[To 20-years anniversary of Chernobyl catastrophe: an attempt to study the vitamin, calcium, iron and selenium status of children and adult population in Slavutich and to correct elicited deficiencies].

The article concisely illustrates the vitamin and mineral state of population of town of Slavutich, including personal of Chernobyl Nuclear Power Station, children of pre-school age and pregnancy women, studied in 1992. Vitamins and minerals deficiency in the main of C and B vitamins and selenium was revealed in all the studied groups. Appropriate measures were developed and introduced to eliminate the detected dusturbances; but however some unsolved problems remained. Taking into account the forthcoming 20th anniversary of Chernobyl disarter, the authors of the come back to considering the obtained data in hope to atlract attention of medical scientific and public to the remained unsolved problems of micronutrient deficiency.

[Effect of diphtheria toxin on the viability of phagocytes and B-lymphocytes in animals sensitive and insensitive to it]. / Vliianie difteriinogo toksina na zhiznesposobnost' fagotsitov i B-limfotsitov chuvstvitel'nykh i nechuvstvitel'nykh k nemu zhivotnykh.

It is well known, that mechanism of diphtheria toxin (DT) action triggers only if toxin penetrates into acid endosome after binding with specific receptor--heparin-binding epidermal grows factor like grows factor (HB-EGF) on the cell surface. We have suggested that DT is capable to penetrate either into B-lymphocytes, which have specific immunoglobulin receptors for DT or into phagocytes, which are able to phagocytosis of DT, because in both of these cases toxin get in endosome with conditions suitable for its activation. To check this hypothesis the comparative studies with insensitive to DT mice lacking specific receptor for DT, and with sensitive to DT guinea pigs were performed. Influence of DT on vitality of phagocytes and B-cells with different specificity from mice and guinea pigs was studied. B-cells were obtained from animals immunized by control antigen--ovalbumine and recombinant diphtheria toxoid--DT without N-terminal 28 aminoacid residues responsible for toxic effect. The results obtained have showed that DT can penetrate into phagocytes and B-cells specific to DT and kill these cells even if they lack classic receptor for DT. This fact evidences that DT is potentially able to inhibit self-directed antibody response and keep from participation of phagocytes in the protection of organism from infection.

[The role of calcium ions in fibrin polymerization]. / Rol' ionov Ca v polimerizatsii fibrina.

The article reviews the literature and authors' own data on the role of Ca ions in blood coagulation, namely, in the process of the formation of highly ordered fibrin clot. It has been shown that the main role of Ca2+ is the timely formation and stabilization of fibrin polymerization sites at all successive stages of the fibrin polymerization process.

Novel monoclonal antibodies against major antigens of Mycobacterium bovis.

MPB70 and MPB83 are among the most characteristically exported proteins defining a strongly expressed phenotype of Mycobacterium bovis. These proteins are known to be homologous to osteoblast-specific factor 2. By in vitro culture of mycobacteria they appear to have a limited species distribution and to be relatively specific for M. bovis. Virtually identical genes are however, present in Mycobacterium tuberculosis. In order to facilitate further research into the immunobiology of these proteins and their potential application for differential diagnosis of tuberculosis as a result of M. bovis, we describe the reactivities of 20 monoclonal antibodies (MoAbs) to these proteins. Immunizing with bovine PPD generated 10 MoAbs. These antibodies reacted preferentially with the soluble MPB70 antigen using reducing conditions in SDS-PAGE with western blotting. Ten MoAbs were generated by immunizing mice with fractions derived from a whole cell sonic extract of M. bovis. These antibodies reacted preferentially with the surface exposed MPB83 lipoglycoprotein.

[Specificity of antibodies to diphtheria toxin subunits in children with various forms of diphtheria infections]. / Spetsifichenost' antitel k sub''edinitsam difteriinogo toksina u detei s razlichnymi formami difteriinoi infektsii.

In order to study the specificity of serum antibodies to separate subunits of diphtheria toxin, SDS-electrophoresis of diphtheria toxin preliminary disintegrated on the subunits via trypsin treatment was performed, followed by immunoblotting assay. 86 blood serum samples of children with diphtheria carriers of toxigenic and non-toxigenic strains of Corynebacterium diphtheriae as well as children with other infectious diseases similar to diphtheria in their clinical manifestation, and healthy ones immunized with DTP-vaccine were tested. A special computer program was written and applied for results processing and assumption. The data obtained showed that there were particular differences in frequency of predominating the antibodies to one or another subunit of diphtheria toxin among various groups of the children. We consider that the different specificity of antibodies of sick children and children-carriers is capable to predetermine the different course of infectious process.

