Pub Date : 2024-10-11DOI: 10.1016/j.toxlet.2024.10.005
T. Kostov , P. Diel , E. Isenmann
Plant steroids such as ecdysterone (ECDY) or diosgenin (DIO) have been associated with anabolic and performance-enhancing effects for years. However, the molecular mechanisms have not yet been extensively studied in skeletal muscle cells. Consequently, the anabolic activity and associated molecular mechanisms of ECDY and DIO alone and in combination were investigated in C2C12 myotubes. Dose-dependent effects of both compounds on myotube diameter, mRNA expression of IGF-1 and PI3KR1 as well as expression of myosin heavy chain (MHC) proteins were analyzed in differentiated C2C12 cells. In addition, the binding affinities to androgen and estrogen receptors were analyzed. Treatment with ECDY and DIO significantly induced hypertrophy of C2C12 myotubes. Partially additive effects were observed. This is supported by the mRNA expression of IGF-1 and PI3KR1 as well as in the expression of MHC. However, no clear statement can be made regarding which combination has the strongest additive effects. Besides the results suggest that, in contrast to ECDY, DIO has antiandrogenic effects and bind on AR. Consequently, it indicate that two different mechanisms of action are activated in ECDY and DIO combinations. However, this must be confirmed in further cell cultures studies and human interventions concerning anti-doping regulations.
多年来，蜕皮激素（ECDY）或薯蓣皂甙（DIO）等植物类固醇一直与合成代谢和提高性能的作用有关。然而，人们尚未在骨骼肌细胞中广泛研究其分子机制。因此，我们在 C2C12 肌管中研究了 ECDY 和 DIO 单独或混合使用时的合成代谢活性和相关分子机制。在分化的 C2C12 细胞中，分析了这两种化合物对肌管直径、IGF-1 和 PI3KR1 的 mRNA 表达以及肌球蛋白重链（MHC）蛋白表达的剂量依赖性影响。此外，还分析了与雄激素和雌激素受体的结合亲和力。用 ECDY 和 DIO 处理可显著诱导 C2C12 肌管肥大。观察到了部分相加效应。IGF-1和PI3KR1的mRNA表达以及MHC的表达都证明了这一点。不过，对于哪种组合具有最强的叠加效应，目前还没有明确的说法。此外，研究结果表明，与 ECDY 不同，DIO 具有抗雄激素作用，并与 AR 结合。因此，这表明 ECDY 和 DIO 组合激活了两种不同的作用机制。不过，这一点必须在进一步的细胞培养研究和有关反兴奋剂条例的人体干预中得到证实。
{"title":"Examination of the anabolic activity and mechanisms of action of the combination of Diosgenin and Ecdysterone in C2C12 myotubes","authors":"T. Kostov , P. Diel , E. Isenmann","doi":"10.1016/j.toxlet.2024.10.005","DOIUrl":"10.1016/j.toxlet.2024.10.005","url":null,"abstract":"<div><div>Plant steroids such as ecdysterone (ECDY) or diosgenin (DIO) have been associated with anabolic and performance-enhancing effects for years. However, the molecular mechanisms have not yet been extensively studied in skeletal muscle cells. Consequently, the anabolic activity and associated molecular mechanisms of ECDY and DIO alone and in combination were investigated in C2C12 myotubes. Dose-dependent effects of both compounds on myotube diameter, mRNA expression of IGF-1 and PI3KR1 as well as expression of myosin heavy chain (MHC) proteins were analyzed in differentiated C2C12 cells. In addition, the binding affinities to androgen and estrogen receptors were analyzed. Treatment with ECDY and DIO significantly induced hypertrophy of C2C12 myotubes. Partially additive effects were observed. This is supported by the mRNA expression of IGF-1 and PI3KR1 as well as in the expression of MHC. However, no clear statement can be made regarding which combination has the strongest additive effects. Besides the results suggest that, in contrast to ECDY, DIO has antiandrogenic effects and bind on AR. Consequently, it indicate that two different mechanisms of action are activated in ECDY and DIO combinations. However, this must be confirmed in further cell cultures studies and human interventions concerning anti-doping regulations.</div></div>","PeriodicalId":23206,"journal":{"name":"Toxicology letters","volume":"401 ","pages":"Pages 181-189"},"PeriodicalIF":2.9,"publicationDate":"2024-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142475461","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-10DOI: 10.1016/j.toxlet.2024.10.007
Hang Luo , Shanshan Zhao , Jing Zi , Yifan Hu , Yuqin Yao , Jingyuan Xiong
