Accumulation of aqueous and nanoparticulate silver by the marine gastropod littorina littorea

The accumulation of Ag by the marine herbivorous gastropod, Littorina littorea, has been studied in a series of exposures in which the metal was added in aqueous form and as nanoparticles, both in the presence and absence of contaminated algal food (Ulva lactuca). Significant accumulation occurred in the gill, kidney, stomach and visceral mass when the snail was exposed to aqueous Ag in the absence of food. Despite the consumption of U. lactuca that had been previously contaminated by Ag, no accumulation was observed from the dietary route. When added as nanoparticles, accumulation of Ag was only measured in the head and gill and only in the absence of contaminated food. These observations suggest that Ag is most bioavailable to L. littorina when in true solution and that Ag measured in external tissues of the snail following exposure to nanoparticles arises from some physical association that does not result in significant transfer of the metal to internal organs. © 2012 Springer Science+Business Media Dordrecht

Intra-specific responses of Cymodocea nodosa to macro-nutrient, irradiance and copper exposure

Excess macro-nutrients, metal contamination and light limitation are three of the most commonly encountered anthropogenic stressors affecting seagrass meadows. In this study, the effects of different combinations of nutrients (N-NO3, P-PO4), copper and irradiance were investigated in shoots of Cymodocea nodosa collected from three meadows in the N. Aegean Sea, one (Nea Karvali) impacted by anthropogenically-derived environmental stressors and two in more pristine condition (Thasos, Brasidas). In a series of laboratory experiments, shoots were exposed to varying nutrient and heavy metal concentrations, as well as varying irradiance levels, for 8days and the effective quantum yield (δF/Fm') and leaf elongation were quantified. Results showed that C. nodosa increased δF/Fm' under high nutrient concentrations (30μΜ N-NO3--2μΜ P-PO43-) but significant differences were only apparent in shoots collected from the oligotrophic-less stressed meadows. Irradiance affected δF/Fm' significantly in all shoots irrespective of source and PO4-P concentration, while higher values were measured under low light conditions and it was identified as the main pathway of eutrophication stress in N. Aegean Cymodocea meadows. Shoots, independently of acclimation were tolerant to copper enrichment, with only the highest copper concentrations (4.7 and 7.9μM) having significant negative effects on δF/Fm'. Shoots from the more pristine meadows were less affected by Cu than those from the highly stressed meadow

Metal accumulation kinetics by the estuarine macroalga, Fucusceranoides

The kinetics of Cu, Cd and Pb accumulation by the macroalga, Fucus ceranoides, was studied under simulated estuarine conditions. Accumulation of Cu and Pb proceeded via a pseudo-first-order reaction that was reversible, suggesting desorption or efflux of accumulated metal, with forward rate constants on the order of 0.1h-1. For both metals, reaction reversibility increased and the equilibrium constant decreased with increasing salinity (from 1 to 33.5) and system response times were <10h throughout. Accumulation of Cd proceeded via a first-order reaction that was irreversible, suggesting little desorption or efflux of metal, with rate constants that decreased with increasing salinity (from 0.023 to 0.015h-1) and reaction half-lives ranging from approximately 30-50h. Inorganic equilibrium speciation calculations suggest that interactions of Cu, Cd and Pb principally involve the respective free ions, but that additional ions (e.g. CdCl+) and biotic processes may also be significant. •Accumulation of Cu, Cd and Pb by the estuarine macroalga, Fucus ceranoides, decreases along a salinity gradient.•Accumulation of Cu and Pb proceeds via a reversible pseudo-first-order reaction.•In contrast, accumulation of Cd proceeds via an irreversible first-order reaction.•Differences in reaction mechanisms are attributed to differences in the ability of metals to be internalised. © 2013 Elsevier Ltd

Extra- and intra-cellular accumulation of platinum group elements by the marine microalga, Chlorella stigmatophora.

To better understand the marine biogeochemistry of the platinum group elements (PGE), Rh(III), Pd(II) and Pt(IV) were added in combination and at ppb concentrations to cultures of the marine microalga, Chlorella stigmatophora, maintained in sea water at 15 °C and under 60 μmol m(-2) s(-1) PAR. The accumulation of PGE was established in short-term (24-h) exposures, and under varying conditions of algal biomass and PGE concentration, and in a longer-term exposure (156-h) by ICP-MS analysis of sea water and nitric acid digests and EDTA washes of the alga. In short-term exposures, and under all conditions, the extent of accumulation by C. stigmatophora was in the order: Rh > Pd >> Pt; and Pd was internalised (or resistant to EDTA extraction) to a considerably greater extent than Rh and Pt. Accumulation isotherms were quasi-linear up to added PGE concentrations of 30 μg L(-1) and all metals displayed a significant reduction in accumulation on a weight-normalised basis with increasing density (biomass) of C. stigmatophora, an effect attributed to the production of exudates able to stabilise metals in sea water through complexation. In the longer-term exposure, kinetic constraints on the reactivities of Rh and, in particular, Pt, resulted in final degrees of accumulation and internalisation by C. stigmatophora that were greatest for Rh and similar between Pd and Pt. Among the PGE, therefore, Rh is predicted to participate in biological removal and transport processes in the marine environment to the greatest extent while decoupling in the biogeochemistries of Pd and Pt is predicted in shorter-term or more transient processes

A simple and effective method for high quality co-extraction of genomic DNA and total RNA from low biomass Ectocarpus siliculosus, the model brown alga.