[Minimal amino acid sequence, recognized by anibodies to the peptide GPQPPQPPQP from the proline-rich region of pertactin]. / Minimal'nye aminokislotnye posledovatel'nosti, raspoznavaemye antitelami k peptidy GPQPPQPPQP iz bogatoi prolinom oblasti pertaktina.

Three antiserum samples obtained from rabbits immunized by the conjugate KLH-10P (keyhole limpet hemocyanine-decapeptide GPQPPQPPQP) were used to study antigenic structure of 10P. Antigenic properties of conjugates 6P (PGPQPP) and 4P (PQPP) with ovalbumine were studied by an indirect immunoassay (ELISA). Also 4P, 6P, PQP and QPP peptides were used for a competition assay. It was found that antibodies to 10P have demonstrated different specificity to short sites. Antibodies recognized such shot peptides as PQP and QPP in the competition assay. The efficiency of serum antibodies reaction with those peptides increased from QPP and PQP to PGPQPP. Only one serum sample had no antibodies to glutaraldehyde. Gly-glutaraldehyde-Gly hapten-like ligand was used to inhibit activity of antibodies to cross-linking agent into two samples. It is allowed to improve analysis of antibodies, which recognize shot sequences PQP and QPP.

Immunochemical characterization of the MPB70/80 and MPB83 proteins of Mycobacterium bovis.

MPB70 and MPB80 (MPB70/80) and MPB83 are closely related antigens which are highly expressed in Mycobacterium bovis. MPB70/80 are soluble secreted antigens, while MPB83 is an exported lipoprotein associated with the bacterial surface. In the present study, these antigens had different mobilities in sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing and nonreducing conditions. These differences may be explained by the fact that MPB70 and MPB83 both have two internal cysteine residues which would create ring structures by disulfide bonding. We analyzed the structures of MPB70/80 and MPB83 by using monoclonal antibodies (MAbs) raised against bovine purified protein derivative or whole M. bovis cells. MAb 1-5C reacted specifically with MPB70 and MPB80, and MAb MBS43 reacted specifically with MPB83, while the other antibodies, including several previously described MAbs, bound all three antigens. MAbs and polyclonal antibodies reacted strongly with reduced protein and less well with nonreduced protein, indicating involvement of linear epitopes. Epitopes of MAbs Bov-1, 2-6B, 1-5C, and 1-1D were mapped by using synthetic peptides of MPB70. Sequence comparison showed the peptide with the 1-5C-reactive epitope to have three residues different from those in the homologous region of MPB83. Exchanges of A for S in position 112 or Q for E in position 116 abolished the reactivity of MAb 1-5C. Polyclonal rabbit antibodies to native purified MPB70 reacted strongly with peptides 6, 7, and 8 of the N-terminal half of mature MPB70. Cattle sera of experimentally M. bovis-infected animals recognized a broader spectrum of peptides. These findings indicate that there is diagnostic potential for these proteins and that there is also a possible role for antibodies in elucidation of the host-mycobacterium relationship involving a surface-bound and exposed lipoprotein, MPB83, and its highly homologous soluble secreted MPB70/80 counterparts.

[Structure and mechanism of functioning of centers of fibrin polymerization]. / Struktura i mekhanizm funktsionirovaniia tsentrov polimerizatsii fibrina.

Fibrinogen takes part in a lot of physiological and pathological processes in the organism. Its most important function consists in preventing blood leakage through the injured wall of the blood vessel. Proteolytic enzyme thrombin generated in the site of the injury splits low-molecular fibrinopeptides A and B from fibrinogen and transforms the latter into the monomeric fibrin which is spontaneously polymerized with formation of a clot of blood closing a lumen of the injured vessel. Fibrin polymerization that completes the stages of blood coagulation is the example of self-assembling of supermolecular biological structures in blood coagulation system. Clearing-up of this process mechanisms is of great theoretical and practical importance, in particular, for clinical medicine connected with treatment of hemorrhages and thromboses.

[Fibrin fragment beta 15-118. Inhibitory and complex-forming properties]. / Fibrinovyi fragment beta 15-118. Ingibitornye i kompleksoobrazuiushchie svoistva.

Peptide beta 15-118 isolated from desAABB-NDSK preserves fibrin polymerization active site "B", inhibits polymerization process at 12 degrees C, eliminates the inhibitory properties of plasmin D-D-fragment but does not influence inhibitory properties of a D-monomer fragment. Complex formation between peptide beta 15-118 and both D- and D-D fragments was electrophoretically demonstrated. Peptide beta 15-118 forms more stable complex with the D-D fragment which does not dissociate in the medium of polymerizing fibrin as the complex of the peptide with monomer D fragment does. Gel filtration data confirm dimerization of D-monomer fragments after their complexing with beta 15-118. This phenomenon suggests that mutual affinity of D-domains in fibrin increases after loci interactions of the "B"-"b" type.

