The HIV Care Cascade and sub-analysis of those linked to but not retained in care: the experience from a tertiary HIV referral service in Dublin Ireland.

BACKGROUND: The HIV Care Cascade model can be used to measure how clinical services align with United Nations' (UN) HIV treatment targets. Previous models have highlighted sequential losses at each step of the Cascade with a significant proportion being not retained in care (NRIC). OBJECTIVE: We aimed to assess the feasibility of meeting the UN targets and assess factors associated with, and calculate the true proportion of those, NRIC. METHODS: All people living with HIV who were linked to our service, one of three specialist HIV care providers in Dublin Ireland, from its establishment in 1993 to 1 December 2014, were included in the cohort and were categorized as linked to care, retained in care (RIC), on antiretroviral therapy (on ART), virally suppressed (HIV RNA <40copies/ml), and NRIC. An analysis of those NRIC was performed to categorize their current status through direct/indirect contact. RESULTS: Of 1000 patients linked to care, 78.7% (n = 787) were RIC, of whom 91.5% (n = 720) were on ART, with 89.9% (n = 644) virally suppressed. Those RIC were more likely older (p = 0.006) and non-IVDU (p < 0.001). Of 213 (21.3%) NRIC, 56 (26.3%) emigrated, 27 (12.7%) transferred care, 15 (7.0%) stopped attending but were contactable, 38 (17.8%) died, and 77 (36.1%) were lost to follow-up. After revision, 10.5% of the cohort was confirmed as NRIC, with 6 of 15 defined as "stopped attending" re-linked to care following direct contact. CONCLUSIONS: Our HIV Care Cascade model demonstrates that the true numbers of patients NRIC may be significantly lower than previously estimated and once RIC, treatment goals approaching the United Nations Programme on HIV and AIDS targets are possible with 91.5% on treatment and almost 90% of those on treatment virally suppressed. That 40% reengaged following direct contact suggests benefit through regular monitoring and direct contact based on the HIV Care Cascade model.

Resilient Nodeless d-Wave Superconductivity in Monolayer FeSe.

Monolayer FeSe exhibits the highest transition temperature among the iron based superconductors and appears to be fully gapped, seemingly consistent with s-wave superconductivity. Here, we develop a theory for the superconductivity based on coupling to fluctuations of checkerboard magnetic order (which has the same translation symmetry as the lattice). The electronic states are described by a symmetry based k·p-like theory and naturally account for the states observed by angle resolved photoemission spectroscopy. We show that a prediction of this theory is that the resultant superconducting state is a fully gapped, nodeless, d-wave state. This state, which would usually have nodes, stays nodeless because, as seen experimentally, the relevant spin-orbit coupling has an energy scale smaller than the superconducting gap.

The effect of initiation of antiretroviral therapy on monocyte, endothelial and platelet function in HIV-1 infection.

OBJECTIVES: Monocyte activation, endothelial dysfunction and platelet activation all potentially contribute to the increased risk of cardiovascular disease (CVD) reported in those with HIV-1 infection. To date, no study has examined how initiation of antiretroviral therapy (ART) affects markers of all three processes. We aimed to compare markers of monocyte, endothelial and platelet function between untreated HIV-positive subjects and HIV-negative controls and to examine the early effects of ART initiation on these markers. METHODS: We measured monocyte [soluble CD14 (sCD14) and sCD163], endothelial [von Willebrand factor (vWF), intercellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM-1)] and platelet [soluble P-selectin (sP-selectin), soluble CD40 ligand (sCD40L) and soluble glycoprotein VI (sGPVI)] biomarkers before and at weeks 4 and 12 post ART initiation in HIV-positive and well-matched HIV-negative controls. RESULTS: We examined 40 subjects, 25 HIV-positive subjects and 15 controls, with a median age of 34 years [interquartile range (IQR) 31, 40 years], of whom 60% were male and 47.5% Caucasian. Pre-ART, all biomarkers (monocyte, endothelial and platelet) were significantly higher in HIV-positive patients versus controls (all P < 0.05) and decreased with ART initiation, except for sCD14, which remained unchanged [median 1680 (IQR 1489, 1946) ng/mL at week 12 versus 1570 (IQR 1287, 2102) ng/mL at week 0; P = 0.7]. Although platelet activation markers reduced to levels comparable to those in controls, endothelial dysfunction markers remained elevated, as did sCD163 [at week 12, median 1005 (IQR 791, 1577) ng/mL in HIV-positive patients versus 621 (IQR 406, 700) ng/mL in controls; P < 0.0001]. CONCLUSIONS: ART initiation resulted in reductions in levels of CVD-associated biomarkers; however, although they improved, markers of endothelial dysfunction and monocyte activation remained elevated. How these persistent abnormalities affect CVD risk in HIV infection remains to be determined.

