- Email: [email protected]
Detecting xenoestrogens with an in vivo assay system
Abstracts / Toxicology Letters 211S (2012) S43–S216
our understanding of the relationship between dietary PCB exposure and health effects, we validated past and concurrent dietary PCB exposure, estimated via food frequency questionnaires (FFQs) against biomarkers of PCBs.We used PCB153, an indicator for PCBs, and created two large recipe-based concentration databases based on >1200 food samples analyzed between 1992 and 2009. The concentrations were extrapolated with 8% per year, reflecting the yearly decline of PCB153 concentrations observed in food and human breast milk, to mirror the PCB153 concentrations in food during the years of the FFQs (1997 and 2004). Serum concentrations of PCBs were analyzed in 201 women (56–85 years of age) from the population-based prospective Swedish Mammography Cohort. To account for within-person variation in FFQ-based PCB153 exposure, intra-class correlation coefficient was estimated from a reproducibility analysis on 165 women who had completed two identical FFQs one year apart. The validity, i.e., the Spearman correlation coefficient between FFQ-based PCB153 exposure (expressed as ng/kg body weight) and serum PCB153 concentrations (adjusted for serum lipids and age, accounting for the within-person variation, was 0.41 (p < 0.001) for concurrent exposure assessment (2004–2006) and 0.34 (p < 0.05) for past exposure assessment (1997). The Spearman correlation between concurrent exposure of PCB153 and other serum PCB congeners (118, 138, 156, 170 and 180) ranged from 0.30 to 0.52 (95% CI; 0.11–0.68). Similar results were obtained for past exposure assessment. We conclude that FFQ-based dietary PCB153 exposure assessed up to nine years earlier, shows reasonable validity in relation to PCB biomarkers, justifying its use in large scale epidemiological studies. doi:10.1016/j.toxlet.2012.03.805
P04: Biomonitoring
P04-01 Perfluorinated alkyl acids in serum from nursing women from Uppsala, Sweden 1996–2010 Anders Glynn 1 , Urs Berger 2 , Anders Bignert 3 , Shahid Ullah 2 , Marie Aune 1 , Sanna Lignell 1 , Per Ola Darnerud 1 Swedish National Food Agency, Sweden, 2 ITM, Stockholm University, Sweden, 3 Swedish Museum of Natural History, Sweden
1
Purpose: We investigated the temporal trends of perfluorinated alkyl acids (PFAAs) in blood serum from nursing women, in order to determine if risk management measures to reduce production, use and emissions of perfluorooctanoate (PFOA) and perfluorooctane sulfonate (PFOS) have affected human exposure to PFAAs. Methods: Serum levels of 13 PFAAs were measured by high performance liquid chromatography coupled to high resolution mass spectrometry. Serum samples from 417 primiparous nursing women (age: 19–41 years), sampled 3 weeks after delivery between 1996 and 2010, were analysed as pooled samples. An effort was made to prepare 3 pooled samples per year with at least 10 individual samples in each pool. Results and conclusions of the study: The results show that the phase out of PFOS and PFOA has resulted in a decreased exposure of primiparas from Uppsala to these compounds. PFOS levels decreased on average 9% per year and PFOA levels 3% per year. However, during the same time period, levels of perfluorinated sulfonates with 4 and 6 carbons and perfluorinated carboxylates with 9–11 carbons increased with 4–14% per year. This shows that the exposure of primiparas from
S49
Uppsala to these compounds increased after the efforts of PFOS and PFOA phase-out were initiated. doi:10.1016/j.toxlet.2012.03.197
