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Assessing estrogenic and anti-estrogenic activity of antimicrobial compounds used in oral care products
S206
Abstracts / Toxicology Letters 280S (2017) S202–S213
the specific respiratory impact of atmospheric ultrafine particles compared to fine particulate matter in a relevant in vivo model of sub-chronic exposure to these pollutants. Quasi-ultrafine (PM0.18 ) and fine (PM2.5 ) particles have been collected in the urban industrial zone of Dunkirk in north France during a 7-month campaign. The physico-chemical study of the collected particles shows that there is no major difference in elemental and surface chemical composition between PM0.18 and PM2.5. BALB/c mice were then exposed intranasally to 10 g of PM0.18 or PM2.5 3 times a week. After 1 or 3-month exposure, broncho alveolar lavages (BAL) were performed and lung tissues were harvested for histological, epigenetic and transcriptomic analyses. Cytological analyses show that both types of particulate fractions can be internalized in lung cells. Cellular analyses of BAL and preliminary transcriptomic data suggest that PM0.18 induced a stronger lung inflammation and mRNA and miRNA deregulations than PM2.5 . Complementary studies are in progress to confirm these first data and to identify the metabolic pathways more specifically associated with the toxicity of ultrafine particles. http://dx.doi.org/10.1016/j.toxlet.2017.07.563 P-06-01-13 Air quality assessment during indoor use of the tobacco heating system THS2.2 Catherine Goujon Ginglinger, Maya Mitova, Serge Maeder, Maurice Smith Research and Development, Philip Morris Products SA, Neuchatel, Switzerland PMI’s heat-not-burn Tobacco Heating System 2.2 (THS 2.2) uses an electronically controlled heating mechanism to precisely heat specially designed tobacco sticks at operating temperatures well below combustion. To address public health concerns about possible presence of polluting substances during indoor use of THS 2.2, a study was designed to compare the environmental aerosol produced by THS 2.2 to background air (no use of any product). A dedicated controlled room to simulate a residential environment with a low ventilation rate (0.5 air changes/h) was used. Twenty three analytes (nicotine, 3-ethenylpyridine, solanesol, respirable suspended particles (RSP) by gravimetric measurement, particulate matter by UV, particulate matter by fluorescence, total volatile organic compounds (TVOCs), acetaldehyde, acrolein, crotonaldehyde, formaldehyde, acrylonitrile, benzene, 1,3-butadiene, isoprene, toluene, glycerin, propylene glycol, N-nitrosonornicotine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, carbon monoxide, nitrogen oxide and combined oxides of nitrogen) were quantified by validated and ISO17025 accredited methods. In comparison to background air, only three compounds can be attributed to the use of THS 2.2: nicotine (1.15 g/m3 ), acetaldehyde (3.44 g/m3 ) and glycerin (10.5 g/m3 ). Their levels are much lower than maximum exposure levels as defined in existing air quality guidelines. Markers of combustion are absent in environmental aerosols of THS 2.2. Based on TVOC data analyses, the chemical composition of the background air and air during use of THS 2.2 were highly similar. In conclusion, using THS 2.2 does not have a negative impact on the overall air quality. http://dx.doi.org/10.1016/j.toxlet.2017.07.564
P-06-01-14 Behavioral effects in adult zebrafish after developmental exposure to carbaryl Daniela Correia, Ana Rita Almeida, Joana Santos, Amadeu M.V.M. Soares, Inês Domingues Department of Biology & CESAM, University of Aveiro, Campus Universitário de Santiago, Aveiro, Portugal In recent years, there is a growing concern to correctly assess the risk of low environmental concentrations of contaminants (e.g. pesticides) to aquatic life. Pesticides can be found in the ng/L and g/L range and, very often, observable effects to non-target organisms like fish, may not be perceptible immediately and long term effects can be underestimated. Hence, our work intends to evaluate the effect of an early (embryonic) exposure to carbaryl in adult fish behaviour. Zebrafish (Danio rerio) embryos were exposed to 4 sublethal concentrations (0, 10, 100 and 1000 