Use of multiple cell and tissue-level biomarkers in mussels collected along two gas fields in the northern Adriatic Sea as a tool for long term environmental monitoring.

As a consequence of the growing global demand of energy supplies, intense oil and gas exploration and exploitation programs have been carried out worldwide especially within the marine environments. The release of oil-derived compounds in the sea from anthropogenic sources both as effluents or accidental spill is perceived as a major environmental concern. An approach based on a combination of biomarkers and the distribution of some classes of environmentally relevant pollutants was used to investigate the occurrence of a stress syndrome in mussels (Mytilus galloprovincialis) collected at three gas platforms placed in two distinct oceanographic districts within the Adriatic Sea. Biological responses were integrated by a ranking algorithm which demonstrated both a range of biological effects reflecting exposure gradients and a temporal related trend in the investigated responses. The overall results demonstrate a moderate to absent pollution from studied gas platforms with low but remarkable biological disturbance in sentinel organisms.

Development of a decision support system to manage contamination in marine ecosystems.

In recent years, contamination and its interaction with climate-change variables have been recognized as critical stressors in coastal areas, emphasizing the need for a standardized framework encompassing chemical and biological data into risk indices to support decision-making. We therefore developed an innovative, expert decision support system (Exp-DSS) for the management of contamination in marine coastal ecosystems. The Exp-DSS has two main applications: (i) to determine environmental risk and biological vulnerability in contaminated sites; and (ii) to support the management of waters and sediments by assessing the risk due to the exposure of biota to these matrices. The Exp-DSS evaluates chemical data, both as single compounds and as total toxic pressure of the mixture, to compare concentrations to effect-based thresholds (TELs and PELs). Sites are then placed into three categories of contamination: uncontaminated, mildly contaminated, and highly contaminated. In highly contaminated sites, effects on high-level ecotoxicological endpoints (i.e. survival and reproduction) are used to determine risk at the organism-population level, while ecological parameters (i.e. alterations in community structure and ecosystem functions) are considered for assessing effects on biodiversity. Changes in sublethal biomarkers are utilized to assess the stress level of the organisms in mildly contaminated sites. In Triad studies, chemical concentrations, ecotoxicological high-level effects, and ecological data are combined to determine the level of environmental risk in highly contaminated sites; chemical concentration and ecotoxicological sublethal effects are evaluated to determine biological vulnerability in mildly contaminated sites. The Exp-DSS was applied to data from the literature about sediment quality in estuarine areas of Spain, and ranked risks related to exposure to contaminated sediments from high risk (Huelva estuary) to mild risk (Guadalquivir estuary and Bay of Cadiz). A spreadsheet-based version of the Exp-DSS is available at the MEECE and DiSIT web sites (www.meece.eu and www.disit.unipmn.it).

The use of protozoa in ecotoxicology: application of multiple endpoint tests of the ciliate E. crassus for the evaluation of sediment quality in coastal marine ecosystems.

Despite an increasing number of surveys describing adverse effects of contaminated sediments on marine organisms, few studies have addressed protists. In this study, the free-crawling marine ciliate Euplotes crassus was evaluated as the test organism for the screening of sediment toxicity using sediments from both coastal and estuarine sites of the Venice Lagoon (Marghera harbour [MH], Valle Millecampi [MV], Murano island [MI] and Lido inlet [LI]). Two endpoints of high ecological value, mortality (Mry) and replication rate (RpR), were assessed in combination with the two sublethal biomarkers of stress, endocytotic rate (Ecy) and lysosomal membrane stability (NRRT). The results showed a significant inhibition of RpR, Ecy and NRRT paralleled by a small and insignificantly increased Mry of the exposed specimens. Our results thus demonstrate that only a combination of mortality and sublethal biomarkers was able to characterise an exposure-related stress syndrome. The suite of biomarkers described here was also able to detect and resolve a pollution-induced stress syndrome at an early stage of pollution. The contamination level of the sediments was assessed using chemical analysis, by estimating bioavailability and by computing a toxic pressure coefficient (TPC) to account for potential additive effects of different pollutants. The observed biological responses were consistent with the contamination levels in sediments, suggesting a high potential for using Protozoa in bioassays to assess environmental risk in coastal marine systems.

The use of multiple endpoints to assess cellular responses to environmental contaminants in the interstitial marine ciliate Euplotes crassus.

