Biomedical subjects

Karl Fent

Publications and source records attributed to Karl Fent.

12 recordsLinked to original sources

PubMed2006

Estrogenic activity of UV filter mixtures.

UV-absorbing chemicals (UV filters) are widely used for protection against UV radiation in sunscreens and in a variety of cosmetic products and materials. Depending on the breadth and factor of UV protection, they are added as single compounds or as a combination thereof. Some UV filters have estrogenic activity, but their activity and interactions in mixtures are largely unknown. In this work, we analyzed 8 commonly used UV filters, which are pure or partial hERalpha agonists, for their estrogenic activity in equieffective mixtures in a recombinant yeast assay carrying the human estrogen receptor alpha (hERalpha). Mixtures of two, four and eight UV filters alone, or in combination with 17 beta estradiol (E2), were assessed at different effect levels and no-observed-effect-concentrations (NOEC). Predictions of the joint effects of these mixtures were calculated by employing the concentration addition (CA) and independent action (IA) model. Most binary mixtures comprising of pure hERalpha agonists showed a synergistic activity at all mixture combinations. Only in combination with benzophenone-1, antagonistic activity was observed at some effect levels. All mixtures of four or eight, pure or pure and partial hERalpha agonists, alone or including E2, showed synergistic activity at concentrations giving an increase of 10% of basal activity (BC10). This occurred even at concentrations that were at the NOEC level of each single compound. Hence, there were substantial mixture effects even though each UV filter was present at its NOEC level. These results show that significant interactions occur in UV filter mixtures, which is important for the hazard and risk assessments of these personal care products.

Benzocaine↗

PubMed2006

The ultraviolet filter 3-benzylidene camphor adversely affects reproduction in fathead minnow (Pimephales promelas).

The ultraviolet (UV) filter 3-benzylidene camphor (3BC) is used in personal care products and in a number of materials for UV protection. 3BC has been shown in vitro and in vivo in fish to be estrogenic, but possible effects on fertility and reproduction are unknown. In this study we evaluate whether 3BC affects reproduction of fish Pimephales promelas. After a preexposure period of 21 days, reproductively mature fathead minnows were exposed to increasing concentrations of 3BC for 21 days in a static-renewal procedure. Actual 3BC concentrations decreased to 23% of initial levels and median concentrations were 0.5, 3, 33, 74, and 285 microg/l. 3BC affected reproduction in a dose-dependent manner with weak effects on fecundity at 3 microg/l, a significant decrease at 74 microg/l, and a cessation of reproduction at 285 microg/l. 3BC was accumulated in fish with an average bioconcentration factor of 313 +/- 151. Dose-dependent demasculinization in secondary sex characteristics of male fish and dose-dependent induction of plasma vitellogenin occurred, which was significant at 74 microg/l and higher. 3BC had a profound and dose-dependent effect on the histology of gonads of male and female fish at 3 microg/l and higher. At 74 and 285 microg/l, oocyte and spermatocyte development was inhibited in male and female gonads. Testes of exposed males had much fewer spermatogenic cysts, and ovaries of exposed females had much fewer mature but more atretic, follicles. This study shows significant effects of the UV filter 3BC on fertility, gonadal development, and reproduction of fish after short-term exposure that may have negative consequences on the population level.

Animals↗

PubMed2006

Cytotoxicity of pharmaceuticals found in aquatic systems: comparison of PLHC-1 and RTG-2 fish cell lines.

There is need for a better understanding of the ecotoxicity of pharmaceuticals present in aquatic systems as only little is known about their potential acute and chronic toxicity to aquatic organisms. In our work, we evaluate the in vitro cytotoxicity of 34 common pharmaceuticals from different classes and with different modes of action using the mitochondrial MTT reduction and neutral red uptake assays in the two fish cell lines, PLHC-1 and RTG-2. Cytotoxicity was found for 21 pharmaceuticals with EC50-values ranging from 2.1microM (1.14mgl(-1)) (doxorubicin) to 8.66mM (1200mgl(-1)) (salicylic acid). There was no significant difference between the MTT and NR assays except for an increase in absorption observed with four pharmaceuticals at low concentrations in the MTT assay with PLHC-1 cells indicating a hormesis effect. The comparison of the cell lines revealed that the PLHC-1 cell line was slightly more sensitive than the RTG-2 cell line, showing cytotoxicity at smaller concentrations. The cytotoxicity of pharmaceuticals showed a correlation with their Log D-values at physiological conditions (pH 7.0). A correlation between the in vitro data and in vivo data was found for Daphnia, but not for fish due to insufficient and heterogeneous data. Our work provides an indication that in vitro cytotoxicity assays with fish cell lines could be suited for the first screening of the acute in vivo toxicity of pharmaceuticals, thereby contributing to the reduction of in vivo experiments. Further investigations with a larger set of pharmaceuticals are needed to strengthen the reliability of the assays and to validate the correlation with in vivo data.

