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The water soluble polysaccharide produced by the green variant of tetrasporic Gigartina skottsbergii was found to be composed of D-galactose and sulfate groups in a molar ratio of 1.0:0.65. (1)H and (13)C NMR spectroscopy studies of the desulfated polysaccharide showed a major backbone structure of alternating 3-linked β-D-galactopyranosyl and 4-linked α-D-galactopyranosyl units, and minor signals ascribed to 3-O-methyl-substitution on the latter unit. Ethylation analysis of the polysaccharide indicated that the sulfate groups are mainly located at position O-2 of 4-linked α-D-galactopyranosyl residue and partially located at positions O-6 of the same unit and at position O-2 of 3-linked β-D-galactopyranosyl residue, and confirmed the presence of 3-O-methyl-galactose in minor amounts (4.4%). The sulfated d-galactan presents a similar structure to λ carrageenan but with much lower sulfation at position O-6 of the α-residue and at position O-2 of β-residue. The antioxidant capacity of the sulfated d-galactan was evaluated by the peroxyl radicals (ORAC method), hydroxyl radicals, chelating activity, and ABTS(+) assays. Kinetic results obtained in these assays were compared with those obtained for the commercial λ carrageenan. The antioxidant activity toward peroxyl radicals was higher for commercial λ carrageenan, this agrees with its higher content of sulfate group. The kinetics of the reaction of both polysaccharides with hydroxyl and ABTS(+) radicals showed a complex mechanism, but the antioxidant activity was higher for the polysaccharide from the green variant of tetrasporic Gigartina skottsbergii., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Anthocyanins in aged Cabernet Sauvignon red wines were studied by HPLC-MS. The major anthocyanin in 6, 7, and 8year old red wine extracts was the dimer vitisin A produced by condensation of malvidin-3-O-glucoside mediated by pyruvic acid. In aged wine, the content of malvidin-3-O-glucoside decreased with age with a concomitant increase of vitisin A. The latter is accompanied by several condensation products with molecular weight up to 1500m/z. Differential pulse voltammetry indicated that aged wines have lower antioxidant capacity than young wine (400mV), but higher than malvidin-3-O-glucoside (483mV). Tafel's plots showed that the electrochemical process occurring in aged wines is different from young wines. Six, seven and eight year old wines show similar behaviour with plots of 234, 177 and 188mV/dec, respectively. These values are higher than the expected 120mV/dec corresponding to a first electronic transfer but smaller compared to the 523mV/dec corresponding to young wine., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The aim of this study was to evaluate the usefulness of oxidative stress biomarkers of pollution in native mussels Mytilus edulis chilensis from the Beagle Channel. Spatial and seasonal variations of catalase, glutathione-S-transferase and lipid peroxidation in gills and digestive gland were analyzed in relation to environmental parameters, heavy metals in sediment and in tissue. Four sites with anthropogenic impact and a control site were selected and monitored during the four seasons of 2007. We found significant differences among sites in concentrations of dissolved nutrients and heavy metals in sediments, with the highest values recorded at sites with anthropogenic pressure. Different patterns were observed between concentrations of metals in tissues and in sediments suggesting differences in bioavailability. There were also significant differences in biomarker responses among sites, despite the strong seasonal variability. Our results showed relatively moderate levels of pollution in the study area as a result of urban influences., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

A coastal system (Ushuaia Bay, Argentina) impacted by anthropogenic activities was studied by the response of local amphipods (Parmorea sp., Gammaridae) to the exposure of coastal sediments in a laboratory assay. Four coastal areas with different loadings of contaminants and one considered as reference were studied. Organic matter, carbohydrates, proteins and heavy metals were measured in sediment samples. Organisms were exposed to sediments for seven days and catalase (CAT), glutathione S-transferase (GST), acetylcholinesterase (AChE) and lipid peroxidation (LPO) were measured afterward. Amphipods exhibited an activation of GST and inhibition of AChE in most impacted areas. Principal Component Analysis (PCA) was conducted in order to associate the biological responses with sediment metal concentration and its eutrophicated status. Levels of Cd and Cr were associated with the inhibition of AChE and with the enhancement of GST. CAT and LPO were enhanced in most areas, but no link was found with the contaminants studied by PCA, suggesting that other parameters present in sediments not included in the PCA affect the amphipods. The most impacted area corresponds to Nautical Club station, with a highly eutrophicated status and high content of metals, where amphipods after the exposure were affected in a biochemical level., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Water quality surrounding the mussel farm of Mytilus edulis chilensis at Brown Bay (Beagle Channel) was evaluated. The levels of five heavy metals in sediment and in gill and digestive gland of mussels were examined to consider potential risks to human health. Cd showed the highest enrichment factor in relation to its level in Earth crust (3.85-21.58), which could be related to an upwelling phenomenon. A seasonal trend was found regarding metal bioaccumulation, being higher in winter than in summer. The bioaccumulation pattern in gill was Zn>Fe>Cu>Cd, meanwhile in digestive gland was Fe>Zn>Cu>Cd. Despite Pb was measured in sediment (15.59-23.91 microg/g dw), it was not available for being incorporated by mussels. In all cases it was below the detection limit (2.37 microg/g dw). With regard to human consumption of mussels from Brown Bay, none of the elements analyzed should cause concern for consumers. Values measured in tissue mussels were below the limit of 10 microg/g dw for Cd and Pb established by SENASA for molluscs. Considering that studied mussels are for human consumption and the relatively high levels of metals in sediment may vary their availability if physical parameters changes, periodical monitoring must be carried out to avoid human risks and to produce food in a responsible manner that complies with the food safety standards., (Copyright (c) 2010 Elsevier Inc. All rights reserved.)

