In vivo exposure of marine mussels to carbamazepine and 10-hydroxy-10,11-dihydro-carbamazepine: Bioconcentration and metabolization.

Aquatic organisms are exposed to pharmaceuticals present in natural waters, but few data are available on the accumulation of these substances in such organisms. The present study evaluated the in vivo bioconcentration of two anticonvulsants--carbamazepine (CBZ) and 10-hydroxy-10,11-dihydro-carbamazepine (10 OH)--in marine mussels (Mytilus galloprovincialis) exposed to nominal 10 µg L(-1) concentrations for one week. The bioconcentration factors (BCFs) were 3.9 and 4.5 L kg(-1) dry weight (dw) for CBZ and 10 OH, respectively. CBZ accumulation reached an average tissue concentration of 29.3 ± 4.8 ng g(-1) dw, and 10 OH accumulated up to 40.9 ± 4.6 ng g(-1) dw in tissues within one week, showing first-order kinetics. BCF obtained with linear QSAR models correctly estimated the CBZ bioconcentration and overestimated the 10 OH bioconcentration to some extent. The detection of two metabolites (carbamazepine-10,11-epoxide and acridine) among the five sought suggested an active metabolism for CBZ. In contrast, none of the 10 OH metabolites were detected in mussels exposed to 10 OH. CBZ showed higher accumulation in the digestive gland, where some relevant metabolites were detected, than in other studied tissues. The implication of those findings on field biomonitoring is discussed.

Detection of emerging contaminants (UV filters, UV stabilizers and musks) in marine mussels from Portuguese coast by QuEChERS extraction and GC-MS/MS.

The UV filters and musk fragrances have come into focus because these compounds are contained and increasingly used not only in sunscreen products but also in many products of daily use, such as cosmetics, skin creams, plastics or varnish. In view of this, the main objective of the present work was to develop and validate a method for the determination of three UV filters, two UV stabilizers and four musks in mussel samples (Mytilus galloprovincialis). The procedure combined a QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) extraction method with an analysis by gas chromatography-tandem mass spectrometry (GC-MS/MS). The methodology allowed the determination of target analytes at trace concentration levels (ng/g), with mean recoveries ranging from 91 to 112%. A monitoring study was conducted in four beaches in the Portuguese coast which are impacted by recreational activities and outflow of treated waste water effluents in rivers. The results are used to assess the occurrence of UV filters in comparison with UV stabilizers and musk fragrances which indicate other activities than bathing.

¿Se cumplen los tratamientos fisioterapéuticos infantiles? La importancia del profesional sanitario / Is there compliance in child physiotherapy treatments? The importance of the health care professional

Objetivos: Estimar el grado de adhesión terapéutica al tratamiento rehabilitador en enfermedad motora del desarrollo de niños entre 0 y 6 años e identificar las variables psicosociales predictoras de la misma. Método Estudio transversal retrospectivo en el que se entrevistó a todos los cuidadores principales de los niños que asistían a tratamiento fisioterapéutico en el Servicio de Fisioterapia del Hospital General Universitario de Alicante y el Centro de Atención Temprana de Alicante. En total, se realizaron 88 entrevistas, de las que solo se analizaron 79 por errores en la cumplimentación. Las variables psicosociales evaluadas fueron: apoyo social, confianza hacia el profesional, autoeficacia percibida y ansiedad. Resultados El modelo de regresión logística binaria realizado para pronosticar la probabilidad de adhesión al tratamiento por el cuidador principal indicó que en un 86,7% de los casos la adhesión al tratamiento estuvo pronosticada por el grado de confianza que los padres tienen en los fisioterapeutas, así como por su percepción de apoyo social. No resultó estadísticamente significativo para otras variables (autoeficacia percibida, ansiedad o las variables sociodemográficas).Conclusiones Este estudio muestra la importancia de variables relacionadas con el entorno socioprofesional frente a variables individuales en la adhesión al tratamiento rehabilitador. Entre ellas, destaca el papel del fisioterapeuta como motivador de conductas de adhesión en los padres o responsables de los niños. Este resultado subraya la necesidad de formación de los fisioterapeutas en habilidades de comunicación y apoyo emocional para mejorar la implicación de los adultos en este tipo de tratamientos para niños con afección del desarrollo