Polycyclic aromatic hydrocarbons (PAHs) exposure is associated with cardiovascular diseases. Toxic effects of PAHs are diverse, while cardiovascular consequences of benzo[b]fluoranthene (B[b]F) are unclear. Here, we reported the impacts of B[b]F on coronary artery and atherosclerosis markers both in mice and umbilical vein endothelial EAhy.926 cells. In mice, we found that B[b]F decreases heart-to-body weight ratio, affects aortic physiology, elevates serum low-density lipoprotein and total cholesterol, increases aortic levels of collagen fiber and atherosclerotic marker vascular cell adhesion molecule-1 (VCAM-1), and downregulates oxidative stress related nuclear factor erythroid 2-related factor 2 (Nrf2). In EAhy.926 cells, we showed that B[b]F inhibits cell proliferation and migration in a dose-dependent manner, induces cell cycle arrest and apoptosis, increases reactive oxygen species, upregulates VCAM-1 level, and suppresses expression of Nrf2. Taken together, our findings reveal that B[b]F exposure may contribute to coronary artery damage and potentially induce atherosclerosis, possibly via the Nrf2-related signaling pathways.
{"title":"Benzo[b]fluoranthene damages coronary artery and affects atherosclerosis markers in mice and umbilical vein endothelial cells","authors":"Hang Luo , Shanshan Zhao , Jing Zi , Yifan Hu , Yuqin Yao , Jingyuan Xiong","doi":"10.1016/j.toxlet.2024.10.007","DOIUrl":"10.1016/j.toxlet.2024.10.007","url":null,"abstract":"<div><div>Polycyclic aromatic hydrocarbons (PAHs) exposure is associated with cardiovascular diseases. Toxic effects of PAHs are diverse, while cardiovascular consequences of benzo[<em>b</em>]fluoranthene (B[<em>b</em>]F) are unclear. Here, we reported the impacts of B[<em>b</em>]F on coronary artery and atherosclerosis markers both in mice and umbilical vein endothelial EAhy.926 cells. In mice, we found that B[<em>b</em>]F decreases heart-to-body weight ratio, affects aortic physiology, elevates serum low-density lipoprotein and total cholesterol, increases aortic levels of collagen fiber and atherosclerotic marker vascular cell adhesion molecule-1 (VCAM-1), and downregulates oxidative stress related nuclear factor erythroid 2-related factor 2 (Nrf2). In EAhy.926 cells, we showed that B[<em>b</em>]F inhibits cell proliferation and migration in a dose-dependent manner, induces cell cycle arrest and apoptosis, increases reactive oxygen species, upregulates VCAM-1 level, and suppresses expression of Nrf2. Taken together, our findings reveal that B[<em>b</em>]F exposure may contribute to coronary artery damage and potentially induce atherosclerosis, possibly via the Nrf2-related signaling pathways.</div></div>","PeriodicalId":23206,"journal":{"name":"Toxicology letters","volume":"401 ","pages":"Pages 150-157"},"PeriodicalIF":2.9,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142432867","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-10DOI: 10.1016/j.toxlet.2024.10.006
Nadja Walle , Adrian A. Doerr , Benjamin Peters , Matthias W. Laschke , Michael D. Menger , Peter H. Schmidt , Markus R. Meyer , Nadine Schaefer
The number of fatal cases involving synthetic cannabinoids (SCs) is increasing. However, interpretation of postmortem (PM) toxicological findings is challenging, due to unknown PM intervals and possible redistribution processes or instabilities. Only sparse data on SCs are available. Therefore, a controlled pig study was performed to determine the PM stability of cumyl-5F-P7AICA under different environmental conditions. Ten pigs inhalatively received 200 µg/kg body weight cumyl-5F-P7AICA each. Six hours later, they were euthanized and biopsies of the main tissues and body fluids were taken. Animals were stored in air or water (n=5 each) and samples were repeatedly taken for three days. Quantification of cumyl-5F-P7AICA and its N-pentanoic acid metabolite (NPA) was performed using standard addition or a fully validated method (blood) followed by LC-MS/MS. Time-dependent concentration changes of cumyl-5F-P7AICA were observed in liver, bile fluid and muscle specimens at both conditions. Concentrations of NPA only changed considerably in duodenum content of animals stored in air. Environment-dependent concentrations changes were only observed for cumyl-5F-P7AICA in kidney and the NPA metabolite in duodenum content. Overall, cumyl-5F-P7AICA and its metabolite seem to be quite stable in PM specimens. Hence, also central blood might be analyzed, if no peripheral blood is available.