A marriage of convenience; a simple food chain comprised of Lemna minor (L.) and Gammarus pulex (L.) to study the dietary transfer of zinc.

Macrophytes contribute significantly to the cycling of metals in aquatic systems, through accumulation during growth and release during herbivory or decomposition. Accumulation of high levels of metals has been extensively documented in Lemnaceae (duckweeds). However, the degree of trophic transfer of metals from Lemnaceae to secondary consumers remains poorly understood. This study demonstrates that zinc accumulated in Lemna minor is bioavailable to the herbivore consumer Gammarus pulex. Overall, the higher the zinc content of L. minor, the more zinc accumulated in G. pulex. Accumulation in G. pulex was such that mortality occurred when they were fed high zinc-containing L. minor. Yet, the percentage of consumed zinc retained by G. pulex actually decreased with higher zinc concentrations in L. minor. We hypothesise that this decrease reflects internal zinc metabolism, including a shift from soluble to covalently bound zinc in high zinc-containing L. minor. Consistently, relatively more zinc is lost through depuration when G. pulex is fed L. minor with high zinc content. The developed Lemna-Gammarus system is simple, easily manipulated, and sensitive enough for changes in plant zinc metabolism to be reflected in metal accumulation by the herbivore, and therefore suitable to study ecologically relevant metal cycling in aquatic ecosystems.

Characterization of associations and development of atazanavir resistance after unplanned treatment interruptions.

OBJECTIVES: Although current guidelines recommend resistance testing prior to antiretroviral therapy (ART) reinitiation after treatment interruptions, virological failure of first-line ritonavir-boosted, protease-inhibitor (PI/r)-containing ART is associated with low emergent PI resistance. In patients experiencing unscheduled treatment interruptions (UTrIs) on ritonavir-boosted atazanavir (ATV/r) ART regimens, we hypothesized low emergence of PI mutations conferring resistance to ATV/r. METHODS: In a retrospective assessment of HIV-infected patients initiating ATV/r-containing ART, using logistic regression we determined factors associated with UTrI, the prevalence of emergent resistance mutations and virological response after ART reinitiation. RESULTS: A total of 202 patients [median age 33 years (interquartile range (IQR) 29-40 years); 52% female; median CD4 count 184 cells/µL (IQR 107-280 cells/µL); median HIV RNA 4.6 log10 HIV-1 RNA copies/mL (IQR 3.2-5.1 copies/mL)] initiated ATV/r between 2004 and 2009; 80 (43%) were ART naïve. One hundred and ten patients (55%) underwent 195 UTrIs after a median (IQR) 25 (10-52) weeks on ART, with a median (IQR) UTrI duration of 10 (3-31) weeks. Fifty-four of 110 patients (49%) underwent more than one UTrI. The commonest reasons for UTrI were nonadherence (52.7%) and drug intolerance (20%). Baseline HIV RNA > 100 000 copies\mL [odds ratio (OR) 3.6; 95% confidence interval (CI) 1.3-9.95] and being HCV positive, an injecting drug user or on methadone (OR 2.4; 95% CI 1.3-4.4) were independently associated with UTrI. In 39 patients with at least two resistance assays during UTrIs, 72 new mutations emerged; four nucleoside reverse transcriptase inhibitor (NRTI), two nonnucleoside reverse transcriptase inhibitor (NNRTI) and 66 protease inhibitor (PI) resistance mutations. All emergent PI resistance mutations were minor mutations. At least 65% of patients were re-suppressed on ATV/r reinitiation. CONCLUSIONS: In this PI-treated cohort, UTrIs are common. All emergent PI resistance mutations were minor and ATV/r retained activity and efficacy when reintroduced, even after several UTrIs, raising questions regarding the need for routine genotypic resistance assays in PI/r-treated patients prior to ART reinitiation after UTrI.

