Role of galloylation and polymerization in cytoprotective effects of polyphenolic fractions against hydrogen peroxide insult

Byproducts and wastes generated by agricultural, food, and forestry industries contain large amounts of polyphenols, which can be potentially used as sources of natural or semisynthetic antioxidants. This study examined and compared the protection against peroxidative damage induced in erythrocytes and 3T3 cell line of polyphenolic fractions from white grape pomace, pine bark, and witch hazel bark. The work pays special attention to the different degrees of polymerization and galloylation of the extracts to contribute to the understanding of their mechanisms of action. Fractions demonstrated different protections against erythrocyte lipid peroxidation, hemolysis, and 3T3 cytotoxicity caused by H(2)O(2). Galloylation is claimed to be related to antioxidant protective capacity, and it is also responsible for the pro-oxidant effect observed at high doses. The results show that not only the percentage of galloylation but also the degree of polymerization are important modulators of their antioxidant capacity. In this sense, it is crucial that novel polyphenolic fractions were prepared attending a value of 3 for the mean degree of polymerization and did not exceed a 30% of galloylation to reach the highest antioxidant capacity with the lowest cytotoxic effects. For this reason, the grape extracts appear to be the best strategy to fight against hydrogen peroxide cell damage.     

Immunomodulatory activity of a new family of antioxidants obtained from grape polyphenols

We examined the potential antioxidant activity and the immunopharmacological activity of new epicatechin conjugates obtained by depolymerization of grape polymeric flavanols in the presence of cysteamine or cysteine and with or without gallate. The compounds studied were (-)-epicatechin (1), cysteinyl-epicatechin (2), cysteamine-epicatechin (3), (-)-epicatechin gallate (4), cysteinyl-epicatechin gallate (5), and cysteamine-epicatechin gallate (6) When incubated with an erythrocyte suspension, flavanols protected the erythrocyte membrane from hemolysis induced by 2,2'-azobis(2-amidinopropane) dihydrochloride, an azo free-radical initiator. All the epicatechin derivatives tested were more efficient as antioxidant than epicatechin. The most potent antioxidant was compound 6. The compounds were tested for their capacity to modulate IL-1beta and IL-6, which are the main cytokine factors influencing the acute phase of the inflammatory response. (-)-Epicatechin and its related compounds inhibited the production of IL-1beta and IL-6 in whole blood incubated in the presence of Escherichia coli lipopolysaccharide. The most efficient inhibitor of cytokine formation was compound 3.     

Estimating the Health Impact of Climate Change With Calibrated Climate Model Output

Studies on the health impacts of climate change routinely use climate model output as future exposure projection. Uncertainty quantification, usually in the form of sensitivity analysis, has focused predominantly on the variability arise from different emission scenarios or multi-model ensembles. This paper describes a Bayesian spatial quantile regression approach to calibrate climate model output for examining to the risks of future temperature on adverse health outcomes. Specifically, we first estimate the spatial quantile process for climate model output using non-linear monotonic regression during a historical period. The quantile process is then calibrated using the quantile functions estimated from the observed monitoring data. Our model also down-scales the gridded climate model output to the point-level for projecting future exposure over a specific geographical region. The quantile regression approach is motivated by the need to better characterize the tails of future temperature distribution where the greatest health impacts are likely to occur. We applied the methodology to calibrate temperature projections from a regional climate model for the period 2041 to 2050. Accounting for calibration uncertainty, we calculated the number of excess deaths attributed to future temperature for three cities in the US state of Alabama.     

Characterization of pigments from different high speed countercurrent chromatography wine fractions

A red wine, made from Cabernet Sauvignon (60%) and Tannat (40%) cultivars, was fractionated by high speed countercurrent chromatography (HSCCC). The biphasic solvent system consisting of tert-butyl methyl ether/n-butanol/acetonitrile/water (2/2/1/5, acidified with 0.1% trifluoroacetic acid) was chosen for its demonstrated efficiency in separating anthocyanins. The different native and derived anthocyanins were identified on the basis of their UV-visible spectra, their elution time on reversed-phase high-performance liquid chromatography (HPLC), and their mass spectra, before and after thiolysis. The HSCCC method allowed the separation of different families of anthocyanin-derived pigments that were eluted in different fractions according to their structures. The hydrosoluble fraction was almost devoid of native anthocyanins. Further characterization (glucose quantification, UV-visible absorbance measurements) indicated that it contained flavanol and anthocyanin copolymers in which parts of the anthocyanin units were in colorless forms. Pigments in the hydrosoluble fraction showed increased resistance to sulfite bleaching and to the nucleophilic attack of water.     

