Ecotoxicological impacts of effluents generated by oil sands bitumen extraction and oil sands lixiviation on Pseudokirchneriella subcapitata.

The exploitation of Athabasca oil sands deposits in northern Alberta has known an intense development in recent years. This development has raised concern about the ecotoxicological risk of such industrial activities adjacent to the Athabasca River. Indeed, bitumen extraction generated large amounts of oil sands process-affected water (OSPW) which are discharged in tailing ponds in the Athabasca River watershed. This study sought to evaluate and compare the toxicity of OSPW and oil sands lixiviate water (OSLW) with a baseline (oil sands exposed to water; OSW) on a microalgae, Pseudokirchneriella subcapitata, at different concentrations (1.9, 5.5, 12.25, 25 and 37.5%, v/v). Chemical analyses of water-soluble contaminants showed that OSPW and OSLW were enriched in different elements such as vanadium (enrichment factor, EF=66 and 12, respectively), aluminum (EF=64 and 15, respectively), iron (EF=52.5 and 17.1, respectively) and chromium (39 and 10, respectively). The toxicity of OSPW on cells with optimal intracellular esterase activity and chlorophyll autofluorescence (viable cells) (72h-IC 50%<1.9%) was 20 times higher than the one of OSW (72h-IC 50%>37.5%, v/v). OSLW was 4.4 times less toxic (IC 50%=8.5%, v/v) than OSPW and 4.5 times more toxic than OSW. The inhibition of viable cell growth was significantly and highly correlated (<-0.7) with the increase of arsenic, beryllium, chromium, copper, lead, molybdenum and vanadium concentrations. The specific photosynthetic responses studied with JIP-test (rapid and polyphasic chlorophyll a fluorescence emission) showed a stimulation of the different functional parameters (efficiency of PSII to absorb energy from photons, size of effective PSII antenna and vitality of photosynthetic apparatus for energy conversion) in cultures exposed to OSPW and OSLW. To our knowledge, our study highlights the first evidence of physiological effects of OSPW and OSLW on microalgae.

Effects of ozone, ultraviolet and peracetic acid disinfection of a primary-treated municipal effluent on the immune system of rainbow trout (Oncorhynchus mykiss).

Municipal sewage effluents are complex mixtures that are known to compromise the health condition of aquatic organisms. The aim of this study was to evaluate the impacts of various wastewater disinfection processes on the immune system of juvenile rainbow trout (Oncorhynchus mykiss). The trout were exposed to a primary-treated effluent for 28 days before and after one of each of the following treatments: ultraviolet (UV) radiation, ozonation and peracetic acid. Immune function was characterized in leucocytes from the anterior head kidney by the following three parameters: phagocytosis activity, natural cytotoxic cells (NCC) function and lymphocyte (B and T) proliferation assays. The results show that the fish mass to length ratio was significantly decreased for the primary-treated and all three disinfection processes. Exposure to the primary-treated effluent led to a significant increase in macrophage-related phagocytosis; the addition of a disinfection step was effective in removing this effect. Both unstimulated and mitogen-stimulated T lymphocyte proliferation in fish decreased dramatically in fish exposed to the ozonated effluent compared to fish exposed to either the primary-treated effluent or to aquarium water. Stimulation of T lymphocytes proliferation was observed with the peracetic acid treatment group. In conclusion, the disinfection strategy used can modify the immune system in fish at the level of T lymphocyte proliferation but was effective to remove the effects on phagocytosis activity.

[Nicorandil-associated anal ulceration]. / Ulcérations anales induites par le nicorandil.

BACKGROUND: Nicorandil is a potassium-channel activator used in the treatment of angina pectoris. The first cases of anal ulcerations induced by nicorandil were published in 2002. CASE REPORT: A 71-year-old man presented with a 2-year history of anal ulcerations occurring within a few months of initiation of treatment with Nicorandil. Histological tests on a biopsy sample showed granulation tissue with non-specific chronic inflammation. Nicorandil was stopped and this resulted in complete healing of the ulcers after three months. DISCUSSION: Nicorandil can induce chronic and extensive anal ulcerations. The pathogenesis is unknown. Patients are usually treated with high doses of nicorandil. Dermatologists should be aware of this rare side-effect which heals after withdrawal of the drug.

Toxicity of nonylphenol on the cnidarian Hydra attenuata and environmental risk assessment.