Objectives: To estimate the level of patient compliance regarding the rehabilitation treatment in 0 to 6 year old children with developmental motor conditions and to identify the predicted psychosocial variables. Method: A retrospective, cross-sectional study was performed by interviewing the caregivers of children undergoing physiotherapy in the General University Hospital of Alicante and the Center for Early Intervention in APSA Alicante, Spain. A total of 88 interviews were performed, 79 of which were analyzed and 9 of which were discarded due to errors in filling out the questionnaires. The psychosocial variables evaluated were social support, confidence in the professional, perceived self-efficacy and anxiety. Results: The binary logistic regression model performed to predict the probability of patient compliance perceived by the principal caregiver indicated that in 86.7% of the cases, patient compliance was predicted by the degree of confidence the parents had in the physiotherapist as well as their perception of social support. No statistically significant results were detected in the other variables (perceived self-efficacy, anxiety or sociodemographic variables).Conclusions: This study has demonstrated the importance of the variables related to the social-professional environment versus the individual variables in the rehabilitating treatment compliance. Of these, it is important to note the role of the physiotherapist as a motivator towards compliance behaviors in the parents or tutors. This results emphasizes the need to educate physiotherapists in communication skills and emotional support in order to improve the involvement of the adults in these types of treatments for children with developmental conditions

Occurrence of venlafaxine residues and its metabolites in marine mussels at trace levels: development of analytical method and a monitoring program.

Coastal areas are subject to growing pressures and impacts because of the increase in human activities. Lipophilic organic contaminants, such as polycyclic aromatic hydrocarbons (PAHs) or polychlorinated biphenyls (PCBs), have been monitored for decades within monitoring programs. However, until now, little information on the detection of so-called "emerging contaminants" such as hydrophilic organic compounds in the marine environment and no data on its metabolites or transformation products in marine organisms is available. In this report, a sensitive analytical methodology for identification and confirmation of venlafaxine (VEN) residues and five of its main metabolites in the marine mussels Mytilus galloprovincialis was validated. The sample preparation procedure was based on the Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) approach. An analytical method was developed to quantify these compounds at trace levels by liquid chromatography coupled to high-resolution mass spectrometry. The method was then applied to marine mussels collected from the Mediterranean Sea in southeastern France. Residues of the antidepressant VEN were occasionally detected at ng/g dw level. In addition, the approach allowed us to identify several transformation products in the analyzed samples. N-desmethylvenlafaxine (NDV) was the most frequently detected metabolite followed by N,O-di-desmethylvenlafaxine (NODDV).

Fast and easy extraction combined with high resolution-mass spectrometry for residue analysis of two anticonvulsants and their transformation products in marine mussels.

Environmental field studies have shown that carbamazepine (Cbz) is one of the most frequently detected human pharmaceuticals in different aquatic compartments. However, little data is available on the detection of this substance and its transformation products in aquatic organisms. This study was thus mainly carried out to optimize and validate a simple and sensitive analytical methodology for the detection, characterization and quantification of Cbz and oxcarbazepine (Ox), two anticonvulsants, and six of their main transformation products in marine mussels (Mytilus galloprovincialis). A modified QuEChERS extraction method followed by analysis with liquid chromatography coupled to high resolution mass spectrometry (HRMS) was used. The analyses were performed using two-stage fragmentation to reveal the different fragmentation pathways that are highly useful for the identification of isomeric compounds, a common problem when several transformation products are analyzed. The developed analytical method allowed determination of the target analytes in the lower ng/g concentration levels. The mean recovery ranged from 67 to 110%. The relative standard deviation was under 11% in the intra-day and 18% in the inter-day analyses, respectively. Finally, the method was applied to marine mussel samples collected from Mediterranean Sea cultures in southeastern France. Residues of the psychiatric drug Cbz were occasionally found at levels up to 3.5ng/g dw. Lastly, in this study, other non-target compounds, such as caffeine, metoprolol, cotinine and ketoprofen, were identified in the real samples analyzed.

