Structure of the core oligosaccharide of a rough-type lipopolysaccharide of Pseudomonas syringae pv. phaseolicola.

The core structure of the lipopolysaccharide (LPS) isolated from a rough strain of the phytopathogenic bacterium Pseudomonas syringae pv. phaseolicola, GSPB 711, was investigated by sugar and methylation analyses, Fourier transform ion-cyclotron resonance ESI MS, and one- and two-dimensional 1H-, 13C- and 31P-NMR spectroscopy. Strong alkaline deacylation of the LPS resulted in two core-lipid A backbone undecasaccharide pentakisphosphates in the ratio approximately 2.5 : 1, which corresponded to outer core glycoforms 1 and 2 terminated with either L-rhamnose or 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo), respectively. Mild acid degradation of the LPS gave the major glycoform 1 core octasaccharide and a minor truncated glycoform 2 core heptasaccharide, which resulted from the cleavage of the terminal Kdo residues. The inner core of P. syringae is distinguished by a high degree of phosphorylation of L-glycero-D-manno-heptose residues with phosphate, diphosphate and ethanolamine diphosphate groups. The glycoform 1 core is structurally similar but not identical to one of the core glycoforms of the human pathogenic bacterium Pseudomonas aeruginosa. The outer core composition and structure may be useful as a chemotaxonomic marker for the P. syringae group of bacteria, whereas a more conserved inner core structure appears to be representative for the whole genus Pseudomonas.     

Structural studies of the capsular polysaccharide and lipopolysaccharide O-antigen of Aeromonas salmonicida strain 80204-1 produced under in vitro and in vivo growth conditions.

Aeromonas salmonicida is a pathogenic aquatic bacterium and the causal agent of furunculosis in salmon. In the course of this study, it was found that when grown in vitro on tryptic soy agar, A. salmonicida strain 80204-1 produced a capsular polysaccharide with the identical structure to that of the lipopolysaccharide O-chain polysaccharide. A combination of 1D and 2D NMR methods, including a series of 1D analogues of 3D experiments, together with capillary electrophoresis-electrospray MS (CE-ES-MS), compositional and methylation analyses and specific modifications was used to determine the structure of these polysaccharides. Both polymers were shown to be composed of linear trisaccharide repeating units consisting of 2-acetamido-2-deoxy-D-galacturonic acid (GalNAcA), 3-[(N-acetyl-L-alanyl)amido]-3,6-dideoxy-D-glucose[3-[(N-acetyl-L-alanyl)amido]-3-deoxy-D-quinovose, Qui3NAlaNAc] and 2-acetamido-2,6-dideoxy-D-glucose (2-acetamido-2-deoxy-D-quinovose, QuiNAc) and having the following structure: [-->3)-alpha-D-GalpNAcA-(1-->3)-beta-D-QuipNAc-(1-->4)-beta-D-Quip3NAlaNAc-(1-]n, where GalNAcA is partly presented as an amide and AlaNAc represents N-acetyl-L-alanyl group. CE-ES-MS analysis of CPS and O-chain polysaccharide confirmed that 40% of GalNAcA was present in the amide form. Direct CE-ES-MS/MS analysis of in vivo cultured cells confirmed the formation of a novel polysaccharide, a structure also formed in vitro, which was previously undetectable in bacterial cells grown within implants in fish, and in which GalNAcA was fully amidated.     

Mesozooplankton interactions with the shelf around the sub-Antarctic Prince Edward Islands archipelago

Mesozooplankton surveys were conducted in April/May for fourconsecutive years (1996–1999) in the vicinity of the PrinceEdward Islands (PEIs), Southern Ocean. The PEIs are locatedin the Polar Frontal Zone, directly in the path of the east-flowingAntarctic Circumpolar Current. Zooplankton were collected byoblique tows using a Bongo net fitted with 300 µm mesh.The abundance, biomass and average size of the mesozooplanktonin the upstream (USR), inter-island (IIR) and downstream (DSR)regions indicated that some groups and species were significantlyaffected by their interaction with the shallow shelf watersof the PEIs. Total mesozooplankton abundance and biomass weretypically highest in the DSR, but no consistent pattern wasevident in the USR and IIR. Copepods, euphausiids and fish weregenerally of a low average size in the IIR. This small sizewas largely attributed to the reduced abundance, or completeabsence, of mesopelagic species from the shelf region. Of totalbiomass, the mesopelagic species Euphausia longirostris, Euphausiasimilis, Pleuromamma abdominalis, Paraeuchaeta biloba and Oncaeaantarctica together contributed an average of 16% to the USR,2% to the IIR and 15% to the DSR. Conversely, epipelagic speciesshowed no consistent pattern of abundance and biomass distributionbetween regions. The low incidence of mesopelagic species overthe island shelf was attributed mainly to reduced advectionof deep water into the shelf region (average depth = 200 m),rather than predation, particularly during the through-flowmode between the islands. This resulted in substantial regionaldifferences in euphausiid community structure. The epipelagicspecies Euphausia vallentini and Thysanoessa vicina completelydominated the IIR, comprising on average 89% of total euphausiidbiomass in this region. However, predation may be importantduring the water-trapping mode between the islands. Advectionof zooplankton into the IIR appeared to be affected by the proximityof the Subantarctic Front (SAF). In 1996, when the SAF was farnorth of the PEIs, reduced current velocities resulted in somedegree of water retention over the shelf and an increased predationimpact. Conversely, when the SAF was close to the PEIs in 1999,more large plankton were transported over the island shelf.High current velocities and productivity associated with theSAF appear to increase the biomass and size of allochthonouszooplankton/nekton advected into the IIR, and consequently mayhave increased the availability of prey to land-based predators.The long-term southward movement of the SAF recently observedin the vicinity of the PEIs may therefore have important implicationsfor the ecosystem of these islands.     