Development of a method for the genetic identification of flatfish species on the basis of mitochondrial DNA sequences

In the present study a method for genetic identification of flatfish species was developed. The technique is based on DNA sequencing of amplified DNA by PCR and subsequent phylogenetic analysis ( FINS). A phylogenetic tree using the cytochrome oxidase subunit I (COI) was constructed and the bootstrap values calculated. The mentioned technique allows the genetic identification of more than 50 flatfish species in fresh, frozen, and precooked products. This analytical system was validated and subsequently applied to 30 commercial samples, obtaining 13 that were incorrectly labeled (43%). Four of the mislabeled samples were whole fish (31%), and nine were fillets (69%). The species with the higher rate of incorrect labeling were Pleuronectes platessa (17%) and Solea solea (10%). Other species incorrectly labeled were Hipoglossus hipoglossus (7%), Reinharditus hippoglossoides, Limanda ferruginea, and Microstomus kitt (3% each species). Therefore, this molecular tool is appropriate to clarify questions related with the correct labeling of commercial products, the traceability of raw materials, and the control of imported flatfish, and also can be applied to questions linked to the control of fisheries.     

Emerging resistant microbiota from an acidic soil exposed to toxicity of Cr, Cd and Pb is mainly influenced by the bioavailability of these metals

Purpose

The effect of pollutants in soil microorganisms is an important issue in order to understand their toxic effects in the environment, as well as for developing adequate bioremediation strategies. In this sense, the main objective of this study was to assess the involvement of the indigenous microbiota of an acidic forest Mediterranean soil by artificial pollution with heavy metals, and to detect and isolate resistant microorganisms that could be useful for bioremediation.

Materials and methods

Samples from a previously unpolluted acidic forest soil were amended with Cr(VI), Cd(II) or Pb(II) at total amounts ranging from 0.1 to 5,000 mg?kg^?1. These soil microcosms were incubated under controlled laboratory conditions for 28 days. Soluble fractions of metals were determined from aqueous extracts. Both activity and composition of the microbial community were assessed, respectively, by respirometric assays and molecular analysis (polymerase chain reaction denaturing gradient gel electrophoresis). The isolation of metal-resistant microorganisms was attempted by culture plating from microcosms incubated with high concentrations of metals. Isolated strains were tested in cultures with minimal medium to check for their metal resistance and their capacity to reduce the presence of toxic Cr(VI).

Results and discussion

A decrease in the soil respirometric activity and changes in the microbial community composition were detected from 10/100 mg?kg^?1 Cr and 1,000 mg?kg^?1 Cd and Pb. Presumably resistant bacterial and fungal populations developed in most of these polluted microcosms; however, the microbiota was severely impaired at the highest additions of Cr. Even though Cr was the most damaging metal in soil microcosms, if the soluble fractions of metals are considered instead of their total added amounts, the comparison among their toxic effects suggests a similar potential toxicity of Cr and Pb. Isolated multiresistant microorganisms were related mainly to Actinobacteria, Firmicutes and Ascomycota. Some of them showed the capacity to reduce Cr(VI) concentrations between 54 % and 70 % of the initial value. These strains were affiliated to several species of Streptomyces and Bacillus.

Conclusions

The combination of respirometric assays with molecular methods has been useful to assess the effect of metals on the soil microbial community, which can greatly be explained by their differential bioavailability. Cultivation-dependent and -independent approaches have proved the presence and development of multiresistant microorganisms in a previously unpolluted soil. Due to their properties, some of the isolated strains are potentially useful for soil bioremediation.     