Estrogen-like and dioxin-like organic contaminants in reclaimed wastewater: transfer to irrigated soil and groundwater.

In semi-arid regions, treated wastewater reuse for irrigation is a common practice since wastewater is considered as a non negligible water resource in these areas. However, treated wastewater contains traces of organic compounds which may contaminate the receiving environment i.e. soil and groundwater. Some of these organic compounds have the ability to bind to estrogen receptor (ER) or dioxin receptor (AhR, aryl hydrocarbon receptor). The fate of these compounds in effluent reused for irrigation, irrigated soils and groundwater is not well addressed yet. In the present study, estrogenic and dioxin-like activities were studied in three media: i) effluents reused for irrigation, ii) soils samples collected from the reclaimed water irrigated plot during six month irrigation, and iii) groundwater sampled before and after irrigation periods. Effluents reused for irrigation exhibited ER and AhR activities at 38.5 +/- 9.9 ng estradiol-equivalent/L (ng E2-EQ/L) and 113.3 +/- 27.7 ng dioxin-equivalent/L (ng TCDD-EQ/L), respectively. Soils showed ER activity (0.05 ng E2-EQ/g) only after 4 months of irrigation. AhR activities detected in all soil samples have not changed during irrigation. In groundwater, ER activities were detected in two piezometers indicating transfer of some estrogenic compounds.

Impact of urban wastewater discharges on the sediments of a small Mediterranean river and associated coastal environment: assessment of estrogenic and dioxin-like activities.

The Mediterranean region includes many small coastal rivers about which little is known concerning organic contaminant loads in their sediment. This study was designed to assess organic contamination in one of these small coastal rivers (Lez River) and associated coastal sediments. Levels of alkylphenols (APs), polycylic aromatic hydrocarbons (PAHs), and polychlorinated biphenyls (PCBs) were measured in sediments of the Lez River and two coastal lagoons impacted by wastewater discharges. In parallel, sediment surrounding a recently built submarine outfall that discharges treated wastewater, from an area encompassing some 450,000 inhabitants, into the sea was monitored a year after the beginning of emission via the outfall. Finally, these sediments were characterized by screening estrogenic, PAH-like and dioxin-like activities using in vitro bioassays. Both chemical analyses and bioassays revealed that wastewater inputs were a source of organic contamination of sediments from the Lez and lagoons, which still persisted 2 years after the discharges were stopped. APs could explain a small proportion of the overall estrogenic activities (up to 31%), suggesting that other estrogenic compounds were also present in the sediments. PAHs explained a great share (83% on average) of the EROD induction potency of the extracts. This survey should be the first step in the long-term monitoring of these sites.

Sludge accumulation in an anaerobic pond and viability of helminth eggs: a case study in Burkina Faso.

Accumulation rates and pathogen concentrations in primary stabilization pond sludges in developing countries are important parameters for adequate sludge management and the safeguarding of public health with sludge reuse in agriculture. An anaerobic pond has been investigated for sludge accumulation rates and helminth egg viability after four years of operation in Burkina Faso. The rate of sludge accumulation was measured at 0.037 m(3)/capita-year or 2.26 kg dry weight/capita-year. An equation describing vertical distribution of total solids in the accumulated sludge was found to be adequately represented by a regression equation. Influent helminth egg concentrations were reduced on average by 90% in the anaerobic pond effluent. Ascaris lumbricoides and Ancylostoma sp. were the most common eggs present in the sludge after four years of operation. The average concentration of helminth eggs in pond sludge was 536 eggs/g TS, and the percentages of viability ranged from 10.8% (47 viable eggs/g TS) to 57.2 (1,772 viable eggs/g TS, with an average rate of 36% (336 viable eggs/g TS). From a sludge depth and section study, egg viability was found to be randomly distributed in the sludge layer.

