[DYNAMICS OF METABOLIC CHARACTERISTICS, MARKERS OF OXIDATIVE STRESS AND VASCULAR WALL DAMAGE DURING TREATMENT OF OBESE PRE-DIABETIC PATIENTS].

We studied lipid and carbohydrate metabolism, markers of oxidative stress (plasma malonic dialdehyde and dienoic conjugates) and vascular wall damage (CRP microalbuminuria, blood desquamated endotheliocytes) before and after 12 week treatment of obese pre-diabetic patients. The study showed positive dynamics of the above metabolic parameters with normalization of carbohydrate metabolism in 60% of the patients and angioprotective effect of prescribed therapy. The proposed treatment can be used to prevent diabetes mellitus.

Rheinheimera japonica sp. nov., a novel bacterium with antimicrobial activity from seashore sediments of the Sea of Japan.

Two gram-negative, aerobic, brown-pigmented, motile rod-shaped bacteria KMM 9512 and KMM 9513(T) were isolated from a sediment sample collected from the Sea of Japan seashore, Russia. On the basis of the 16S rRNA gene sequence analysis, novel strains KMM 9512 and KMM 9513(T) positioned within the genus Rheinheimera (class Gammaproteobacteria) as a separate subline adjacent to Rheinheimera baltica DSM 14885(T) sharing highest gene sequence similarities of 98.6-97.6 % to their closest phylogenetic relatives, Rheinheimera muenzenbergensis LMG 27269(T), R. baltica DSM 14885(T), Rheinheimera aquimaris JCM 14331(T), Rheinheimera nanhaiensis KACC 14030(T), and Rheinheimera pacifica KMM 1406(T). Strains KMM 9512 and KMM 9513(T) belong to the same separate genospecies on the basis of the 16S rRNA gene sequence similarity (99.8 %) and their DNA relatedness to each other (89 %) and to closely related Rheinheimera species (25-53 %). The major isoprenoid quinone was Q-8, polar lipids consisted of phosphatidylserine, phosphatidylglycerol, phosphatidylethanolamine, unknown aminolipids, unknown aminophospholipids, unknown phospholipids, and unknown lipids, and major fatty acid were C16:0, C16:1 ω7c, C17:1 ω8c, C12:0 3-OH followed by C17:0 and C18:1 ω7c in both strains. Strains KMM 9512 and KMM 9513(T) revealed a remarkable antagonistic activity toward a number of gram-positive and gram-negative microorganisms. On the basis of phylogenetic analysis, DNA-DNA hybridization results, and phenotypic differences, strains KMM 9512 and KMM 9513(T) are proposed to be classified as a novel species of the genus Rheinheimera, Rheinheimera japonica sp. nov. The type strain of this species is KMM 9513(T) = NRIC 0918(T).

Structure of an acidic O-specific polysaccharide of the marine bacterium Pseudoalteromonas agarivorans KMM 232 (R-form).

An acidic O-specific polysaccharide containing L-rhamnose, 2-acetamido-2-deoxy-D-galactose, 2,6-dideoxy-2-(N-acetyl-L-threonine)amino-D-galactose, and 2-acetamido-2-deoxy-D-mannuronic acid was obtained by mild acid degradation of the lipopolysaccharide of the marine bacterium Pseudoalteromonas agarivorans KMM 232 (R-form) followed by gel-permeation chromatography. The polysaccharide was subjected to Smith degradation to give a modified polysaccharide with trisaccharide repeating unit containing L-threonine. The initial and modified polysaccharides were studied by sugar analysis and 1H- and 13C-NMR spectroscopy, including COSY, TOCSY, ROESY, and HSQC experiments, and the structure of the branched tetrasaccharide repeating unit of the polysaccharide was established.

Oceanisphaera litoralis gen. nov., sp. nov., a novel halophilic bacterium from marine bottom sediments.

A polyphasic taxonomic study was performed to characterize a new bacterial isolate, designated KMM 3654(T), from a marine bottom sand sample. The strain was Gram-negative, encapsulated, aerobic, moderately halophilic and grew between 0.5 and 10 % NaCl and at 4-42 degrees C. Its DNA G+C content was 56.4 mol%. Isolate KMM 3654(T) was phylogenetically closely related to members of the genus Oceanimonas, showing 96.7 and 95.6 % 16S rRNA gene sequence similarity to Oceanimonas doudoroffii DSM 7028(T) and Oceanimonas baumannii ATCC 700832(T), respectively. Strain KMM 3654(T) shared some physiological and chemotaxonomic properties with these two Oceanimonas species, but differed from them in morphology, growth at 4 degrees C, urease activity, weak phenol degradation and utilization of phenylacetate. On the basis of phenotypic and phylogenetic evidence, Oceanisphaera litoralis gen. nov., sp. nov. is proposed, with the type strain KMM 3654(T) (=DSM 15406(T)).

