Biomedical subjects

Hélène Budzinski

Publications and source records attributed to Hélène Budzinski.

13 recordsLinked to original sources

PubMed2006

Alteration of steroid hormone profile in juvenile turbot (Psetta maxima) as a consequence of short-term exposure to 17alpha-ethynylestradiol.

The synthetic estrogen 17alpha-ethynylestradiol (EE2) is among the most potent xenoestrogens found in treated sewage. Adverse effects have been reported in fish exposed to this chemical, such as vitellogenin induction or alteration of gonad structure. In this study, the effects of EE2 on sex steroid profile in juvenile turbot (Psetta maxima) were investigated in a 15-day in vivo experiment. An analytical method was developed for the simultaneous gas chromatography/mass spectrometry determination of 14 steroids in gonads, plasma and bile. The impact of EE2 on the endocrine status of juvenile turbot was sex-dependent, since males were much more sensitive than females. Even at very low concentration (3.5 ng l(-1)), EE2 depressed androgen production as well as plasma levels in males. Androstenedione and 11-ketotestosterone were highly affected by EE2-exposure, since their production was suppressed in EE2-treated males. Furthermore, testis estrogen production and conjugation increased as a consequence of EE2 exposure. Overall, the ratio of androgens to estrogens in EE2-exposed males dropped in testis and plasma and reached values observed in females. These results indicate that juvenile male turbots are susceptible to hormonal imbalance as a consequence of short-term exposure to environmentally relevant EE2 levels.

Animals↗

PubMed2006

Alteration of steroid hormone balance in juvenile turbot (Psetta maxima) exposed to nonylphenol, bisphenol A, tetrabromodiphenyl ether 47, diallylphthalate, oil, and oil spiked with alkylphenols.

In recent years, concern has been raised about the ability of some classes of environmental contaminants to disrupt the endocrine system of both humans and wildlife. In this study, juvenile turbots (Psetta maxima) were exposed under laboratory conditions to selected waterborne contaminants: oil, oil spiked with alkylphenols, bisphenol A, diallylphthalate, tetrabrominated diphenyl ether 47, and p-nonylphenol as a positive control for "estrogenic-type" effects. This work focused on sex steroids, because these hormones play a key role in the reproduction process. Analytical procedures, involving the off-line coupling of solid phase extraction and gas chromatography/mass spectrometry, were developed for the determination of 12 endogenous sex steroids levels in fish plasma, bile, and gonads. Because of the sexual immaturity of the fish used in this study, however, only six steroids could be detected in juvenile turbots. Bisphenol A and p-nonylphenol exhibited the highest potency towards steroids dynamics, lowering the ratio of androgens to estrogens in all three studied matrices. However, these two chemicals had different modes of action, because p-nonylphenol induced a decrease of androstenedione and 11-ketotestosterone levels, whereas bisphenol A exposure led to an elevation of estrone level. Overall, these two chemicals seemingly disrupted the activity of some steroidogenesis enzymes, leading to serious hormonal imbalance in juvenile turbot.

Animals↗

PubMed2006

Molecular diversity studies of bacterial communities of oil polluted microbial mats from the Etang de Berre (France).

The biodiversity of microbial mats inhabiting the oil-contaminated lagoon Etang de Berre was determined by molecular approaches. The fingerprint of denaturing gradient gel electrophoresis (DGGE) and automatic ribosomal intergenic spacer analysis (ARISA) of mats exposed to different pollution levels showed specific microbial communities for each site but similar diversity richness. Species composition of the mats were compared by constructing 16S rRNA libraries. Amplified rDNA restriction analysis (ARDRA) of clone libraries confirmed their similar level of diversity richness. Phylogenetic analysis of the 16S rRNA sequences showed that the classes gamma and alpha of Proteobacteria were abundantly present in both sites whereas phylotypes related to the delta-Proteobacteria and to the uncultured WS3 group were mainly found in the site with the highest pollution. Identification of the species involved in oil degradation by combining culture-based approaches and DGGE, showed that enrichment cultures were constituted by members of the Rhodobacterales and species related to Rhodococcus, Sphingomonas, Xanthomonas and Microbacterium, all of them known for their ability to degrade hydrocarbons. Our findings suggest that oil pollution has not affected the biodiversity richness of the mats. However, the populations involved in hydrocarbon degradation represent a minor fraction of the mat communities in the Etang de Berre.