{"title":"Are the postmortem concentration changes of the synthetic cannabinoid cumyl-5F-P7AICA and its N-pentanoic acid metabolite dependent on the environmental conditions? – A systematic study following pulmonary administration to pigs","authors":"Nadja Walle , Adrian A. Doerr , Benjamin Peters , Matthias W. Laschke , Michael D. Menger , Peter H. Schmidt , Markus R. Meyer , Nadine Schaefer","doi":"10.1016/j.toxlet.2024.10.006","DOIUrl":"10.1016/j.toxlet.2024.10.006","url":null,"abstract":"<div><div>The number of fatal cases involving synthetic cannabinoids (SCs) is increasing. However, interpretation of postmortem (PM) toxicological findings is challenging, due to unknown PM intervals and possible redistribution processes or instabilities. Only sparse data on SCs are available. Therefore, a controlled pig study was performed to determine the PM stability of <em>cumyl</em>-5F-P7AICA under different environmental conditions. Ten pigs inhalatively received 200 µg/kg body weight <em>cumyl</em>-5F-P7AICA each. Six hours later, they were euthanized and biopsies of the main tissues and body fluids were taken. Animals were stored in air or water (n=5 each) and samples were repeatedly taken for three days. Quantification of <em>cumyl</em>-5F-P7AICA and its <em>N</em>-pentanoic acid metabolite (NPA) was performed using standard addition or a fully validated method (blood) followed by LC-MS/MS. Time-dependent concentration changes of <em>cumyl</em>-5F-P7AICA were observed in liver, bile fluid and muscle specimens at both conditions. Concentrations of NPA only changed considerably in duodenum content of animals stored in air. Environment-dependent concentrations changes were only observed for <em>cumyl</em>-5F-P7AICA in kidney and the NPA metabolite in duodenum content. Overall, <em>cumyl</em>-5F-P7AICA and its metabolite seem to be quite stable in PM specimens. Hence, also central blood might be analyzed, if no peripheral blood is available.</div></div>","PeriodicalId":23206,"journal":{"name":"Toxicology letters","volume":"401 ","pages":"Pages 170-180"},"PeriodicalIF":2.9,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142442058","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Caco-2 cells, a human colorectal adenocarcinoma cell line, are widely used to model small intestinal epithelial cells in the drug development process because they can predict drug absorption with high accuracy. However, Caco-2 cells have several issues. First, Caco-2 cells have little expression of cytochrome P450 3A4 (CYP3A4), which is a major drug-metabolizing enzyme in the human intestine. We previously developed Caco-2 cells whose expression of CYP3A4 can be controlled using doxycycline (Dox) (CYP3A4-Caco-2 cells) (Ichikawa et al., Sci. Rep, 2021). However, since the Tet-On system was used to regulate CYP3A4 expression in these cells, there was concern about drug-drug interactions. The second issue is that in the human small intestine, carboxylesterase 2 (CES2) is more highly expressed than carboxylesterase 1 (CES1), while in Caco-2 cells CES1 is more highly expressed. The third issue is the low level expression of uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1), a phase II drug-metabolizing enzyme. In this study, we used genome-editing technology to establish CES1-knockout Caco-2 cells whose CYP3A4 and UGT1A1 expression can be regulated by the Tet-Off system. These cell lines would be useful in pharmaceutical researches, including intestinal toxicological studies, as an in vitro model for orally administered drugs.
