylmuramidase of virulent strains greatly differed from those obtained from nonvirulent ones: the lysis rate of M+ variants was less than that of M- variants; M- strains gave rather steep kinetic curves of lysis, while those of M+ strains were more declivous. In this study group A S. pyogenes cultures isolated from healthy and sick children in a summer camp were used. The study revealed that cultures, newly isolated from healthy and sick children, were heterogeneous with respect to the M+ and M- state of group A S. pyogenes strains, the amino acid composition of pepsin fragments of M-proteins in the cultures of streptococcus M 1 under study being also heterogeneous.

[Streptococcal group A carriage: new data on the characteristics of the biology of the causative agent and of the immune response of the host]. / Nositel'stvo streptokokka gruppy A: novye dannye po kharakteristike biologii vozbuditelia i immunnogo otveta khoziaina.

Carriership of group A streptococcus (A-STR) was registered with significantly greater frequency among children of preschool and early school age (29.8%) than among adults (4.0%). Cultures of A-STR isolated from carriers were characterized by their low capacity for secreting erythrogenic toxin of type A (ET-A), lipoteichoic acid and by lower content of surface proteins than those isolated from tonsillitis and scarlet fever patients. Highly hydrophobic strains of A-STR prevailed in long-time carriers and patients. Elevated immunological characteristics for group-specific polysaccharide A and ET-A were more frequently detected in children than in adults, which correlated with frequency of A-STR carriership in these groups and the capacity of the infective agent to produce ET-A. Some specific features in the character of humoral immune response in carriers, manifested mainly by defects of local immunity (a low level of SIgA to polysaccharide A), were established.

[An explosive outbreak of strep throat morbidity in an adult organized collective]. / Eksplozivnaia vspyshka zabolevaemosti anginoi v organizovannom vzroslom kollektive.

Altogether 162 cases of tonsillitis were registered in two military units during the period of May 11-16. The disease took an acute course with short-time fever, symptoms of acute intoxication, sore throat, pronounced inflammatory changes in tonsils and swelling of regional (submaxillary and anterocervical) lymph nodes. In some of the patients (1.1%) sickness and vomiting and in 0.2% diarrhea were registered. In 6.7% of the patients scarlatiniform eruptions were observed on days 2-3 of the disease. In the course of the bacteriological examination of the patients group A streptococci, serovar T II, were isolated. The dynamic study of paired sera showed a considerable increase in the number of patients with a high level of antibodies to streptolysin 0 and group A polysaccharide. All patients were fed at the same canteen. The factor of the transmission of this infection could be butter, stored without observation of the required temperature conditions and apportioned by the soldiers of the kitchen police. Experimental study revealed that group A streptococci are capable for proliferation and accumulation in butter.

[The biochemical and immunological properties of group A type M 29 streptococci cultured in the presence of a bovine blood serum preparation]. / Biokhimicheskie i immunologicheskie svoistva streptokokkov gruppy A tipa M 29, kul'tivirovannykh v prisutstvii preparata iz syvorotki krovi krupnogo rogatogo skota.

The influence of the preparation of cattle blood serum on group A streptococcus, type M 29, has been studied. The study has revealed that the addition of 17% of dialysis water obtained from a fraction of cattle blood serum to the standard culture medium (3% Todd-Hewitt broth) produces changes in the amino acid composition of the cell walls of M+ variant without altering the antiphagocytic resistance of the mutant thus obtained. The dialysate of the pepsin digest of the cell walls of the mutant contains Fc-receptors and receptors to fibrinogen, while the initial strain contains only receptors to fibrinogen which are, in this case, the pepsin fragments of M protein. The study has revealed similarity in the amino acid compositions of these proteins (receptors to fibrinogens) of phenotypes M+ and M2+. Thus, our data confirm that the initial strain and the mutant belong to different phenotypes of group A streptococcus, type M 29.

[The modification of the cell wall proteins in group A streptococci type M 29 under the influence of spermidine contained in the culture medium]. / Modifikatsiia belkov kletochnykh stenok streptokokkov gruppy A tipa M 29 pod deistviem spermidina, soderzhashchegosia v srede kul'tivirovaniia.

The addition of spermidine into growth medium used for the cultivation of group A streptococci, type M 29, leads to changes in the amino acid composition of cell walls and surface proteins isolated by the method of E. H. Beachey et al. The separation of surface proteins into fibrinogen-binding proteins and fibrinogen receptors by affinity chromatography techniques on cellulose with covalently bound fibrinogen indicates that the proportion of these proteins in pepsin extracts obtained from different strains varies. Both spermidine and avirulent strains have similar content of fibrinogen-binding proteins, although these proteins are absent in virulent strains. Different amounts of fibrinogen receptors are extracted from all strains. As shown in the enzyme immunoassay, fibrinogen receptors contain no group-specific polysaccharide A, Fc-receptors and interact with total antiserum to group A streptococci, type M 29 [correction of 28]. Fibrinogen receptors isolated from the strains under study have been found to have similar amino acid composition. On the basis of these results we believe that neither receptor capacity to fibrinogen nor amino acid composition is indicative of the protective properties of protein M.