Animals↗

PubMed2006

Multiple hormonal activities of UV filters and comparison of in vivo and in vitro estrogenic activity of ethyl-4-aminobenzoate in fish.

UV filters have been detected in surface water, wastewater and fish, and some of them are estrogenic in fish. At present, little is known about their additional hormonal activities in different hormonal receptor systems despite their increasing use and environmental persistence. Besides estrogenic activity, UV filters may have additional activities, both agonistic and antagonistic in aquatic organisms. In our study, we investigate a series of UV filters for multiple hormonal activities in vitro in human receptor systems and evaluate the predictive value of these findings for the activity in fish in vitro and in vivo. First we systematically analysed the estrogenic, antiestrogenic, androgenic, and antiandrogenic activity of 18 UV filters and one metabolite in vitro at non-cytotoxic concentrations with recombinant yeast systems carrying either a human estrogen (hERalpha) or androgen receptor (hAR). All 19 compounds elicited hormonal activities, surprisingly most of them multiple activities. We found 10 UV-filters having agonistic effects towards the hERalpha. Surprisingly, we identified for the first time six UV filters with androgenic activities and many of them having pronounced antiestrogenic and antiandrogenic activities. As much as 17 compounds inhibited 4,5-dihydrotestosterone activity in the hAR assay, while 14 compounds inhibited estradiol activity in the hERalpha assay, indicating antiandrogenic and antiestrogenic activity, respectively. In particular, the antiandrogenic activities of phenyl- and benzyl salicylate, benzophenone-1 and -2, and of 4-hydroxybenzophenone were higher than that of flutamide, a known hAR antagonist. In a second series of experiments, we investigated the predictive power of the hERalpha assay for aquatic organisms by further investigating the estrogenic UV filter ethyl 4-aminobenzoate (Et-PABA) in vitro and in vivo in fish. Et-PABA showed estrogenic activity in a recombinant yeast system carrying the rainbow trout estrogen receptor (rtERalpha) with higher activity than in the hERalpha assay. In addition, Et-PABA induced vitellogenin after 14 days of exposure in juvenile fathead minnows at 4394mug/L. Our study shows estrogenic activity of this UV filter in fish both in vitro and in vivo. In conjunction with in vitro human receptor-based systems our results give a more detailed picture about distinct hormonal activities of UV filters occurring in aquatic systems. We conclude that receptor-based assays are important for in vitro assessment of UV-filters prior to or concurrently with in vivo assays, which ultimately provide data for the environmental risk assessment of these important personal care products.

Androgen Antagonists↗

PubMed2006

Estrogenic activity of pharmaceuticals and pharmaceutical mixtures in a yeast reporter gene system.

Pharmaceuticals enter aquatic environments in unchanged form or as metabolites. Little is known about their potential hormonal activity, which is of particular interest due to potential long-term effects on fertility and reproduction in aquatic organisms. Moreover, there is a need to assess the combined activity of pharmaceutical mixtures. In this study, 37 pharmaceuticals have been analysed in vitro for estrogenic activity using a recombinant yeast system expressing the human estrogen receptor alpha. Six pharmaceuticals belonging to different therapeutic classes, cimetidine, fenofibrate, furosemide, paracetamol, phenazone and tamoxifen, exhibited weak estrogenic activity. Furosemide showed an almost full concentration-response curve, whereas the other compounds showed low efficacy. The half-maximal activities of the pharmaceuticals were in the range of 0.66-25.53 mM. Furthermore, binary mixtures of furosemide and 17beta-estradiol (E2), and furosemide and phenazone, and mixtures of up to five active pharmaceuticals were assessed for their combinatory activity at different equipotent concentrations. The estrogenic activity of binary mixtures of furosemide with E2 and phenazone, respectively, followed the model of concentration addition (CA). Mixtures of other pharmaceuticals often deviated from the CA model, because extrapolations become inaccurate with only partial and non-parallel concentration-response curves having low efficacy. This demonstrates that full and parallel concentration-response curves are a prerequisite for accurate predictions of mixture activity. Our study demonstrates for the first time weak estrogenic activity in vitro of some common pharmaceuticals and their mixtures.

