Biomass,composition and activity of organism assemblages along a salinity gradient in sea ice subjected to river discharge in the Baltic Sea

A study was undertaken to examine the activity and composition of the seasonal Baltic Sea land-fast sea-ice biota along a salinity gradient in March 2003 in a coastal location in the SW coast of Finland. Using a multi-variable data set, the less well-known algal and protozoan communities, and algal and bacterial production in relation to the physical and chemical environment were investigated. Also, the first coincident measurements of bacterial production and dissolved organic matter (DOM) in a sea-ice system are reported. Communities in sea ice were clearly autotrophy-dominated with algal biomass representing 79% of the total biomass. Protozoa and rotifers made up 18% of biomass in the ice and bacteria only 3%. Highest biomasses were found in mid-transect bottom ice. Water column assemblages were clearly more heterotrophic: 39% algae, 12% bacteria and 49% for rotifers and protozoa. Few significant correlations existed between DOM and bacterial variables, reflecting the complex origin of ice DOM. Dynamics of dissolved organic carbon, nitrogen and phosphorus (DOC, DON and DOP) were also uncoupled. A functional microbial loop is likely to be present in the studied ice. Existence of an under-ice freshwater plume affects the ecosystem functioning: Under-ice water communities are influenced directly by river-water mixing, whereas the ice system seems to be more independent—the interaction mainly taking place through the formation of active bottom communities.     

Solid-supported 2'-O-glycoconjugation of oligonucleotides by azidation and click reactions

2'-O-[(2-Bromoethoxy)methyl]cytidine and 2'-O-[(2-azidoethoxy)methyl]cytidine have been prepared and introduced as appropriately protected 3'-phosphoramidite (1) and 3'-(H-phosphonate) (2) building blocks, respectively, into 2'-O-methyl oligoribonucleotides. The support-bound oligonucleotides were subjected to two consecutive conjugations with alkynyl-functionalized monosaccharides. The first saccharide was introduced by a Cu(I) promoted click reaction with 2 and the second by azidation of the 2-bromoethoxy group of 1 followed by the click reaction. The influence of the 2'-glycoconjugations on hybridization with DNA and 2'-O-methyl RNA targets was studied. Two saccharide units within a 15-mer oligonucleotide had a barely noticeable effect on the duplex stability, while introduction of a third one moderately decreased the melting temperature.     

Attenuated expression of tenascin-c in ovalbumin-challenged STAT4-/- mice

Background

Asthma leads to structural changes in the airways, including the modification of extracellular matrix proteins such as tenascin-C. The role of tenascin-C is unclear, but it might act as an early initiator of airway wall remodelling, as its expression is increased in the mouse and human airways during allergic inflammation. In this study, we examined whether Th1 or Th2 cells are important regulators of tenascin-C in experimental allergic asthma utilizing mice with impaired Th1 (STAT4-/-) or Th2 (STAT6-/-) immunity.

Methods

Balb/c wildtype (WT), STAT4-/- and STAT6-/- mice were sensitized with intraperitoneally injected ovalbumin (OVA) followed by OVA or PBS airway challenge. Airway hyperreactivity (AHR) was measured and samples were collected. Real time PCR and immunohistochemistry were used to study cytokines and differences in the expression of tenascin-C. Tenascin-C expression was measured in human fibroblasts after treatment with TNF-α and IFN-γ in vitro.

Results

OVA-challenged WT mice showed allergic inflammation and AHR in the airways along with increased expression of TNF-α, IFN-γ, IL-4 and tenascin-C in the lungs. OVA-challenged STAT4-/- mice exhibited elevated AHR and pulmonary eosinophilia. The mRNA expression of TNF-α and IFN-γ was low, but the expression of IL-4 was significantly elevated in these mice. OVA-challenged STAT6-/- mice had neither AHR nor pulmonary eosinophilia, but had increased expression of mRNA for TNF-α, IFN-γ and IL-4. The expression of tenascin-C in the lungs of OVA-challenged STAT4-/- mice was weaker than in those of OVA-challenged WT and STAT6-/- mice suggesting that TNF-α and IFN-γ may regulate tenascin-C expression in vivo. The stimulation of human fibroblasts with TNF-α and IFN-γ induced the expression of tenascin-C confirming our in vivo findings.

