Evaluation of secretory activity in carp pancreatocytes (Cyprinus carpio L. ) when using lactobacillus-based chelate and probiotic supplemental feed complexes / Avaliação da atividade secretora em pancreatócitos de carpa (Cyprinus carpio L. ) ao usar quelato à base de lactobacilos e complexos suplementares de alimentação probiótica

The experimental research was carried out on the juvenile carp (Cyprinus carpio L.). The impact from supplemental feeds consisting of variable concentrations of chelate compounds, biogenic trace elements and probiotic lactobacillus-based product Bacillus subtilis VKPM B-2335 was evaluated. Optical and qualitative parameters of the lactobacillus base were studied in order to identify the major group of substances potentially able to influence the end result. The purpose of this research was to identify changes in the structure of the zymogen granules and their dimensions at which supplemental feeds produce a stimulating effect on the synthesis of zymogens in exogenous cells of the secretory part of pancreas. At the outcome of the study, for the first time, it was possible to prove that the integrated action of chelates and lactobacillus-based probiotics complemented each other. Metal chelate compounds contributed to enlargement of the zymogen granules, if compared to the control values. The bacterial products accelerated production of the zymogen granules in acinar cells diffusely located in carp hepatopancreas.

A pesquisa experimental foi realizada em carpas juvenis (Cyprinus carpio L.). O impacto de suplementos alimentares consistindo em concentrações variáveis de compostos quelatos, oligoelementos biogênicos e produto probiótico baseado em lactobacilos Bacillus subtilis VKPM B-2335 foi avaliado. Parâmetros ópticos e qualitativos da base de lactobacilos foram estudados a fim de identificar o maior grupo de substâncias potencialmente capazes de influenciar o resultado final. O objetivo dessa pesquisa foi identificar mudanças na estrutura dos grânulos de zimogênio e suas dimensões nas quais alimentos suplementares produzem um efeito estimulante na síntese de zimogênios em células exógenas da parte secretora do pâncreas. No resultado do estudo, pela primeira vez, foi possível comprovar que a ação integrada dos quelatos e dos probióticos à base de lactobacilos se complementava. Compostos quelatos metálicos contribuíram para o aumento dos grânulos de zimogênio, se comparados aos valores de controle. Os produtos bacterianos aceleraram a produção dos grânulos de zimogênio nas células acinares localizadas difusamente no hepatopâncreas da carpa.

Evaluation of secretory activity in carp pancreatocytes (Cyprinus carpio L.) when using lactobacillus-based chelate and probiotic supplemental feed complexes

Abstract The experimental research was carried out on the juvenile carp (Cyprinus carpio L.). The impact from supplemental feeds consisting of variable concentrations of chelate compounds, biogenic trace elements and probiotic lactobacillus-based product Bacillus subtilis VKPM B-2335 was evaluated. Optical and qualitative parameters of the lactobacillus base were studied in order to identify the major group of substances potentially able to influence the end result. The purpose of this research was to identify changes in the structure of the zymogen granules and their dimensions at which supplemental feeds produce a stimulating effect on the synthesis of zymogens in exogenous cells of the secretory part of pancreas. At the outcome of the study, for the first time, it was possible to prove that the integrated action of chelates and lactobacillus-based probiotics complemented each other. Metal chelate compounds contributed to enlargement of the zymogen granules, if compared to the control values. The bacterial products accelerated production of the zymogen granules in acinar cells diffusely located in carp hepatopancreas.

Resumo A pesquisa experimental foi realizada em carpas juvenis (Cyprinus carpio L.). O impacto de suplementos alimentares consistindo em concentrações variáveis de compostos quelatos, oligoelementos biogênicos e produto probiótico baseado em lactobacilos Bacillus subtilis VKPM B-2335 foi avaliado. Parâmetros ópticos e qualitativos da base de lactobacilos foram estudados a fim de identificar o maior grupo de substâncias potencialmente capazes de influenciar o resultado final. O objetivo dessa pesquisa foi identificar mudanças na estrutura dos grânulos de zimogênio e suas dimensões nas quais alimentos suplementares produzem um efeito estimulante na síntese de zimogênios em células exógenas da parte secretora do pâncreas. No resultado do estudo, pela primeira vez, foi possível comprovar que a ação integrada dos quelatos e dos probióticos à base de lactobacilos se complementava. Compostos quelatos metálicos contribuíram para o aumento dos grânulos de zimogênio, se comparados aos valores de controle. Os produtos bacterianos aceleraram a produção dos grânulos de zimogênio nas células acinares localizadas difusamente no hepatopâncreas da carpa.