Land-use and socio-economic correlates of plant invasions in European and North African countries

Biological invasions are causing significant ecological impacts and economical costs world-wide. Human activities are the primary cause of this conservation and environmental problem. We conducted a stepwise regression analysis between several land-use and socio-economic parameters and the density of alien species in European and North African countries. We found that the parameters that best explained the density of alien plants were the Human Development Index and imports. The extent of terrestrial transport networks and the percentage of protected areas were the land-use variables that best accounted for the density of alien plants. If we are going to influence policies dealing with biological invasions, more emphasis should be placed on the analysis of land-use and socio-economic determinants at different spatial scales and their changes throughout time.     

Effects of resource availability on condensed tannins and nitrogen in two Quercus species differing in leaf life span

Seedlings of Quercus ilex and Q. cerrioides, an evergreen and a winter-deciduous oak co-occurring in western-Mediterranean forests, were grown at two light regimes (8 and 36% of photosynthetically active radiation), at two water regimes (500 and 800 mm) and with two nutrient availabilities (standard substrate and 7% increase in soil N). The concentrations of soluble condensed tannins (CT) and nitrogen in the leaves were analyzed to test the phenotypic plasticity of these commonly related parameters in two con-generic species with contrasting leaf habit. Q. ilex contains seven times more CT and a few less N than Q. cerrioides. Light increased CT, whereas neither fertilization nor water had an effect on CT. N concentration was decreased by light, increased by fertilization and not affected by water treatment. Plant growth was increased by light but not affected by fertilization or water treatment. CT were negatively correlated with N concentration. CT of the evergreen species exhibited greater plasticity than the deciduous one as reflected by a steeper negative correlation among nitrogen and CT concentrations in Q. ilex. Given the antiherbivory activity of CT, this implies that in less shaded environments, e.g. canopy aperture by disturbances, leaf tissue quality for herbivores will be much more reduced in Q. ilex than in Q. cerrioides. Higher leaf CT in Q. ilex and its higher plasticity to light availability may explain the higher browsing by sheep in Q. cerrioides than in Q. ilex resprouts, as well as the low recruitment rates of seedlings of the former species, reported in other studies.     

Carbon Isotope Discrimination, Gas Exchange and Stem Growth of Four Euramerican Hybrid Poplars under Different Watering Regimes

Seasonal changes in carbon isotope discrimination (Δ) and gas exchange traits were assessed in four Populus×euramericana clones differing in growth potential. Measurements were made during the second year after establishment in the field under two watering regimes, which were defined by the time-span between flood irrigations, hence resulting in different dry-down cycles: high irrigation (conservative schedule currently applied in the Ebro Valley, Spain) and low irrigation (equivalent to about a one-fourth reduction in water inputs). Net CO₂ assimilation rate (A), stomatal conductance (g_s), intrinsic water-use efficiency (A/g_s) and other related photosynthetic traits (leaf nitrogen concentration, leaf greenness and leaf mass per area) were measured prior to watering, and Δ was analysed in water-soluble leaf extracts (Δ_s) and bulk leaves (Δ_l). Stem growth was monitored over 3 years starting at the year of establishment (1998). Data were subjected to a repeated measures ANOVA over time for a randomised block split-plot design across watering regimes. Significant differences between watering regimes were detected using a long-term estimate of photosynthetic performance such as Δ_l, in agreement with changes in soil water status and evapotranspirative demand. However, the lack of significant genotype×watering regime interactions for gas exchange traits and Δ_s suggested that water shortage imposed by low irrigation was not sufficient to reveal physiological adaptations to drought. In this regard, the reduction in water inputs brought about by low irrigation did not reduce tree growth for any of the clones, suggesting that the current irrigation scheme employed in the region is superfluous to the water consumption needs of poplars. Genotypic variation was detected in gas exchange traits, Δ_s, Δ_l and stem growth under both watering treatments. Significant correlations with stem volume for Δ_s (r = −0.60, p<0.05) and A (r = + 0.61, p<0.05) suggested that growth was improved by higher water-use efficiency (the ratio of carbon fixed to water lost, as inferred by Δ_s) due to variation in A rather than in g_s. This observation corroborated the expectation derived from current theories that a lower Δ is related to higher stem volume, as a result of changes in net CO₂ assimilation rates.