{"title":"Generation and application of CES1-knockout Tet-Off-regulated CYP3A4 and UGT1A1-expressing Caco-2 cells","authors":"Michika Murata , Kentaro Okada , Masaki Takahashi , Yukiko Ueyama-Toba , Sumito Ito , Hiroyuki Mizuguchi","doi":"10.1016/j.toxlet.2024.10.003","DOIUrl":"10.1016/j.toxlet.2024.10.003","url":null,"abstract":"<div><div>Caco-2 cells, a human colorectal adenocarcinoma cell line, are widely used to model small intestinal epithelial cells in the drug development process because they can predict drug absorption with high accuracy. However, Caco-2 cells have several issues. First, Caco-2 cells have little expression of cytochrome P450 3A4 (CYP3A4), which is a major drug-metabolizing enzyme in the human intestine. We previously developed Caco-2 cells whose expression of CYP3A4 can be controlled using doxycycline (Dox) (CYP3A4-Caco-2 cells) (Ichikawa et al., Sci. Rep, 2021). However, since the Tet-On system was used to regulate CYP3A4 expression in these cells, there was concern about drug-drug interactions. The second issue is that in the human small intestine, carboxylesterase 2 (CES2) is more highly expressed than carboxylesterase 1 (CES1), while in Caco-2 cells CES1 is more highly expressed. The third issue is the low level expression of uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1), a phase II drug-metabolizing enzyme. In this study, we used genome-editing technology to establish <em>CES1</em>-knockout Caco-2 cells whose CYP3A4 and UGT1A1 expression can be regulated by the Tet-Off system. These cell lines would be useful in pharmaceutical researches, including intestinal toxicological studies, as an in vitro model for orally administered drugs.</div></div>","PeriodicalId":23206,"journal":{"name":"Toxicology letters","volume":"401 ","pages":"Pages 158-169"},"PeriodicalIF":2.9,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142393544","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-03DOI: 10.1016/j.toxlet.2024.10.004
Chanhee Kim , Zhaohan Zhu , W. Brad Barbazuk , Rhonda L. Bacher , Christopher D. Vulpe
Physiologically relevant in vitro models are a priority in predictive toxicology to replace and/or reduce animal experiments. The compromised toxicant metabolism of many immortalized human liver cell lines grown as monolayers as compared to in vivo metabolism limits their physiological relevance. However, recent efforts to culture liver cells in a 3D environment, such as spheroids, to better mimic the in vivo conditions, may enhance the toxicant metabolism of human liver cell lines. In this study, we characterized the dynamic changes in the transcriptome of HepG2/C3A hepatocarcinoma cell spheroids maintained in a clinostat system (CelVivo) to gain insight into the metabolic capacity of this model as a function of spheroid size and culture time. We assessed morphological changes (size, necrotic core), cell health, and proliferation rate from initial spheroid seeding to 35 days of continuous culture in conjunction with a time-course (0, 3, 7, 10, 14, 21, 28 days) of the transcriptome (TempO-Seq, BioSpyder). The phenotypic characteristics of HepG2/C3A growing in spheroids were comparable to monolayer growth until ∼Day 12 (Day 10–14) when a significant decrease in cell doubling rate was noted which was concurrent with down-regulation of cell proliferation and cell cycle pathways over this time period. Principal component analysis of the transcriptome data suggests that the Day 3, 7, and 10 spheroids are pronouncedly different from the Day 14, 21, and 28 spheroids in support of a biological transition time point during the long-term 3D spheroid cultures. The expression of genes encoding cellular components involved in toxicant metabolism and transport rapidly increased during the early time points of spheroids to peak at Day 7 or Day 10 as compared to monolayer cultures with a gradual decrease in expression with further culture, suggesting the most metabolically responsive time window for exposure studies. Overall, we provide baseline information on the cellular and molecular characterization, with a particular focus on toxicant metabolic capacity dynamics and cell growth, of HepG2/C3A 3D spheroid cultures over time.