Fate of endocrine disrupters in waste stabilization pond systems.

The performance in the removal of estrogenicity from wastewater was studied in three wastewater treatment plants (WWTPs). Different treatment processes were evaluated: stabilization ponds and trickling filter. Sampling was performed from the input to the output of the treatment systems. The total estrogenic activity was determined with MCF-7-derived cell lines which express the endogenous estrogen receptor alpha. The two wastewater stabilization ponds with long retention time had high removal of estrogenicity (90% to 95%). Trickling filters despite being effective at removing organic load were less effective in removing estrogenicity (42%), and post tertiary ponds enhanced estrogenicity removal.

Polycyclic aromatic hydrocarbon metabolites and 7-ethoxyresorufin O-deethylase activity in caged European eels.

This study investigated the contribution of two biomarkers, bile polycyclic aromatic hydrocarbon (PAH) metabolites and 7-ethoxyresorufin O-deethylase (EROD), activity in the assessment of PAH contaminated sites. European eels (Anguilla anguilla) were caged in a freshwater stream upstream and downstream from local industrial effluent outlets. Bile PAH metabolites were recorded as fluorescent aromatic compounds by synchronous fluorescence spectroscopy and as a marker for total PAH metabolism: 1-hydroxypyrene (1-OH Pyr) was isolated by high-pressure liquid chromatography and quantified. After 14 and 28 days of caging, EROD activity, bile fluorescence (synchronous fluorometric measurement), and 1-OH Pyr concentrations in bile were higher at the downstream site than at the upstream site. This increase was similar after 2 and 4 weeks of caging. During a reversibility study, EROD activity, bile fluorescence, and 1-OH Pyr concentrations decreased, and this trend was similar for the three markers. These results suggest that PAHs could be the main factor responsible for EROD induction in eels caged at the downstream site.

Impact assessment of various rearing systems on fish health using multibiomarker response and metal accumulation.

European sea bass were reared in three different systems: one flow-through (FTS), one recirculating (RAS), and one recirculating with a high-rate algae pond (RAS + HRAP). After 1 year of rearing, the final fish weight was 15% lower in the RAS compared to the FTS. The accumulation of a growth-inhibiting substance in the RAS is the main hypothesis explaining this difference. As in environmental risk assessment, fish bioaccumulation markers and biomarkers were used to demonstrate exposure to and effects of the rearing water in the three rearing systems. Thirty fish per system were sacrificed before their condition factor (CF) and liver somatic index (LSI) were calculated. Nine biomarkers, including ethoxyresorufin-O-deethylase (EROD) and superoxide dismutase (SOD), were measured in liver and twelve metals including As, Cd, Cu, Pb, Cr, and Zn, for which there are regulations regarding human consumption, were measured in liver and muscle. In all systems, CF and LSI were not significantly different and no correlation was found with biomarker activity or metal concentration. EROD and SOD activities were significantly increased in RAS. Accumulation of seven and four metals in muscle and liver, respectively, was significantly higher in the RAS relative to FTS. The HRAP prevented metal accumulation except for chromium and arsenic. Eight metal concentrations were significantly higher in liver than in muscle. Concentrations of toxic metals were similar to reported values and below FAO/WHO recommended values for human consumption.

Estrogenic activity of cosmetic components in reporter cell lines: parabens, UV screens, and musks.