The O-antigen determines the specificity of bacterial serotype, a sort of bacterial fingerprinting. In this work we report the extraction, purification and characterization of the O-antigen of two pathogenic bacteria, Escherichia coli O16 and Salmonella typhimurium. Molecular fingerprints found in the vibrational spectra represent a powerful analytical technique for identification (or differentiation) of molecular moieties in complex systems such as pathogens. In addition, advantages of vibrational Raman scattering are unique thanks to the high sensitivity and specificity achieved via surface-enhanced Raman scattering (SERS). SERS is used here to take advantage of characteristic vibrational frequency differences of O-antigens, thus allowing bacterial differentiation. Characteristic fundamental vibrational modes associated with the monosaccharide N-acetylglucosamine and deformations of the O-antigen chains provide the main spectroscopic differences between the O-antigens of E. coli O16 and S.typhimurium.

A homology model of Saccharomyces cerevisiae phosphoenolpyruvate (PEP) carboxykinase (ATP + oxaloacetate right arrow over left arrow ADP + PEP + CO(2)) in complex with its substrates shows that the isobutyl group of Leu460 is in close proximity to the adenine ring of the nucleotide, while the carboxyl group of Glu299 is within hydrogen-bonding distance of the ribose 2'OH. The Leu460Ala mutation caused three-fold and seven-fold increases in the K (m) for ADPMn(-) and ATPMn(2-), respectively, while the Glu299Ala mutation had no effect. Binding studies showed losses of approximately 2 kcal mol(-1) in the nucleotide binding affinity due to the Leu460Ala mutation and no effect for the Glu299Ala mutation. PEP carboxykinase utilized 2'deoxyADP and 2'deoxyATP as substrates with kinetic and equilibrium dissociation constants very similar to those of ADP and ATP, respectively. These results show that the hydrophobic interaction between Leu460 and the adenine ring of the nucleotide significantly contributed to the nucleotide affinity of the enzyme. The 2'deoxy nucleotide studies and the lack of an effect of the Glu299Ala mutation in nucleotide binding suggest that the possible hydrogen bond contributed by Glu299 and the ribose 2'OH group may not be relevant for nucleotide binding.

Phosphoenolpyruvate (PEP) carboxykinases catalyse the reversible formation of oxaloacetate (OAA) and ATP (or GTP) from PEP, ADP (or GDP) and CO(2). They are activated by Mn(2+), a metal ion that coordinates to the protein through the epsilon-amino group of a lysine residue, the N(epsilon-2)-imidazole of a histidine residue, and the carboxylate from an aspartic acid residue. Neutrality in the epsilon-amino group of Lys213 of Saccharomyces cerevisiae PEP carboxykinase is expected to be favoured by the vicinity of ionised Lys212. Glu272 and Glu284, located close to Lys212, should, in turn, electrostatically stabilise its positive charge and hence assist in keeping the epsilon-amino group of Lys213 in a neutral state. The mutations Glu272Gln, Glu284Gln, and Lys212Met increased the activation constant for Mn(2+) in the main reaction of the enzyme up to seven-fold. The control mutation Lys213Gln increased this constant by ten-fold, as opposed to control mutation Lys212Arg, which did not affect the Mn(2+) affinity of the enzyme. These observations indicate a role for Glu272, Glu284, and Lys212 in assisting Lys213 to properly bind Mn(2+). In an unexpected result, the mutations Glu284Gln, Lys212Met and Lys213Gln changed the nucleotide-independent OAA decarboxylase activity of S. cerevisiae PEP carboxykinase into an ADP-requiring activity, implying an effect on the OAA binding characteristics of PEP carboxykinase., (Copyright 2010 Elsevier Masson SAS. All rights reserved.)

Mussels coming from a mussel farm at Brown Bay (Beagle Channel) were transplanted to four sites inside Ushuaia Bay for 2 and 4 weeks. The objective of this study was to assess the quality of coastal waters of Ushuaia Bay by measuring catalase activity, lipid peroxidation, total lipid content, bioaccumulation of heavy metals and condition index in transplanted mussel Mytilus edulis chilensis. Biomarkers except condition index showed significant differences among exposure times as well as among tissues. Digestive gland presented the highest catalase activity, malondialdehyde level and total lipid content. Digestive gland also was the main target tissue of accumulation of iron and copper, while gill accumulated the highest levels of zinc. A principal component analyzes with the whole set of data allowed to separate stations based on physicochemical conditions and biochemical responses of each studied area., ((c) 2009 Elsevier Inc. All rights reserved.)