P04-02 Biological monitoring of aflatoxin (AFB1) in workers of swine and poultry production Susana Viegas, Luisa Veiga, Ana Almeida, Elisabete Carolino, Carla Viegas ESTeSL-IPL, Portugal Keywords: Aflatoxin B1; Biological monitoring; Occupational exposure; Poultry; Swine production Although a great body of literature exists concerning the ingestion of food contaminated with aflatoxin, there are still few studies regarding mycotoxin inhalation in occupational settings. Since mycotoxins are relatively non-volatile, inhalation exposure is cause by inhalation of airborne fungal particulates or fungi-contaminated substrates that contain aflatoxin. We intend to know if there is occupational exposure to aflatoxin in Portuguese poultry and swine production. A total of 19 individuals (11 swine; 8 poultry) agreed and provided blood samples during the course of this investigation. Measurement of AFB1 was performed by ELISA. The samples were treated with pronase (Merck), wash in a Column C18 and purification was made with immunoaffinity columns (R. biopharma), specific for AFB1. It was applied statistical test (Mann–Whitney) to verified statistical difference in AFB1 results between the two settings. Results varied with concentrations from
P04-03 Detecting xenoestrogens with an in vivo assay system Wenjau Lee 1 , Chi-Wei Kan 1 , Chung-Kai Su 1 , Kataaki Okubo 2 , Yoshitaka Nagahama 3 Chang Jung Christian University, Taiwan, 2 The University of Tokyo, Japan, 3 National Institute for Basic Biology, Japan 1
Purpose: As xenoestrogens may interfere with the reproductive health of humans and wildlife, it is essential to investigate their effects and monitor their presence in the environment. Since the gonadotropin-releasing hormone (GnRH) is critical for the initiation and maintenance of the reproductive function, we developed an assay system to directly assess the effects of xenoestrogens, using a transgenic medaka (Oryzias latipes) in which the gene for the green fluorescent protein (GFP) was placed under the control of the gnrh3 promoter, one of the three paralogous GnRH genes. As
S50
Abstracts / Toxicology Letters 211S (2012) S43–S216
medaka embryos are transparent, the fluorescent expression can be easily observed in vivo. Methods: We exposed freshly spawned medaka embryos to bisphenol A (BPA), nonylphenol (NP), 17estrodiol (E2), or a water sample from a polluted river, and observed the embryos daily. After the embryos hatched, the gene expression levels of the GnRHs, GnRH receptors, and estrogen receptors (ERs) were measured with qRT-PCR. Results and Conclusions: We found that the exposures significantly decreased the fluorescent intensity, and slowed down the head growth and the eye development. The chemicals also lowered the heart rate, lengthened the time to hatch, suppressed the expression of the three GnRH genes, and upregulated the ER␣ mRNA level. Importantly, the GnRH3 mRNA level was significantly correlated with the fluorescence intensity of the GnRH3 neurons. We concluded that BPA, NP, and E2 had significant effects on the embryonic development, and the gnrh3-GFP transgenic medaka can be an ideal assay system for xenoestrogens and environmental water samples. doi:10.1016/j.toxlet.2012.03.199
P04-04 Heavy metals of Yenic¸a˘ ga Lake (Turkey) and its potential sources: Soil, sediment, water Yasemin Saygi 1 , Sibel Atasagun 2
and macrophyta species) and investigate the ecologic effects in the Karabo˘gaz Lake. The lake is a part of Kızılırmak Delta which has 6 km length and 2 km width and is one of the most valuable wetland systems in the northern Turkey as a “Ramsar Site” by Ministry of Environment in 1998. Bird Life International included the area in the Important Bird Areas (IBA) list of Europe (Grimmet and Jones 1989). The lake and catchment area (2710 ha) included “Special Protected Area”. Agricultural facilities are so intensive at the field and irrigation water is attaining the lake via drainage channel at the south side of the lake. Despite these problems, any study was performed on pesticide residues accumulation in Karabo˘gaz Lake. During our project studies, some physical and chemical analyses (salinity, EC, TDS, pH, dissolved oxygen, temperature, depth) of lake water investigated by seasonally periodic sampling throughout the year. In these studies, multiresidue analyses carried out in the water, sediment, Cyprinus carpio and macrophyta species (Chara vulgaris and Potamegaton pectinatus) using QUECHERS methods and Shimadzu QP 2010 GC-MS. According to our study results; especially pyrethrins, carbamate insecticides and fungicides are the most common chemicals. As a bioaccumulation data, macrophyta species contain mainly pyrethrins and also piperonyl butoxide (PBO), the other samples contain mainly Molinate, Tebufenozide, Propamocarb and the other common pesticides at the field. doi:10.1016/j.toxlet.2012.03.201