{"title":"Time-course characterization of whole-transcriptome dynamics of HepG2/C3A spheroids and its toxicological implications","authors":"Chanhee Kim , Zhaohan Zhu , W. Brad Barbazuk , Rhonda L. Bacher , Christopher D. Vulpe","doi":"10.1016/j.toxlet.2024.10.004","DOIUrl":"10.1016/j.toxlet.2024.10.004","url":null,"abstract":"<div><div>Physiologically relevant <em>in vitro</em> models are a priority in predictive toxicology to replace and/or reduce animal experiments. The compromised toxicant metabolism of many immortalized human liver cell lines grown as monolayers as compared to <em>in vivo</em> metabolism limits their physiological relevance. However, recent efforts to culture liver cells in a 3D environment, such as spheroids, to better mimic the <em>in vivo</em> conditions, may enhance the toxicant metabolism of human liver cell lines. In this study, we characterized the dynamic changes in the transcriptome of HepG2/C3A hepatocarcinoma cell spheroids maintained in a clinostat system (CelVivo) to gain insight into the metabolic capacity of this model as a function of spheroid size and culture time. We assessed morphological changes (size, necrotic core), cell health, and proliferation rate from initial spheroid seeding to 35 days of continuous culture in conjunction with a time-course (0, 3, 7, 10, 14, 21, 28 days) of the transcriptome (TempO-Seq, BioSpyder). The phenotypic characteristics of HepG2/C3A growing in spheroids were comparable to monolayer growth until ∼Day 12 (Day 10–14) when a significant decrease in cell doubling rate was noted which was concurrent with down-regulation of cell proliferation and cell cycle pathways over this time period. Principal component analysis of the transcriptome data suggests that the Day 3, 7, and 10 spheroids are pronouncedly different from the Day 14, 21, and 28 spheroids in support of a biological transition time point during the long-term 3D spheroid cultures. The expression of genes encoding cellular components involved in toxicant metabolism and transport rapidly increased during the early time points of spheroids to peak at Day 7 or Day 10 as compared to monolayer cultures with a gradual decrease in expression with further culture, suggesting the most metabolically responsive time window for exposure studies. Overall, we provide baseline information on the cellular and molecular characterization, with a particular focus on toxicant metabolic capacity dynamics and cell growth, of HepG2/C3A 3D spheroid cultures over time.</div></div>","PeriodicalId":23206,"journal":{"name":"Toxicology letters","volume":"401 ","pages":"Pages 125-138"},"PeriodicalIF":2.9,"publicationDate":"2024-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142378339","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-03DOI: 10.1016/j.toxlet.2024.10.002
Lathan Ball , H. Carter , C. Baker , R. Porter
Polycylic Aromatic Hydrocarbons (PAHs) are produced during the incomplete burning of organic materials. PAH sources include vehicle exhaust, tobacco smoke and waste incineration. Environmental and occupational exposures to PAHs are known to occur. Cancer is a significant endpoint of PAH exposure and several occupations associated with high PAH exposure have been classified by IARC as carcinogenic to humans (Group 1).
Pyrene is a common component of PAH mixtures and metabolism of pyrene leads to the excretion of 1-hydroxypyrene glucuronide (1-OHPyrG) in urine. Laboratory measurement of urinary 1-OHPyrG is employed in occupational and environmental biomonitoring programmes. The production of an anti-1-OHPyrG monoclonal antibody would allow the development of a PAH biomonitoring ELISA facilitating large scale laboratory screening and routine testing.
The development of a lateral flow immunoassay and the production of a field test (point of use test) would greatly increase the value of biomonitoring. A novel Lateral Flow has been developed which employs an anti-1-OHPyrG sheep monoclonal antibody (Mab) to capture the PAH metabolite. The captured metabolite is visualised through a second Mab raised against the Mab-1-OHPyrG immune complex. This sandwich assay provides a positive correlation between the assay signal and biomarker concentration.
A Smartphone camera allows signal measurement and a carefully considered ‘app’ provides result interpretation and data analysis. Results are provided in an exposed/not-exposed format. Performance of the lateral flow was confirmed through a comparative study and field trial. The development of a lateral flow test provides "real-time" analysis to occupational health professionals. On-site screening allows the immediate confirmation of safe working practice, provides immediate reassurance to those involved in potentially hazardous activities and greatly increases the efficacy of biomonitoring.