To assess the hypothesis that gap junctions (GJs) participate on leukocyte-endothelium interactions in the inflammatory response, we compared leukocyte adhesion and transmigration elicited by cytokine stimulation in the presence or absence of GJ blockers in the hamster cheek pouch and also in the cremaster muscle of wild-type (WT) and endothelium-specific connexin 43 (Cx43) null mice (Cx43e(-/-)). In the cheek pouch, topical tumor necrosis factor-alpha (TNF-alpha; 150 ng/ml, 15 min) caused a sustained increment in the number of leukocytes adhered to venular endothelium (LAV) and located at perivenular regions (LPV). Superfusion with the GJ blockers 18-alpha-glycyrrhetinic acid (AGA; 75 microM) or 18-beta-glycyrrhetinic acid (50 microM) abolished the TNF-alpha-induced increase in LAV and LPV; carbenoxolone (75 microM) or oleamide (100 microM) reduced LAV by 50 and 75%, respectively, and LPV to a lesser extent. None of these GJ blockers modified venular diameter, blood flow, or leukocyte rolling. In contrast, glycyrrhizin (75 microM), a non-GJ blocker analog of AGA, was devoid of effect. Interestingly, when AGA was removed 90 min after TNF-alpha stimulation, LAV started to rise at a similar rate as in control. Conversely, application of AGA 90 min after TNF-alpha reduced the number of previously adhered cells. In WT mice, intrascrotal injection of TNF-alpha (0.5 microg/0.3 ml) increased LAV (fourfold) and LPV (threefold) compared with saline-injected controls. In contrast to the observations in WT animals, TNF-alpha stimulation did not increase LAV or LPV in Cx43e(-/-) mice. These results demonstrate an important role for GJ communication in leukocyte adhesion and transmigration during acute inflammation in vivo and further suggest that endothelial Cx43 is key in these processes.

The distribution of urocanic acid (UCA) isomers between aqueous solutions and n-octanol, egg yolk phosphatidylcholine (eggPC) liposomes or bovine serum albumin (BSA) has been evaluated. Regarding its partitioning between water and n-octanol, the behaviour of both isomers is very similar, and the amount incorporated to the organic solvent is mostly determined by the fraction of the compound that, in the aqueous phase, is present as uncharged species. This implies that the highest hydrophobicity occurs near the isoelectric point. cis- and trans-UCA are readily incorporated into eggPC unilamellar liposomes. A simple pseudophase treatment of ultrafiltration data renders a binding constant of 0.20+/-0.04mL/mg for the trans isomer at pH 7.4. The binding constant decreases, by a factor two, at pH 5.0, suggesting that the negatively charged species is more favourably bound to the liposomes than the neutral species, which is mostly present as zwitterions. The cis-isomer, at both pHs, is less incorporated to the bilayers. trans-UCA and cis-UCA readily bind to BSA at pH 7.4, with binding constants of 3400M(-1) and 6900M(-1), respectively. This result suggests that, as in the octanol/water partitioning, hydrophobic interactions predominate and the degree of binding is determined by the fraction present as uncharged species. A smaller binding constant at pH 5.0 indicates that the charge of the protein is also plying a relevant role.

Acute toxicity of copper, cadmium, and zinc on isopod Exosphaeroma gigas was evaluated at 20 per thousand and 30 per thousand salinity. Six concentrations were assayed to estimate effective concentration of the toxicant that affects 50% of tested animal (EC50), while physiological responses and bioaccumulation were determined at 0.42 and 1.95 mg L(-1) of each metal. The following toxicity orders were obtained: Cd>or=Cu>Zn at 20 per thousand salinity and Cu>Zn>or=Cd at 30 per thousand salinity. Copper treatments showed a reduction in oxygen consumption at 30 per thousand salinity, while an opposite trend was observed at 20 per thousand salinity. Zinc caused dissimilar effects, while in cadmium significant reduction was only registered in 0.42(20 per thousand) mg L(-1). Ammonia excretion was generally higher in treatments than control at 20 per thousand salinity. At 30 per thousand salinity, excretion did not change in relation to control or diminished. In general terms, O:N atomic ratios indicated a preponderant protein metabolism. Bioaccumulation of assayed metals was higher at lowest salinity and increased with increasing toxic concentrations.

Column leaching experiments were conducted to test the ability of Chilean volcanic soils in retaining the mineral constituents and metals in sewage sludge and sludge ash that were incorporated into the soils. Small or negligible amounts of the total content of Pb, Fe, Cr, Mn, Cd, and Zn (0 to <2%), and more significant amounts of mineral constituents such as Na (7-9%), Ca (7-13%), PO4 (4-10%), and SO4 (39-46%) in the sludge and sludge ash were readily soluble. When they were incorporated on the surface layer of the soils and leached with 12 pore volumes of water over a 3 month period of time, less than 0.1% of the total amount of heavy metals and PO4 in the sludge and sludge ash were collected in the drainage water. Cation exchange selectivity, specific anion adsorption and solubility are the processes that cause the reduction of leaching. The volcanic soils were capable of retaining the mineral constituents, P, and metals in applied sewage sludge and sludge ash and gradually released them as nutrients for plant growth.