Drug Interactions↗

PubMed2006

Comparison of in vitro and in vivo estrogenic activity of UV filters in fish.

In this work, we evaluate whether in vitro systems are good predictors for in vivo estrogenic activity in fish. We focus on UV filters being used in sunscreens and in UV stabilization of materials. First, we determined the estrogenic activity of 23 UV filters and one UV filter metabolite employing a recombinant yeast carrying the estrogen receptor of rainbow trout (rtERalpha) and made comparisons with yeast carrying the human hERalpha for receptor specificity. Benzophenone-1 (BP1), benzophenone-2 (BP2), 4,4-dihydroxybenzophenone, 4-hydroxybenzophenone, 2,4,4-trihydroxy-benzophenone, and phenylsalicylate showed full dose-response curves with maximal responses of 81-115%, whereas 3-benzylidene camphor (3BC), octylsalicylate, benzylsalicylate, benzophenone-3, and benzophenone-4 displayed lower maximal responses of 15-74%. Whereas the activity of 17beta-estradiol was lower in the rtERalpha than the hERalpha assay, the activities of UV filters were similar or relatively higher in rtERalpha, indicating different relative binding activities of both ER. Subsequently, we analyzed whether the in vitro estrogenicity of eight UV filters is also displayed in vivo in fathead minnows by the induction potential of vitellogenin after 14 days of aqueous exposure. Of the three active compounds in vivo, 3BC induced vitellogenin at lower concentrations (435 microg/l) than BP1 (4919 microg/l) and BP2 (8783 microg/l). The study shows, for the first time, estrogenic activities of UV filters in fish both in vitro and in vivo. Thus we propose that receptor-based assays should be used for in vitro screening prior to in vivo testing, leading to environmental risk assessments based on combined, complementary, and appropriate species-related assays for hormonal activity.

Animals↗

PubMed2005

Ecotoxicology of human pharmaceuticals.

Low levels of human medicines (pharmaceuticals) have been detected in many countries in sewage treatment plant (STP) effluents, surface waters, seawaters, groundwater and some drinking waters. For some pharmaceuticals effects on aquatic organisms have been investigated in acute toxicity assays. The chronic toxicity and potential subtle effects are only marginally known, however. Here, we critically review the current knowledge about human pharmaceuticals in the environment and address several key questions. What kind of pharmaceuticals and what concentrations occur in the aquatic environment? What is the fate in surface water and in STP? What are the modes of action of these compounds in humans and are there similar targets in lower animals? What acute and chronic ecotoxicological effects may be elicited by pharmaceuticals and by mixtures? What are the effect concentrations and how do they relate to environmental levels? Our review shows that only very little is known about long-term effects of pharmaceuticals to aquatic organisms, in particular with respect to biological targets. For most human medicines analyzed, acute effects to aquatic organisms are unlikely, except for spills. For investigated pharmaceuticals chronic lowest observed effect concentrations (LOEC) in standard laboratory organisms are about two orders of magnitude higher than maximal concentrations in STP effluents. For diclofenac, the LOEC for fish toxicity was in the range of wastewater concentrations, whereas the LOEC of propranolol and fluoxetine for zooplankton and benthic organisms were near to maximal measured STP effluent concentrations. In surface water, concentrations are lower and so are the environmental risks. However, targeted ecotoxicological studies are lacking almost entirely and such investigations are needed focusing on subtle environmental effects. This will allow better and comprehensive risk assessments of pharmaceuticals in the future.

Adrenergic beta-Antagonists↗

PubMed2004

Ecotoxicological effects at contaminated sites.

