Ecological features of trace elements tolerant microbes isolated from sewage sludge of urban wastewater treatment plant.

Worldwide wastewater treatment plants generate enormous amounts of sewage sludge, and their further disposal depends on the treatment technologies applied and spontaneously occurring microbiological processes. From different ages urban sewage sludge, 12 strains of bacteria with simultaneous tolerance to two or more trace elements: Co, Ni, Cu, Zn, Cd and Pb at concentration of 3-5 mmol were isolated and identified by PCR of target genes and Sanger sequencing methods. The isloated metal(loids) tolerant strains belong to the species, i.e., Serratia fonticola, Rhodococcus qingshengii, Pseudomonas fragi, Pseudomonas extremaustralis, Pseudomonas cedrina, Stenotrophomonas maltophilia, Serratia liquefaciens and Citrobacter freundii. The ecological features of the isolated strains were studied. The optimal growth temperatures for most strains was 15-30°C at pH range of 5-9, although some strains grew at 7°C (Pseudomonas fragi SS0-4, Serratia fonticola SS0-9 and Serratia fonticola SS12-11). Satisfactory growth of two strains (Serratia fonticola SS0-1and Citrobacter freundii SS60-12) was noted in an acidic medium at pH 4. Most of the strains grew in the NaCl concentration range of 1-5%. The isolated bacteria resistant to high concentrations of trace elements can be used for the effective mineralization of sewage sludge and for the decontamination of wastewater.

Structure and gene cluster of the O-polysaccharide of Yersinia rohdei H274-36/78.

A branched O-polysaccharide was obtained by mild acid degradation of the lipopolysaccharide of Yersinia rohdei H274-36/78 and found to contain d-rhamnose, d-mannose, and 3,6-dideoxy-4-C-[(S)-1-hydroxyethyl]-d-xylo-hexose called yersiniose A (Yer). Partial acid hydrolysis of the O-polysaccharide eliminated Yer residues to give a modified linear polysaccharide. Studies by sugar analysis and 1H and 13C NMR spectroscopy, including computational NMR analysis, enabled structure elucidation of a hexasaccharide repeating unit of the O-polysaccharide having two Yer residues attached as monosaccharide side chains. The O-antigen gene cluster of Y. rohdei H274-36/78 located between JUMPStart and galF genes contained putative genes for synthesis of precursors of two O-antigen constituents, GDP-d-Man and GDP-d-Rha, whereas genes responsible for synthesis of CDP-Yer were within the chromosome outside the O-antigen gene cluster. Glycosyltransferase genes and ABC 2 transporter genes were present in the O-antigen gene cluster, and hence the structure established is consistent with the polysaccharide synthesis gene content of the genome.

Erratum to: "Structural Relationships Between Genetically Closely Related O-Antigens of Escherichia coli and Shigella spp." [Biochemistry (Moscow), 81, 600 (2016)].

This corrects the article DOI: 10.1134/S0006297916060067.

Zinc and lead detoxifying abilities of humic substances relevant to environmental bacterial species.

The effect of humic substances (HS) and their different fractions (humic acids (HA) and hymatomelanic acids (HMA)) on the toxicity of zinc and lead to different strains of bacteria was studied. All tested bacteria demonstrated a lower resistance to zinc than lead showing minimum inhibitory concentrations of 0.1 - 0.3mM and 0.3-0.5mM, respectively. The highest resistance to lead was characteristic of Pseudomonas chlororaphis PCL1391 and Rhodococcus RS67, while Pseudomonas chlororaphis PCL1391 showed the greatest resistance to zinc. The combined fractions of HS and HA alone reduced zinc toxicity at all added concentrations of the organic substances (50 - 200mgL-1) to all microorganisms, while hymatomelanic acids reduced zinc toxicity to Pseudomonas chlororaphis PCL1391 at 200mgL-1 organic concentration only. The HS fractions imparted similar effects on lead toxicity also. This study demonstrated that heavy metal toxicity to bacteria could be reduced through complexation with HS and their fractions. This was particularly true when the metal-organic complexes held a high stability, and low solubility and bioavailability.