Halomonas halocynthiae sp. nov., isolated from the marine ascidian Halocynthia aurantium.

The marine bacterium strain KMM 1376T was isolated from gill tissue of the ascidian Halocynthia aurantium, an inhabitant of the coastal waters of the Sea of Japan. Strain KMM 1376T is an aerobic, gram-negative, non-motile, encapsulated, non-pigmented, slightly halophilic bacterium that is characterized by fimbria-like structures, growth in 0.5-15% NaCl at 7-35 degrees C and absence of acid production from many carbohydrates. The DNA G+C content is 54 mol%. The main fatty acids are C16:0, C16.1omega7c and C18:1omega9c. Comparative 16S rDNA sequence analysis of strain KMM 1376T revealed 92.7-95.3% sequence similarity to members of Halomonas. Based on phenotypic and phylogenetic characteristics, it is proposed that the novel isolate be classified as Halomonas halocynthiae sp. nov., with the type strain KMM 1376T (= DSM 14573T).

[Degradation of fucoidan by the marine proteobacterium Pseudoalteromonas citrea]. / Osobennosti degradatsii fukoidana morskimi proteobakteriiami Pseudoalteromonas citrea.

It was found that Pseudoalteromonas citrea strains KMM 3296 and KMM 3298 isolated from the brown algae Fucus evanescens and Chorda filum, respectively, and strain 3297 isolated from the sea cucumber Apostichopus japonicus are able to degrade fucoidans. The fucoidanases of these strains efficiently degraded the fucoidan of brown algae at pH 6.5-7.0 and remained active at 40-50 degrees C. The endo-type hydrolysis of fucoidan resulted in the formation of sulfated alpha-L-fucooligosaccharides. The other nine strains of P. citrea studied (including the type strain of this species), which were isolated from other habitats, were not able to degrade fucoidan.

Chemical characterization of lipid A from some marine proteobacteria.

Lipids A from type and wild strains of marine Proteobacteria belonging to Alteromonadaceae (Alteromonas (1 species), Idiomarina (1 species), and Pseudoalteromonas (8 species) genera) and Vibrionaceae (Shewanella (1 species) and Vibrio (1 species) genera) families and Marinomonas genus (1 species) were isolated by hydrolysis of their respective lipopolysaccharides with 1% acetic acid. Based on thin-layer chromatography data, the lipids A studied had low heterogeneity and generated family-specific patterns varying in numbers of bands and their chromatographic mobility. Total chemical analysis of the compounds showed that they contained glucosamine, phosphate, and fatty acids with decanoate (I. zobellii KMM 231(T) lipid A) or dodecanoate (lipids A of the other bacteria) and 3-hydroxy alkanoates as the major fatty acid components. Unlike terrestrial bacterial lipids A, lipids A of marine Proteobacteria had basically monophosphoryl (except V. fluvialis AQ 0002B lipid A with its two phosphate groups) and pentaacyl (except S. alga 48055 and V. fluvialis AQ 0002B lipids A which were found to have six residues of fatty acids per molecule of glucosamine disaccharide) structural types, low toxicity, and may be useful as potential endotoxin antagonists.

O-specific polysaccharide of the marine bacterium "Alteromonas marinoglutinosa" NCIMB 1770.

The O-specific polysaccharide of the marine bacterium "Alteromonas marinoglutinosa" NCIMB 1770 was obtained by mild acid degradation of the corresponding lipopolysaccharide and found to contain D-galactose, N-acetyl-D-glucosamine, and N-acetyl-D-mannosamine residues in equimolar ratio. Based on methylation analysis, periodate oxidation, and 13C-NMR spectroscopy data of native and modified polysaccharides, the following structure of the trisaccharide repeating unit of the O-specific polysaccharide was established: [structure: see text]

Retrieval of the species Alteromonas tetraodonis Simidu et al. 1990 as Pseudoalteromonas tetraodonis comb. nov. and emendation of description.

