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  • 1.
    Engwall, Magnus
    et al.
    Örebro University, School of Science and Technology.
    Venizelos, Nikolaos
    Örebro University, School of Health and Medical Sciences.
    Westman, Ola
    Örebro University, School of Science and Technology.
    Larsson, Maria
    Örebro University, School of Science and Technology.
    Nordén, Marcus
    Örebro University, School of Science and Technology.
    Hollert, Henner
    Rheinisch-Westfälische Technische Hochschule (RWTH), Aachen, Germany.
    Johansson, Jessica
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Polycyclic aromatic hydrocarbons (PAHs) reduce hepatic beta-oxidation of fatty acids in chick embryos2013In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 20, no 3, p. 1881-1888Article in journal (Refereed)
    Abstract [en]

    Polycyclic aromatic hydrocarbons (PAHs) are widespread fused-ring contaminants formed during incomplete combustion of almost all kind of organic materials from both natural and anthropogenic sources. Some PAHs have been shown to be carcinogenic to humans, and a wide range of PAHs are found in wildlife all around the globe including avian species. The purpose of this project was to assess the effects of a standard mixture of 16 PAHs (United States Environmental Protection Agency) on the hepatic fatty acid beta-oxidation in chicken embryos (Gallus gallus domesticus) exposed in ovo. The hepatic beta-oxidation was measured using a tritium release assay with [9,10-H-3]-palmitic acid (16:0) as substrate. Treated groups were divided into groups of 0.05, 0.1, 0.3, 0.5, and 0.8 mg PAHs/kg egg weight. The hepatic beta-oxidation was reduced after exposure in ovo to the 16 PAHs mixture compared to control. The mechanisms causing reduced fatty acid oxidation in the present study are unclear, however may be due to deficient membrane structure, the functionality of enzymes controlling the rate of fatty acid entering into the mitochondria, or complex pathways connected to endocrine disruption. To the best of our knowledge, this is the first time a PAH-caused reduction of hepatic beta-oxidation of fatty acids in avian embryos has been observed. The implication of this finding on risk assessment of PAH exposure in avian wildlife remains to be determined.

  • 2.
    Nordén, Marcus
    Örebro University, School of Science and Technology.
    Comparative avian developmental toxicity of PFAAs2013Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    Perfluoroalkyl acids (PFAAs) are persistent organic pollutants that can commonly be found in environmental matrixes and wildlife from all over the globe. The PFAAs have been used in applications such as water and dirt repelling treatments for textiles, oil-resistant paper coatings and fire-fighting foams. Four studies were designed to evaluate the occurrence of PFAAs in Swedish populations of birds, the developmental toxicity of different PFAAs and species sensitivity differences as well as possible modes of action for the toxicity. The studied species were domestic chicken, and the wild species great cormorant and herring gull. Cormorant and gull eggs were collected from bird colonies in Lake Vänern, Sweden. Chemical analyses were performed on some of the eggs to determine the occurrence of 15 PFAAs in the eggs. The other eggs and eggs of domestic chicken were incubated and injected with solutions of the PFAAs PFOS, PFOA, PFBS and PFUnDA. The eggs were candled every 1-3 days to determine viability. High levels of PFAAs, mainly PFOS followed by PFUnDA, were found in the herring gull and great cormorant eggs. PFOS was found at concentrations up to 1163 ng/g and 771 ng/g in cormorant and herring gull, respectively. In the toxicity tests, chicken was found to be more sensitive than the wild species and cormorant was in general the least sensitive species. PFOA was found to be the most toxic of the chemicals followed by PFOS, PFBS and PFUnDA in decreasing order. Comparing these results with the levels of these chemicals found in the eggs of herring gull and great cormorant, PFOS is the chemical of most concern. Although PFOA had the highest toxicity, the levels found in the eggs were very low. In an additional study, the hepatic β-oxidation in developing chicken embryos after in ovo exposure to PFOS was studied with a tritium release assay. PFOS was found to increase the β-oxidation of palmitic acid at PFOS concentrations 3-7 times lower than the average egg levels in cormorant and herring gull. Therefore the occurrence of effects on the fatty acid metabolism cannot be ruled out. The doses of effect on embryo survival in the toxicity and the levels found in the herring gulls and cormorants gives a small margin of safety for the wild populations. Continued environmental monitoring and further studies on the toxicity of PFAAs that occur at high environmental concentrations is important.