Bacteria↗

PubMed2005

Solid-phase extraction and purification for the quantification of polycyclic aromatic hydrocarbon metabolites in fish bile.

An analytical protocol including solid-phase extraction and purification is described for the individual quantification of polycyclic aromatic hydrocarbon metabolites (hydroxylated PAHs) in liquid biological matrices such as plasma and bile. The method consists in an enzymatic deconjugation followed by a solid-phase extraction on a C18 cartridge and by a cleanup on an NH2 cartridge. Extracts are then submitted to a derivatization step before gas chromatography/mass spectrometry (GC/MS) analysis. The quantification of PAH metabolites is ensured by adding an internal standard, 1-hydroxypyrene deuterated, at the beginning of the protocol. Recoveries obtained for the entire protocol were for the major part of the compounds between 96 and 70%. However, recoveries were not so satisfying concerning 2-hydroxybiphenyl and especially 3-hydroxybenzo(a)pyrene, with 62 and 36% respectively. Finally, the protocol was applied to different fish bile samples and showed its good applicability to fish bile samples. The NH2 cleanup step has been proved to be a very selective purification step, necessary to remove most of the bile pigments before GC/MS injection. Different PAH metabolites could be detected in those natural samples and their quantification allowed us to distinguish different levels of fish exposure.

Animals↗

PubMed2005

Analysis of polycyclic aromatic hydrocarbons adsorbed on particles of atmospheric interest using pressurised fluid extraction.

Pressurised fluid extraction (PFE) was used for the measurement of 13 polycyclic aromatic hydrocarbons (PAHs) adsorbed on different types of particles: two model particles (PAH-coated silica, PAH-coated graphite) and two natural atmospheric particles (urban dust and diesel exhaust, from NIST reference materials). Samples were analysed by gas chromatography coupled to mass spectrometry. Extraction efficiency was evaluated with internal standard recovery yields and was shown to depend on the nature of the particle, on the structure of the analytes and on the PAH concentration. Extraction conditions (toluene, 130 degrees C, 130 bar, 2 x 8-min static cycles) were optimised to extract PAHs when strongly interacting with solid matrices and were validated by the analysis of two PAH-certified materials.

Adsorption↗

PubMed2005

Determination of steroidal hormone profiles along the Jalle d'Eysines River (near Bordeaux, France).

Steroidal estrogens have been shown to be the main contributors to the estrogenic activity observed in aquatic systems contaminated with sewage treatment work effluents. Although the occurrence of steroid hormones in the environment has received a great deal of attention, little is known about their fate in aquatic systems. In the presentwork, concentrations of conjugated and unconjugated hormonal steroids (estrone, 17beta-estradiol, estriol, 17alpha-ethynylestradiol, mestranol, progesterone, norethindrone, and D-norgestrel) were determined in the effluent of the Eysines sewage treatment plant (near Bordeaux, France), and along the receiving river, the Jalle d'Eysines River. Sampling was undertaken in summer and in winter conditions, to study both the temporal and the spatial distributions of steroids in this river. Only unconjugated natural estrogens were detected in the effluent. Estrone was the dominant compound (detected in all effluent samples, 17.1-71.0 ng x L(-1)), while estradiol and its metabolite estriol were detected only once, at much lower levels (4.4 and 2.9 ng x L(-1), respectively). Levels of estrogens were clearly raised above the detection limits downstream of the Eysines STP effluent discharge. Seasonal variations of estrone degradation rates were observed. In summer, the apparent decay rates of estrogen levels exceeded that of dilution, indicating high removal rates from the water column: 50% of the initial amount of this steroid was degraded within 1.7 km downstream of the effluent discharge. In winter, however, estrone levels did not significantly decrease over a 10 km reach downstream of the effluent discharge (1.9-1.8 ng x L(-1)). Steroids were determined in the particulate material of the river, but levels were below the detection limits (0.4-1.9 ng x g(-1)), indicating that sorption was not a major sink of estrogens. Therefore, it is likely that biodegradation plays a major role in the removal of steroids from the river and the different decay rates are probably related to differences in bacterial activity within the river.