Conclusions

Expression of tenascin-C is significantly attenuated in the airways of STAT4-/- mice, which may be due to the impaired secretion of TNF-α and IFN-γ in these mice.     

Scales of horizontal patchiness in chlorophyll a, chemical and physical properties of landfast sea ice in the Gulf of Finland (Baltic Sea)

Horizontal variation of first-year landfast sea ice properties was studied in the Gulf of Finland, the Baltic Sea. Several scales of variation were considered; a number of arrays with core spacings of 0.2, 2 and 20 m were sampled at different stages of the ice season for small-scale patchiness. Spacing between these arrays was from hundreds of meters to kilometers to study mesoscale variability, and once an onshore–offshore 40-km transect was sampled to study regional scale variability. Measured variables included salinity, stable oxygen isotopes (¹⁸O), chlorophyll a (chl-a), nutrients and dissolved organic carbon. On a large scale, a combination of variations in the under-ice water salinity (ice porosity), nutrient supply and the stage of ice development control the build-up of ice algal biomass. At scales of hundreds of meters to kilometers, there was significant variability in several parameters (salinity, chl-a, snow depth and ice thickness). Analyses of the data from the arrays did not show evidence of significant patchiness at scales <20 m for algal biomass. The results imply that the sampling effort in Baltic Sea ice studies should be concentrated on scales of hundreds of meters to kilometers. Using the variations observed in the study area, the estimate for depth-integrated algal biomass in landfast sea ice in the Gulf of Finland (March 2003) is 5.5±4.4 mg chl-a m^–2.     

Hydrolysis of a mRNA 5'-cap model substrate, 5',5'-ApppA by di- and trinuclear zinc(II) complexes of a polyamino-polyol ligand

Copper(II) and zinc(II) complexes of a polyamino-polyol ligand 1,3,5-trideoxy-1,3,5-tris(methylamino)-cis-inositol (tmci) have been investigated as potential candidates for the selective elimination of the 5'-cap structure of mRNA. A cap-model compound ApppA has been utilised as substrate for studying the effect of the different metal ion complex catalysts on the hydrolysis of the triphosphate bridge. Kinetic experiments have been performed by the variation of pH, metal-to-ligand ratio and total concentrations of the metal ion and ligand. The zinc(II) complexes of tmci have been proved to possess a remarkable activity for the hydrolysis of ApppA. The observed rate enhancement compared to the uncatalysed reaction was found to be 12,000-fold, in the presence of 4.5mM zinc(II) and 1.5mM tmci at pH approximately 7.5. In contrast with the copper(II) containing systems, an extra product has also been formed during the cleavage process, beside the expected AMP and ADP. According to the ESI-MS characterisation of the samples, the additional product is a covalent phosphoester adduct of AMP and the ligand. The formation of this species is initiated by a nucleophilic attack of a zinc(II)-bound alcohol or alkoxo group on one of the alpha phosphate groups of ApppA, which leads to the formation of a phosphodiester bond. In an alternative pathway, the substrate is cleaved into AMP and ADP. According to the pH-potentiometric studies, performed with the tmci-zinc(II) system, di- and trinuclear complexes are responsible for the accelerated ApppA hydrolysis. The copper(II)-tmci 2:1 system showed only a modest kinetic activity. The rate acceleration significantly increased when threefold excess of copper(II) was applied. Although, the detailed investigations above pH approximately 6.6 have been prevented by precipitate formation during the addition of the substrate into the reaction solution, the activity of the copper(II)-tmci 3:1 system does not exceed that of the zinc(II) complexes. Due to the specific mechanism leading to the covalent extra product, the zinc(II) complexes of tmci provide a comparable rate enhancement for ApppA hydrolysis to the widely studied lanthanide or copper(II) species, in spite of the fact that they are stronger Lewis acids.     