mg/L) of carbaryl during 4 days and then kept in standard cultivation conditions until adulthood. Two behaviour tests were then performed to assess anxiety-like behavior (novel tank test and light/dark stimulus) plus a feeding test. Our data showed that zebrafish pre-exposed to carbaryl, remained longer periods in the bottom layer of the aquarium suggesting a decrease of the exploratory behavior. Further, fish preexposed to 1000 mg/L of carbaryl showed lower swimming activity (distance moved and time spent swimming) than non-exposed fish and presented reduced thigmotaxis during the dark stimulus when compared to control, suggesting repression of anxiety-like behavior. Pre-exposed fish also took more time to feed than non-exposed fish. Our results show that developmental exposure to carbaryl has long lasting effects on adult behaviour highlighting the importance of the environmental conditions during early life stages of organisms. Moreover, this work shows the importance of assessing the effects of chemicals in relevant exposure scenarios that take into account long-term effects. http://dx.doi.org/10.1016/j.toxlet.2017.07.565 P-06-01-15 Assessing estrogenic and anti-estrogenic activity of antimicrobial compounds used in oral care products Klára Petru˚ 1,2 , Martin Ezechiáˇs 1 , Lucie Linhartová 1 , Tomáˇs Cajthaml 1,2 1
Institute of Microbiology Academy of Sciences of the Czech Republic, v.v.i., Prague, Czech Republic 2 Institute for Environmental Studies, Faculty of Science, Charles University, Prague, Czech Republic The potential triclosan for hormone disruption triggered its removal from personal care product lines (e.g. mouthwashes, toothpastes). There is a strong demand for finding new effective and safe substitutes to be used in oral care hygiene and other consumer products, including cosmetics. Even though the in vitro efficacy of new/renewed antiseptics are well-documented, much less is known about their adverse health effect on humans or environmental impact. Estrogenic and anti-estrogenic potential of 10 selected antimicrobial compounds commonly used in oral care hygiene was assessed. Both human T47D breast carcinoma cell line using altered secretion of cytokine CXCL12 and functional yeast-luciferase reporter gene assay were employed to confirm receptor-binding activities.
Abstracts / Toxicology Letters 280S (2017) S202–S213
None of the tested compounds exhibited estrogenic activity. However, two antiseptics possessing quaternary ammonium moiety (octenidine and cetylpiridin) and one monoterpene derivative (thymol) revealed anti-estrogenic effects and inhibited the estrogen response in concentration-dependent manner for both, the T47D cell line and yeast assay. An inhibition effect expressed as IC50 increased as follows, thymol < cetylpiridinium < octenidine. The results highlight the endocrine disrupting effects of antiseptic compounds commonly used in personal care products and provide better insight into the health and environmental risk evaluation. Further evaluation and monitoring of these compounds in the environment should be considered. http://dx.doi.org/10.1016/j.toxlet.2017.07.566 P-06-01-16 Transcriptional and physiological effects of the anticancer drug tamoxifen on reproduction of daphnia magna
S207
on non-target organisms have required. In particular, pharmaceuticals used for human health have the relative low toxicity compared with toxicants; however, prolong or overdose exposure of these pharmaceuticals could lead to adverse toxicological effects. Acetaminophen (APAP) has frequently detected in aquatic environment but there is a lack of knowledge about toxicity by APAP although it was reported serious physiological effect such as reduced reproduction, low growth rate and abnormal behavior on aquatic organism. In order to develop reliable and sensitive methods for evaluating APAP toxicity using the daphnia model, the present study focused on cytochrome P450 (CYP)-based metabolizing system because it is a first-line defense system and a major cause of APAP side effects. We analyzed total metabolites extracted from D. magna-exposed to APAP. Also, several molecular and biochemical indicators associated with the toxic metabolite were identified, including strong activation of a certain CYP gene, glutathione (GSH) depletion, ROS/RNS production and thioredoxin reductase (TrxR) activity. http://dx.doi.org/10.1016/j.toxlet.2017.07.568