Environmental Monitoring↗

PubMed2005

Development of an analytical procedure for determination of selected estrogens and progestagens in water samples.

An analytical procedure has been developed for determination of eight selected natural and synthetic hormonal steroids in surface water and in effluent samples. Several methodological points have been investigated and are discussed; they include the choice of the solid-phase extraction sorbent, the influence of flow rate on recovery, the breakthrough volume for a given sorbent (Env+ and Oasis HLB), sample clean up, and sample storage. As regards the latter point, it was found that when no preservative was added to effluent from a sewage-treatment plant, severe loss of steroids occurred-85% of progesterone and about 30% of both estrone and estradiol were found to be degraded in 24 h. The procedure developed was applied to samples from the Seine river estuary. Sex steroids were not detected in surface water; estrone was the most commonly detected steroid in sewage-treatment plant effluent, with levels ranging from 1.8 to 8.3 ng L(-1). Synthetic estrogens (ethynylestradiol and mestranol) and progestagens (levonorgestrel and norethindrone) were never detected, whatever the sampling season. Overall, for 162 out of 168 measurements levels were below the detection limits of the developed procedure.

Clinical Laboratory Techniques↗

PubMed2005

One-year monitoring survey of organic compounds (PAHs, PCBs, TBT), heavy metals and biomarkers in blue mussels from the Arcachon Bay, France.

Marine mussels Mytilus sp. were transplanted on a monthly basis in cages over one year to oyster farms and harbours in the Arcachon Bay (France) in order to assess the water quality of the bay. Contaminant levels (organotin compounds, trace metals, PCBs and PAHs) were measured in tissues of transplanted mussels and mussels from a reference station, along with physiological parameters of the mussels (condition indexes, lipid content and dry weight). Four biomarkers (AChE: acetylcholinesterase activity, GST: gluthathione S-transferase activity, CAT: catalase activity and TBARS: thiobarbituric acid-reactive substance content) were also monitored. The remote stations monitored (oyster parks) exhibited no accumulation pattern of pollutants. Their respective concentrations therefore constitute a background level of the contamination in the bay ([TBT]= 30 ng Sn g(-1) dw, [SigmaHAPs]= 100 ng g(-1) dw, [SigmaPCBs]= 35 ng g(-1) dw). The elevated chemical contamination of the largest harbour of the bay, the Arcachon harbour, can be interpreted in terms of persistence of organotin compounds ([SigmaOTs]= 1500-2000 ng Sn g(-1) dw) and PAHs ([SigmaHAPs]= 4500-5000 ng g(-1) dw) in sediments and, to a lesser extent, of direct inputs of copper ([Cu]= 20 microg g(-1) dw in harbours versus 7 in oyster parks) and petrogenic PAHs ([methylphenanthrenes]= 1600 ng g(-1) dw in the dockyard versus 170 at the gas stations), related to the use of copper-based antifouling paints and to dockyard activity, respectively. However, the Arcachon Bay presents a low contamination level by PCBs and metals, including harbour stations. Furthermore, higher levels of other PAHs (particularly alkyl PAHs such as methylphenanthrenes/1600 ng g(-1) dw) not included in the 16 PAHs from the EPA priority list (usually studied in biomonitoring programmes/1500 ng g(-1) dw) in the Arcachon harbour underline the need to integrate these compounds in biomonitoring of highly PAH-polluted areas such as harbours in order to avoid misinterpretation of the biological responses observed. Biomarker responses were not able to discriminate the different chemical contamination levels recorded in the Arcachon Bay and rather reflected changes in environmental factors. Furthermore, the strong intraspecies variability of biological responses could be due to genetic differences of mussels from the Arcachon Bay. It is the first time that such an integrated monitoring is performed in the Arcachon Bay, also taking into account seasonal variations of chemical contents and biomarkers levels in mussel tissues.