    List of papers
    1. High levels of perfluoroalkyl acids in eggs and embryo livers of great cormorant (Phalacrocorax carbo sinensis) and herring gull (Larus argentatus) from Lake Vanern, Sweden
    Open this publication in new window or tab >>High levels of perfluoroalkyl acids in eggs and embryo livers of great cormorant (Phalacrocorax carbo sinensis) and herring gull (Larus argentatus) from Lake Vanern, Sweden
    2013 (English)In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, ISSN 1614-7499, Vol. 20, no 11, p. 8021-8030Article in journal (Refereed) Published
    Abstract [en]

     In the eggs and developing chick livers in the two wild bird species, great cormorant and herring gull, the concentrations of a range of 15 perfluoroalkyl acids (PFAAs) were determined. Eggs of the two species were collected from Lake Vanern, Sweden, and analysed either as undeveloped egg (whole egg or separated into yolk and albumen) or incubated until start of the hatching process when the chick liver was removed and analysed. High levels of PFAAs were found in all matrixes except albumen. The predominant PFAA was perfluorooctane sulfonate (PFOS), which was found in the mug/g wet weight (ww) range in some samples of cormorant whole egg, yolk and liver and herring gull egg yolk and liver. The average concentration in yolk was 1,506 ng/g ww in cormorant and 589 ng/g ww in herring gull. The average liver concentrations of PFOS were 583 ng/g ww in cormorant and 508 ng/g ww in herring gull. At these concentrations, biochemical effects in the developing embryo or effects on embryo survival cannot be ruled out. For perfluoroalkyl carboxylates (PFCAs), the liver/egg and liver/yolk concentration ratios increased with PFCA chain length in cormorant but not in herring gull, indicating that chain length could possibly affect egg-to-liver transfer of PFCAs and that species differences may exist.

    National Category
    Natural Sciences
    Research subject
    Biology; Enviromental Science
    Identifiers
    urn:nbn:se:oru:diva-29083 (URN)10.1007/s11356-013-1567-3 (DOI)000325811600047 ()
    Funder
    Formas
    Available from: 2013-05-21 Created: 2013-05-21 Last updated: 2017-12-06Bibliographically approved
    2. Developmental toxicity of PFOS and PFOA in great cormorant (Phalacrocorax carbo sinensis), herring gull (Larus argentatus) and chicken (Gallus gallus domesticus)
    Open this publication in new window or tab >>Developmental toxicity of PFOS and PFOA in great cormorant (Phalacrocorax carbo sinensis), herring gull (Larus argentatus) and chicken (Gallus gallus domesticus)
    (English)Manuscript (preprint) (Other academic)
    Abstract [en]

    Perfluoroalkyl acids (PFAAs) are found globally in environmental samples and have been studied in various species. In this study we compare the sensitivity of three avian species to the toxic effects of perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA). Eggs of great cormorant (Phalacrocorax carbo sinensis) and herring gull (Larus argentatus) and the domestic White Leghorn chicken (Gallus gallus domesticus) were exposed in ovo by injection into the air sac. Effects on embryo survival were observed following exposure to PFOS and PFOA in chicken and herring gull. Chicken was found to be the most sensitive species with 50% reduced embryo survival at 8.5 μg/g egg for PFOS and 2.5 μg/g egg for PFOA. Cormorant was shown to be the least sensitive species. The difference in sensitivity between chicken and herring gull was a factor of 2.7 for PFOS and 3.5 for PFOA. Between chicken and great cormorant the sensitivity difference was 2.6 for PFOS and 8.2 for PFOA. Effects on embryo survival were seen at egg injection doses of PFOS close to levels found in environmental samples from wild birds, indicating that PFOS could be having effects in highly exposed populations of birds. This study also shows that there are differences in species sensitivity to PFOS and PFOA that should be taken into consideration in avian wildlife risk assessment.