The brown seaweed Ectocarpus siliculosus is an emerging model species distributed worldwide in temperate coastal ecosystems. Over 1500 strains of E. siliculosus are available in culture from a broad range of geographic locations and ecological niches. To elucidate the molecular mechanisms underlying its capacity to cope with different environmental and biotic stressors, genomic and transcriptomic studies are necessary; this requires the co-isolation of genomic DNA and total RNA. In brown algae, extraction of nucleic acids is hindered by high concentrations of secondary metabolites that co-precipitate with nucleic acids. Here, we propose a reliable, rapid and cost-effective procedure for the co-isolation of high-quality nucleic acids using small quantities of biomass (25-, 50- and 100 mg) from strains of E. siliculosus (RHO12; LIA4A; EC524 and REP10-11) isolated from sites with different environmental conditions. The procedure employs a high pH extraction buffer (pH 9.5) which contains 100 mM Tris-HCl and 150 mM NaCl, with the addition of 5 mM DTT and 1% sarkosyl to ensure maximum solubility of nucleic acids, effective inhibition of nuclease activity and removal of interfering contaminants (e.g. polysaccharides, polyphenols). The use of sodium acetate together with isopropanol shortened precipitation time and enhanced the yields of DNA/RNA. A phenol:chlorophorm:isoamyl alcohol step was subsequently used to purify the nucleic acids. The present protocol produces high yields of nucleic acids from only 25 mg of fresh algal biomass (0.195 and 0.284 µg mg(-1) fresh weigh of RNA and DNA, respectively) and the high quality of the extracted nucleic acids was confirmed through spectrophotometric and electrophoretic analyses. The isolated RNA can be used directly in downstream applications such as RT-PCR and the genomic DNA was suitable for PCR, producing reliable restriction enzyme digestion patterns. Co-isolation of DNA/RNA from different strains indicates that this method is likely to have wider applications for intra- and inter-specific studies on other brown algae

DNA methylation profiling to assess pathogenicity of BRCA1 unclassified variants in breast cancer

Germline pathogenic mutations in BRCA1 increase risk of developing breast cancer. Screening for mutations in BRCA1 frequently identifies sequence variants of unknown pathogenicity and recent work has aimed to develop methods for determining pathogenicity. We previously observed that tumor DNA methylation can differentiate BRCA1-mutated from BRCA1-wild type tumors. We hypothesized that we could predict pathogenicity of variants based on DNA methylation profiles of tumors that had arisen in carriers of unclassified variants. We selected 150 FFPE breast tumor DNA samples [47 BRCA1 pathogenic mutation carriers, 65 BRCAx (BRCA1-wild type), 38 BRCA1 test variants] and analyzed a subset (n=54) using the Illumina 450K methylation platform, using the remaining samples for bisulphite pyrosequencing validation. Three validated markers (BACH2, C8orf31, and LOC654342) were combined with sequence bioinformatics in a model to predict pathogenicity of 27 variants (independent test set). Predictions were compared with standard multifactorial likelihood analysis. Prediction was consistent for c.5194-12G>A (IVS 19-12 G>A) (P>0.99); 13 variants were considered not pathogenic or likely not pathogenic using both approaches. We conclude that tumor DNA methylation data alone has potential to be used in prediction of BRCA1 variant pathogenicity but is not independent of estrogen receptor status and grade, which are used in current multifactorial models to predict pathogenicity

Coping with metastatic melanoma: the last year of life.

BackgroundFew longitudinal studies have concurrently investigated cognitive appraisal, coping and psychological adjustment in patients with terminal cancer. This study aimed to (i) consider patterns of change in these variables during the last year of life and (ii) consider covariates associated with patients' psychological adjustment.Methods and patientsQuestionnaires were sent to a cohort of stage IV melanoma patients seen at the Sydney Melanoma Unit between 1991 and 1996, approximately every 3 months, for up to 2 years. A sub-sample of 110 patients completed at least one questionnaire in the last year of life. Repeated measures linear regression was used to model cognitive appraisal, coping and psychological adjustment.ResultsIn the last year of life, patients' cognitive appraisal of their disease remained relatively stable, whereas their use of active coping strategies increased (p=0. 04). There was some deterioration in psychological adjustment, particularly in patients' ability to minimize the impact of cancer on daily life (p=0.03), but this effect did not remain significant when patients' level of tiredness was included in the model. Cognitive appraisal, coping style and quality of life indicators were all associated with psychological adjustment.ConclusionThese findings suggest that while patients work hard to actively cope with their disease, they experience increasing levels of tiredness, and deterioration in their mood and ability to function in their daily lives

Recognition without identification, erroneous familiarity, and déjà vu

Déjà vu is characterized by the recognition of a situation concurrent with the awareness that this recognition is inappropriate. Although forms of déjà vu resolve in favor of the inappropriate recognition and therefore have behavioral consequences, typical déjà vu experiences resolve in favor of the awareness that the sensation of recognition is inappropriate. The resultant lack of behavioral modification associated with typical déjà vu means that clinicians and experimenters rely heavily on self-report when observing the experience. In this review, we focus on recent déjà vu research. We consider issues facing neuropsychological, neuroscientific, and cognitive experimental frameworks attempting to explore and experimentally generate the experience. In doing this, we suggest the need for more experimentation and amore cautious interpretation of research findings, particularly as many techniques being used to explore déjà vu are in the early stages of development.PostprintPeer reviewe