Structure of the capsular polysaccharide and the O-side-chain of the lipopolysaccharide from Acetobacter methanolicus MB 58/4 (IMET 10945), and of oligosaccharides resulting from their degradation by the bacteriophage Acml.

The capsular polysaccharide (CPS) and the O-side-chain of the lipopolysaccharide (LPS) of Acetobacter methanolicus MB 58/4 (IMET 10945) have been shown to contain the same disaccharide repeating unit, namely, ----2)-beta-D-Galf-(1----3)-beta-D-Galp-(1----. Degradation of the CPS and the LPS with the bacteriophage Acml gave fragments built up of 1-5 repeating units; the octasaccharide preponderated. The phage-associated depolymerase proved to be a beta-D-galactofuranoside hydrolase.

[Synthesis of N-glycoproteins with a native type of carbohydrate-peptide bond]. / Sintez N-glikoproteinov s prirodnym tipom uglevod-peptidnoi sviazi.

Reaction of beta-oligoglycosylamines obtained from carbohydrate chains of N-glycoproteins (ovalbumin, ovomucoid, riboflavin-binding glycoprotein from hen egg white, and asialofetuin) with bovine serum albumin, lysozyme, and poly(L-Asp) in presence of water-soluble carbodiimide gave rise to a series of glycoconjugates, modelling natural N-glycoproteins. Carbohydrate-peptide bond was shown to be of N-glycosylamide type with participation of Asp and Glu residues. The method allows one to obtain synthetic N-glycoproteins from oligomannoside, complex and hybrid oligosaccharide chains, and may find application both in biochemistry and biotechnology for improvement of physico-chemical properties of unglycosylated proteins.

Enzyme immunoassay (EIA) test systems for the detection of Salmonella O-antigen.

Four EIA (enzyme immunoassay) test systems for the detection of salmonella O-antigens in various biological tissues were studied. To find the optimum test system two types of affinity purified antibodies were used to coat the microtitre plates: (i) monospecific antibodies isolated on immunoadsorbent bearing synthetic O-antigen factor 4 (O:4; salmonella serogroup B) as ligand, (ii) antibodies specific for lipopolysaccharide (LPS) B isolated from the IgG fraction of hyperimmune sera. The use of affinity purified antibodies led to an increase in the sensitivity of 'sandwich' EIA by an order of magnitude and to improved specificity. Competitive EIA was ten to twenty times less sensitive. It is demonstrated for the first time that salmonella O-antigens can be detected in the sera of animals within a day after challenge. For patients with salmonellosis, O-antigen could be detected after the fifth day of illness, but in the urine and faeces only (not in the blood serum), in 30%-70% of cases. This substantially improves the identification of salmonellosis from among other enteric infections. In competitive EIA, inhibition of standard antibodies by the sera under study was caused not by the presence of O-antigen but by antibodies homologous to the coating antigen. This resulted in "blocking' of the latter and led to false-positive results. The results obtained enable the optimum EIA technique to be selected to improve the serological diagnosis of salmonellosis with both synthetic salmonella O-antigens and LPS.

[Structure of carbohydrate chains of the riboflavin-binding glycoprotein of chicken egg protein. II. 1H-NMR (500 MHz) spectroscopy of the major neutral oligosaccharides]. / Struktura uglevodnykh tsepei riboflavinsviazyvaiushchego glikoproteina belka kurinogo iaitsa. II. Spektroskopiia 1H-YaMR (500 MGts) glavnykh neitral'nykh oligosakharidov.