{"title":"The development of rapid immunoassays for the urinary analysis of 1-hydroxypyrene glucuronide facilitate both laboratory and on-site polycyclic aromatic hydrocarbon biomonitoring","authors":"Lathan Ball , H. Carter , C. Baker , R. Porter","doi":"10.1016/j.toxlet.2024.10.002","DOIUrl":"10.1016/j.toxlet.2024.10.002","url":null,"abstract":"<div><div>Polycylic Aromatic Hydrocarbons (PAHs) are produced during the incomplete burning of organic materials. PAH sources include vehicle exhaust, tobacco smoke and waste incineration. Environmental and occupational exposures to PAHs are known to occur. Cancer is a significant endpoint of PAH exposure and several occupations associated with high PAH exposure have been classified by IARC as <em>carcinogenic to humans</em> (Group 1).</div><div>Pyrene is a common component of PAH mixtures and metabolism of pyrene leads to the excretion of 1-hydroxypyrene glucuronide (1-OHPyrG) in urine. Laboratory measurement of urinary 1-OHPyrG is employed in occupational and environmental biomonitoring programmes. The production of an anti-1-OHPyrG monoclonal antibody would allow the development of a PAH biomonitoring ELISA facilitating large scale laboratory screening and routine testing.</div><div>The development of a lateral flow immunoassay and the production of a field test (<em>point of use test</em>) would greatly increase the value of biomonitoring. A novel Lateral Flow has been developed which employs an anti-1-OHPyrG sheep monoclonal antibody (Mab) to capture the PAH metabolite. The captured metabolite is visualised through a second Mab raised against the Mab-1-OHPyrG immune complex. This sandwich assay provides a positive correlation between the assay signal and biomarker concentration.</div><div>A Smartphone camera allows signal measurement and a carefully considered ‘app’ provides result interpretation and data analysis. Results are provided in an exposed/not-exposed format. Performance of the lateral flow was confirmed through a comparative study and field trial. The development of a lateral flow test provides \"real-time\" analysis to occupational health professionals. On-site screening allows the immediate confirmation of safe working practice, provides immediate reassurance to those involved in potentially hazardous activities and greatly increases the efficacy of biomonitoring.</div></div>","PeriodicalId":23206,"journal":{"name":"Toxicology letters","volume":"401 ","pages":"Pages 116-124"},"PeriodicalIF":2.9,"publicationDate":"2024-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142376087","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Increasing number of studies suggested that environmental deleterious impacts (such as estrogen-like endocrine disruptors, EDCs, unhealthy diet) during early human development affect the risk of developing non-communicable diseases including prostate cancer (PCa) later in life. To test if the combination of EDCs and unhealthy induces adult prostate lesions, we developed an experimental model of adult male Sprague Dawley rats exposed during gestation (from day 7) to weaning to high fat diet (HFD 60 % fat), or to a xenoestrogen (estradiol benzoate, EB, 2.5 µg/d) from post-natal days 1–5, or to a combination of both. EB and EB+HFD exposures induced decreased prostate weight in adult rats along with inflammatory status. A white blood cell infiltrate was observed after EB exposure and more dramatic lesions were observed with the combined exposure, along with a gland destruction. The lesions, following EB or EB+HFD exposure, are associated with elevated mRNA levels for TNFa, IL6 and CCL2/MCP1 pro-inflammatory cytokines while the levels of the anti-inflammatory IL10 cytokine remained unchanged. This activation of NLRP3 and elevated levels of CASP1 were observed following EB or EB+HFD exposures associated with elevated mRNA levels for IL1b, substrates for the NLRP3 complex. HFD exposure alone has mild if not pro-inflammatory effects in adult prostate. In conclusion, we showed that developmental combined exposure to EB and HFD programmed prostate inflammatory lesions in adult prostate. Since proliferative inflammatory atrophy and chronic inflammation of prostate may drive cell to become cancer cells, our model might be useful for study onset of PCa.