Mina Cho System Toxicology Research Center, Korea Institute of Toxicology, Daejeon, Republic of Korea Tamoxifen (TAM), a selective estrogen receptor modulator, has been used as an anti-cancer drug for mammalian breast and endometrial cancer. By increasing TAM usage, it has frequently reported that TAM is a potential endocrine disruptor capable of interfering with reproduction in non-target organisms. However, the mode of action of TAM on endocrine system is still unknown. In this study, we performed a 21-d chronic toxicity test using a crustacean, Daphnia magna, and also investigated the transcriptional modulation of major genes related to endocrine, molting, development and reproduction (e.g., Dm-HR96, EcR, Cyp314, vtg and vmo1) after TAM exposure for 24 h, 48 h and 72 h. As a result, we observed the decrease in total number of eggs per individual at a concentration-dependent manner, and the expression of oogenesis-related genes was inhibited by TAM exposure in timeand dose-dependent. Also, the expression of HR96, a putative toxicant receptor, and molting-related genes is also downregulated in time- and dose-dependent. In order to know the presence of CYP19A-like gene that is a key gene to regulate estrogen system, we measured aromatase activity in D. magna exposed to TAM. Our findings suggested that TAM could regulate reproductive behavior by reducing molecular mechanisms involved in oogenesis and molting process. This research supports that D. magna would be a useful alternative model for rapidly evaluating the reproductive effects induced by drug toxicity. http://dx.doi.org/10.1016/j.toxlet.2017.07.567 P-06-01-17 Novel approach for evaluating pharmaceuticals toxicity using daphnia model: Analysis of the mode of cytochrome P450-generated metabolite action after acetaminophen exposure Ryeook Kim System Toxicology Research Center, Korea Institute of Toxicology, Daejeon, Republic of Korea By increasing concerns for pharmaceuticals in aquatic environments, useful tools capable of monitoring toxicological effects
P-06-01-18 Investigation on the relationship between oxidative stress and cognitive neuropsychology status in patients with MS Ali Akbar Malekirad 1,2 , Hakimeh Nikpour 3 , Fardin Faraji 4 , Sima Abbasi 5 1
Department of Biology, Payame Noor University, Tehran, Islamic Republic of Iran 2 Toxicology and Diseases Group, Pharmaceutical Sciences Research Center, Tehran University of Medical Sciences, Tehran, Islamic Republic of Iran 3 Department of Psychology, Payame Noor University, Tehran, Islamic Republic of Iran 4 Department of Neurology, Arak University of Medical Sciences, Arak, Islamic Republic of Iran 5 Department of Literature, Payame Noor University, Tehran, Islamic Republic of Iran Oxidative stress by changing the amount of reactive oxygen species (ROS) and antioxidant parameters can cause several neurological disorders such as multiple sclerosis (MS). MS patients often have cognitive impairments and often impact on speech, finding the word and remembering the pronunciation of words. Therefore, we decided to investigate the relationship between oxidative stress and cognitive neuropsychology status of patients with MS. In this clinical trial of 58 non-depressed patients with Multiple Sclerosis in collaboration with the MS Society of Markazi province were randomly selected. Blood samples are taken and oxidative stress factors and cognitive function, it was measured (memory, inhibitory control and selective attention, decision-making, planning, sustained attention, social cognition and cognitive flexibility). Manifold correlation test was used to analyze the data. The results showed that there is a significant relationship between oxidative stress indices on the one hand, memory, inhibitory control, selective attention, decision-making, planning, Sustained attention, social cognition and cognitive flexibility on the other hand. Because of the relationship between cognitive dysfunction and oxidative stress in patients with MS and patients suffering from this disorder, according to mental status and perceptions patients is essential and it seems that reduce oxidative stress can affect cognitive decline. http://dx.doi.org/10.1016/j.toxlet.2017.07.569