This paper presents the results of investigations on the suitability of Euplotes crassus, an interstitial marine ciliate, to be used as model organism in ecotoxicology and thereafter to evaluate the toxicity of estuarine and coastal sediments upon laboratory exposure. Nowadays, anthropogenic activities have resulted in accumulation of metals and organic pollutants in the environment as well as in the food chain hence leading to serious ecological and human health problems. This may pose a risk to benthic and epibenthic organisms and it is crucial to discover toxicity tests that will identify adverse effects of sediment-associated chemicals on benthic organisms. Due to their nature as a eukaryotic cell/organism and their position in the food web, ciliated protozoa are suitable models for evaluating the effects of pollution on aquatic communities. Lethal and sublethal effects of exposure to inorganic and organic pollutants were tested on the cell mortality, replication rate, lysosomal membrane stability and endocytosis rate of E. crassus. Increasing nominal concentrations of individual and mixtures of mercury, copper, and benzo(a)pyrene were investigated in this study as they might be bioavailable in naturally occurring polluted sites. A significant decrease in the mean replication rate (p<0.05) was found after 24h exposures to m/µM concentrations of all tested pollutants. At the same time, significant decreases of lysosomal membrane stability (p<0.05) were observed for Cu (5 µM), Hg (10 nM), and B(a)P (200 nM). Among the entire suite of tests, endocytosis rate test demonstrated the highest sensitivity. Exposures to binary mixtures of all studied pollutants were performed showing both inorganic-organic and inorganic-inorganic additive and/or antagonist effects. Moreover, medium salinity was also varied to mimic estuarine-like environmental conditions linking biological response to ionic strengths. Under these conditions significant increases of both endocytosis rate and lysosomal membrane stability were observed and related to the increment of some Hg- and Cu-related toxic complexes. The studied biomarkers were always able to discriminate between the effects of organic and inorganic pollutants. Together with the short time and simplicity of the test procedures, results obtained in this study indicate that E. crassus is a promising and convenient bioindicator for evaluating the toxicity of different environmental matrixes like pore water, sediments and wastewaters--polluted by metals and organic pollutants.

Integration of biochemical, histochemical and toxicogenomic indices for the assessment of health status of mussels from the Tamar Estuary, U.K.

The aim of this study was to examine whether a combination of biochemical, histopathological and toxicogenomic data could be used as a valuable tool for the assessment of biological risk associated with pollutants within the Tamar River and Estuary, S.W. England, U.K. Accordingly, biochemical and histopathological biomarkers (protein carbonyls, lipofuscin, neutral lipids, lysosomal stability [N-acetyl-ß-hexosaminidase and neutral red], lysosomal volume, ferric reducing antioxidant power [FRAP] and malonaldehyde [MDA]) and gene expression profiles were assessed in 5 sites from the Tamar River and Estuary (Neal Point, Town Quay, Wilcove, Cremyll Ferry and Whitsand; and a reference site, Trebarwith Strand, N. Cornwall). PAHs were measured in mussel tissue and sediment and metals were measured in mussel tissue only. Data from the biomarkers was integrated into a Mussel Expert System (MES) model to produce a simple assessment of mussel stress. Clear gradients of mussel toxicity were identified by the biomarkers (with the exception of neutral lipids) with the highest impacted animals found furthest up the Tamar, whilst the MES was unable to identify a gradient of effect. Gene expression profiles also indicated a gradient of stress with the greatest number of significantly up- or down- regulated genes found at the uppermost 2 sites. The MES did, however, determine that mussels from all sites, except the reference site, were highly stressed; a conclusion that could not be inferred from the biomarker data alone. It is concluded that the MES is a valuable tool that permits integration and interpretation of complex sets of biomarker data by identifying the biological meaning of biomarker changes.

Acute effects of benzo[a]pyrene on digestive gland enzymatic biomarkers and DNA damage on mussel Mytilus galloprovincialis.

In the present study, mussel (Mytilus galloprovincialis) digestive gland biotransformation and detoxification responses to acute exposure to the polycyclic aromatic hydrocarbon benzo[a]pyrene (B[a]P) were investigated. Mussels were exposed to a sublethal dose of B[a]P (75 nM; 19 microg/l per animal) for 24, 48 and 72h. The following biological responses were measured in the digestive gland tissues: (1) B[a]P hydroxylase (BPH) activity, as phase I biotransformation parameter; (2) glutathione S-transferase (GST) activity as a phase II conjugation enzyme, (3) catalase (CAT) activity as potential biomarker of oxidative stress, (4) acetylcholinesterase (AChE) activity as an indication of possible neurotoxicity response. DNA damage was assessed over time using the single cell gel electrophoresis comet assay and the micronuclei test. BPH and GST activities showed an increasing trend over exposure period. CAT activity showed a symmetrical bell shape response with a maximum at 48h. AChE activity was significantly depressed after 48 and 72h exposure to B[a]P. Comet assay and micronuclei test in digestive gland cells suggest that B[a]P exposure induced significant DNA damage with a maximum response after 72h exposure.