A polyphasic taxonomy study was undertaken of three strains of Pseudoalteromonas haloplanktis subsp. tetraodonis (Simidu et al. 1990) Gauthier et al. 1995. DNA was prepared from each of the strains and genomic relatedness was measured by DNA-DNA hybridization. Strains KMM 458T and IAM 14160T shared 99% genetic relatedness, but were only 48-49% related to the type strain of Pseudoalteromonas haloplanktis subsp. haloplanktis, IAM 12915T. The third strain, P. haloplanktis subsp. tetraodonis A-M, showed 83% genetic similarity with P. haloplanktis subsp. haloplanktis IAM 12915T and 32% with KMM 458T. From these results, it is concluded that strains KMM 458T and IAM 14160T comprise a separate species, originally described as Alteromonas tetraodonis, whereas strain A-M belongs to the species Pseudoalteromonas haloplanktis. Based on phenotypic and chemotaxonomic data, genomic fingerprint patterns, DNA-DNA hybridization data and phylogenetic analysis of 16S rRNA, it is proposed that the species Alteromonas tetraodonis be retrieved and recognized as Pseudoalteromonas tetraodonis comb. nov. (type strain IAM 14160T = KMM 458T).

Structure of an acidic polysaccharide from a marine bacterium Pseudoalteromonas distincta KMM 638 containing 5-acetamido-3,5,7,9-tetradeoxy-7-formamido-L-glycero-L-manno-nonulosonic acid.

An acidic polysaccharide was obtained from the lipopolysaccharide of Pseudoalteromonas distincta strain KMM 638, isolated from a marine sponge, and found to contain D-GlcA, D-GalNAc, 2-acetamido-2,6-dideoxy-D-glucose (D-QuiNAc) and two unusual acidic amino sugars: 2-acetamido-2-deoxy-D-galacturonic acid (D-GalNAcA) and 5-acetamido-3,5,7,9-tetradeoxy-7-formamido-L-glycero-L-manno-nonulosonic acid (Pse5Ac7Fo, a derivative of pseudaminic acid). Oligosaccharides were derived from the polysaccharide by partial acid hydrolysis and mild alkaline degradation and characterised by electrospray ionisation (ESI) MS and 1H and 13C NMR spectroscopy. Based on these data and NMR spectroscopic studies of the initial and O-deacetylated polysaccharides, including quaternary carbon detection, 2D COSY, TOCSY, ROESY, H-detected 1H,13C HMQC and HMBC experiments, the following structure of the branched pentasaccharide repeating unit was established: [structure: see text].

Reclassification of Alteromonas distincta Romanenko et al. 1995 as Pseudoalteromonas distincta comb. nov.

The 16S rRNA gene of Alteromonas distincta KMM 638T was amplified, cloned and sequenced. The nucleotide sequence was aligned with sequences of representative strains of Alteromonas, Moritella, Pseudoalteromonas and Shewanella. Results of phylogenetic analysis, using neighbour-joining and Fitch-Margoliash methods, clearly indicated that this species should be assigned to the genus Pseudoalteromonas. On the basis of polyphasic data obtained from previous work and this study, it is proposed that the species Alteromonas distincta be reclassified as Pseudoalteromonas distincta comb. nov. with type strain KMM 638T (= ATCC 700518T).

Idiomarina gen. nov., comprising novel indigenous deep-sea bacteria from the Pacific Ocean, including descriptions of two species, Idiomarina abyssalis sp. nov. and Idiomarina zobellii sp. nov.

Two bacterial strains, KMM 227T and 231T, were isolated from seawater samples collected from the north-western Pacific Ocean at a depth of 4000-5000 m and were characterized using polyphasic taxonomy. Both were Gram-negative, psychrotolerant, heterotrophic, aerobic and required NaCl for growth (0.6-15.0%). The temperature for growth was 4-30 degrees C. Both strains were rod-shaped, with a single flagellum. However, strain KMM 231T revealed a single long fimbrium. Cellular fatty acids detected in the isolates were predominantly odd-numbered and iso-branched, with 15 and 17 carbons (ca. 70%). Also present were saturated and monounsaturated straight-chain fatty acids. Results of phylogenetic analyses, employing three tree-making methods, strongly indicated that the two strains formed a distinct lineage within a clade containing the genera Alteromonas, Colwellia and Pseudoalteromonas, in the gamma-Proteobacteria. The two strains shared 16S rDNA sequence similarity of 96.9% and genomic DNA relatedness of 27%; the latter was determined by dot-blot hybridization. The strains were differentiated by the presence of fimbria, production of chitinase, ability to grow on 15% NaCl and BIOLOG profiles. Given the polyphasic evidence accumulated in this study, it is proposed that the two deep-sea isolates be classified in the genus Idiomarina gen. nov., as Idiomarina abyssalis sp. nov. (type strain is KMM 227T) and Idiomarina zobellii sp. nov. (type strain is KMM 231T).

[Microbiological monitoring of causative agents of sapronoses in the water of the Bogatinskoe water reservoir]. / Mikrobiologicheskii monitoring vozbuditelei sapronozov v bode Bogatinskogo vodokhranilishcha.