Acetylcholinesterase↗

PubMed2004

Simultaneous determination of the antiretroviral agents: amprenavir, lopinavir, ritonavir, saquinavir and efavirenz in human peripheral blood mononuclear cells by high-performance liquid chromatography-mass spectrometry.

A selective and accurate assay for the simultaneous quantitation of four protease inhibitors (PIs) (amprenavir (APV), lopinavir (LPV), ritonavir (RTV) and saquinavir (SQV)) and a non-nucleoside reverse transcriptase inhibitor (NNRTI) (efavirenz, EFV) in human peripheral blood mononuclear cells using high-performance liquid chromatography-mass chromatography (LC/MS) has been developed and validated. After liquid-liquid extraction, the antiretroviral agents were separated within 15 min. The calibration curves of each drug showed a good linearity in a range of concentration between 2 and 200 ng/3 x 10(6) cells for amprenavir, lopinavir, efavirenz, 1.60 and 128 ng/3 x 10(6) cells for ritonavir and saquinavir. Mean intra- and inter-assay coefficients of variation over the ranges of the standard curves were less than 15% and mean extraction recoveries ranged 88.7-112.1%. The limits of quantification were 2 ng/3 x 10(6) cells for amprenavir, lopinavir, efavirenz, 1 ng/3 x 10(6) cells for ritonavir and 1.6 ng/3 x 10(6) cells for saquinavir. This novel LC/MS assay, which provides an excellent method for simultaneous intra-cellular determination of amprenavir, lopinavir, ritonavir, saquinavir and efavirenz in human peripheral blood mononuclear cells, could be successfully applied for therapeutic drug monitoring and pharmacokinetic studies.

Chromatography, High Pressure Liquid↗

PubMed2004

Virological, intracellular and plasma pharmacological parameters predicting response to lopinavir/ritonavir (KALEPHAR study).

OBJECTIVES: To assess the impact of HIV-1 protease mutations and intracellular and plasma lopinavir minimum concentrations (Cmin) on virological success or failure on lopinavir/ritonavir-containing highly active antiretroviral therapy (HAART). DESIGN: HIV-1-infected HAART-experienced patients included in an observational study, received lopinavir/ritonavir (400/100 mg twice a day) plus two to three nucleoside reverse transcriptase inhibitors (NRTI) or one NRTI plus one non-NRTI. A viral load less than 50 copies/ml at month 6 defined virological success. METHODS: Intracellular and plasma lopinavir concentrations were determined by high-pressure liquid chromatography with mass-spectrometry detection. Reverse transcriptase and protease genes were sequenced at baseline and the time of virological failure. RESULTS: When the 38 patients started the lopinavir/ritonavir-based regimen, baseline median (25-75th percentile) values were: CD4 cell count 218 cells/microl (133-477); plasma HIV-1-RNA load 5.3 log10 copies/ml (3.8-5.1); number of lopinavir mutations four per protease gene (two to six). Univariate analysis associated virological success or failure at month 6 (21/38 patients) with the number of baseline lopinavir mutations, intracellular and plasma lopinavir Cmin, and the genotype inhibitory quotient (GIQ) at months 1 and 6. Multivariate analysis showed that the number of baseline lopinavir mutations and intracellular and plasma lopinavir Cmin were independently associated with virological success or failure. We defined the most discriminating intracellular and plasma lopinavir Cmin efficacy thresholds (8 and 4 microg/ml, respectively) and GIQ thresholds (1 and 3, respectively). CONCLUSION: The monitoring of lopinavir/rironavir-based HAART efficacy should include the number of baseline lopinavir/ritonavir mutations, intracellular and plasma lopinavir Cmin and GIQ calculation.