Contamination sites pose significant environmental hazards for terrestrial and aquatic ecosystems. They are important sources of pollution and may result in ecotoxicological effects on terrestrial, groundwater and aquatic ecosystems. At severely contaminated sites, acute effects occur, but the core problem lies in long-term chronic effects. Ecotoxicological effects occur at all levels of biological organization, from the molecular to the ecosystem level. Not only certain organisms may be affected, but the ecosystems as a whole, both terrestrial and aquatic, in its function and structure. Contaminants at large contaminated sites often share critical properties such as high acute and/or chronic toxicity, high environmental persistence, often high mobility leading to contamination of groundwater, and high lipophilicity leading to bioaccumulation in food webs. Contaminants present at polluted sites occur as mixtures, therefore interactions between individual compounds are of importance. The bioavailability is a key factor for ecotoxicological effects of contaminants. This is demonstrated by a case study on organotins. Organotins belong to the most toxic pollutants known so far for aquatic life. Widespread contamination of harbor sediments occurs globally due to the ongoing use of organotins in antifouling paints in large ships. In lake sediments, tributyl- and triphenyltin are very persistent and bioavailable to biota even after a long time. Bioavailability of these compounds is dependent on pH and organic matter. Organotins are accumulated in sediments, but remobilization occurs when contaminated sediments are disturbed and dredged. A key question in dealing with contaminated sites is the assessment and evaluation of the toxicity of contaminants to the environment. Usually, established OECD tests and whole effluent toxicity tests are performed for an ecotoxicological evaluation and for hazard assessment. However, these assays are often expensive, laborious and sometimes not sensitive enough. As a consequence, we have used fast and reliable in vitro systems such as fish cell lines for the evaluation of sediments and landfill leachates contaminated by polychlorinated hydrocarbons (PAH). Determination of cytotoxicity as a measure for acute toxicity, and induction of cytochrome P4501A (CYP1A) as a biomarker of exposure and toxicity were found to be important measures, which can be used for hazard and risk assessment. We have developed a concept for the ecotoxicological evaluation of PAH contamination based on induction equivalents, which can be applied for aquatic and terrestrial ecosystems. One of the key question and present gaps, however, includes the long-term chronic ecotoxicological effects on soil and aquatic biota, which are largely unknown.

Animals↗

PubMed2003

Ecotoxicological problems associated with contaminated sites.

Contamination sites pose significant environmental hazards for terrestrial and aquatic ecosystems. They are important sources of pollution and may result in ecotoxicological effects. At severely contaminated sites acute effects occur, but the core problem lies in possible long-term chronic effects. Ecotoxicological effects occur at all levels of the biological organization, from the molecular to the ecosystem level. Not only certain organisms may be affected, but the ecosystems as a whole in its function and structure. Contaminants at large contaminated sites often share critical properties such as toxicity, high environmental persistence, often high mobility prone to contamination of groundwater, and high lipophilicity resulting in bioaccumulation in food webs. Contaminants present at polluted sites occur as mixtures; therefore, interactions between individual compounds may be of importance. The bioavailability is a key factor responsible for ecotoxicological effects of contaminants: only the bioavailable fraction induces ecotoxicological effects, as shown for organotin compounds. Organotins belong to the most toxic pollutants known so far for aquatic life. Widespread contamination of harbor sediments occurs globally due to the ongoing use of organotins in antifouling paints on large ships. In lake sediments, tributyl- and triphenyltin are very persistent and bioavailable to biota even after a long time. The bioavailability of organotins is dependent on the pH and the content of organic matter. Organotins accumulate in sediments, but remobilization occurs during disturbance and dredging. A key question in dealing with contaminated sites is whether, and to what extent ecotoxicity occurs. Usually, established OECD tests and whole effluent toxicity tests are performed for an ecotoxicological evaluation and for risk assessment. However, these assays are often expensive, laborious and sometimes not sensitive enough. As a consequence, we have used rapid and inexpensive in vitro systems such as fish cell lines for the evaluation of sediments and landfill leachates, which were contaminated by polycyclic aromatic hydrocarbons (PAHs). The determination of cytotoxicity as a measure for acute toxicity, and the induction of cytochrome P4501A (CYP1A) as a biomarker of exposure and effects were found to be important measures, which can be used for hazard and risk assessment. We have developed a concept for the ecotoxicological evaluation of PAH contamination based on induction equivalents, which can be applied for aquatic and terrestrial ecosystems. One of the key question and present gaps, however, includes the long-term chronic ecotoxicological effects of single compounds and mixtures on soil and aquatic biota at contaminated sites. This should be addressed in the future.

Animals↗

PubMed2002

Effects of long-term nonylphenol exposure on gonadal development and biomarkers of estrogenicity in juvenile rainbow trout Oncorhynchus mykiss.