The impact of catchment conifer plantation forestry on the hydrochemistry of peatland lakes.

The hydrochemistry of 26 small blanket bog lakes was examined to assess the impact of conifer plantation forestry on lake water chemistry. Lakes were selected from three distinct catchment land use categories: i) unplanted blanket bog only present in the catchment, ii) mature (closed-canopy) conifer plantation forests only present in the catchment and iii) catchments containing mature conifer plantation forests with recently clearfelled areas. All three catchment land uses were replicated across two geologies: sedimentary (sandstone) and igneous (granite). Lakes with afforested catchments across both geologies had elevated concentrations of phosphorus (P), nitrogen (N), total dissolved organic carbon (TDOC), aluminium (Al) and iron (Fe), with the highest concentrations of each parameter recorded from lakes with catchment clearfelling. Dissolved oxygen was also significantly reduced in the afforested lakes, particularly the clearfell lakes. Analysis of runoff from a nearby recently clearfelled site revealed high biological and chemical oxygen demands, consistent with at least part of the elevated concentrations of TDOC emanating from clearfelled sites having higher biochemical lability. Inorganic fertilisers applied at the start of the forest cycle, the decay of the underlying peat soil and accumulated surface tree litter, and leachate from felled trees are the likely sources of the elevated concentrations of plant nutrients, TDOC, heavy metals and major ions, with excessive peat soil disturbance during clearfelling likely exacerbating the runoff into lakes. Our study has demonstrated a clear, deleterious impact of conifer plantations on the water quality draining from blanket bog catchments, with major implications for the management of afforested peatlands.

Diversity and bioactive potential of endospore-forming bacteria cultured from the marine sponge Haliclona simulans.

AIMS: Despite the frequent isolation of endospore-formers from marine sponges, little is known about the diversity and characterization of individual isolates. The main aims of this study were to isolate and characterize the spore-forming bacteria from the marine sponge Haliclona simulans and to examine their potential as a source for bioactive compounds. METHODS AND RESULTS: A bank of presumptive aerobic spore-forming bacteria was isolated from the marine sponge H. simulans. These represented c. 1% of the total culturable bacterial population. A subgroup of thirty isolates was characterized using morphological, phenotypical and phylogenetic analysis. A large diversity of endospore-forming bacteria was present, with the thirty isolates being distributed through a variety of Bacillus and Paenibacillus species. These included ubiquitous species, such as B. subtilis, B. pumilus, B. licheniformis and B. cereus group, as well as species that are typically associated with marine habitats, such as B. aquimaris, B. algicola and B. hwajinpoensis. Two strains carried the aiiA gene that encodes a lactonase known to be able to disrupt quorum-sensing mechanisms, and various isolates demonstrated protease activity and antimicrobial activity against different pathogenic indicator strains, including Clostridium perfringens, Bacillus cereus and Listeria monocytogenes. CONCLUSIONS: The marine sponge H. simulans harbours a diverse collection of endospore-forming bacteria, which produce proteases and antibiotics. This diversity appears to be overlooked by culture-dependent and culture-independent methods that do not specifically target sporeformers. SIGNIFICANCE AND IMPACT OF STUDY: Marine sponges are an as yet largely untapped and poorly understood source of endospore-forming bacterial diversity with potential biotechnological, biopharmaceutical and probiotic applications. These results also indicate the importance of combining different methodologies for the comprehensive characterization of complex microbial populations such as those found in marine sponges.