Alkylphenols and their derivatives, alkylphenol polyethoxylates (APEs), are synthetic chemicals of concern owing to their endocrine properties. Nonylphenol (NP) is a critical APE metabolite because of its recalcitrance to biodegradation, toxicity, and ability to bio-accumulate in aquatic organisms. Studies of NP effects in vertebrates demonstrated estrogenic disrupting properties in fish, birds, reptiles, and mammal cells in which NP displaces the natural estrogen from its receptor. Less is known on its toxicity toward invertebrates. Effects on reproduction have been reported, but toxicity on development has been poorly documented thus far. We investigated NP toxicity on survival and regeneration of the freshwater coelenterate Hydra attenuata. Hydra is known for its regenerative capacity and its sensitivity to chemical pollution. It has been used for over 20 years to screen for teratogenicity of chemicals (Johnson et al. (1982) Teratog Carcinog Mutagen 2:263-276). Our results showed that hydra appeared as one of the most sensitive species to acute and chronic toxicity of NP compared to several freshwater invertebrates. Regeneration was disrupted at NP concentrations lower than those affecting survival. Toxicity thresholds of NP for aquatic vertebrates and invertebrates are also reported and discussed in the context of environmental risk assessment and of water quality objectives recommended for surface waters in industrialized countries. NP levels have decreased during the last decade because of a voluntary agreement of surfactant producers and users. At present, concentrations of NP found in surface waters are far below 1 microg/L in Europe, but can reach several microg/L when wastewater treatment plant inefficiency occurs.

Effects of municipal effluents on serotonin and dopamine levels in the freshwater mussel Elliptio complanata.

Sex differentiation and gametogenesis represent critical steps in the reproductive process and are subject to hormonal control by serotonin, dopamine and steroids such as estradiol-17beta and testosterone. The purpose of this study sought to examine the endocrine-disrupting activity that a primary-treated municipal effluent might have on the metabolism of biogenic amine levels. First, serotonin receptors transfected in Chinese hamster ovary (CHO) cells were used to screen for the presence of serotonin receptor agonist or antagonist. Second, one group of Elliptio complanata mussels were exposed to single compounds likely to be found in municipal wastewaters and another group was exposed in situ to the municipal effluent plume for 90 days in experimental cages. Results showed that solid phase C-8 extracts of surface water downstream a municipal effluent could activate the transport of serotonin by receptors at a distance of at least 5 km from its outfall thereby indicating the presence of serotonin mimics in the effluent dispersion plume. Levels of serotonin and monoamine oxidase (MAO) activity in nerve ganglia of mussels exposed for 90 days to the municipal effluent were, respectively, reduced and increased at a distance 10-km downstream. Injections of estradiol-17beta and nonylphenol in mussels decreased the levels of serotonin and dopamine, but increased MAO activity in the gonad and nerve ganglia. Exposure to estrogenic chemicals present in municipal effluents may therefore alter the normal metabolism of serotonin and dopamine, both of which are involved in sexual differentiation in bivalves and fish. Chemicals acting through E2 receptor-mediated pathways and serotonin receptors are likely to cause the observed effects.

Sex alteration in soft-shell clams (Mya arenaria) in an intertidal zone of the Saint Lawrence river (Quebec, Canada).

The purpose of this study was to verify whether any changes in sex ratio might occur in soft-shell clams (Mya arenaria) located in an intertidal harbor zone located at the mouth of the Saguenay Fjord in the Saint Lawrence estuary (Baie Sainte-Catherine (BSC), Québec, Canada) likely to be contaminated by organotin compounds. Bivalves were harvested at the BSC harbor site and from two reference sites. Condition index (weight to length ratio), gonado-somatic index, sex ratio, vitellin-like proteins, organotin concentrations in gonad tissue, maturation stages of the gonads, the number of estradiol-17beta binding sites and the capacity of female gonad extracts to produce estradiol-17beta were determined in collected animals. Results showed that sex ratio in clams was significantly skewed toward males. Moreover, the condition and gonad-somatic indices, vitellin-like proteins in female gonads and the capacity of female gonads to produce estradiol-17beta were significantly reduced at the harbor site with respect to the reference sites. Maturation status of male gonads was clearly delayed at the harbor site. Additionally, gonad tissue contained tributyltin (TBT) at an average level of 109+/-18 ngSn/gdry wt. at the harbor site while organotins were not detected from the reference sites. Finally, female gonads had a higher number of unoccupied estradiol binding sites at the harbor site indicating low levels of this steroid in this tissue. Overall, this paper is first to report that clams collected in the vicinity of a TBT contaminated harbor are subject to masculinizing effects which seems to be consistent with biological effects that organotins are known to exert toward some other marine invertebrates.