Stochastic boundary molecular dynamics simulation of L-ribose in the active site of Actinoplanes missouriensis xylose isomerase and its Val135Asn mutant with improved reaction rate

We used molecular dynamics simulations to study how a non-natural substrate, L-ribose, interacts with the active site of Actinoplanes missouriensis xylose isomerase. The simulations showed that L-ribose does not stay liganded in the active site in the same way as D-xylose, in which the oxygens O2 and O4 are liganded to the metal M1. The oxygen O4 of L-ribose moved away from the metal M1 to an upside down position. Furthermore, the distances of the carbons C1 and C2 of L-ribose to the catalytic metal M2 were higher than in the case of D-xylose. These findings explain the extremely low reaction rate of xylose isomerase with L-ribose. The mutation V135N close to the C5-OH of the substrate increased the reaction efficiency 2- to 4-fold with L-ribose. V135N did not affect the reaction with D-xylose and L-arabinose, whereas the reaction with D-glucose was impaired, probably due to a hydrogen bond between Asn-135 and the substrate. When L-ribose was the substrate, Asn-135 formed a hydrogen bond to Glu-181. As a consequence, O4 of L-ribose stayed liganded to the metal M1 in the V135N mutant in molecular dynamics simulations. This explains the decreased K(m) of the V135N mutant with L-ribose.     

CD2AP contributes to cell migration and adhesion in cultured gastric epithelium

The potential association of CD2AP with the adherens junction protein E-cadherin, co-localization with the actin cytoskeleton, and involvement in cell migration was investigated in cultured rat gastric mucosal cells. In stationary cells, CD2AP was localized perinuclearly while E-cadherin was expressed along cell-cell contacts and F-actin formed a branched network and adhesion belts. In migrating cells, CD2AP appeared as thread-like accumulations in the leading edges, colocalizing with F-actin and occasionally with E-cadherin. Intracellular injection of anti-CD2AP significantly retarded the migration speed of the cells suggesting a crucial role for CD2AP in mucosal cell migration, possibly as a scaffolding protein between cell membrane proteins and actin cytoskeleton. Co-immunoprecipitation assays revealed that CD2AP and E-cadherin are in a complex in HGF stimulated cells. It is concluded that CD2AP interacts with E-cadherin and co-localizes with F-actin in the leading edge of migrating cells, and significantly contributes to cell migration in restituting gastric epithelium.     

Topical superantigen exposure induces epidermal accumulation of CD8+ T cells, a mixed Th1/Th2-type dermatitis and vigorous production of IgE antibodies in the murine model of atopic dermatitis

Patients with atopic dermatitis (AD) have repeated cutaneous exposure to both environmental allergens and superantigen-producing strains of Staphylococcus aureus. We used a murine model of AD to investigate the role of staphylococcal enterotoxin B (SEB) in the modulation of allergen-induced skin inflammation. Mice were topically exposed to SEB, OVA, a combination of OVA and SEB (OVA/SEB), or PBS. Topical SEB and OVA/SEB exposure induced epidermal accumulation of CD8+ T cells and TCRVbeta8+ cells in contrast to OVA application, which induced a mainly dermal infiltration of CD4+ cells. SEB and OVA/SEB exposure elicited a mixed Th1/Th2-associated cytokine and chemokine expression profile within the skin. Restimulation of lymph node cells from OVA- and OVA/SEB-exposed mice with OVA elicited strong production of IL-13 protein, whereas substantial amounts of IFN-gamma protein were detected after SEB stimulation of cells derived from SEB- or OVA/SEB-exposed mice. Topical SEB treatment elicited vigorous production of SEB-specific IgE and IgG2a Abs and significantly increased the production of OVA-specific IgE and IgG2a Abs. The present study shows that topical exposure to SEB provokes epidermal accumulation of CD8+ T cells, a mixed Th2/Th1 type dermatitis and vigorous production of specific IgE and IgG2a Abs, which can be related to the chronic phase of atopic skin inflammation.