Structure and gene cluster of a tyvelose-containing O-polysaccharide of an entomopathogenic bacterium Yersinia entomophaga MH96T related to Yersinia pseudotuberculosis.

An O-polysaccharide was isolated from the lipopolysaccharide of an entomopathogenic bacterium Yersinia entomophaga MH96T by mild acid hydrolysis and studied by 2D NMR spectroscopy. The following structure of the branched tetrasaccharide repeating unit of the polysaccharide was established: where Tyv indicates 3,6-dideoxy-d-arabino-hexose (tyvelose). The structure established is consistent with the gene content of the O-antigen gene cluster. The O-polysaccharide structure and gene cluster of Y. entomophaga are related to those of some Y. pseudotuberculosis serotypes.

Structural Relationships Between Genetically Closely Related O-Antigens of Escherichia coli and Shigella spp.

Gene clusters for biosynthesis of 24 of 34 basic O-antigen forms of Shigella spp. are identical or similar to those of the genetically closely related bacterium Escherichia coli. For 18 of these relatedness was confirmed chemically by elucidation of the O-antigen (O-polysaccharide) structures. In this work, structures of the six remaining O-antigens of E. coli O32, O53, O79, O105, O183 (all related to S. boydii serotypes), and O38 (related to S. dysenteriae type 8) were established using (1)H and (13)C NMR spectroscopy. They were found to be identical to the Shigella counterparts, except for the O32- and O38-polysaccharides, which differ in the presence of O-acetyl groups. The structure of the E. coli O105-related O-polysaccharide of S. boydii type 11 proposed earlier is revised. The contents of the O-antigen gene clusters of the related strains of E. coli and Shigella spp. and different mechanisms of O-antigen diversification in these bacteria are discussed in view of the O-polysaccharide structures established. These data illustrate the value of the O-antigen chemistry and genetics for elucidation of evolutionary relationships of bacteria.

Synthetic neoglycoconjugates of cell-surface phosphoglycans of Leishmania as potential anti-parasite carbohydrate vaccines.

Leishmania are a genus of sandfly-transmitted protozoan parasites that cause a spectrum of debilitating and often fatal diseases in humans throughout the tropics and subtropics. During the parasite life cycle, Leishmania survive and proliferate in highly hostile environments. Their survival strategies involve the formation of an elaborate and dense cell-surface glycocalyx composed of diverse stage-specific glycoconjugates that form a protective barrier. Phosphoglycans constitute the variable structural and functional domain of major cell-surface lipophosphoglycan and secreted proteophosphoglycans. In this paper, we discuss structural aspects of various phosphoglycans from Leishmania with the major emphasis on the chemical preparation of neoglycoconjugates (neoglycoproteins and neoglycolipids) based on Leishmania lipophosphoglycan structures as well as the immunological evaluation for some of them as potential anti-leishmaniasis vaccines.

[The influence of vitamin B group on monooxygenase activity of cytochrome P450 3A4: pharmacokinetics and electro analysis of catalytic properties].

It was shown that vitamin B group permit to shorten the longitude of diclofenak therapy and to reduce the daytime dose of this drug. All three schemes of diclofenac treatment - only diclofenac, diclofenac plus 2 tablets of Gitagamp (mixture of vitamin B group), and diclofenac plus 4 tablets of Gitagamp--gave maximum value of diclofenal in blood through 1 hour after treatment. In the case of diclofenak treatment without vitamins Cmax corresponds to 1137.2 +/- 82.4 ng/ml, with 2 tablets of Gitagamp--Cmax 1326.7 +/- 122.5 ng/ml, and with 4 tablets--Cmax 2200.4 +/- 111.3 ng/ml. Positive influence of vitamin B group on the decrease of pain syndrome was shown. Pharmacodynamics and pharmacokinetics data were confirmed in electrochemical experiments with cytochrome P450 3A4. For enzyme immobilization screen printed graphite electrodes modified with gold nanoparticles and synthetic membrane-like compound didodecyldimethylammonium bromide (DDAB/Au) were used. Electrochemical analysis reviled the influence of vitamin B group on metabolism of non steroid anti inflammation drug diclofenac catalyzed by cytochrome P450 3A4. Riboflavin was the most effective inhibitor of diclofenac hydroxylation by cytochrome P450 3A4 as was compared at 300 M concentration of vitamin B group (B1, B2, B6). These data confirmed the opportunity of pharmacokinetic parameters regulation and the level of pharmacodynamic effects by the influence of vitamin B group on the catalytic activity of cytochrome P450 3A4.