The immune response to cytochrome c in BALB/c mice is delayed due to inability of their non-specific antigen-presenting cells to provide its immunodominant epitope.

The antibody response to horse cytochrome c (cyt.c) in BALB/c mice developed slowly and a substantial production of IgG antibodies was observed only 26-30 days after immunization. Lymph node cells (LNC) of unimmunized mice proliferated weakly in response to both native cyt.c and its synthetic peptides. On day 8 after immunization, LNC could not be stimulated with native cyt.c and peptide 92-104. However, they did proliferate in response to cyt.c peptides 1-6, 1-13, 2-13, 14-22, 46-56, 57-77, 61-77 and 61-69 which are closely related in horse and mouse cyt.c. On day 26, both native cyt.c and the peptides, including 92-104, were equally active in stimulating LNC proliferation. Both plastic-adherent and cyt.c-specific cells panned from day 8 cells enhanced the response of unprimed cells to native cyt.c. Elimination of B cells demonstrated that primary recognition of cyt.c was mediated, at least partly, by non-specific antigen-presenting cells (APC) while later B cells of additional specificities were involved. It is concluded that immunization with horse cyt.c initiated an autoimmune response resulting in T-dependent anergy. Peptide determinants processed by non-specific APC stimulated corresponding autoreactive T cells. Specific B cells which appeared as a result of the response maturation processed successfully the immunodominant epitope and finally mediated proliferative and antibody responses.

[Study of the polymerization of fibrin using monoclonal antibodies 2D-2A and their Fab-fragments]. / Issledovanie polimerizatsii fibrina s pomoshch'iu monoklonal'nykh antitel 2D-2A i ikh Fab-fragmentov.

It has been shown that monAb's 2d-2a and their Fab-fragments are specific and effective inhibitors of fibrinogen clotting. Only one IgG molecule of monAb's 2d-2a can bind with one of their epitopes situated around peptide bond B beta Arg14-Gly15 in dimer fibrinogen molecule reducing the rate of protofibril lateral association and clot turbidity with only one fibrinopeptide B splitting off per fibrinogen molecule by thrombin. But two molecules of Fab-fragments of monAb's 2d-2a join to both of their epitopes and inhibit fibrinogen clotting dramatically without clot formation and with no fibrinopeptide B splitting off. These data suggest that the site of fibrin protofibril lateral coalescence is localized in NH2-terminal part of fibrin (ogen) B beta-chain, i.e. central E-domain of fibrin molecule takes part in protofibril lateral association. The mutual space orientation of NH2-terminal regions of fibrinogen B beta-chains is discussed.

Selective killing of tumor cells in vitro by immunotoxin composed of antitumor antibiotic streptonigrin and polyclonal specific antibodies.

Streptonigrin N-hydroxysuccinimide ester (STN-COONSu) was obtained by carbodiimide synthesis. Poly-L-lysine (PLL) was loaded with STN-COONSu and conjugated to polyclonal rabbit immunoglobulin G (IgG) activated with sodium periodate. Non-specific IgG and IgG against Ehrlich carcinoma cells were used to construct non-specific and specific immunotoxins. Immunotoxins contained 100 molecules of streptonigrin per 1 molecule of IgG. The streptonigrin concentration that caused 50% of inhibition of [3H]thymidine incorporation in Ehrlich carcinoma cells (IC50) was 0.8 micrograms/ml for specific immunotoxin, 16 micrograms/ml for non-specific immunotoxin, and 20 micrograms/ml for the poly-L-lysine-streptonigrin conjugate (PLL-STN) used as the initial water-soluble form of antibiotic. Our results demonstrate that the toxicity for target cells of streptonigrin conjugated to specific IgG was 25 times higher than that of the initial water soluble form of antibiotic. This specific immunotoxin was non-toxic for non-target cells.

Analysis of immunogenic properties of nonapeptide TVGRGDPHQ from Bordetella pertussis filamentous hemagglutinin.

Immunogenic properties of TVGRGDPHQ nonapeptide which is correspondent to the region 1094-1102 of B. pertussis filamentous hemagglutinin (FHA) were studied. The conjugate of bovine serum albumin with nonapeptide was used for immunization of BALB/c and CBA mice. Antisera of the both lines of mice cross-reacted with a number of antigens, but using affinity chromatography peptide and FHA specific antibodies were extracted. Affinity purified rabbit antibodies to TVGRGPHQ which recognize FHA were also obtained. Therefore the antibodies to the peptide which placed RGD-containing region responsible for macrophage CR3-integrin interaction are capable to distinguish the native antigen. Thus these data are an additional evidence for the nonapeptide use as a component of synthetic vaccine against whooping-cough.