    Keywords
    PFOS, PFOA, chicken, herring gull, great cormorant, developmental toxicity
    National Category
    Biological Sciences
    Research subject
    Biology
    Identifiers
    urn:nbn:se:oru:diva-32655 (URN)
    Available from: 2013-12-06 Created: 2013-12-06 Last updated: 2017-10-17Bibliographically approved
    3. Developmental toxicity of PFBS and PFUnDA in great cormorant (Phalacrocorax carbo sinensis),herring gull (Larus argentatus) and chicken (Gallus gallus domesticus)
    Open this publication in new window or tab >>Developmental toxicity of PFBS and PFUnDA in great cormorant (Phalacrocorax carbo sinensis),herring gull (Larus argentatus) and chicken (Gallus gallus domesticus)
    (English)Manuscript (preprint) (Other academic)
    Abstract [en]

    Perfluorinated alkyl acids (PFAAs) are found in a wide range of biological matrixes due to their persistent nature and widespread and longtime use. Among the most commonly found in wildlife are perfluorooctane sulfonate (PFOS) and perfluoroundecanoic acid (PFUnDA). PFOS and its precursors have been replaced by shorter chained compounds such as perfluorobutane sulfonate (PFBS). PFOS and perfluoroctanoic acid (PFOA) are generally the most studied and studies on wild species of birds are scarce. In this study chicken, great cormorant and herring gull eggs were exposed to solutions of PFBS and PFUnDA. The eggs were incubated and candled to monitor embryo survival. PFBS significantly reduced survival in all species at 17.3 μg/g anion dose. Chicken LD50 was calculated to 29 μg/g. The cormorant was less sensitive. PFUnDA did not affect embryo survival in any of the species but did increase liver somatic index and decrease heart somatic index in chicken. Chemical analysis confirmed that the liver absorption of the chemicals correlated to the injected dose. Chicken was found to accumulate more of the PFAAs in the liver than the wild species. Comparing observed effects and environmental levels, PFBS and PFUnDA are of minor environmental concern compared to PFOS. However, due to their persistent nature and the possibility of cumulative effects of several PFAAs, continued monitoring and moretoxicological studies are needed.

    Keywords
    PFBS, PFUnDA
    National Category
    Biological Sciences
    Research subject
    Biology
    Identifiers
    urn:nbn:se:oru:diva-32656 (URN)
    Available from: 2013-12-06 Created: 2013-12-06 Last updated: 2017-10-17Bibliographically approved
    4. Perfluorooctane sulfonate increases β-oxidation of palmitic acid in chicken liver
    Open this publication in new window or tab >>Perfluorooctane sulfonate increases β-oxidation of palmitic acid in chicken liver
    2012 (English)In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 19, no 5, p. 1859-1863Article in journal (Refereed) Published
    Abstract [en]

    Purpose: Perfluorooctane sulfonate (PFOS) belongs to a group of chemicals called perfluoroalkyl acids that have been extensively used in various applications such as stain and oil resistant treatments for fabrics, fire-fighting foams, and insecticides. These chemicals present an environmental and health risk being present in many samples both in wildlife and humans. In this study, we investigate the effect of PFOS on fatty acid β-oxidation in developing chicken embryos.

    Methods: Fertilized chicken eggs were exposed in ovo to PFOS at day 4 of incubation. On day 10, the eggs were dissected and livers were incubated in vitro with (3)H-palmitic acid for 2 h. The media were collected, and after clean up, the amount of tritiated water was measured with liquid scintillation counting to determine the rate of palmitic acid β-oxidation.

    Results: PFOS was found to induce fatty acid β-oxidation at doses starting from a lowest observed effect level (LOEL) of 0.1 μg/g egg weight. Maximum induction of 77 % compared to control was seen at 0.3 μg/g.

    Conclusions: The administered doses in which effects are seen are around and even lower than the levels that can be found in wild populations of birds. General population human levels are a factor of two to three times lower than the LOEL value of this study. The environmental contamination of PFOS therefore presents a possibility of effects in wild populations of birds.