The results of the microbiological monitoring of potential causative agents of sapronoses in the water of the Bogatinskoye reservoir revealed that in the summer period of 1998 the mass accumulation of virulent Aeromonas sobria (up to 25% of the total number of heterotrophic bacteria) took place. The autumn period was characterized by a decrease in the number of A. sobria and the detection of bacteria of the genus Vibrio (up to 22%) with V. mimicus and V. metschnikovii identified among them in the water ecosystems of the southern regions of the Maritime Territory.

Structure of sulfated O-specific polysaccharide of the marine bacterium Pseudoalteromonas marinoglutinosa KMM 232.

A sulfated O-specific polysaccharide containing D-mannose, L-rhamnose, and the sulfate group was obtained by mild acid hydrolysis of lipopolysaccharide (S-form) of the marine bacterium Pseudoalteromonas marinoglutinosa KMM 232. Based on analysis of methylation and 13C-NMR spectroscopy of native and desulfated polysaccharides, the following structure of disaccharide repeat unit in the O-specific polysaccharide has been established: [scheme]. This is the first report of a sulfated O-specific polysaccharide isolated from gram-negative bacteria.

[Structure of teichoic acids from marine microorganisms Bacillus subtilis and Bacillus licheniformis]. / Struktura teikhoevykh kislot iz morskikh mikroorganizmov Bacillus subtilis i B. licheniformis.

Teichoic acids from the cell walls of marine bacilli Bacillus subtilis CMM (Collection of Marine Microorganisms) 234 (R-1) and B. licheniformis CMM 454 (1-1G-2) were isolated and characterized. These teichoic acids were found to have identical structures and are composed of the glucose, ribitol, and phosphoric acid residues. On the basis of 13C NMR and 31P NMR spectra of the teichoic acids and the products of their dephosphorylation, we established the following structure for the biopolymer: poly[-->5)-4-O-beta-D-glucopyranosylribitol-(1-phospho].

Structure of a highly acidic O-specific polysaccharide of lipopolysaccharide of Pseudoalteromonas haloplanktis KMM 223 (44-1) containing L-iduronic acid and D-QuiNHb4NHb.

An acidic O-specific polysaccharide was obtained by mild acid degradation of the lipopolysaccharide isolated by phenol-water extraction of Pseudoalteromonas haloplanktis strain KMM 223 (44-1). L-Iduronic acid (IdoA) was found to be a component of the polysaccharide and identified by NMR spectroscopy and after carboxyl-reduction followed by acid hydrolysis and acetylation, by GLC-MS as 2,3,4-tri-O-acetyl-1,6-anhydroidose. On the basis of 1H and 13C NMR spectroscopic studies, including 1D NOE, 2D NOESY, HSQC and HMBC experiments, the following structure of the branched pentasaccharide repeating unit of the polysaccharide was established: -->4)-beta-D-GlcpAI-(1-->4)-beta-D-GlcpAII-(1-->3)-beta-D-++ +QuipNHb4NHbII- (1-->2)-alpha-L-IdopA-(-->4 increases 1 alpha-D-QuipNAc4NAcI where QuiNAc4NAc and QuiNHb4NHb are 2,4-diacetamido-2,4,6-trideoxyglucose and 2,4,6-tri-deoxy-2,4- di[(S)-3-hydroxybutyramido]glucose, respectively. This is the first report of L-iduronic acid in a lipopolysaccharide and of D-QuiNHb4NHb in nature.

[Structure of glycerophosphate-containing O-specific polysaccharide from Pseudoalteromonas sp. KMM 639]. / Struktura glitserofosfatsoderzhashchego O-spetsificheskogo polisakharida Pseudoalteromonas sp. KMM 639.

On the basis of acid hydrolysis, dephosphorylation, methylation, and 13C NMR spectroscopy data, the O-specific polysaccharide of Pseudoalteromonas sp. KMM 639 was shown to be a glycerophosphate-containing polymer built of repeating disaccharide units of the following structure: [formula: see text]

Evidence for several types of biologically active substances in north Pacific sea anemones.

1. Twenty-two sea anemone samples from seven species were collected in Aleutian and Comandorskiye Islands from sub-littoral region (> 50m depth). 2. Water-ethanol extracts of sea anemones were tested using various test-systems after ethanol evaporation. 3. All sea anemones extracts inhibited DNA and most of them inhibited RNA synthesis in Ehrlich carcinoma tumor cells. 4. Extracts of most sea anemones species showed high hemolytic activity. 5. The extracts proved to be nontoxic or display low toxicity being i.p. injected into mice. 6. Some extracts precipitated virus of aleutian disease of mink. 7. None of the extracts showed activity toward Gram +ve, Gram -ve bacteria or yeast.