Reductive cleavage of riboflavin-binding glycoprotein from hen egg white (RF-GPw) with LiBH4/tert-BuOH followed by NaBH4/NaOH treatment gave rise to oligosaccharide alditols, fractionated by a successive HPLC on muBondapak C18 and Zorbax NH2 columns. Seven main individual oligosaccharide alditols were isolated and their structure was investigated by 1H NMR 500-MHz spectroscopy. The structure and relative content of the main oligosaccharide chains were proved to be identical in RF-GPw and ovomucoid. Structure of polypeptide chains and their molecular weight, number of glycosylation sites and their structure had little or no effect on the glycosylation pattern in both glycoproteins. HPLC of the oligosaccharide alditols from another egg white glycoprotein, ovotransferrin, also revealed its high microheterogeneity and close resemblance to those of ovomucoid and RF-GPw.

[Specificity of enzymes of biosynthesis of Salmonella anatum O-antigen for polyprenyl derivatives of different chain length and saturation]. / Spetsifichnost' fermentov biosinteza O-antigena Salmonella anatum k proizvodnym poliprenolov raznoi dliny tsepi i nasyshchennosti.

Polyprenyl phosphates of different structure were prepared and their ability to serve as sugar acceptors in the biosynthesis of O-specific polysaccharides of Salmonella anatum was investigated. It was demonstrated that C30-C80-polyprenyl phosphates with unsaturated alpha-isoprene unit were as active as natural acceptor (undecaprenyl phosphate) in this enzymic system. C15- and C100-polyprenyl phosphates of this series were less effective in O-antigen repeating unit formation. Citronellyl and dolichyl phosphates (derivatives of C10- and C105-polyprenols, respectively, with saturated alpha-isoprene unit) were poor substrates. For polymerization of repeating units, the polyprenol chain-length is of utmost importance: its shortening results in a marked drop in the efficiency of respective compounds as substrates.

[Conformation analysis of oligosaccharide determinants of group substances with Le specificity]. / Konformatsionnyi analiz determinantnykh oligosakharidov gruppovykh veshchestv so spetsifichnost'iu Le.

Theoretical conformational analysis of the tri- and tetrasaccharides determining group specificity Lea, Leb, and H has been carried out. It is shown that these oligosaccharides are structurally rigid systems and in aqueous solution are present predominantly (statistical weight greater than 95%) in the only conformation.

Specific method for the fragmentation of the polypeptide chain of glycoproteins. Distribution of carbohydrate chains on the peptide core of blood-group-specific glycoprotein.

A method for specific fragmentation of the polypeptide backbone of glycoproteins at the glycosylated serine and threonine residues has been developed. The fragmentation includes beta-elimination of the carbohydrate chains, followed by bromination of the resulting enamine groups, and cleavage of the brominated amino acid residues by alkaline sodium borohydride. The method was employed for fragmentation of the peptide core of pig blood-group substance H. Essentially all the serine and threonine residues were shown to be O-glycosylated, and rather frequently either adjacent or separated by a single amino acid (mainly alanine). When they were separated by two or three amino acid residues, proline was preponderant.

Characterization of the primary structure and the microheterogeneity of the carbohydrate chains of porcine blood-group H substance by 500-MHz 1H-NMR spectroscopy.

In blood-group H substance from pig stomach cell linings, carbohydrate structures are known to occur which are O-glycosidically linked via GalNAc to Ser or Thr of the polypeptide backbone. They have in common the Gal(beta 1 leads to 3)GalNAc core unit, which bears one or more N-acetyllactosamine branches. The latter are terminated either by Fuc in (alpha 1 leads to 2) linkage or by GlcNAc in (alpha 1 leads to 4) linkage to Gal. Both terminal sequences are considered to be porcine blood-group H antigenic determinants. Employing 500-MHz 1H-NMR spectroscopy, the spectral features of the oligosaccharide-alditols corresponding to these chains were established. Insight could be gained into the microheterogeneity displayed by some of the alditol fractions as to the distribution of the terminating Fuc and GlcNAc residues over the various branches. Such information can hardly be obtained using other approaches.