    Place, publisher, year, edition, pages
    Heidelberg, Germany: Springer Berlin/Heidelberg, 2012
    Keywords
    Perfluorooctane sulfonate, PFOS, exposure, chicken, palmitic acid, β-oxidation, tritium
    National Category
    Medical and Health Sciences Biological Sciences Biomedical Laboratory Science/Technology
    Research subject
    Biology; Biomedical Laboratory Science
    Identifiers
    urn:nbn:se:oru:diva-24853 (URN)10.1007/s11356-012-0869-1 (DOI)000305884600053 ()22441698 (PubMedID)2-s2.0-84863225035 (Scopus ID)
    Funder
    Swedish Research Council Formas
    Available from: 2012-08-23 Created: 2012-08-23 Last updated: 2017-12-07Bibliographically approved
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  • 3.
    Nordén, Marcus
    et al.
    Örebro University, School of Science and Technology.
    Berger, Urs
    Department of Applied Environmental Science (ITM), Stockholm University, Sweden.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Developmental toxicity of PFBS and PFUnDA in great cormorant (Phalacrocorax carbo sinensis),herring gull (Larus argentatus) and chicken (Gallus gallus domesticus)Manuscript (preprint) (Other academic)
    Abstract [en]

    Perfluorinated alkyl acids (PFAAs) are found in a wide range of biological matrixes due to their persistent nature and widespread and longtime use. Among the most commonly found in wildlife are perfluorooctane sulfonate (PFOS) and perfluoroundecanoic acid (PFUnDA). PFOS and its precursors have been replaced by shorter chained compounds such as perfluorobutane sulfonate (PFBS). PFOS and perfluoroctanoic acid (PFOA) are generally the most studied and studies on wild species of birds are scarce. In this study chicken, great cormorant and herring gull eggs were exposed to solutions of PFBS and PFUnDA. The eggs were incubated and candled to monitor embryo survival. PFBS significantly reduced survival in all species at 17.3 μg/g anion dose. Chicken LD50 was calculated to 29 μg/g. The cormorant was less sensitive. PFUnDA did not affect embryo survival in any of the species but did increase liver somatic index and decrease heart somatic index in chicken. Chemical analysis confirmed that the liver absorption of the chemicals correlated to the injected dose. Chicken was found to accumulate more of the PFAAs in the liver than the wild species. Comparing observed effects and environmental levels, PFBS and PFUnDA are of minor environmental concern compared to PFOS. However, due to their persistent nature and the possibility of cumulative effects of several PFAAs, continued monitoring and moretoxicological studies are needed.

  • 4.
    Nordén, Marcus
    et al.
    Örebro University, School of Science and Technology.
    Berger, Urs
    Department of Applied Environmental Science (ITM), Stockholm University, Sweden.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Developmental toxicity of PFOS and PFOA in great cormorant (Phalacrocorax carbo sinensis), herring gull (Larus argentatus) and chicken (Gallus gallus domesticus)Manuscript (preprint) (Other academic)
    Abstract [en]

    Perfluoroalkyl acids (PFAAs) are found globally in environmental samples and have been studied in various species. In this study we compare the sensitivity of three avian species to the toxic effects of perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA). Eggs of great cormorant (Phalacrocorax carbo sinensis) and herring gull (Larus argentatus) and the domestic White Leghorn chicken (Gallus gallus domesticus) were exposed in ovo by injection into the air sac. Effects on embryo survival were observed following exposure to PFOS and PFOA in chicken and herring gull. Chicken was found to be the most sensitive species with 50% reduced embryo survival at 8.5 μg/g egg for PFOS and 2.5 μg/g egg for PFOA. Cormorant was shown to be the least sensitive species. The difference in sensitivity between chicken and herring gull was a factor of 2.7 for PFOS and 3.5 for PFOA. Between chicken and great cormorant the sensitivity difference was 2.6 for PFOS and 8.2 for PFOA. Effects on embryo survival were seen at egg injection doses of PFOS close to levels found in environmental samples from wild birds, indicating that PFOS could be having effects in highly exposed populations of birds. This study also shows that there are differences in species sensitivity to PFOS and PFOA that should be taken into consideration in avian wildlife risk assessment.