[One center's experience of coronary angioplasty with a 5 French catheter guide by a femoral approach at the same time as coronary angiography]. / Expérience monocentrique de l'angioplastie coronaire par cathéter guide 5 French menée par voie fémorale dans le même temps que l'examen coronarographique.

The object of this study was to assess the feasibility of so-called ad hoc 5 F percutaneous transluminal coronary angioplasty (PTCA). This monocentric register included 200 consecutive procedures (233 lesions) of 5F PTCA by a femoral approach after a bolus of standard heparin (50 to 70 IU/kg). The population included 15.4% of stable angina, 29.4% of unstable angina, 11% acute phase, 13.5% post-revascularisation angina and 30.7% post-infarction cases. A successful procedure was defined as a good angiographic result without ischaemic complications. A failed 5F procedure was defined by the need to fall back on a 6F PTCA. The peripheral vascular complications were recorded. The lesions were stented in 77.4% of cases including 13.4% of direct stenting. There were 200 successful procedures (87%). The failures (N = 26) were mainly explained by the inability to cross chronic obstruction (N = 11). The ischaemic complications included 2 coronary bypasses (2 retrograde dissections of the left anterior descending artery) and 7 enzymatic increases without ECG changes. Fall back to 6F PTCA was required in 4 cases (1.7%) always because of the instability of the 5F catheter guide before the procedure. The quality of coronary contrast was estimated to be good. The vascular complication rate was low with 2% of communicating haematomas (N = 4). Therefore, 5F PTCA is feasible with failure and complication rates comparable to those reported with catheters of larger dimensions. One of its principal advantages is "ad hoc" angioplasty after 5F coronary angiography.

Delayed gametogenesis of Mya arenaria in the Saguenay fjord (Canada): a consequence of endocrine disruptors?

The Saguenay fjord located in Canada on the north coast of the Saint-Lawrence Estuary, is well known for its multiple contaminations following a linear distribution upstream to downstream. Mya arenaria is well established in the fjord and potentially exposed to persistent sediment contamination as an endobenthic bivalve. From May to October 1997, clams energy storage and utilization in the gonad was shown to be closely linked with reproduction. Wherever the sampling site location in the fjord, a seasonal pattern is evident for lipid and glycogen levels in the gonad, this, reflecting the vitellogenic process. Nevertheless, in comparison with energy status and gametogenesis of clams collected on the north coast of the Saint-Lawrence Estuary, clams located in the fjord have to face limited nutritive conditions resulting in a single and shorter reproductive period. Although environmental factors could explain differences in physiological condition and reproductive status observed between clams from the fjord and the estuary, our results can discriminate clams from the upper part of the fjord, according to a delayed gametogenesis, concomitant with significant higher gonad glycogen concentrations observed in June 1996 and 1997. A persistent dysfunction of a vitellogenic process is suspected to be due to the exposure to anti-estrogenic contaminants.

Evidence of coprostanol estrogenicity to the freshwater mussel Elliptio complanata.

Coprostanol (5 beta (H)-cholestan-3 beta ol) is a reduced metabolite of cholesterol produced by micro-organisms found in the intestinal tract of mammals. This substance abounds in urban effluents and is accumulated by organisms living in the vicinity of municipal effluent outfalls. In an earlier study, freshwater mussels exposed to contaminated river water for 62 days accumulated large quantities of coprostanol (Cop) in their soft tissues (16 micrograms/g dry wt.). Moreover, these mussels were found to have elevated levels of vitellin in their hemolymphs, suggesting estrogenic effects. Although municipal wastewaters are known to contain other estrogenic compounds capable of inducing Vn synthesis in mussels, the estrogenic potential of coprostanol was singled out for examination. To this end, mussels were first injected with concentrations of coprostanol via the abductor muscle route, and allowed to stand in aerated water for 72 h at 15 degrees C. The levels of Vn in mussel hemolymph were assayed using the organic alkali-labile phosphate method. A competitive estradiol-binding assay was then devised to measure the ability of coprostanol to compete in the binding of fluorescein-labeled estradiol-albumin to cytosolic proteins. Coprostanol partially reversed the binding of labeled estradiol-albumin to cytosolic proteins with an EC50 of 1 mM. In addition, injections of coprostanol and estradiol-17 beta led to increased levels of vitellins in the hemolymph of treated mussels. Moreover, incubation of cop in gonad homogenate extracts in the presence of NADPH led to the formation of two compounds, as determined by high-performance thin-layer chromatography. One of these compounds appears to be the C17 oxidation product of coprostanol, whose polarity is similar to that of estradiol. The results present evidence that coprostanol is estrogenic to freshwater mussels.