Abstracts / Toxicology Letters 280S (2017) S202–S213
the specific respiratory impact of atmospheric ultrafine particles compared to fine particulate matter in a relevant in vivo model of sub-chronic exposure to these pollutants. Quasi-ultrafine (PM0.18 ) and fine (PM2.5 ) particles have been collected in the urban industrial zone of Dunkirk in north France during a 7-month campaign. The physico-chemical study of the collected particles shows that there is no major difference in elemental and surface chemical composition between PM0.18 and PM2.5. BALB/c mice were then exposed intranasally to 10 g of PM0.18 or PM2.5 3 times a week. After 1 or 3-month exposure, broncho alveolar lavages (BAL) were performed and lung tissues were harvested for histological, epigenetic and transcriptomic analyses. Cytological analyses show that both types of particulate fractions can be internalized in lung cells. Cellular analyses of BAL and preliminary transcriptomic data suggest that PM0.18 induced a stronger lung inflammation and mRNA and miRNA deregulations than PM2.5 . Complementary studies are in progress to confirm these first data and to identify the metabolic pathways more specifically associated with the toxicity of ultrafine particles. http://dx.doi.org/10.1016/j.toxlet.2017.07.563 P-06-01-13 Air quality assessment during indoor use of the tobacco heating system THS2.2 Catherine Goujon Ginglinger, Maya Mitova, Serge Maeder, Maurice Smith Research and Development, Philip Morris Products SA, Neuchatel, Switzerland PMI’s heat-not-burn Tobacco Heating System 2.2 (THS 2.2) uses an electronically controlled heating mechanism to precisely heat specially designed tobacco sticks at operating temperatures well below combustion. To address public health concerns about possible presence of polluting substances during indoor use of THS 2.2, a study was designed to compare the environmental aerosol produced by THS 2.2 to background air (no use of any product). A dedicated controlled room to simulate a residential environment with a low ventilation rate (0.5 air changes/h) was used. Twenty three analytes (nicotine, 3-ethenylpyridine, solanesol, respirable suspended particles (RSP) by gravimetric measurement, particulate matter by UV, particulate matter by fluorescence, total volatile organic compounds (TVOCs), acetaldehyde, acrolein, crotonaldehyde, formaldehyde, acrylonitrile, benzene, 1,3-butadiene, isoprene, toluene, glycerin, propylene glycol, N-nitrosonornicotine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, carbon monoxide, nitrogen oxide and combined oxides of nitrogen) were quantified by validated and ISO17025 accredited methods. In comparison to background air, only three compounds can be attributed to the use of THS 2.2: nicotine (1.15 g/m3 ), acetaldehyde (3.44 g/m3 ) and glycerin (10.5 g/m3 ). Their levels are much lower than maximum exposure levels as defined in existing air quality guidelines. Markers of combustion are absent in environmental aerosols of THS 2.2. Based on TVOC data analyses, the chemical composition of the background air and air during use of THS 2.2 were highly similar. In conclusion, using THS 2.2 does not have a negative impact on the overall air quality. http://dx.doi.org/10.1016/j.toxlet.2017.07.564
P-06-01-14 Behavioral effects in adult zebrafish after developmental exposure to carbaryl Daniela Correia, Ana Rita Almeida, Joana Santos, Amadeu M.V.M. Soares, Inês Domingues Department of Biology & CESAM, University of Aveiro, Campus Universitário de Santiago, Aveiro, Portugal In recent years, there is a growing concern to correctly assess the risk of low environmental concentrations of contaminants (e.g. pesticides) to aquatic life. Pesticides can be found in the ng/L and g/L range and, very often, observable effects to non-target organisms like fish, may not be perceptible immediately and long term effects can be underestimated. Hence, our work intends to evaluate the effect of an early (embryonic) exposure to carbaryl in adult fish behaviour. Zebrafish (Danio rerio) embryos were exposed to 4 sublethal concentrations (0, 10, 100 and 1000 mg/L) of carbaryl during 4 days and then kept in standard cultivation conditions