[Rhizosphere bacteria Pseudomonas aureofaciens and Pseudomonas chlororaphis oxidizing naphthalene in the presence of arsenic].

Rhizosphere strains of P. aureofaciens BS1393(pBS216, pKS1) and P. chlororaphis PCL1391(pBS216, pKS1), exhibiting the ability to stimulate the growth of plants and protect them from phytopathogens, have been obtained. In these strains, plasmid pBS216 ensures naphthalene degradation and plasmid pKS1 confers resistance to arsenic. In the presence of arsenic and naphthalene, the number of living cells and the growth rate of the arsenic-resistant strains were higher than those of the arsenic-sensitive strains BS1393(pBS216) and PCL1391(pBS216). During the cultivation of the resistant strains, arsenic had no inhibitory effect on the activity of the key enzymes of naphthalene biodegradation, except for catechol-2,3-dioxygenase. In a model system containing plant-microbial associations, strains BS1393(pBS216, pKS1) and PCL1391(pBS216, pKS1) degraded as much as 97% of added naphthalene in the presence of arsenic.

[Synthesis of alpha-D-xylopyranosyl phosphate].

Synthesis of alpha-D-xylopyranosyl phosphate is carried out by a modified MacDonald method.

[Effect of natural and genetically modified rhizospheric Pseudomonas aureofaciens bacteria on accumulation of arsenic by plants]. / Vliianie prirodnykh i geneticheski modifitsirovannykh rizosfernykh bakterii Pseudomonas aureofaciens na nakoplenie mysh'iaka rasteniiami.

Gene constructions rendering bacteria resistant to arsenic and capable of dissolving phosphates and/or arsenates were created by cloning ars operon and the gene of citrate synthase from a chromosome of the strain Pseudomonas aeruginosa PAO1. Genetically modified variants of the strain Pseudomonas aureofaciens BS1393 have been constructed, which are resistant to high concentrations of arsenic and dissolve poorly soluble phosphates and/or arsenates. Recombinant strains P. aureofaciens BS1393(pUCP22::arsRBC) and P. aureofaciens BS1393(pUCP22::gltA) exerted positive effects on the survival of sorgo (Sorghum saccharatum L.) and its ability to accumulate arsenic.

[A synthesis of dolichyl fluorophosphate]. / Sintez dolikholftorfosfata.

An improved method for the synthesis of dolichyl H-phosphonate was developed using 2-chloro-4H-1,3,2-benzodioxaphosphorin-4-one (salicyl chlorophosphite) as a reagent. Dolichyl phosphorofluoridate was for the first time synthesized from dolichyl H-phosphonate by its treatment with chlorotrimethylsilane, oxidation with iodine, and subsequent interaction with fluoride ion in pyridine.

[Immune disorders in congenital hydronephrosis complicated by obstructive pyelonephritis]. / Immunnye nausheniia pri vrozhdennom gidronefroze, oslozhnennom obstruktivnym pielonefritom.

Congenital hydronephrotic transformation complicated by obstructive pyelonephritis with intact renal function in children exhibited association with some antigens of the major histocompatibility system and their combinations, changes in immunity and nonspecific resistance. Marked and stable shifts in immune reactivity give grounds for inclusion of immunomodulating drugs in combined treatment of the above patients.

[Synthesis of dolicyl- and polyprenyl sulfates]. / Sintez dolikhil- i poliprenilsul'fatov.

Dolichyl and polyprenyl sulfates were synthesized as analogues of dolichyl and polyprenyl phosphates by the interaction of dolichols and polyprenols with the pyridine-sulfuric anhydride complex.