Occurrence of compounds estrogenic to freshwater mussels in surface waters in an urban area.

Estrogens play a major role in the sexual differentiation, gonad development, and oocyte growth of most oviparous organisms. They also stimulate vitellogenesis, the formation of high-density glycolipophosphoprotein that serves as an energy source for the developing embryo. Surface waters from the St. Lawrence River, obtained in the vicinity of an urban area (Montreal, Quebec, Canada), were studied with respect to their estrogenic potential to the freshwater mussel Elliptio complanata. Estrogenicity was measured in water extracts by means of a competitive assay of estradiol binding to cytosolic proteins and by the vitellin-inducing ability of mussel hemolymph following direct extract injection. Surface-water samples drawn downstream of a municipal outfall plume and in a river draining a large farming and agricultural area had high levels of total and fecal coliform bacteria. High levels of estrogen competitors were also found and were able to induce vitellins in injected mussels. Moreover, the estrogen-competing potential of the extracts was found to be significantly correlated with total and fecal coliform bacteria (R = 0.9, p < 0.01) and with the levels of vitellins in the hemolymph (R = 0.62, p = 0.03). The results indicate that water samples drawn from within the municipal effluent plume and from a river draining an agricultural area are estrogenic to freshwater mussels. Thus, the environmental inputs of estrogens are likely to be associated with human sewage and pesticide products.

Evaluation of estrogenic effects of municipal effluents to the freshwater mussel Elliptio complanata.

Municipal effluents are an important source of estrogens to the aquatic environment. The purpose of this study was to examine the estrogenicity of municipal effluents to the indigenous freshwater mussel, Elliptio complanata. First, estradiol-binding sites in gonad homogenates were characterized to determine the binding affinity and specificity of estrogens. Mussels were exposed to increasing concentrations of a municipal effluent for 96 h at 15 degrees C. In another experiment, mussels were placed in cages and submerged for 62 days at 1.5 km upstream and 5 km downstream of a municipal effluent plume in the St. Lawrence River. Mussels were harvested for assessment of vitellogenin-like proteins in the hemolymph and determination of total lipid, carbohydrate and protein in the gonad. The presence of specific estrogen-binding sites was found in both male and female gonads. Binding of estradiol to cytosol proteins reached saturation, yielding a dissociation constant of 0.4 nM. Vitellogenin (Vg) levels increased significantly in both the hemolymph and the gonad after exposure to the effluent. Moreover, females appeared to be more sensitive than males to producing Vg. Mussels exposed in situ to contaminated surface waters had higher levels of Vg at the downstream site, again, females had higher levels of Vg than did males. On the other hand, lipid and sugar levels in male gonads were significantly increased at the downstream site. Moreover, mussels at the downstream site had decreased shell growth length and increased total and soft tissue weights. We conclude that municipal effluents contain bio-available xenoestrogens at levels sufficient to elicit effects in freshwater mussels.

Immediate femoral sheath removal after 6 French routine coronary angioplasty with a weight-adjusted low-dose heparin: results of a prospective registry.