Development of an expert system for the integration of biomarker responses in mussels into an animal health index.

Biomarkers on sentinel organisms are utilised worldwide in biomonitoring programs. However, the lack of effective interpretational capacity has hampered their uptake for use for assessment of risk in environmental management. The aim of the present study was to develop and test an objective decision-support or expert system capable of integrating biomarker results into a five-level health-status index. The expert system is based on a set of rules derived from available data on responses to natural and contaminant-induced stress of marine mussels. Integration of parameters includes: level of biological organization; biological significance; mutual interrelationship; and qualitative trends in a stress gradient. The system was tested on a set of biomarker data obtained from the field and subsequently validated with data from previous studies. The results demonstrate that the expert system can effectively quantify the biological effects of different levels of pollution. The system represents a simple tool for risk assessment of the harmful impact of contaminants by providing a clear indication of the degree of stress syndrome induced by pollutants in mussels.

Study of GH sensitivity in chilean patients with idiopathic short stature.

We hypothesized that some children with idiopathic short stature in Chile might bear heterozygous mutations of the GH receptor. We selected 26 patients (3 females, 23 males) from 112 patients who consulted for idiopathic short stature at the University of Chile. Their chronological age was 8.3 +/- 1.9, and bone age was 6.1 +/- 1.0 yr. Their height was -3.0 +/- 0.7 SDS; IGF-I, -1.2 +/- 1.1 SD; IGF binding protein 3, -0.7 +/- 2.0 SDS; and GH binding protein, 0.4 +/- 0.8 SDS. Patients were admitted, and blood samples were obtained every 20 min to determine GH concentrations overnight. Coding sequences and intron-exon boundaries of exons 2-10 of GH receptor gene were amplified by PCR and subsequently analyzed through single-strand conformational analysis. Mean serum GH concentration, over 12-h, was 0.20 +/- 0.08 nM; pulse amplitude, 0.40 +/- 0.15 nM; number of peaks, 5.8 +/-1.5 peaks/12 h; peak value of GH during the 12-h sampling, 1.03 +/- 0.53 nM; and area under the curve, 151.4 +/- 56.1 nM/12 h. There were positive correlations between mean GH vs. area under the curve (P < 0.001) and GH peak (P < 0.01). The single-strand conformational analysis of the GH receptor gene showed abnormal migration for exon 6 in 9 patients and for exon 10 in 9 patients, which (by sequence analysis) corresponded to 2 polymorphisms of the GH receptor gene: an A-to-G transition in third position of codon 168 in exon 6 and a C-to-A transversion in the first position of codon 526 in exon 10. We further sequenced all coding exons and intron-exon boundaries in the most affected patients (nos. 6, 9, 11, 14, 15, 16, and 23). This analysis revealed a C-to-T transition in codon 161 of exon 6 in patient 23, which results in an amino acid change (Arg to Cys) in an heterozygous form in the patient and his father. In conclusion, the results of our study suggest that, in Chilean patients with idiopathic short stature, GH receptor gene mutations are uncommon, although we cannot exclude mutations that were missed by single-strand conformational analysis or mutations within introns or in the promoter regions of the GH receptor gene.

[Improved survival in the treatment of ruptured abdominal aortic aneurysm]. / Miglioramento degli indici di sopravvivenza nel trattamento degli aneurismi rotti dell'aorta addominale.

Personal experience in the management of 70 ruptured abdominal aortic aneurysm over the past 5 years is reported dividing patients into 2 groups, before and after establishment of intensive care unit: survival was 30.76% on the first, 66.66% in the second. Factors that allowed that result are discussed.

[Introduction to total parenteral nutrition]. / Introduzione alla nutrizione parenterale totale.

Total parenteral nutrition constitutes a recent resource of intensive therapy that can be applied to a great number of clinical cases. Its realization however requires precise knowledge of metabolic problems and of possible technical complications. As it is not exempt from risks it must be applied with specific competence and in carefully chosen cases.

[The disseminated intravascular coagulation syndrome. Physiopathological and clinical aspects]. / Sindrome da coagulazione intravascolare disseminata. Aspetti fisiopathologici e clinici.

On the basis of the data expressed in the world medical literature the Authors examine the syndrome of disseminated intravascular coagulation, discussing the etiopathogenetic motives, the most significant physiopathologic aspects and the diagnostic-differential and therapeutic problems. They conclude pointing out the necessity of including the eventuality of D.I.C. among the diagnostic possibilities, in as much as the early identification of the syndrome and an adequate therapeutic treatment, employed at the right time, can consent, in not few cases, to save the patient's life.