[The low-molecular protein of the cell wall in streptococci group A devoid of serological type specificity]. / Nizkomolekuliarnyi belok kletochnoi stenki streptokokkov gruppy A, lishennyi tipovoi serologicheskoi spetsifichnosti.

The scheme for the isolation and purification of low-molecular cell-wall protein without type specificity, including the extraction of the cell walls of group A streptococci, type M 29, with 1% solution of Triton X-100, the separation of the extract by ion-exchange chromatography in DEAE-trisacryl M with the subsequent two-stage gel filtration in superfine Sephadex G-50, is described. The isolated protein had a molecular weight of 4,000 daltons and contained no admixtures of group-specific polysaccharide A, phosphorus, nucleic acids and Fc receptors and interacted with antisera to group A streptococcal cells of heterologous type M in the enzyme immunoassay (EIA). Purified protein was characterized by a high content of glycine. The antigenic determinants of immobilized protein, recognized by antibodies in EIA, were sensitive to the action of trypsin and resistant to the action of pepsin, papain, pronase E and sodium periodate.

[The pharyngeal beta-hemolytic streptococcal carrier state among blood donors]. / Glotochnoe nositel'stvo beta-gemoliticheskikh streptokokkov sredi donorov krovi.

459 blood donors aged 18-50 years were examined in 1987-1988 in Moscow. Among them, carrier state with respect to beta-hemolytic streptococci was detected in 107 donors (23.3%). The number of carriers gradually decreased with the increase of age of the examined donors. Group C streptococci occurred least of all (6.9%). Group A beta-hemolytic streptococci were isolated in 16.7% of the carriers. The isolation rate of streptococci from blood achieved its maximum in autumn and winter months and did not depend on preceding diseases, unhealthy working conditions, the rhesus factor and, with the exception of group A streptococci, the blood group. Among tonsillectomized donors carrier state with respect to beta-hemolytic streptococci occurred 2.2 times less frequently than among donors who had not undergone tonsillectomy. Carrier state with respect to beta-hemolytic streptococci was accompanied by higher levels of salivary sIgA antibodies to polysaccharide A, serum antibodies to polysaccharide A and circulating polysaccharide A. All beta-hemolytic streptococci were sensitive to erythromycin. All groups of streptococci showed the highest percentage of cultures resistant to gentamicin and tetracycline. In 100% of cases group A streptococci were sensitive to benzylpenicillin, methicillin, ampicillin, erythromycin and lincomycin.

[A study of the variability of group A type M29 streptococcus on exposure to biologically active substances]. / Izuchenie izmenchivnosti streptokokka gruppy A tip M29 pri vozdeistviina nego biologicheski aktivnykh veshchestv.

Biologically active substances spermidine and the compounds contained in the low-molecular fraction of cattle blood serum change the phenotype of type 29 group A Streptococcus. Amino acid analysis and enzymatic destruction of cell walls have demonstrated changes of the streptococcal biological characteristics and cell wall structural organization. These changes may result from loss of supervariable component of the M-protein when compounds from cattle blood serum low-molecular fraction are added to culture medium and by unbalanced growth of Streptococcus after spermidine addition to growth medium.

[Heterogeneity of streptococcus group A cell wall protein components]. / Geterogennost' belkovykh komponentov kletochnoi stenki streptokokka gruppy A.

Cell wall surface proteins of group A streptococcus (M 29) were isolated by mild chemical extraction with 1 M hydroxylamine pH 6.0 (37 degrees C). The proteins were purified by ammonium sulfate fractionation, gel filtration on Sephadex G-150 and ion-exchange chromatography on DEAE-Trisacryl M. Using two independent methods (disc electrophoresis in 7.5% PAAG pH 8.9 and high pressure gel filtration), it was shown that after chromatography on Sephadex G-150 the original protein fraction contains up to 8 protein components, while SDS-PAAG electrophoresis performed according to Laemmli revealed up to 25 protein components in the same fraction. During SDS-PAAG electrophoresis six protein fractions performed after ion-exchange chromatography were resolved into 40 protein components whose molecular masses vary from 13 to 80 kDa. Possible reasons for the heterogeneity of surface proteins of group A streptococcus cell wall are discussed.

[The physicochemical and biochemical characteristics of the cell walls in M+ and M- variants of Streptococcus group A type 29]. / Fiziko-khimicheskaia i biokhimicheskaia kharakteristika kletochnykh stenok M+ i M--variantov streptokokka gruppy A tipa 29.

The amino acid composition of cell walls and surface proteins, isolated from virulent (M+) and avirulent (M-) streptococcal strains (group A, type 29) has been determined by the method of E. H. Beachey et al. The kinetics of the lysis and proteolysis of streptococcal cell walls with muramidase and protease obtained from Actinomyces levoris and streptolysin has been studied. The constants describing the progress rates of these processes has been determined; their values in case of both lysis and proteolysis are higher in virulent strains than in avirulent ones.

