[An evaluation of the protective activity of a multicomponent vaccine made from opportunistic microorganisms in the oral method of immunization in a model of local staphylococcal infection]. / Otsenka protektivnoi aktivnosti polikomponentnoi vaktsiny iz uslovno-patogennykh mikroorganizmov pri oral'nom metode immunizatsii na modeli lokal'noi stafilokokkovoi infektsii.

The protective activity of a multicomponent vaccine against S. aureus, prepared from Staphylococcus, Klebsiella, Proteus and Escherichia coli antigens, intended for oral administration, has been studied on the model of a local staphylococcal infection. The study has revealed that the vaccine, when administered orally to mice, prevents the development of gangrene in the paw, the death of the animals, and essentially decreases the local focus of infection. The optimum treatment schedule consists of five administrations of the preparation in doses of 2 mg.

[The antigenic activity of vaccines made from opportunistic microorganisms in an experiment to immunize animals by oral and combined methods]. / Izuchenie antigennoi aktivnosti vaktsin iz uslovno-patogennykh mikroorganizmov v éksperimente immunizatsii zhivotnykh oral'nymi i kombinirovannymi metodami.

The results of the study of the antigenic activity of multicomponent vaccine consisting of staphylococcal, Klebsiella, Proteus, Escherichia coli antigens and staphylococcal monovaccine, introduced into rabbits and guinea pigs by different routes, are presented. As shown in this study, the multicomponent vaccine introduced orally in 5 administrations stimulated the production of antibodies to all components, but the intensity of antibody formation to each of them was different. Antibodies to E. coli antigen were found to be at the lowest level. Staphylococcal antigen, introduced orally both as the component of the multicomponent vaccine and as the monovaccine, ensured pronounced stimulation of the production of antistaphylococcal antibodies, not lower than that observed after subcutaneous injection. The comparative study of the activity of the multicomponent vaccine introduced by combined routes, oral-subcutaneous and subcutaneous-oral, showed the advantage of the former one.

[The specific activity of a multicomponent vaccine made from the antigens of opportunistic microorganisms]. / Spetsificheskaia aktivnost' polikomponentnoi vaktsiny iz antigenov uslovno-patogennykh mikroorganizmov.

Multicomponent vaccine prepared from the antigens of 4 representatives of opportunistic microflora possesses high specific activity. The passive hemagglutination (PHA) test with the use of associated diagnosticum showed that antibody titers in the sera of immunized rabbits increased 10- to 10(4)-fold in comparison with the titers observed prior to immunization. The PHA test with the use of the antigens contained in the vaccine revealed the accumulation of antibodies to each of the 4 components of the preparation in the blood sera of immunized rabbits. When stored at 4 degrees C, the vaccine was shown to retain its specific activity for 5 years (the term of observation).

[Isolation and characteristics of new restriction endonucleases from Haemophilus influenzae]. / Vydelenie i kharakteristika novykh éndonukleaz restriktsii iz Haemophilus influenzae.

Various strains of Haemophilus influenzae have been examined for the presence of site-specific endonuclease activities, and eleven restriction endonucleases have been isolated from seven strains. For all the endonucleases recognition sequences were determined, for three of them cleavage sites being identified. The enzymes proved to be isoschizomers of known endonucleases, viz. Hin1 I, Hin8 I--Acy I; Hin1 II, Hin8 II--Nla III; Hin2 I, Hin5 I--Hpa II; Hin3 I--Cau II; Hin5 II--Asu I; Hin5 III--Hind III; Hin6 I, Hin7 I--Hha I. Restriction endonucleases Hin1 I, Hin1 II and Hin6 I recognize nucleotide [formula: see text] sequences 5'GRCGPYC, 5'CATG, 5'GCGC, respectively, and cleave them as indicated by arrows.

[The method of isolation of Sau 6782-type methylases]. / Sposob polucheniia metilaz Sau 6782-tipa.