1
Hacettepe University, Turkey, 2 Ankara University, Turkey
Yenic¸a˘ga Lake is located in the Western Black Sea region in Turkey. The quality of surface water of Yenic¸a˘ga Lake has diminished over the last decades as a result of increased anthropogenic factors. Today another important environmental threat for the lake is the artesian wells that its water discharges through creeks or superficial drainage into the lake and it has rich with heavy metals. The objective of the study was to designate of distribution and accumulation of heavy metals (Al, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Mo, Cd, Sn, Ba, Pb) in the water, sediments, plankton of Yenic¸a˘ga Lake and its potential sources (creeks, sewage, artesian well, soil). The sampling was carried out from March 2008 to February 2009 as monthly intervals at three stations in the lake. Element analyses were performed by ICP-MS. Results showed that Al, As, Mn, Pb, Fe concentration in lake water and/or its feeding sources were above the recommended water standards (WHO, EC, EPA, TS-266). The accumulation order of trace metals were Fe > Al > Mn > Zn > Ba > Ni > Cr > As > Cu > Pb > Co > Mo > Sn > Cd in the lake, creeks sediment and soil samples. The similar results suggest that the accumulation of heavy metals in the sediment is a natural process. Metals accumulated in the lake are naturally mixed from the soil. However, the presence of heavy metals in the analysis of artesian well water and sewage reveals that the transportation occurs also from the groundwater into the lake. doi:10.1016/j.toxlet.2012.03.200
P04-05 Determination of multiresidual pesticide bioaccumulation in aquatic food chain in the Karabogaz Lake (Samsun, Turkey) Zeynep Yurtkuran, Yasemin Saygi, Oner Kocak Hacettepe University, Turkey The purpose of the study is that determination of multiresidual pesticide analysis in the food chain (water, sediment, some fish
P04-06 Determination of lymphocyte DNA damage by use of comet assay in painters Zeliha Kayaalti 1 , Esma Soylemez 1 , Aysegül Bacaksiz 1 , Emrah Dural 1 , Hinc¸ Yilmaz 2 , Tülin Soylemezoglu 1 Ankara University, Turkey, 2 Ankara Occupational Diseases Hospital, Turkey
1
Painters are commonly exposed to an extensive variety of substances like organic solvents such as toluene, white spirits, xylene and other volatile paint components throughout their working lifetime. Also, workers in paint manufacture are classified as an occupation that increases certain cancers risk. The physicochemical properties of solvents can cause disturbances to cellular structures, including damage to DNA. The Comet assay is a rapid and sensitive method for measuring DNA single-strand breaks. The aim of this study was to determine lymphocyte DNA damage in painters. For this purpose, 70 whole blood samples (35 painters and 35 control) were studied by comet assay and evaluated according to four image-analysis parameters including tail intensity (TI), tail DNA (DNAt), tail moment (TM) and olive tail moment (OTM). Also, urinary hippuric acid rate which is used as a marker for toluene exposure was also measured in painters. As a result, mean of TI, DNAt, TM and OTM in organic solventexposed group had higher than in control group. TI, DNAt and TM were statistically significant between the exposed and nonexposed groups (p < 0.05), but OTM was not (p > 0.05). Moreover, smoking habit was statistical compared with DNA damage parameters and hippuric acid rates in exposed group. While statistically significant association was determined with hippuric acid rates (p < 0.05), there was not any statistically association with DNA damage parameters (p > 0.05). The study is suggested that organic solvents exposure can induce lymphocyte DNA damage in painters