{"title":"Combined developmental exposure to estrogenic endocrine disruptor and nutritional imbalance induces long term adult prostate inflammation through inflammasome activation","authors":"Katia Gharieb , Nezli Doumandji , Wafa Bensalem , Rachel Paul Bellon , Lilia Inoubli , Bénazir Siddeek , Alexandra Traverse-Glehen , Myriam Decaussin-Petrucci , Michele Trabucchi , Mohamed Benahmed , Claire Mauduit","doi":"10.1016/j.toxlet.2024.10.001","DOIUrl":"10.1016/j.toxlet.2024.10.001","url":null,"abstract":"<div><div>Increasing number of studies suggested that environmental deleterious impacts (such as estrogen-like endocrine disruptors, EDCs, unhealthy diet) during early human development affect the risk of developing non-communicable diseases including prostate cancer (PCa) later in life. To test if the combination of EDCs and unhealthy induces adult prostate lesions, we developed an experimental model of adult male Sprague Dawley rats exposed during gestation (from day 7) to weaning to high fat diet (HFD 60 % fat), or to a xenoestrogen (estradiol benzoate, EB, 2.5 µg/d) from post-natal days 1–5, or to a combination of both. EB and EB+HFD exposures induced decreased prostate weight in adult rats along with inflammatory status. A white blood cell infiltrate was observed after EB exposure and more dramatic lesions were observed with the combined exposure, along with a gland destruction. The lesions, following EB or EB+HFD exposure, are associated with elevated mRNA levels for TNFa, IL6 and CCL2/MCP1 pro-inflammatory cytokines while the levels of the anti-inflammatory IL10 cytokine remained unchanged. This activation of NLRP3 and elevated levels of CASP1 were observed following EB or EB+HFD exposures associated with elevated mRNA levels for IL1b, substrates for the NLRP3 complex. HFD exposure alone has mild if not pro-inflammatory effects in adult prostate. In conclusion, we showed that developmental combined exposure to EB and HFD programmed prostate inflammatory lesions in adult prostate. Since proliferative inflammatory atrophy and chronic inflammation of prostate may drive cell to become cancer cells, our model might be useful for study onset of PCa.</div></div>","PeriodicalId":23206,"journal":{"name":"Toxicology letters","volume":"402 ","pages":"Pages 1-14"},"PeriodicalIF":2.9,"publicationDate":"2024-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142378338","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-27DOI: 10.1016/j.toxlet.2024.09.006
Jeff B. Willey , Chun Lei Liang , Tyler Pollock , Cheryl Khoury , Errol M. Thomson , Mike Walker , Annie St-Amand
Exposure load (EL) is an indicator of multiple chemical exposures based on human biomonitoring data. We used EL methodology and human biomonitoring health-based guidance values (HB2GVs) as exposure thresholds to create a new metric called Cumulative Health Risk from Exposure Load (CHREL). HB2GVs are derived by calculating the concentration of a biomarker consistent with a health protective exposure guidance value. CHREL analysis was conducted using Canadian Health Measures Survey (CHMS) cycle 3 and 4 biomonitoring data. Based on 18 chemicals, more than half of the Canadian population had an estimated CHRELTOTAL of 1 or more, indicative of chemical exposures potentially above selected exposure guidance values. Females had a significantly lower CHRELTOTAL compared to males, 12–19 year olds had a lower CHRELTOTAL compared to older age groups (significant compared to 40–59 year olds), and nonsmokers had a significantly lower CHRELTOTAL than smokers. Small segments of the population had a CHRELLIVER or a CHRELNERV of 1 or more, indicating exposures potentially above guideline levels for chemicals affecting the liver or nervous system. CHRELCANC was calculated based on 6 chemicals with HB2GVs derived for cancer endpoints. At the 10−5 risk level, most people had an estimated CHRELCANC of 3, indicative of multiple chemicals that may exceed negligible cancer risk. The most important contributors to exposures above HB2GVs were inorganic arsenic, mercury, acrylamide, xylenes, benzene and triclosan. Keeping certain assumptions, uncertainties and limitations in mind, the CHREL indicator can be used to obtain a picture of potential cumulative health risks from combined chemical exposures in a population, and as a comparative measure between subpopulations, including vulnerable subgroups.