Diversity and antimicrobial activity of Pseudovibrio spp. from Irish marine sponges.

AIMS: To evaluate the diversity and antimicrobial activity present among Pseudovibrio spp. isolated from marine sponges. METHODS AND RESULTS: Seventy-three bacterial isolates from the marine sponges Polymastia boletiformis, Axinella dissimilis and Haliclona simulans were identified as Pseudovibrio spp. using phylogenetic analysis of 16S rRNA gene sequences. Genetic diversity among these isolates was estimated using random amplification of polymorphic DNA (RAPD), and 33 RAPD types were identified among the 73 Pseudovibrio isolates. These Pseudovibrio spp. were assayed for the production of compounds with antimicrobial activity against various clinically relevant pathogens. Sixty-two (85%) of the isolates showed activity against at least one of the pathogens tested, including Escherichia coli, Salmonella enterica serotype Typhimurium, methicillin-resistant Staphylococcus aureus (MRSA), and Clostridium difficile. PCR screens of the Pseudovibrio isolates also revealed the presence of potential antibiotic-producing polyketide synthase genes. CONCLUSIONS: Marine sponges harbour a diverse population of Pseudovibrio spp., the majority of which demonstrate antimicrobial activity. The identification of several different antimicrobial activity spectra suggests that the Pseudovibrio isolates may produce a suite of antimicrobial compounds. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study in which an extended population of Pseudovibrio isolates from marine sponges has been analysed and establishes the little-studied Pseudovibrio as a potentially important genus in the search for antimicrobial compounds of clinical relevance.

Psychometric comparison of standard and computerized administration of the Alzheimer's Disease Assessment Scale: Cognitive Subscale (ADASCog).

BACKGROUND: The Alzheimer's Disease Assessment Scale ­ Cognitive Subscale (ADAS-Cog) has become the de facto gold standard for assessing the efficacy of anti-dementia treatments. However, manual administration of the ADAS-Cog is subject to procedural inconsistencies, including scoring and transcription errors, which can introduce unwanted variance and compromise data quality within and across sites and trials. To address such concerns, a computerized version was developed that integrates, rather than replaces, the examiner, standardizes administration, and uses electronic data capture at the point of patient contact. The examiner can control administration and pacing, pause or repeat digitized instructions, score verbal report and overt behavioral performance, and freely interact with the subject. PURPOSE: To conduct psychometric comparisons of traditional, paper-based administration of the standard ADAS-Cog (sADAS) with examiner- assisted administration of the computerized ADAS-Cog (cADAS). METHODS: Eighty-eight patients (39M; 49F) with mild to moderate Alzheimer's disease were tested on three occasions with each version over a period of one year with one month between paired visits. RESULTS: Intraclass Correlation Coefficients (ICC) comparisons between sADAS and cADAS were significant for total score (ICC=0.96) and all subscores (ICCs ranged 0.78-0.93), with no significant differences on paired t-tests. The mean ICCs across cADAS scores for test-retest reliability for short-term (mean ICC=0.96) and long-term (mean ICC=0.91) comparisons were significantly higher than across sADAS scores (mean ICCs were 0.87 and 0.84, respectively). CONCLUSIONS: These results indicate that examiner-assisted, computerized administration is equivalent to traditional, paper-based administration, and shows significantly greater test-retest reliability.

Tenderness enhancement of beef from Bos indicus and Bos taurus cattle following electrical stimulation.