our understanding of the relationship between dietary PCB exposure and health effects, we validated past and concurrent dietary PCB exposure, estimated via food frequency questionnaires (FFQs) against biomarkers of PCBs.We used PCB153, an indicator for PCBs, and created two large recipe-based concentration databases based on >1200 food samples analyzed between 1992 and 2009. The concentrations were extrapolated with 8% per year, reflecting the yearly decline of PCB153 concentrations observed in food and human breast milk, to mirror the PCB153 concentrations in food during the years of the FFQs (1997 and 2004). Serum concentrations of PCBs were analyzed in 201 women (56–85 years of age) from the population-based prospective Swedish Mammography Cohort. To account for within-person variation in FFQ-based PCB153 exposure, intra-class correlation coefficient was estimated from a reproducibility analysis on 165 women who had completed two identical FFQs one year apart. The validity, i.e., the Spearman correlation coefficient between FFQ-based PCB153 exposure (expressed as ng/kg body weight) and serum PCB153 concentrations (adjusted for serum lipids and age, accounting for the within-person variation, was 0.41 (p < 0.001) for concurrent exposure assessment (2004–2006) and 0.34 (p < 0.05) for past exposure assessment (1997). The Spearman correlation between concurrent exposure of PCB153 and other serum PCB congeners (118, 138, 156, 170 and 180) ranged from 0.30 to 0.52 (95% CI; 0.11–0.68). Similar results were obtained for past exposure assessment. We conclude that FFQ-based dietary PCB153 exposure assessed up to nine years earlier, shows reasonable validity in relation to PCB biomarkers, justifying its use in large scale epidemiological studies. doi:10.1016/j.toxlet.2012.03.805
P04: Biomonitoring
P04-01 Perfluorinated alkyl acids in serum from nursing women from Uppsala, Sweden 1996–2010 Anders Glynn 1 , Urs Berger 2 , Anders Bignert 3 , Shahid Ullah 2 , Marie Aune 1 , Sanna Lignell 1 , Per Ola Darnerud 1 Swedish National Food Agency, Sweden, 2 ITM, Stockholm University, Sweden, 3 Swedish Museum of Natural History, Sweden
1
Purpose: We investigated the temporal trends of perfluorinated alkyl acids (PFAAs) in blood serum from nursing women, in order to determine if risk management measures to reduce production, use and emissions of perfluorooctanoate (PFOA) and perfluorooctane sulfonate (PFOS) have affected human exposure to PFAAs. Methods: Serum levels of 13 PFAAs were measured by high performance liquid chromatography coupled to high resolution mass spectrometry. Serum samples from 417 primiparous nursing women (age: 19–41 years), sampled 3 weeks after delivery between 1996 and 2010, were analysed as pooled samples. An effort was made to prepare 3 pooled samples per year with at least 10 individual samples in each pool. Results and conclusions of the study: The results show that the phase out of PFOS and PFOA has resulted in a decreased exposure of primiparas from Uppsala to these compounds. PFOS levels decreased on average 9% per year and PFOA levels 3% per year. However, during the same time period, levels of perfluorinated sulfonates with 4 and 6 carbons and perfluorinated carboxylates with 9–11 carbons increased with 4–14% per year. This shows that the exposure of primiparas from
S49
Uppsala to these compounds increased after the efforts of PFOS and PFOA phase-out were initiated. doi:10.1016/j.toxlet.2012.03.197