Environmental pollutants with estrogenic activity including nonylphenol (NP) have the potential to alter gonadal development and reproduction of wild fish. To investigate the estrogenic action of environmentally relevant concentrations of NP, rainbow trout (Oncorhynchus mykiss) were continuously exposed during the embryonic, larval and juvenile life stage to 1.05 and 10.17 microg/l NP for 1 year, and sexual differentiation, vitellogenin (VG), VG mRNA, and zona radiata protein (ZRP) expression were examined after that period. The applied NP concentrations did not affect mortality and hatching rates, and did not have an influence on the body weight of 1-year-old fish. No occurrence of testis-ova was observed and sex-ratios of NP exposed groups of fish were unchanged when compared with control groups. The induction of VG and ZRP expression was a more sensitive reaction to the presence of NP than the formation of testis-ova and the reversal of sex. Increased VG expression in trout liver occurred already at 1.05 microg/l NP, whereas VG mRNA levels, quantified by competitive RT-PCR, were not significantly elevated in NP exposed fish. ZRP contents were significantly higher at 10.17 microg/l NP. Since induction of VG did not occur in all fish exposed to 1 or 10 microg/l NP and ZRP induction did not occur in all fish exposed to 10 microg/l, some individuals may be more affected by exposure to NP than others. This study demonstrates that NP concentrations typically found in sewage treatment effluents and some rivers do not affect sexual differentiation in rainbow trout, but induce VG and ZRP expression in the liver of exposed fish.

Animals↗

PubMed2002

Development of a fish reporter gene system for the assessment of estrogenic compounds and sewage treatment plant effluents.

This study reports on the development and application of a fish-specific estrogen-responsive reporter gene assay. The assay is based on the rainbow trout (Oncorhynchus mykiss) gonad cell line RTG-2 in which an acute estrogenic response is created by cotransfecting cultures with an expression vector containing rainbow trout estrogen receptor a complementary DNA (rtERalpha cDNA) in the presence of an estrogen-dependent reporter plasmid and an estrogen receptor (ER) agonist. In a further approach, RTG-2 cells were stably transfected with the rtERalpha cDNA expression vector, and clones responsive to 17beta-estradiol (E2) were selected. The estrogenic activity of E2, 17alpha-ethinylestradiol, 4-nonylphenol, nonylphenoxy acetic acid, 4-tert-octylphenol, bisphenol A, o,p'-DDT, p,p'-DDT, o,p'-2,2-bis(chlorophenyl)-1,1-dichloroethylene (o,p'-DDE), p,p'-DDE, o,p'-2,2-bis(chlorophenyl)-1,1-di-chloroethane (o,p'-DDD), p,p'-DDD, and p,p'-2,2-bis(chlorophenyl)acetic acid (p,p'-DDA) was assessed at increasing concentrations. All compounds except o,p'-DDT, p,p'-DDE, and p,p'-DDA showed logistic dose-response curves, which allowed the calculation of lowest-observed-effect concentrations and the concentrations at which half-maximal reporter gene activities were reached. To check whether estrogen-responsive RTG-2 cells may be used to detect the estrogenic activity of environmental samples, an extract from a sewage treatment plant (STP) effluent was assessed and found to have estrogenic activity corresponding to the transcriptional activity elicited by 0.05 nM of E2. Dose-response curves of nonylphenol, octylphenol, bisphenol A, and o,p'-DDD revealed that the RTG-2 reporter gene assay is more sensitive for these compounds when compared to transfection systems recombinant for mammalian ERs. These differences may have an effect on the calculation of E2 equivalents when estrogenic mixtures of known constitution, or environmental samples, such as STP effluents, are assessed.

Animals↗

PubMedDate not supplied

Assessment of hormonal activity of UV filters in tadpoles of frog Xenopus laevis at environmental concentrations.

Residues of UV filters used as ultraviolet absorbers in sunscreens, cosmetics and in light protection have been found in surface water and fish. Recently some of them were shown bearing estrogenic activity, however, very little is known about possible adverse effects to aquatic life. Here we investigate whether two UV filters, 4-methylbenzylidene camphor (4-MBC) and 3-benzylidene camphor (3-BC), interfere with the thyroid and sex hormone system during frog metamorphosis. Xenopus laevis tadpoles were exposed to 1, 5 and 50 microg/L of 4-MBC and 3-BC, respectively, for 35 days (NF stage 52-66). The rate of metamorphosis was not affected, and no obvious differences in body and tail length compared to controls were observed. Neither 4-MBC, nor 3-BC led to effects on the sex ratio or gross gonad morphology of X. laevis at stage 66. Our results indicate that these UV filters do not negatively affect the thyroid system and sex ratio of frogs at environmental concentrations.

Animals↗