High voltage electrical stimulation (1130 V peak, 14.28 bidirectional half sinusoidal pulses/s) or low voltage stimulation (45 V peak, 36 alternating square wave pulses/s) was used on cattle: (1) low voltage stimulation applied for 10 or 40 s with fast and slow chilling or high voltage stimulation for 60 s with normal chilling, applied to 100% Bos taurus cattle, (2) low voltage stimulation (40 s) and high voltage stimulation (60 s) with normal chilling applied to mixed Bos indicus and B.taurus cattle, (3) high voltage stimulation (54 s) with normal chilling applied to B. taurus and B. indicus cattle of 0-100% B. indicus composition, and (4) high voltage stimulation (60 s) applied to 100% B. taurus and 100% B. indicus cattle. All stimulation parameters enhanced the tenderness of steaks from M. longissimus thoracis et lumborum (LTL) aged at 1°C up to 28 days compared with non stimulated LTL. Short low voltage stimulation of 10s was marginally more effective than no stimulation and longer durations of 40s were very effective and high voltage stimulation was most effective. The shear force values for non stimulated B. indicus LTL are much greater than for B. taurus, but following high voltage stimulation LTL of B. indicus were similar to B. taurus and all had lower shear force values than from non stimulated carcasses. Thus adequate electrical stimulation removes any toughness of LTL related to B. indicus genetic composition.

Shellfish toxicity: human health implications of marine algal toxins.

Five major human toxic syndromes caused by the consumption of shellfish contaminated by algal toxins are presented. The increased risks to humans of shellfish toxicity from the prevalence of harmful algal blooms (HABs) may be a consequence of large-scale ecological changes from anthropogenic activities, especially increased eutrophication, marine transport and aquaculture, and global climate change. Improvements in toxin detection methods and increased toxin surveillance programmes are positive developments in limiting human exposure to shellfish toxins.

The effects of salinity on the Manila clam (Ruditapes philippinarum) using the neutral red retention assay with adapted physiological saline solutions.

This study investigated the internal osmotic regulatory capabilities of the Manila clam (Ruditapes philippinarum) following in vivo exposure to a range of salinities. A second objective was to measure the health status of the Manila clam following exposure to different salinities using the neutral red retention (NRR) assay, and to compare results using a range of physiological saline solutions (PSS). On exposure to seawater of differing salinities, the Manila clam followed a pattern of an osmoconformer, although they seemed to partially regulate their circulatory haemolytic fluids to be hyperosmotic to the surrounding aqueous environment. Significant differences were found when different PSS were used, emphasizing the importance of using a suitable PSS to reduce additional osmotic stress. Using PSS in the NRR assay that do not exert additional damage to lysosomal membrane integrity will help to more accurately quantify the effects of exposure to pollutants on the organism(s) under investigation.

Effects of contaminated sediment from Cork Harbour, Ireland on the cytochrome P450 system of turbot.

Hatchery-reared juvenile turbot (Scophthalmus maximus L.) were exposed for 3 weeks, under laboratory conditions, to inter-tidal sediments collected from polluted sites in Cork Harbour (Whitegate and Agahda) and a reference site at Ballymacoda Co., Cork, Ireland. The potential of the sediment exposure to induce cytochrome P450 activities and CYP1A1 in the fish was assessed. Chemical analysis revealed that the sediments originating from the reference and harbour sites were contaminated principally with PAHs-the harbour sites having double the levels of those at the reference site. Following 3 weeks exposure to the sediments western blotting demonstrated a strong immunogenic response for CYP1A1 in the liver, but not for gill or intestine. P450 activities were generally significantly higher than those exposed to reference site sediment. Liver was the most responsive tissue with significantly greater P450 activities compared with gill and intestinal tissues.

Evidence of a highly complex phylogeographic structure on a specialist river bird species, the dipper (Cinclus cinclus).

This study details the phylogeographic pattern of the white-throated dipper (Cinclus cinclus), a Palearctic, temperate, passerine bird that is exclusively associated with flowing water. Our results reveal a complex phylogeographic structure with at least five distinct lineages for the Western Palearctic region. As for many species of the Western Palearctic fauna and flora, this genetic structure is probably linked to the isolation of populations in different southern refuges during glacial periods. Furthermore, the isolation of populations in Scandinavia and/or Eastern regions, but also in Morocco and probably in Corsica, was accentuated by ecological and biogeographic barriers during Quaternary interglacial periods. During glacial periods, Italy, Sicily and the Balkano-Carpathian region acted as major refuge zones for the dipper. At the end of the last ice age, Western Europe was repopulated by dippers from an Italian refuge, while Eastern Europe was recolonised by Balkano-Carpathian birds. A large contact zone between these two lineages was evidenced and extends from Luxembourg to Hungary. Finally, our results indicate the need to clarify the taxonomic status of the dipper, especially concerning the European subspecies whose validity appears uncertain.