[The determination of neurospecific proteins in the blood of patients with closed craniocerebral trauma and their diagnostic significance]. / Opredelenie neirospetsificheskikh belkov v krovi bol'nykh s zakrytoi cherepno-mozgovoi travmoi i ikh diagnosticheskoe znachenie.

Data are reported on the content of neurospecific proteins in the blood serum of 87 patients with closed head injuries. It has been established that the method of competent EIA can be used for measuring the content of neurospecific proteins. It has also been shown that their content in the blood depended on the severity of injury: the more severe injury the higher the protein content. Some aspects have been revealed of the blood content of protein 14-3-2 and total myelin protein. The authors discuss a possible diagnostic importance of these proteins in the differential diagnosis of the severity of closed head injuries and in the determination of lesions of the neuroglia and neurons both in mild and grave trauma.

[The dynamics of the humoral immune response to mycobacterial antigens in mice]. / Dinamika gumoral'nogo immunnogo otveta na antigeny mikobakterii u myshei.

The main regularities of humoral immune response to mycobacterial antigens have been studied in experiments on BALB/c mice immunized with live and thermoinactivated Mycobacterium tuberculosis, var. bovis. As shown in this study, the maximum level of serum antibodies to mycobacterial antigen is achieved in two weeks after immunization irrespective of the dose and viability or mycobacteria, then follows a decrease in the antibody level. The absence of uniformity in the dependence of primary immune response and the formation of immunological memory on the dose and viability of mycobacteria has been demonstrated.

[Chromatographic analysis and immunobiologic properties of tuberculins]. / Khromatograficheskii analiz i immunobiologicheskie svoistva tuberkulinov.

Immunobiologic activities of tuberculin preparations and their components have been comparatively studied using gel filtration and high-performance liquid chromatography (HPLC) techniques. It is shown that high-molecular weight fraction of protein-purified derivate tuberculin (PPD) had higher activity as compared to nonfractionated preparation in skin tests on guinea pigs sensitized with Mycobacterium bovis BCG as well as in enzyme-linked immunosorbent assay with affinity purified rabbit antibodies against PPD. Using the preparative HPLC-technique we failed to isolate a component of PPD having greater tuberculin test potency than nonfractionated preparation.

[The formation of delayed hypersensitivity to mycobacterial antigens]. / Formirovanie giperchuvstvitel'nosti zamedlennogo tipa k mikobakterial'nym antigenam.

In experiments on guinea pigs and BALB/c mice delayed hypersensitivity to mycobacterial antigens was induced by the sensitization of the animals with live BCG or killed Mycobacterium bovis or M. avium in incomplete Freund's adjuvant. In the study of the dynamics of the development of skin reactivity to tuberculin some advantages of the sensitization of guinea pigs with live mycobacteria were revealed, while after the revaccination of the animals no development of secondary cell-mediated immune response was observed. The immunization of guinea pigs with atypical mycobacteria prior to their sensitization with BCG was found to lead to the development of higher skin reactivity to allergen prepared from atypical mycobacteria than skin reactivity to tuberculin.

The study of fibrin polymerization with monoclonal antibodies.

Three kinds of monoclonal antibody (Mab) of different specificity have been obtained against the N-terminal disulphide knots of fibrinogen and fibrin. Their effects on different phases of fibrin polymerization have been studied. These antibodies were shown to be directed against different epitopes of the B beta(1-53) fragment of the fibrinogen molecule. The different Mab had different effects both on the rate of protofibril lateral aggregation and on the final turbidity of fibrin clots. The Mab were of three specificities: (1) those from clone 2d-2a inhibited the rate of lateral aggregation of protofibrils and decreased the turbidity of the final clot; (2) those from clone B-4C accelerated the polymerization step but did not affect clot turbidity: and (3) those from clone D-IB did not have any effect on either fibrin polymerization or final clot turbidity. The localization of the epitopes recognized by all three kinds of Mab and analysis of our own data and those of others allow us to conclude that one of the active loci involved in protofibril lateral association is situated in the B beta(15-53) fragment of the fibrinogen molecule. Fibrinopeptide B does not need to be split off for this site to function. Fibrin polymerization can occur when one of the two sites of protofibril lateral aggregation in dimeric fibrin molecules is blocked by Mab, and the final clot turbidity is then reduced. The splitting off of one of the two fibrinopeptides B in fibrinogen molecules by thrombin can take place even when the second B beta(Arg14-Gly15) bond is blocked by an antibody molecule.(ABSTRACT TRUNCATED AT 250 WORDS)