Synthesis and antifibrillatory activity of nibentan and its analogues.

A series of 1,5-diaminopentane derivatives, structurally related to nibentan, was synthesized and tested for antifibrillatory activity. Improved modifications of some known chemical syntheses were proposed. (+/-)-N-[5-(Diethylamino)-1-(4-nitrophenyl)pentyl]-benzamide hydrochloride, (+/-)-N-[5-(diethylamino)-1-(4-nitrophenyl)pentyl]-4-nitrobenzamide hydrochloride and (+/-)-N-[5-(diethylamino)-1-(4-hydroxyphenyl)pentyl]-4-nitrobenzamide hydrochloride were more potent than nibentan and possessed a longer duration of action (up to 5 h in comparison with 60-90 min for nibentan). The antifibrillatory activity of (+/-)-N-[5-(diethylamino)-1-(4-methoxyphenyl)pentyl]-4-nitrobenzamide hydrochloride was comparable to that of nibentan but exceeded the potency of D-sotalol and sematilide.

Dolichyl sulphate and H-phosphonate: enzymatic reactions with activated sugars.

Two phosphate-modified analogues of dolichyl phosphate were evaluated as substrates or inhibitors of the reactions catalyzed by mammalian microsomal enzymes. Dolichyl H-phosphonate could serve as an efficient acceptor for mannosyl and glucosyl transfer. The reaction products were chromatographically different from those formed from dolichyl phosphate. Lower activity of the H-phosphonate was observed for the reaction of N-acetylglucosaminyl phosphate transfer from UDP-GlcNAc. Dolichyl sulphate was shown not to serve as a substrate for the transfer of mannosyl (from GDP-Man), glucosyl (from UDP-Glc) or N-acetylglucosaminyl phosphate (from UDP-GlcNAc) residues in the presence of rat liver microsomes. Weak inhibitory properties of this analogue were demonstrated.

Fucosylation of synthetic oligosaccharides--plant xyloglucan fragments--by mammalian and amphibian fucosyltransferases.

Liver and oviduct of frogs Rana temporaria and Xenopus laevis were shown to possess the activity of beta-galactoside alpha 1-2-fucosyltransferase. Extracts of these tissues as well as porcine and rat submaxillary gland homogenates catalyze the fucosylation of synthetic plant xyloglucan fragments, the disaccharide D-Gal(beta 1-2)D-Xyl and the tetrasaccharide D-Gal(beta 1-2)D-Xyl(alpha 1-6)D-Glc(beta 1-4)D-Glc; the active preparations can be used for enzymatic synthesis of biologically active oligosaccharides.

[Partial purification of glycosyltransferases participating in the biosynthesis of Salmonella anatum and S. kentucky O-antigens using high performance gel chromatography]. / Chastichnaia ochistka glikoziltransferaz, uchastvuiushchikh v biosinteze O-antigenov Salmonella antatum i S. kentucky s pomoshch'iu vysokoéffektivnoi gel'-khromatografii.

The solubilized glycosyltransferases which catalyse the biosynthesis of Salmonella anatum and S. kentucky O-specific polysaccharides were partially purified by HPLC on Superose 12. Two mannosyl transferases from S. kentucky were separated by gel chromatography; these transferases were found useful for chemical-enzymic synthesis of polyprenylpyrophosphate derivatives of trisaccharides Tal-Man-Gal and Man-Tal-Gal.

[Incorporation of paratose residue into Salmonella O-specific polysaccharides using enzymatic reactions]. / Vvedenie ostatka paratozy v O-spetsificheskie polisakharidy salmonell s pomoshch'iu fermentativnykh reaktsii.

The block mechanism of O-specific polysaccharides biosynthesis was demonstrated for Salmonella nitra (serogroup A) and S. haifa (serogroup B). Due to the moderate specificity of glycosyl transferases from S. nitra, S. typhimurium, S. haifa and S. kentucky (serogroup C3) towards the 3,6-dideoxyhexose structure a paratose residue can be incorporated into the polysaccharide chain instead of an abequose residue, and vice versa.