Impacts of microcystins on the feeding behaviour and energy balance of zebra mussels, Dreissena polymorpha: a bioenergetics approach.

Microcystins are produced by bloom-forming cyanobacteria and pose significant health and ecological problems. To investigate the impacts of these biotoxins on the physiology of the zebra mussels, Dreissena polymorpha, a series of short-term feeding experiments were conducted in the laboratory. We used five microalgal diets consisting of single-cell suspensions of the green algae, Chlorella vulgaris, the diatom, Asterionella formosa, the cryptophyte, Cryptomonas sp. and two strains of the toxic cyanobacterium, Microcystis aeruginosa (strains CCAP 1450/06 and CCAP 1450/10). A sixth diet was a mixture of the diatom and the CCAP 1450/10 cyanobacterial strain. The low-toxicity strain CCAP 1450/06 contained 7.4 microg l(-1) of the MC-LR variant while the very toxic strain CCAP 1450/10 contained 23.8 microg l(-1) of MC-LR and 82.9 microg l(-1) of MC-LF. A flow-through system was designed to measure the following feeding parameters: clearance, filtration, ingestion and absorption rates. Ultimately the scope for growth (SFG) was determined as a net energy balance. We observed that mussels cleared the cyanobacterial species containing MC-LF (mean+/-95% confidence interval) at a significant lower rate (498+/-82 ml h(-1) g(-1) for the single cell suspension and 663+/-100 ml h(-1) g(-1) for the mixture diet) than all of the non-toxic species and the cyanobacterium containing MC-LR (all above 1l h(-1) g(-1)). The same pattern was observed with all the feeding parameters, particularly absorption rates. Furthermore, MC-LF caused an acute irritant response manifested by the production of 'pseudodiarrhoea', unusually fluid pseudofaeces, rich in mucus and MC-LF-producing Microcystis cells, ejected through the pedal gape of the mussels. This overall response therefore demonstrates selective rejection of MC-LF-producing cyanobacteria by zebra mussels, enhancing the presence of the very toxic MC-LF-producing M. aeruginosa in mixed cyanobacterial blooms and in the benthos. Finally, we observed that the SFG (mean+/-95% confidence interval) of mussels feeding on M. aeruginosa containing MC-LF was significantly lower (34.0+/-18.8 J h(-1) g(-1) for the single cell suspension and 83.1+/-53.0 J h(-1) g(-1) for the mixture diet) than for mussels ingesting non-toxic diets, except for C. vulgaris (all above 200 J h(-1)g(-1)). This reveals a sublethal, stressful effect of microcystins (particularly MC-LF) on the feeding behaviour and energy balance of the zebra mussel.

Two-dimensional electrophoresis analysis of glutathione affinity-selected proteins from the clam Tapes semidecussatus: Evidence for tissue-specific expression of redox proteins.

Proteins from gill, digestive gland and mantle of the clam Tapes semidecussatus were selected on glutathione (GSH) agarose to simplify proteomic comparison. Analysis by two-dimensional gel electrophoresis (2D SDS PAGE) revealed tissue-specific patterns of protein expression with some spots common to all tissues. Immunoblotting of gill and digestive gland separations identified some spots as glutathione transferases (GSTs). In gill and digestive gland several spots were immunoblotted with Pi class GSTs indicating multiple isoenzymes. Selected spots were excised, digested with trypsin and analyzed by reversed phase C-18 high performance liquid chromatography and tandem mass spectrometry. This confirmed that gill and digestive gland share some GST isoenzymes. Our results suggest that this clam expresses a complex tissue-specific pattern of GSH-binding proteins, which may reflect different redox requirements in each tissue.

A test battery approach for the ecotoxicological evaluation of estuarine sediments.

