No abstract

Two-dimensional gel electrophoresis (2-DE) is a unique method of large-scale protein characterisation, and is a powerful approach in the study of protein expression. In the present work, the experimental conditions for 2-DE of foot proteins from the mussel Mytilus galloprovincialis Lmk were established; the technique was performed with intertidal and cultured mussels using Melanie 3 software for data analysis. This powerful technique enabled the visualisation of a total of 750 protein spots consistently expressed in the foot. The intensity of 92 selected spots was compared between intertidal and cultured mussels, and statistically significant differences were detected in the expression of 45 (48.9%) of the 92 proteins analysed. In 31 of these proteins, intensity was higher in the cultured stock than in the intertidal mussels, while in 14 proteins spot intensity was higher in the latter. Using mass spectrometry (MS) combined with sequence database searching, 6 of the most prominent differentially expressed proteins were analysed. Of these, 1 was identified as being Heat-shock Protein 70, and 2 were shown to be cytoskeleton-associated proteins, myosin and actin. Heat-shock Protein 70, which is known to be involved in cellular transport and chaperoning and associated with stress situations, was more highly expressed in intertidal mussels living in littoral areas than in cultured mussels. These findings are discussed in connection with the molecular changes involved in the adaptation of mussels to different ecological conditions. KEY WORDS: Mytilus galloprovincialis · Two-dimensional gel electrophoresis · Proteome · Foot proteinsResale or republication not permitted without written consent of the publisher

Vascular damage occurring after cerebral ischemia may lead to a worse outcome in patients with ischemic stroke, as it facilitates edema formation and hemorrhagic transformation. There are several phases in the development of vascular injury (acute, subacute and chronic) and different mediators act in each one. Therapeutic options to avoid vascular injury must be focused on acting in each phase. However, even though experimental studies have demonstrated the benefit of therapeutic interventions both in the acute and chronic phases of cerebral ischemia, only the chronic phase offers a therapeutic window sufficiently wide enough to provide vascular protection in clinical practice. Several drugs including erythropoietin and HMG-CoA reductase inhibitors (statins), antihypertensive (angiotensin modulators), antibiotics (minocycline) and antihyperglycemic drugs (thiazolidinediones) have been proved to provide vascular protection in patients with ischemic stroke. Anti-inflammatory, antioxidant, and antiapoptotic actions are responsible for the vascular protective effect related to these drugs.

Two-dimensional electrophoresis (2-DE) has been used to measure the degree of genetic variability of the marine mussel Mytilus galloprovincialis. Genetic polymorphisms were detected in 33 of a total of 86 polypeptides scored among the most abundant proteins from foot samples in 38 individuals. Estimates of average heterozygosity were 0.10170.018 and 0.11470.021 in a natural and a cultured population, respectively, from the NW of the Iberian Peninsula. These are the highest estimates of average heterozygosity reported by 2-DE in an animal species to date. We consider that these data throw open the question of the level of genetic variability detectable by two-dimensional electrophoresis. Multilocus genotype data were used to infer haplotypic frequencies by means of the EM algorithm in order to detect linkage disequilibrium between loci coding abundant proteins. Significant associations were found in 22.7% of the 406 two-locus pairs analysed. Also, clusters of loci in which all pairwise combinations exhibit statistically significant associations were detected and physical linkage between some of these loci is postulated from the linkage disequilibrium data.

The aim of this study was to assess the efficacy of dairy industry sewage sludge as a fertilizer for an acid soil, and to evaluate the leaching of its nutrients and pollutants. The sludge was applied to pots containing an acid pasture soil (pH 4.4, organic matter 9.6%) in which Lolium multiflorum was then sown. Two different doses of sewage sludge (80 and 160 cubic m ha-1) and a dose of 15:15:15 NPK (675 kg ha-1) were compared. Changes in soil properties (leachate and soil fraction) and the production and nutritional status of the crop were monitored for nine weeks. The addition of the sludge led to substantial increases in crop production, which was attributed to improvements in crop nutrient concentrations (mainly N and P) and soil properties (pH, total N and available P). The sludge-treated soils lost less NO3–, Ca, Mg and K through leaching than those treated with the mineral fertilizer. The amounts of nutrient accumulated in the crop showed, however, that the input of K and to a lesser extent of Mg, were insufficient to cover requirements. Supplements of these elements are therefore necessary.