In this work, the estrogenic effects of three classes of substances included in cosmetic formulations-parabens, ultraviolet (UV) screens, and musk fragrances-were studied. Their estrogenic activity was measured with the use of three reporter cell lines: HELN, HELN ERalpha, and HELN ERbeta. These three cell lines allowed for the measurement of estrogenic activity toward estrogen receptors alpha and beta (ERalpha and ERbeta, while taking nonspecific interactions into account. Eight of the 15 substances tested showed specific estrogenic activity with the following degree of potency on ERalpha butylparaben > propylparaben > homosalate = octyl-dimethyl-PABA = 4-methyl-benzylidenecamphor = octyl-methoxycinnamate > ethylparaben = galaxolide. Among these active substances, parabens activated ERalpha and ERbeta similarly, UV screens activated ERalpha moderately and had almost no effect on ERbeta, and fragrances did not activate ERbeta. Methylparaben, ethylparaben, musk moskene, celestolide, and cashmeran did not activate estrogenic responses up to 10(-5) M. Musk ketone and benzophenone-3 were not considered estrogenic at 10(-5) M.

Effects of human pharmaceuticals on cytotoxicity, EROD activity and ROS production in fish hepatocytes.

Pharmaceuticals are found in the aquatic environment but their potential effects on non-target species like fish remain unknown. This in vitro study is a first approach in the toxicity assessment of human drugs on fish. Nine pharmaceuticals were tested on two fish hepatocyte models: primary cultures of rainbow trout hepatocytes (PRTH) and PLHC-1 fish cell line. Cell viability, interaction with cytochrome P450 1A (CYP1A) enzyme and oxidative stress were assessed by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrasodium bromide tetrazolium (MTT), 7-ethoxyresorufin-o-deethylase (EROD) and dichlorofluorescein (DCFH-DA) assays, respectively. The tested drugs were clofibrate (CF), fenofibrate (FF), carbamazepine (CBZ), fluoxetine (FX), diclofenac (DiCF), propranolol (POH), sulfamethoxazole (SFX), amoxicillin (AMX) and gadolinium chloride (GdCl(3)). All substances were cytotoxic, except AMX at concentration up to 500 microM. The calculated MTT EC(50) values ranged from 2 microM (CF) to 651 microM (CBZ) in PLHC-1, and from 53 microM (FF) to 962 microM (GdCl(3)) in PRTH. CF, FF, and FX were the most cytotoxic drugs and induced oxidative stress before being cytotoxic. Compared to hepatocytes from human and dog, fish hepatocytes seemed to be more susceptible to the peroxisome proliferators (PPs) CF and FF. In PLHC-1 cells none of the tested drugs induced the EROD activity whereas POH appeared as a weak EROD inducer in PRTH. Moreover, in PRTH, SFX, DiCF, CBZ and to a lesser extend, FF and CF inhibited the basal EROD activity at clearly sublethal concentrations which may be of concern at the biological and chemical levels in a multipollution context.

Estrogenic activity in water and sediments of a French river: contribution of alkylphenols.

Alkylphenols, known to possess estrogenic activity, have been found in the aquatic environment. In this study, we focused on the contribution of alkylphenols to total estrogenic activity in sediment and water extracts of French rivers. Four sites representing rural, agricultural, urban, and industrial watersheds were studied. The concentrations of alkylphenols in water and sediment were quantified by GC/MS. Estrogen-responsive reporter cell lines (MELN) have been used for investigating estrogenic activity at these sites. These observed activities were compared with activities mediated by known concentrations of alkylphenols. In water, the concentration of alkylphenols, from 0.06 to 0.550 microg x L(-1) and from < 0.001 microg x L(-1) to 0.077 microg x L(-1) for nonylphenols and 4t-octylphenol, respectively, were too low to contribute to the observed estrogenic activity. In sediment of the industrial, rural, and urban sites, the observed estrogenic activities could be explained in great part by the alkylphenol concentrations from 0.26 to 2.87 microg x g(-1) and from 0.005 microg x g(-1) to 0.49 microg x g(-1) for nonylphenols and 4t-octylphenol, respectively. In the agricultural site, the alkylphenols (0.022 microg x g(-1) of nonylphenols) poorly contribute to the observed estrogenic activity. Other compounds, such as natural and synthetic hormones, present in water and sediments could act additively in the overall activity.