Adult↗

PubMed2004

Production of highly 13C-labeled polyphenols in Vitis vinifera cell bioreactor cultures.

The use of Vitis vinifera cells grown in a 2 l-stirred tank bioreactor for producing isotopically 13C-labeled phenolic substances is presented. Several culture parameters were optimized to achieve characteristics of growth and polyphenol metabolism similar to that recorded in shake flasks. Administration of [1-13C]L-phenylalanine (3 mM) to grape cell suspension cultures led to the production of 13C-labeled stilbenes (trans- and cis-piceids), catechins (catechin and epicatechin) and anthocyanins (delphinidin-, cyanidin-, petunidin-, peonidin- and malvidin-3-O-beta-glucosides). Incorporation of [1-13C]L-phenylalanine into polyphenols was measured by means of 13C satellites in the proton NMR spectrum and EA-IRMS. The enrichment of labeling obtained for all the compounds (between 40 and 65%) is sufficient to investigate their absorption and metabolism in humans.

Bioreactors↗

PubMed2003

Microcosm tributyltin bioaccumulation and multibiomarker assessment in the blue mussel Mytilus edulis.

Blue mussels, Mytilus edulis, were exposed to two different concentrations of tributyltin (TBT) in seawater, 1,000 ng Sn/L (C1 experiment) and 10 ng Sn/L (C2 experiment), for 4 d, in order to evaluate the bioaccumulation of TBT by mussels Mytilus edulis in microcosms and to test the ability of a multimarkers analysis to determine the effects of TBT on the biochemical parameters in mussels. Tissue burdens of Mytilus edulis were 204 +/- 7 and 2,120 +/- 4 ng Sn/g TBT after the 4-d tests for the C2 and C1 experiments, respectively. Analyses of dissected organs and/or tissues demonstrated that TBT accumulated to the greatest extent in gills in the C1 experiment and in the digestive gland in the C2 experiment. Bioconcentration factors (BCFs) were 12,100 +/- 300 and 2,000 +/- 10 for mussels exposed in the C2 and C1 experiments, respectively. The four biomarkers used in this work were acetylcholinesterase (AChE), glutathione S-transferase (GST), catalase (CAT) activities, and thiobarbituric acid-reactive substances (TBARS) contents. No significant changes were observed in the measured enzyme activities or in TBARS concentration after the 4-d TBT exposure.

Acetylcholinesterase↗

PubMed2002

The effects of elutriates from PAH and heavy metal polluted sediments on Crassostrea gigas (Thunberg) embryogenesis, larval growth and bio-accumulation by the larvae of pollutants from sedimentary origin.

The release, bio-availability and toxicity of contaminants, when sediments are resuspended have been examined, studying concurrently their effects on the embryogenesis and on the larval growth of the Crassostrea gigas larvae and their bio-accumulation in those organisms. Three characteristic sediments have been selected (one contaminated by PAHs, a second by heavy metals and the last by the both pollutants). The organisms were directly exposed to elutriates obtained from each sediment or fed on algae (Isochrysis galbana) contaminated with the same elutriates. The elutriates used in this study show contamination levels similar to those observed in some polluted coastal and estuary environments. The larval growth test has appeared to be more sensitive that the embryotoxicity test. The biological effects and the contaminant bio-accumulation were more pronounced when larvae were directly exposed to different elutriates. In the case of PAHs, the contamination of algae was sufficient to lead to effect on the larval growth of the Crassostrea gigas. In all cases, a fraction of contaminants adsorbed on suspended particles was bio-available and accumulated by the larvae. This study has shown that resuspending polluted sediments constitutes a threat to pelagic organisms and than the C. gigas larval growth may be proposed as a test to protect the most sensitive areas.

Animals↗