This study assesses the feasibility and safety of immediate sheath removal after coronary angioplasty with the use of 6 French (Fr) guiding catheters by the femoral route and weight-adjusted low-dose heparin (100 IU/kg). We prospectively evaluated such a strategy among a single-center cohort of 261 consecutive patients undergoing routine percutaneous transluminal coronary angioplasty (PTCA). Immediate sheath withdrawal was performed in cases when post-PTCA residual coronary stenosis was less than 30%, with or without stenting. One hundred eighty-two (70%) of the enrolled patients were eligible for immediate sheath removal. When compared with non-eligible patients (sheath removal 4 hours or more post-PTCA), we observed a reduction of hematoma occurrence (15% vs. 30%; p < 0.01), time to manual hemostasis of the puncture site (13.8 +/- 7 vs. 19.7 +/- 12 minutes; p < 0.0001), and time to hospital discharge (2.2 +/- 1.9 vs. 2.8 +/- 1.8 days; p < 0.02), while ischemic event rate was similar (1 vs. 2 non-Q wave myocardial infarction; 2 vs. 1 repeat PTCA for out-of-lab acute vessel closure). In conclusion, a good angiographic result at completion of PTCA using a 6 Fr sheath, even without stenting, makes an immediate sheath removal feasible at no increased risk and with a potential reduction in minor bleeding complications.

Sorption of organic genotoxins to particulate matter in industrial effluents.

In an earlier work [White PA et al. (1996): Environ Mol Mutagen 27:116-139] we examined the genotoxicity of dichloromethane extracts from a variety of industrial effluent samples. in this companion work, we used the SOS Chromotest to investigate the sorption of the extracted genotoxins to effluent suspended particulate matter. The affinity of the genotoxins for particulate matter is expressed as a genotoxicity sorption partition coefficient (Kd-genotox). The results indicate that industries known for their emission of combustion by-products, such as polycyclic aromatic hydrocarbons, often have high Kd-genotox values (>/= 10(6). These include metal refining and founding industries as well metal surface treatment facilities. In contrast, Kd-genotox values for pulp and paper mills and sewage treatment facilities are several orders of magnitude lower (70%) were obtained for metal surface treatment and inorganic and organic chemical production facilities. Low values (>30%) were obtained for sewage treatment facilities and pulp and paper mills. The results also demonstrate the effect of variations in the concentration of available particulate matter on the genotoxicity of both aqueous and particulate extracts. The results suggest that the sorptive properties of the particulate matter itself are reduced when the concentration of particulate matter is very high (>1,000 mg per 1). The use of sorption partition information in inferring the physical-chemical nature of the putative genotoxins and the implications of the results for assessing the hazard posed to aquatic biota by industrial genotoxins are discussed.

A miniaturized Ames mutagenicity assay employing bioluminescent strains of Salmonella typhimurium.

Increased awareness of the role of environmental factors in carcinogenesis has led to an emphasis on preventing or minimizing exposure to genotoxicants. This is presently promoting the development of simple, rapid, cost-effective mutagenicity screening assays. We have developed a test system based on the well-known Salmonella mutagenicity assay. The lux genes, which permit cells to emit light through bioluminescence, were introduced into Salmonella typhimurium strain TA98. These bacteria were exposed for 48 h to chemicals or complex mixtures in 48-well microplates containing an appropriate liquid medium. Cells were subsequently centrifuged and resuspended in buffer. The final postexposure revertant biomass was then estimated using a microluminometer. Replication trials confirmed methodological reproducibility. Clear dose-response relationships were obtained with the direct frameshift mutagens 4NQO and 2NF. Mutagenicity threshold effect concentrations found for these compounds were comparable to those reported in the literature. Industrial effluents and environmental extracts (effluents, suspended solids) were also tested and results compared well with those of the SOS Chromotest. While further validation of this new adaptation of the Ames test will be required, it appears at this time that it could be well suited for routine screening of xenobiotics and environmental samples.

Technique for tricuspid annuloplasty with a flexible linear reducer: medium-term results.

The surgical treatment for tricuspid incompetence requires a choice between reconstruction or replacement with a prosthetic valve. The conservative approach offers many advantages in functional tricuspid incompetence. Our technique for annuloplasty utilizes a flexible linear reducer (FLR) to support a selective reduction of the distended portion of the anulus. The insertion of the septal leaflet is always left free, and physiological flexibility of the tricuspid anulus is assured. The repaired valve is sturdy because of the prosthetic support, and inserting the FLR is simple and poses no risk to the His bundle. The medium-term results (mean follow-up 10 months) in 20 consecutive patients (all with a complete postoperative reinvestigation) are satisfactory. Because this procedure is simple and harmless, it frequently can be applied for direct correction of functional tricuspid incompetence during polyvalvular operation.