Synthesis of the tetrasaccharide repeating-unit of the O-specific polysaccharide from Salmonella senftenberg

The oligosaccharide β-d-Man-(1 → 4)-α-l-Rha (1 → 3)-d-Gal-(6 ← 1)-α-d-Glc, which is the repeating unit of the O-specific polysaccharide chain of the lipopolysaccharide from Salmonella senftenberg, was obtained by glycosylation of benzyl 2,4-di-O-benzyl-6-O-(2,3,4-tri-O-benzyl-6-O-p-nitrobenzoyl-α-d-glucopyranosyl)-β-d-galactopyranoside or benzyl 2-O-acetyl-6-O-(2,3,4-tri-O-benzyl-6-O-p-nitrobenzoyl-α-d-glucopyranosyl)-β-d-galactopyranoside with 3-O-acetyl-4-O-(2,3,4,6-tetra-O-acetyl-β-d-mannopyranosyl)-β-l-rhamnopyranose 1,2-(methyl orthoacetate) followed by removal of protecting groups.     

Study of the microcirculatory bed by a method of demonstrating lactate dehydrogenase]

The method of detection of lactate dehydrogenase with some modification was used to study the microcirculatory bed in total preparations of serous membranes and plane sections of organs of any square surface. The employment of non-fixed material, the short time (from 15 to 60 min) necessary to obtain preparations of any size and simultaneous determination of the localization and the degree of the activity of the enzyme--are, to the authors opinion, undoubtful advantages of the given method, which can be an addition to the well-known impregnation method of determination of the microcirculatory bed.     

Expression of the recombinant antibacterial peptide sarcotoxin IA in Escherichia coli cells

Sarcotoxin IA is an antibacterial peptide that is secreted by a meat-fly Sarcophaga peregrina larva in response to a hypodermic injury or bacterial infection. This peptide is highly toxic against a broad spectrum of both Gram-positive and Gram-negative bacteria and lethal to microbes even at nanomolar concentrations. However, research needs as well as its potential use in medicine require substantial amounts of highly purified sarcotoxin. Because heterologous expression systems proved to be inefficient due to sarcotoxin sensitivity to intracellular proteases, here we propose the biosynthesis of sarcotoxin precursors in Escherichia coli cells that are highly sensitive to the mature peptide. To optimize its biosynthesis, sarcotoxin was translationally fused with proteins highly expressed in E. coli. A fusion partner and the position of sarcotoxin in the chimeric polypeptide were crucial for protecting the sarcotoxin portion of the fusion protein from proteolysis. Released after chemical cleavage of the fusion protein and purified to homogeneity, sarcotoxin displayed antibacterial activity comparable to that previously reported for the natural peptide.     

A large proportion of mediastinal and perirenal visceral fat of Siberian adult people is formed by UCP1 immunoreactive multilocular and paucilocular adipocytes

Journal of Physiology and Biochemistry - Many deleterious consequences for health of excessive fat accumulation are due to visceral fat. Browning of visceral fat is mainly cold dependent and has...     

Trophic-based diversification in benthivorous charrs (Salvelinus) dwelling littoral zones of Northern lakes

Charrs of the genus Salvelinus form distinct trophic morphs living in sympatry in numerous postglacial lakes. Here we demonstrate that charrs can diversify into amphipod foraging specialists and sedentary macrobenthos consumers in the shallow-water ecosystems. This pattern was revealed in three out of six lakes under exploration supported by differences in stomach content, trophic-transmitted parasite, and stable isotope ratio analyzes. The body shape and growth rate comparison indicates that this kind of trophic-based diversification emerges at a juvenile stage and is maintained throughout the whole life. The restriction in gene flow found between the morphs allows to propose the possibility for the hereditable-based specialization to evolve. We found that those diversification phenomena are possible only in the lakes situated in vicinity of the ocean coastline, while no evidence of this split was found for inland mountain lakes. We suggest that the trophic-based diversification in the littoral ecosystems is accounted for the heterogeneity in the ecological conditions and food resources’ distribution linked to coastal wind action. This phenomenon was previously reported for the charr in Lake Fjellfrosvatn, Scandinavia, so it seems to be some universal yet poorly described kind of disruptive selection pressure for northern latitude fishes.