  • 5.
    Nordén, Marcus
    et al.
    Örebro University, School of Science and Technology.
    Berger, Urs
    Stockholms universitet, Stockholm, Sweden.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    High levels of perfluoroalkyl acids in eggs and embryo livers of great cormorant (Phalacrocorax carbo sinensis) and herring gull (Larus argentatus) from Lake Vanern, Sweden2013In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, ISSN 1614-7499, Vol. 20, no 11, p. 8021-8030Article in journal (Refereed)
    Abstract [en]

     In the eggs and developing chick livers in the two wild bird species, great cormorant and herring gull, the concentrations of a range of 15 perfluoroalkyl acids (PFAAs) were determined. Eggs of the two species were collected from Lake Vanern, Sweden, and analysed either as undeveloped egg (whole egg or separated into yolk and albumen) or incubated until start of the hatching process when the chick liver was removed and analysed. High levels of PFAAs were found in all matrixes except albumen. The predominant PFAA was perfluorooctane sulfonate (PFOS), which was found in the mug/g wet weight (ww) range in some samples of cormorant whole egg, yolk and liver and herring gull egg yolk and liver. The average concentration in yolk was 1,506 ng/g ww in cormorant and 589 ng/g ww in herring gull. The average liver concentrations of PFOS were 583 ng/g ww in cormorant and 508 ng/g ww in herring gull. At these concentrations, biochemical effects in the developing embryo or effects on embryo survival cannot be ruled out. For perfluoroalkyl carboxylates (PFCAs), the liver/egg and liver/yolk concentration ratios increased with PFCA chain length in cormorant but not in herring gull, indicating that chain length could possibly affect egg-to-liver transfer of PFCAs and that species differences may exist.

  • 6.
    Nordén, Marcus
    et al.
    Örebro University, School of Science and Technology.
    Halldin, Krister
    Karolinska Institute, Stockholm, Sweden.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Gene expression analysis of chicken during development subsequent to PFOS and PFOA exposure2010In: In Vivo, ISSN 0258-851X, E-ISSN 1791-7549, Vol. 24, no 3, p. 358-358Article in journal (Refereed)
    Abstract [en]

    Perfluorinated compounds have been manufactured for over50 years and have a wide range of applications due to theirsurfactant properties. They have been used as fabricprotectors to repel water and dirt, fire-fighting foams, nonstickcoatings, insecticides and all weather clothing. PFCsare widespread in the environment and are found globally inwildlife. Among the most abundant are perfluorooctanesulfonate (PFOS) and perfluorooctanoic acid (PFOA). Thelevels in common guillemot eggs from the Baltic Sea areamong the highest found in the Scandinavian environment.Previous studies show effects on embryo survival of chickenat levels close to levels found in the Baltic guillemot. Toinvestigate the possible mechanisms of action, microarraygene expression analysis was conducted. Chicken eggs wereincubated at 37.5 degrees and 60% relative humidity. PFOSor PFOA was administered to eggs on day 4 of incubationby injection into the air cell of the egg. 1 μl injectionsolution per gram egg was used. The injection solutionscontained PFOS or PFOA dissolved at differentconcentrations in sterile water with 5% or 2.5% dimethylsulfoxide respectively. The doses were 10 and 3 mg/kg forPFOS and 1.6 and 0.5 mg/kg for PFOA. The vehiclesolutions were used as control treatment. At day 19 theembryos were sacrificed and the liver was extracted. Four livers per dose and each vehicle control were used. RNA wasextracted from the livers using Qiagen RNeasy Mini Kit.cDNA was synthesized from mRNA and used for microarrayanalysis on Agilent two-color chicken microarrays.