until adulthood. Two behaviour tests were then performed to assess anxiety-like behavior (novel tank test and light/dark stimulus) plus a feeding test. Our data showed that zebrafish pre-exposed to carbaryl, remained longer periods in the bottom layer of the aquarium suggesting a decrease of the exploratory behavior. Further, fish preexposed to 1000 mg/L of carbaryl showed lower swimming activity (distance moved and time spent swimming) than non-exposed fish and presented reduced thigmotaxis during the dark stimulus when compared to control, suggesting repression of anxiety-like behavior. Pre-exposed fish also took more time to feed than non-exposed fish. Our results show that developmental exposure to carbaryl has long lasting effects on adult behaviour highlighting the importance of the environmental conditions during early life stages of organisms. Moreover, this work shows the importance of assessing the effects of chemicals in relevant exposure scenarios that take into account long-term effects. http://dx.doi.org/10.1016/j.toxlet.2017.07.565 P-06-01-15 Assessing estrogenic and anti-estrogenic activity of antimicrobial compounds used in oral care products Klára Petru˚ 1,2 , Martin Ezechiáˇs 1 , Lucie Linhartová 1 , Tomáˇs Cajthaml 1,2 1
Institute of Microbiology Academy of Sciences of the Czech Republic, v.v.i., Prague, Czech Republic 2 Institute for Environmental Studies, Faculty of Science, Charles University, Prague, Czech Republic The potential triclosan for hormone disruption triggered its removal from personal care product lines (e.g. mouthwashes, toothpastes). There is a strong demand for finding new effective and safe substitutes to be used in oral care hygiene and other consumer products, including cosmetics. Even though the in vitro efficacy of new/renewed antiseptics are well-documented, much less is known about their adverse health effect on humans or environmental impact. Estrogenic and anti-estrogenic potential of 10 selected antimicrobial compounds commonly used in oral care hygiene was assessed. Both human T47D breast carcinoma cell line using altered secretion of cytokine CXCL12 and functional yeast-luciferase reporter gene assay were employed to confirm receptor-binding activities.
Abstracts / Toxicology Letters 280S (2017) S202–S213
None of the tested compounds exhibited estrogenic activity. However, two antiseptics possessing quaternary ammonium moiety (octenidine and cetylpiridin) and one monoterpene derivative (thymol) revealed anti-estrogenic effects and inhibited the estrogen response in concentration-dependent manner for both, the T47D cell line and yeast assay. An inhibition effect expressed as IC50 increased as follows, thymol < cetylpiridinium < octenidine. The results highlight the endocrine disrupting effects of antiseptic compounds commonly used in personal care products and provide better insight into the health and environmental risk evaluation. Further evaluation and monitoring of these compounds in the environment should be considered. http://dx.doi.org/10.1016/j.toxlet.2017.07.566 P-06-01-16 Transcriptional and physiological effects of the anticancer drug tamoxifen on reproduction of daphnia magna
S207
on non-target organisms have required. In particular, pharmaceuticals used for human health have the relative low toxicity compared with toxicants; however, prolong or overdose exposure of these pharmaceuticals could lead to adverse toxicological effects. Acetaminophen (APAP) has frequently detected in aquatic environment but there is a lack of knowledge about toxicity by APAP although it was reported serious physiological effect such as reduced reproduction, low growth rate and abnormal behavior on aquatic organism. In order to develop reliable and sensitive methods for evaluating APAP toxicity using the daphnia model, the present study focused on cytochrome P450 (CYP)-based metabolizing system because it is a first-line defense system and a major cause of APAP side effects. We analyzed total metabolites extracted from D. magna-exposed to APAP. Also, several molecular and biochemical indicators associated with the toxic metabolite were identified, including strong activation of a certain CYP gene, glutathione (GSH) depletion, ROS/RNS production and thioredoxin reductase (TrxR) activity. http://dx.doi.org/10.1016/j.toxlet.2017.07.568