[The epidemic process of an acute streptococcal respiratory infection in large organized collectives of children]. / Epidemicheskii protsess ostroi respiratornoi streptokokkovoi infektsii v krupnykh detskikh organizovannykh kollektivakh.

The study of the specific features of the development of the epidemic process of scarlet fever, tonsillitis, and acute respiratory diseases (ARD) in two large organized groups of children revealed the presence of some differences which depended on the character of prophylactic measures taken in these groups. Thus, in the absence of prophylaxis with bicillin a pronounced increase in the level of carriership, accompanied by an increase in the infective capacity of carriers, was noted. This resulted in a high level and unfavorable dynamics of morbidity in scarlet fever, tonsillitis, and ARD. On the contrary, the use of prophylaxis with bicillin ensured the stability of the level of carriership, while the infective capacity of carriers was not pronounced. At the same time a rise in ARD morbidity was insignificant, and morbidity in scarlet fever and tonsillitis was reduced to nil.

[Study of the composition of cell wall of group A Streptococcus after hydrolysis using muramidase from Streptomyces levoris]. / Izuchenie sostava kletochnoi stenki streptokokka gruppy A pri posledovate'lnom gidrolize muramidazoi Streptomyces levoris.

The aim of the experiment was to study the lysis products of cell walls of group A streptococci resulting from exposure to N-acetylmuramidase. It was shown that for isolating surface proteins free of polysaccharide and peptidoglycan fragments it was necessary to treat the streptococcal cell walls with endo-beta-N-acetylmuramidase for no more than 30 minutes. Prolonged hydrolysis with muramidase led to the presence of polysaccharide and the peptidoglycan fragments in the protein fractions, intracellular wall proteins covalently bound to the peptidoglycan fragments and polysaccharide being also released.

[The effect of blood serum components on the growth, biochemical and biological properties of streptococcus]. / Vliianie komponentov syvorotki krovi na rost, biokhimicheskie i biologicheskie svoistva streptokokka.

The components of cattle blood serum, added to the medium for the cultivation of group A streptococci, considerably decrease the period of adaptation and increase the balanced growth rate of streptococci, which is manifested by changes in the surface structures of the cell wall: the absence or modification of protein M. Streptococci grown under these conditions lose their capacity for phagocytosis, and from the cell walls obtained from these streptococci no surface protein M can be isolated by pepsin treatment. Nevertheless, the ratio of the main cell-wall components (proteins, polysaccharide and peptidoglycan), the amino acid composition, as well as the resistance of the cell walls to the action of trypsin and endo-N-acetylmuramidase are the same in M+ and Mx variants, that makes it possible to infer that the modification of protein M or the inhibition of its synthesis occurs during the growth of streptococci in the presence of blood serum components.

[Hydrophobic quality and the level of surface proteins in the cell wall of cultures of Streptococcus pyogenes isolated from clinically healthy children from an organized collective]. / Gidrofobnos't i kolichestvennoe sederzhanie poverkhnostnykh belkov kletochnoi stenki ku'ltur Streptococcus pyogenes, vydelennykh ot klinicheski zdorovykh detei v organizovannom kollektive.

The Str. pyogenes cultures isolated from clinically healthy children were found to have different hydrophobic properties and quantities of surface cell wall proteins, and to exhibit pronounced variability in these signs in time. Most studied cultures lacked a correlation between their hydrophobic properties and the quantity of surface proteins of the cell wall.

[Changes in the cell wall composition and structure of Streptococcus pyogenes during batch culture]. / Izmenenie sostava i struktury kletochnykh stenok Streptococcus pyogenes pri periodicheskom kul'tivirovanii.

Changes in S. pyogenes cells in the process of batch cultivation have been studied. The composition of S. pyogenes cell walls has been studied by amino acid analysis; besides, their resistance to enzymatic hydrolysis and the electric conductivity of cell-wall lysates have been determined at different phases of the growth of S. pyogenes. The molar amino acid composition, expressed in percent, is unrelated to the growth phase, while the content of amino acids in preparations changes in the process of growth and reaches its maximum in the middle and in the end of the logarithmic phase. At the same time the electric conductivity of cell-wall lysates reaches the minimum level at these growth stages. The authors suggest that additional electrically charged compositions are formed in the cell walls at the beginning of the logarithmic and stationary phases. A considerable increase in the initial rate of cell-wall lysis with muramidase has been found to occur at the end of the logarithmic phase. This difference in the initial rate in the initial rates of lysis of S. pyogenes cell walls at different growth phases decreases after previous treatment of the cell walls with streptolytin possessing proteolytic activity. Analysis of these data leads to a conclusion on the "loose" structure of the outer protein layer of the cell wall at the end of the logarithmic phase of the growth curve.