  • 7.
    Nordén, Marcus
    et al.
    Örebro University, School of Science and Technology.
    Westman, Ola
    Örebro University, School of Science and Technology.
    Venizelos, Nikolaos
    Örebro University, School of Health and Medical Sciences, Örebro University, Sweden.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Perfluorooctane sulfonate increases β-oxidation of palmitic acid in chicken liver2012In: Environmental science and pollution research international, ISSN 0944-1344, E-ISSN 1614-7499, Vol. 19, no 5, p. 1859-1863Article in journal (Refereed)
    Abstract [en]

    Purpose: Perfluorooctane sulfonate (PFOS) belongs to a group of chemicals called perfluoroalkyl acids that have been extensively used in various applications such as stain and oil resistant treatments for fabrics, fire-fighting foams, and insecticides. These chemicals present an environmental and health risk being present in many samples both in wildlife and humans. In this study, we investigate the effect of PFOS on fatty acid β-oxidation in developing chicken embryos.

    Methods: Fertilized chicken eggs were exposed in ovo to PFOS at day 4 of incubation. On day 10, the eggs were dissected and livers were incubated in vitro with (3)H-palmitic acid for 2 h. The media were collected, and after clean up, the amount of tritiated water was measured with liquid scintillation counting to determine the rate of palmitic acid β-oxidation.

    Results: PFOS was found to induce fatty acid β-oxidation at doses starting from a lowest observed effect level (LOEL) of 0.1 μg/g egg weight. Maximum induction of 77 % compared to control was seen at 0.3 μg/g.

    Conclusions: The administered doses in which effects are seen are around and even lower than the levels that can be found in wild populations of birds. General population human levels are a factor of two to three times lower than the LOEL value of this study. The environmental contamination of PFOS therefore presents a possibility of effects in wild populations of birds.

  • 8.
    Westman, Ola
    et al.
    Örebro University, School of Science and Technology.
    Nordén, Marcus
    Örebro University, School of Science and Technology.
    Larsson, Maria
    Örebro University, School of Science and Technology.
    Venizelos, Nikolaos
    Örebro University, School of Science and Technology.
    Hollert, Henner
    RWTH Aachen University, Aachen, Germany.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Reduced beta-oxidation in avian following exposure of polycyclic aromatic hydrocarbons2012In: Toxicology Letters, ISSN 0378-4274, E-ISSN 1879-3169, Vol. 211, no Supplement, p. s86-s87Article in journal (Refereed)
  • 9.
    Westman, Ola
    et al.
    Örebro University, School of Science and Technology.
    Nordén, Marcus
    Örebro University, School of Science and Technology.
    Venizelos, Nikolaos
    Örebro University, School of Health and Medical Sciences.
    Engwall, Magnus
    Örebro University, School of Science and Technology.
    Effects of perfluorinated compounds on hepatic fatty acid oxidation in avian embryos using a tritium release assay2010In: In Vivo, ISSN 0258-851X, E-ISSN 1791-7549, Vol. 24, no 3, p. 363-364Article in journal (Refereed)
    Abstract [en]

    Objective: Due to high persistence and bioaccumulationperfluorinated compounds (PFCs) are found globally in varioustypes of wildlife samples and also human samples¹. Accordingto our laboratory studies, perfluorooctane sulfonate (PFOS) hascaused early mortality in chicken embryos at doses close toconcentrations found in eggs of the Baltic guillemot². We havedesigned a method in which hepatic embryonic tissue fromchicken (Gallus domesticus) is used to investigate the effectsof PFCs on the β-oxidation of fatty acids.Materials and Methods: The embryos were exposed inovo to PFCs. On day 10 embryo livers were incubated invitro with tritiated fatty acids. Fatty acid oxidation was thencalculated from the tritium released into water, using ascintillation counter.Results: Our studies suggest a small but significant increaseof the β-oxidation of fatty acids in chicken embryonic livertissue in vitro after in ovo exposure to PFOS. The β-oxidationwas significantly induced after embryo exposure to 1 mg/kgPFOS (p=0.003) and 10 mg/kg PFOS (p=0.04), being 39%and 34% higher, respectively compared to control.Conclusion: The results show that in ovo exposure incombination with an in vitro method using a tritium releaseassay to detect effects on the β-oxidation of fatty acids inavian embryo hepatic tissue could be a useful method inelucidating possible mechanisms behind avian developmental toxicity.

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