Reporter cell lines to study the estrogenic effects of xenoestrogens.

In order to characterize the estrogenic activity of chemicals, we established complementary in vitro recombinant receptor-reporter gene assays in stably transfected MCF-7 and HeLa cells. MCF-7 cells which express the endogenous estrogen receptor alpha (ER alpha) were stably transfected with only an estrogen-regulated luciferase gene. These cells enable the detection of compounds which bind to ER alpha or interfere with the induction of ER alpha mediated gene expression. Furthermore, HeLa cells, which do not express endogenous ERs, were transfected with an ER alpha or an ER beta construct together with an estrogen-regulated luciferase gene, or a chimeric GAL4-ER alpha receptor and the corresponding luciferase reporter gene. Finally, we tested these four cellular models as tools to check the estrogenic activities of several potential xenoestrogens and to detect estrogenic activity in wastewater sewage treatment effluents. In all of the models, nonylphenol mixture (NPm), 4n-nonylphenol (4nNP), 2,4'-DDE, 4,4'-DDE and wastewater sewage treatment effluent were active, while PCB mixture (Aroclor 1254), PCB 77, atrazine and lindane (gamma hexachlorocyclohexane) were inactive. Dioxin partially activates the estrogen receptor in MCF-7 cells while in HeLa-derived cell lines, it decreased the estrogenic-induced expression of luciferase.

Laboratory and field-caging studies on hepatic enzymatic activities in European eel and rainbow trout.

Caging field experiments were conducted on young European eel (Anguilla anguilla) and on rainbow trout (Onchorynchus mykiss) for the determination of a biomonitoring protocol of freshwater streams. Hepatic monooxygenase and conjugation enzyme activities were measured as biomarkers of exposure to some organic pollutants. Laboratory studies were performed to compare the responsiveness of these two species concerning their enzymatic activities. The induction of monooxygenase activity, measured as the deethylation of 7-ethoxyresorufin (EROD activity) was examined by beta-naphtoflavone (beta-NF) treatment. Dose-response study revealed that EROD activity increased significantly from 1 mg beta-NF/kg bw and was on a plateau at 50 mg beta-NF/kg bw for both species. No significant effect on glutathione S-transferase (GST) activity was observed. During a time-course study, EROD activity increased, starting from the second day for both species. In European eel, this enzymatic activity decreased significantly from the seventh day, whereas no significant decrease was observed for rainbow trout after 14 days. During a 2-week field study, EROD and GST activities were measured in fish held in cages upstream and downstream of a polluted area. For both species, the induction of EROD activity was observed in the polluted site after 14 days of caging; GST activity was not significantly affected. Monooxygenase activity appeared to be a sensitive tool for biomonitoring freshwater streams for both species.

Reliability and feasibility of pregnancy home-use tests: laboratory validation and diagnostic evaluation by 638 volunteers.

All 27 home-use tests sold in France in 1989 for the self-diagnosis of pregnancy were evaluated. The kits were first tested by qualified clinical chemistry technologists. Eleven kits with 100% specificity and 100% sensitivity were retained for the diagnostic study. Each of 638 laywomen was given a kit and asked to perform the assay with a coded urine specimen containing either no human chorionic gonadotropin (hCG) or an hCG concentration adjusted to the claimed detection limit (1 DL) or twice the detection limit (2 DL). After testing, each participant filled out a detailed questionnaire. The results showed a diagnostic specificity of 86-100% for 10 kits but a diagnostic sensitivity of 85-100% for only 5 kits at 2 DL and for only 2 at 1 DL. Among the 478 positive urine samples distributed, 230 were falsely interpreted as negative. The main explanation for such a high percentage of false-negative results was difficulty in understanding the explanatory leaflets accompanying the kits and hence in reading the results, regardless of the socioeconomic situation of the participant. We conclude that pregnancy home-use tests should be subjected to rigorous analytical controls and evaluated by a panel of potential users before being released on the market.