Heterogeneous profile of methylases was found in S. aureus 6782 strain. A procedure was developed for isolation of individual methylases from Sau 6782 free of Sau 6782 restrictases and unspecific nucleases by means of hydrophobic chromatography on phenyl-Sepharose. Ion exchange, affinity chromatographies and gel filtration were also used for isolation of the Sau 6782 methylases. But this technique was of limited suitability for isolation of individual methylating enzymes of the Sau 6782 type because it did not allow to separate completely these methylases from contaminating enzymes of DNA degradation. Effects of salts, glycerol and Triton X-100 on activity of total preparation of methylases Sau 6782 were studied. Na+ and K+ chlorides, ammonium sulfate at concentrations 0.4 M and highen as well as Triton X-100 inhibited reversibly the methylase activity followed by complete reduction up to initial level after dialysis. Glycerol at concentration 60% activated Sau 6782 methylases by 50% and stabilized the enzyme.

[Design of a nutrient medium for bacteria of the genus Haemophilus--producers of restriction endonucleases]. / Konstruirovanie pitatel'noi sredy dlia bakterii roda Haemophilus--produtsentov restriktiruiushchikh éndonukleaz.

A culture medium for the cultivation of hemophilic bacteria, containing acidic casein hydrolysate, aminopeptide and fodder yeast extract, has been proposed. The growth-stimulating properties of this medium have been studied on 5 strains producing restrictases differing in their specificity. In growing these producer strains in a model ANKUM-2 fermenter with the supply of carbohydrate substrates (glucose, sucrose, glycerin) the yield of biomass, considered to be high for hemophilic bacteria (10-14 g of humid substance from 1 liter of the medium), has been achieved. As shown on H. influenzae Rc B-2297 used as an example, an increase in the yield of microbial biomass leads to a decrease in restrictase specific activity.

[Activity of restriction endonuclease Sso II in Escherichia coli strains transformed by Shigella sonnei 47 plasmids]. / Aktivnost' restriktsionnoi éndonukleazy Sso II v shtammakh Escherichia coli, transformirovannykh plazmidami Shigella sonnei 47.

The inverse dependence of activity of restriction endonuclease SsoII preparations on the number of low molecular mass plasmids of Shigella sonnei transforming Escherichia coli recipient cells producing the enzyme has been shown. Escherichia coli strain producing efficiently one of two Shigella sonnei 47 restriction endonucleases SsoII has been isolated. The producer strain harbours two of the nine Shigella sonnei 47 plasmids. One of them P4 codes for SsoII+ phenotype while the other P9 determines the plasmids conjugation transfer. Biochemical and physiological characteristics of the producer strain XS13 are identical to the ones of the recipient Escherichia coli strain PS200. XS13 is unable to induce keratoconjunctivitis in guinea pigs in pathogenicity test.

[Purification and substrate specificity of BcmI restriction endonuclease]. / Ochistka i opredelenie substratnoi spetsifichnosti restriktsionnoi éndonukleazy BcmI.

A strain producing the new specific restriction endonuclease BcmI has been found in the Bacillus generum. The enzyme has been purified by chromatography on the blue sepharose, phosphocellulose PII, heparin sepharose. The analogous purification has been obtained when the blue sepharose has been substituted for the orange sepharose, the home produced sorbent. The BcmI enzyme has been shown by the substrate specificity definition to be an isoschizomer of the restriction endonuclease ClaI.

[Restriction endonuclease Sau 6782]. / Restriktiruiushchaia éndonukleaza Sau 6782.

Data are described on identification, isolation and purification of restricting endonuclease Sau 6782 as well as on estimation of the enzyme recognition site. Conditions were developed for growing of Staphylococcus aureus 6782 strain, which enabled to produce a maximal yield of the restricting activity containing minimal level of nucleases. The procedure for isolation and purification of restrictase Sau 6782 involved affinity chromatography on Blue Sepharose and cation exchange chromatography on phosphocellulose PII. The enzyme preparation obtained was free from impurities of unspecific nucleases. The yield of the Sau 6782 restrictase constituted 1,000 un from 1 g of the culture cells. Restrictase Sau 6782 recognized the nucleotide sequence 5'...GATC...3' and was the isoshizomere of the Sau 3A enzyme.