P04-02 Biological monitoring of aflatoxin (AFB1) in workers of swine and poultry production Susana Viegas, Luisa Veiga, Ana Almeida, Elisabete Carolino, Carla Viegas ESTeSL-IPL, Portugal Keywords: Aflatoxin B1; Biological monitoring; Occupational exposure; Poultry; Swine production Although a great body of literature exists concerning the ingestion of food contaminated with aflatoxin, there are still few studies regarding mycotoxin inhalation in occupational settings. Since mycotoxins are relatively non-volatile, inhalation exposure is cause by inhalation of airborne fungal particulates or fungi-contaminated substrates that contain aflatoxin. We intend to know if there is occupational exposure to aflatoxin in Portuguese poultry and swine production. A total of 19 individuals (11 swine; 8 poultry) agreed and provided blood samples during the course of this investigation. Measurement of AFB1 was performed by ELISA. The samples were treated with pronase (Merck), wash in a Column C18 and purification was made with immunoaffinity columns (R. biopharma), specific for AFB1. It was applied statistical test (Mann–Whitney) to verified statistical difference in AFB1 results between the two settings. Results varied with concentrations from
P04-03 Detecting xenoestrogens with an in vivo assay system Wenjau Lee 1 , Chi-Wei Kan 1 , Chung-Kai Su 1 , Kataaki Okubo 2 , Yoshitaka Nagahama 3 Chang Jung Christian University, Taiwan, 2 The University of Tokyo, Japan, 3 National Institute for Basic Biology, Japan 1
Purpose: As xenoestrogens may interfere with the reproductive health of humans and wildlife, it is essential to investigate their effects and monitor their presence in the environment. Since the gonadotropin-releasing hormone (GnRH) is critical for the initiation and maintenance of the reproductive function, we developed an assay system to directly assess the effects of xenoestrogens, using a transgenic medaka (Oryzias latipes) in which the gene for the green fluorescent protein (GFP) was placed under the control of the gnrh3 promoter, one of the three paralogous GnRH genes. As
S50
Abstracts / Toxicology Letters 211S (2012) S43–S216
medaka embryos are transparent, the fluorescent expression can be easily observed in vivo. Methods: We exposed freshly spawned medaka embryos to bisphenol A (BPA), nonylphenol (NP), 17estrodiol (E2), or a water sample from a polluted river, and observed the embryos daily. After the embryos hatched, the gene expression levels of the GnRHs, GnRH receptors, and estrogen receptors (ERs) were measured with qRT-PCR. Results and Conclusions: We found that the exposures significantly decreased the fluorescent intensity, and slowed down the head growth and the eye development. The chemicals also lowered the heart rate, lengthened the time to hatch, suppressed the expression of the three GnRH genes, and upregulated the ER␣ mRNA level. Importantly, the GnRH3 mRNA level was significantly correlated with the fluorescence intensity of the GnRH3 neurons. We concluded that BPA, NP, and E2 had significant effects on the embryonic development, and the gnrh3-GFP transgenic medaka can be an ideal assay system for xenoestrogens and environmental water samples. doi:10.1016/j.toxlet.2012.03.199
P04-04 Heavy metals of Yenic¸a˘ ga Lake (Turkey) and its potential sources: Soil, sediment, water Yasemin Saygi 1 , Sibel Atasagun 2
and macrophyta species) and investigate the ecologic effects in the Karabo˘gaz Lake. The lake is a part of Kızılırmak Delta which has 6 km length and 2 km width and is one of the most valuable wetland systems in the northern Turkey as a “Ramsar Site” by Ministry of Environment in 1998. Bird Life International included the area in the Important Bird Areas (IBA) list of Europe (Grimmet and Jones 1989). The lake and catchment area (2710 ha) included “Special Protected Area”. Agricultural facilities are so intensive at the field and irrigation water is attaining the lake via drainage channel at the south side of the lake. Despite these problems, any study was performed on pesticide residues accumulation in Karabo˘gaz Lake. During our project studies, some physical and chemical analyses (salinity, EC, TDS, pH, dissolved oxygen, temperature, depth) of lake water investigated by seasonally periodic sampling throughout the year. In these studies, multiresidue analyses carried out in the water, sediment, Cyprinus carpio and macrophyta species (Chara vulgaris and Potamegaton pectinatus) using QUECHERS methods and Shimadzu QP 2010 GC-MS. According to our study results; especially pyrethrins, carbamate insecticides and fungicides are the most common chemicals. As a bioaccumulation data, macrophyta species contain mainly pyrethrins and also piperonyl butoxide (PBO), the other samples contain mainly Molinate, Tebufenozide, Propamocarb and the other common pesticides at the field. doi:10.1016/j.toxlet.2012.03.201