Mina Cho System Toxicology Research Center, Korea Institute of Toxicology, Daejeon, Republic of Korea Tamoxifen (TAM), a selective estrogen receptor modulator, has been used as an anti-cancer drug for mammalian breast and endometrial cancer. By increasing TAM usage, it has frequently reported that TAM is a potential endocrine disruptor capable of interfering with reproduction in non-target organisms. However, the mode of action of TAM on endocrine system is still unknown. In this study, we performed a 21-d chronic toxicity test using a crustacean, Daphnia magna, and also investigated the transcriptional modulation of major genes related to endocrine, molting, development and reproduction (e.g., Dm-HR96, EcR, Cyp314, vtg and vmo1) after TAM exposure for 24 h, 48 h and 72 h. As a result, we observed the decrease in total number of eggs per individual at a concentration-dependent manner, and the expression of oogenesis-related genes was inhibited by TAM exposure in timeand dose-dependent. Also, the expression of HR96, a putative toxicant receptor, and molting-related genes is also downregulated in time- and dose-dependent. In order to know the presence of CYP19A-like gene that is a key gene to regulate estrogen system, we measured aromatase activity in D. magna exposed to TAM. Our findings suggested that TAM could regulate reproductive behavior by reducing molecular mechanisms involved in oogenesis and molting process. This research supports that D. magna would be a useful alternative model for rapidly evaluating the reproductive effects induced by drug toxicity. http://dx.doi.org/10.1016/j.toxlet.2017.07.567 P-06-01-17 Novel approach for evaluating pharmaceuticals toxicity using daphnia model: Analysis of the mode of cytochrome P450-generated metabolite action after acetaminophen exposure Ryeook Kim System Toxicology Research Center, Korea Institute of Toxicology, Daejeon, Republic of Korea By increasing concerns for pharmaceuticals in aquatic environments, useful tools capable of monitoring toxicological effects
P-06-01-18 Investigation on the relationship between oxidative stress and cognitive neuropsychology status in patients with MS Ali Akbar Malekirad 1,2 , Hakimeh Nikpour 3 , Fardin Faraji 4 , Sima Abbasi 5 1
Department of Biology, Payame Noor University, Tehran, Islamic Republic of Iran 2 Toxicology and Diseases Group, Pharmaceutical Sciences Research Center, Tehran University of Medical Sciences, Tehran, Islamic Republic of Iran 3 Department of Psychology, Payame Noor University, Tehran, Islamic Republic of Iran 4 Department of Neurology, Arak University of Medical Sciences, Arak, Islamic Republic of Iran 5 Department of Literature, Payame Noor University, Tehran, Islamic Republic of Iran Oxidative stress by changing the amount of reactive oxygen species (ROS) and antioxidant parameters can cause several neurological disorders such as multiple sclerosis (MS). MS patients often have cognitive impairments and often impact on speech, finding the word and remembering the pronunciation of words. Therefore, we decided to investigate the relationship between oxidative stress and cognitive neuropsychology status of patients with MS. In this clinical trial of 58 non-depressed patients with Multiple Sclerosis in collaboration with the MS Society of Markazi province were randomly selected. Blood samples are taken and oxidative stress factors and cognitive function, it was measured (memory, inhibitory control and selective attention, decision-making, planning, sustained attention, social cognition and cognitive flexibility). Manifold correlation test was used to analyze the data. The results showed that there is a significant relationship between oxidative stress indices on the one hand, memory, inhibitory control, selective attention, decision-making, planning, Sustained attention, social cognition and cognitive flexibility on the other hand. Because of the relationship between cognitive dysfunction and oxidative stress in patients with MS and patients suffering from this disorder, according to mental status and perceptions patients is essential and it seems that reduce oxidative stress can affect cognitive decline. http://dx.doi.org/10.1016/j.toxlet.2017.07.569