The purpose of this study was to evaluate the overall sensitivity and applicability of a number of bioassays representing multiple trophic levels, for the preliminary ecotoxicological screening (Tier I) of estuarine sediments. Chemical analyses were conducted on sediments from all sampling sites to assist in interpreting results. As sediment is an inherently complex, heterogeneous geological matrix, the toxicity associated with different exposure routes (solid, porewater and elutriate phases) was also assessed. A stimulatory response was detected following exposure of some sediment phases to both the Microtox and algal bioassays. Of the bioassays and endpoints employed in this study, the algal test was the most responsive to both elutriates and porewaters. Salinity controls, which corresponded to the salinity of the neat porewater samples, were found to have significant effects on the growth of the algae. To our knowledge, this is the first report of the inclusion of a salinity control in algal toxicity tests, the results of which emphasise the importance of incorporating appropriate controls in experimental design. While differential responses were observed, the site characterised as the most polluted on the basis of chemical analysis was consistently ranked the most toxic with all test species and all test phases. In terms of identifying appropriate Tier I screening tests for sediments, this study demonstrated both the Microtox and algal bioassays to be more sensitive than the bacterial enzyme assays and the invertebrate lethality assay employing Artemia salina. The findings of this study highlight that salinity effects and geophysical properties need to be taken into account when interpreting the results of the bioassays.

Assessing the potential of fish cell lines as tools for the cytotoxicity testing of estuarine sediment aqueous elutriates.

In the present study, we assess the potential of fish cell lines (CHSE, EPC and RTG-2) to be used as screening tools for the ecotoxicological assessment of estuarine sediments. The processing of sediment to a form suitable for in vitro exposure is an inherent problem when using cell cultures. The approach employed in this study was to prepare aqueous elutriate extracts from whole sediments, which were subsequently used to reconstitute powdered media. This procedure allowed the exposure of cell cultures to concentrations of up to and including 100% of the original aqueous sample. Cytotoxicity was assessed using multiple endpoint measurements. Cell viability was quantified using the neutral red and alamar blue colorimetric assays, which specifically assess lysosomal and mitochondrial function, respectively. In addition, the total protein content of the cells was measured using the coomassie blue assay. Initial tests were conducted to ensure that any resultant cytotoxicity was due to sample contaminants and not osmotic stress. In addition, elutriate samples were spiked with a model toxicant to verify the ability of the cell lines to detect and respond to bioavailable contaminants. Chemical analyses were conducted on sediments from all sampling sites to assist in interpreting any observed cytotoxicity. A differential response was observed for the cytotoxicity assays following exposure treatments, which emphasises the importance of employing multiple endpoints for the determination of toxicity. Of the three cell lines utilised in this study, RTG-2 cells were the most suitable for the testing of estuarine aqueous elutriate samples on the basis of tolerance to osmolality effects. Slight toxicity was observed following exposure to the aqueous elutriates tested in this study using RTG-2 cells and the alamar blue assay. In order to fully evaluate the overall sensitivity of this cell line, further research is warranted using an extensive range of test sites incorporating more polluted sediments.

An assessment of the pollutant status of surficial sediment in Cork Harbour in the South East of Ireland with particular reference to polycyclic aromatic hydrocarbons.

Surface sediment from three polluted sites within Cork Harbour, Ireland, and from a relatively clean reference site were collected and analysed for polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs), brominated flame retardants (BFRs), organotins (OTs), and heavy metals. PAHs were determined to be the most abundant class of contaminant. Concentrations of the sum (Sigma) of the 21 PAHs measured from the Harbour sites (2877.70 ng g(-1), 1000.7 ng g(-1) and 924.40 ng g(-1) dry weight respectively) were significantly higher than that of the sediment from the reference site (528.30 ng g(-1) dry weight). An inner harbour site, Douglas being the more contaminated of the three harbour sites. A similar pattern was observed with the other contaminants however, these compounds, with the exception of the heavy metals, all tended to be detected at concentrations on or below detection limits.