1
Hacettepe University, Turkey, 2 Ankara University, Turkey
Yenic¸a˘ga Lake is located in the Western Black Sea region in Turkey. The quality of surface water of Yenic¸a˘ga Lake has diminished over the last decades as a result of increased anthropogenic factors. Today another important environmental threat for the lake is the artesian wells that its water discharges through creeks or superficial drainage into the lake and it has rich with heavy metals. The objective of the study was to designate of distribution and accumulation of heavy metals (Al, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Mo, Cd, Sn, Ba, Pb) in the water, sediments, plankton of Yenic¸a˘ga Lake and its potential sources (creeks, sewage, artesian well, soil). The sampling was carried out from March 2008 to February 2009 as monthly intervals at three stations in the lake. Element analyses were performed by ICP-MS. Results showed that Al, As, Mn, Pb, Fe concentration in lake water and/or its feeding sources were above the recommended water standards (WHO, EC, EPA, TS-266). The accumulation order of trace metals were Fe > Al > Mn > Zn > Ba > Ni > Cr > As > Cu > Pb > Co > Mo > Sn > Cd in the lake, creeks sediment and soil samples. The similar results suggest that the accumulation of heavy metals in the sediment is a natural process. Metals accumulated in the lake are naturally mixed from the soil. However, the presence of heavy metals in the analysis of artesian well water and sewage reveals that the transportation occurs also from the groundwater into the lake. doi:10.1016/j.toxlet.2012.03.200
P04-05 Determination of multiresidual pesticide bioaccumulation in aquatic food chain in the Karabogaz Lake (Samsun, Turkey) Zeynep Yurtkuran, Yasemin Saygi, Oner Kocak Hacettepe University, Turkey The purpose of the study is that determination of multiresidual pesticide analysis in the food chain (water, sediment, some fish
P04-06 Determination of lymphocyte DNA damage by use of comet assay in painters Zeliha Kayaalti 1 , Esma Soylemez 1 , Aysegül Bacaksiz 1 , Emrah Dural 1 , Hinc¸ Yilmaz 2 , Tülin Soylemezoglu 1 Ankara University, Turkey, 2 Ankara Occupational Diseases Hospital, Turkey
1
Painters are commonly exposed to an extensive variety of substances like organic solvents such as toluene, white spirits, xylene and other volatile paint components throughout their working lifetime. Also, workers in paint manufacture are classified as an occupation that increases certain cancers risk. The physicochemical properties of solvents can cause disturbances to cellular structures, including damage to DNA. The Comet assay is a rapid and sensitive method for measuring DNA single-strand breaks. The aim of this study was to determine lymphocyte DNA damage in painters. For this purpose, 70 whole blood samples (35 painters and 35 control) were studied by comet assay and evaluated according to four image-analysis parameters including tail intensity (TI), tail DNA (DNAt), tail moment (TM) and olive tail moment (OTM). Also, urinary hippuric acid rate which is used as a marker for toluene exposure was also measured in painters. As a result, mean of TI, DNAt, TM and OTM in organic solventexposed group had higher than in control group. TI, DNAt and TM were statistically significant between the exposed and nonexposed groups (p < 0.05), but OTM was not (p > 0.05). Moreover, smoking habit was statistical compared with DNA damage parameters and hippuric acid rates in exposed group. While statistically significant association was determined with hippuric acid rates (p < 0.05), there was not any statistically association with DNA damage parameters (p > 0.05). The study is suggested that organic solvents exposure can induce lymphocyte DNA damage in painters