{"title":"Cumulative Health Risk from Exposure Load (CHREL): Looking at multi-chemical exposures through the lens of biomonitoring guidance values","authors":"Jeff B. Willey , Chun Lei Liang , Tyler Pollock , Cheryl Khoury , Errol M. Thomson , Mike Walker , Annie St-Amand","doi":"10.1016/j.toxlet.2024.09.006","DOIUrl":"10.1016/j.toxlet.2024.09.006","url":null,"abstract":"<div><div>Exposure load (EL) is an indicator of multiple chemical exposures based on human biomonitoring data. We used EL methodology and human biomonitoring health-based guidance values (HB2GVs) as exposure thresholds to create a new metric called Cumulative Health Risk from Exposure Load (CHREL). HB2GVs are derived by calculating the concentration of a biomarker consistent with a health protective exposure guidance value. CHREL analysis was conducted using Canadian Health Measures Survey (CHMS) cycle 3 and 4 biomonitoring data. Based on 18 chemicals, more than half of the Canadian population had an estimated CHREL<sub>TOTAL</sub> of 1 or more, indicative of chemical exposures potentially above selected exposure guidance values. Females had a significantly lower CHREL<sub>TOTAL</sub> compared to males, 12–19 year olds had a lower CHREL<sub>TOTAL</sub> compared to older age groups (significant compared to 40–59 year olds), and nonsmokers had a significantly lower CHREL<sub>TOTAL</sub> than smokers. Small segments of the population had a CHREL<sub>LIVER</sub> or a CHREL<sub>NERV</sub> of 1 or more, indicating exposures potentially above guideline levels for chemicals affecting the liver or nervous system. CHREL<sub>CANC</sub> was calculated based on 6 chemicals with HB2GVs derived for cancer endpoints. At the 10<sup>−5</sup> risk level, most people had an estimated CHREL<sub>CANC</sub> of 3, indicative of multiple chemicals that may exceed negligible cancer risk. The most important contributors to exposures above HB2GVs were inorganic arsenic, mercury, acrylamide, xylenes, benzene and triclosan. Keeping certain assumptions, uncertainties and limitations in mind, the CHREL indicator can be used to obtain a picture of potential cumulative health risks from combined chemical exposures in a population, and as a comparative measure between subpopulations, including vulnerable subgroups.</div></div>","PeriodicalId":23206,"journal":{"name":"Toxicology letters","volume":"401 ","pages":"Pages 139-149"},"PeriodicalIF":2.9,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142354500","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-24DOI: 10.1016/j.toxlet.2024.09.008
Dat Thanh Dinh , Gilang Putra Bahari , Qi Xu , Cheng-Hao Wei , Dar-Ren Chen , Wei-Chung Hsieh , Po-Hsiung Lin
The primary goals of this study were to investigate the formation of abasic sites (AP sites) induced by methyl methanesulfonate (MMS) and hydrogen peroxide (H2O2), and to characterize specific types of these pro-mutagenic DNA lesions in calf thymus DNA (CT-DNA), and BEAS-2B human lung normal cell line. Furthermore, these profiles were compared with those observed in leukocytes derived from healthy controls (HC), breast cancer patients (BCP) before treatment, and 5-year survivors. Results indicated that both H2O2 and MMS induced the concentration- and time-dependent formation of AP sites in CT-DNA. To characterize the specific types of AP sites induced by H2O2 or MMS, we performed AP site cleavage assay using putrescine, T7 exonuclease (T7 Exo), and exonuclease III (Exo III). Results showed that the AP sites induced by H2O2 in CT-DNA were predominantly 5’-and 3’-nicked AP sites and no intact AP sites were detected. By contrast, the majority of AP sites generated by MMS in CT-DNA are not excisable and are classified as residual and intact AP sites. Similar approaches were performed in human BEAS-2B cells and comparable observations were confirmed in the cell-based model. Further investigation indicated that the profile of the AP sites observed in Taiwanese HC is identical to that of BEAS-2B cells treated with H2O2 whereas the pattern of AP sites detected in BCP is similar to that of CT-DNA exposed to H2O2, suggesting that these AP sites were produced primarily through reactive oxygen species (ROS) generation. More than 70 % of the AP sites in leukocytes derived from BCP were 5’-nicked and residual AP sites. Furthermore, the characteristics of the AP sites detected in 5-year survivors are comparable with the ones in HC by using putrescine cleavage assay. Overall, we speculate that deficiency in the DNA repair cascade may play a role in mediating the formation of specific types of AP sites detected in BCP.
这项研究的主要目的是调查甲磺酸甲酯（MMS）和过氧化氢（H2O2）诱导的消旋位点（AP 位点）的形成，并描述小牛胸腺 DNA（CT-DNA）和 BEAS-2B 人肺部正常细胞系中这些促突变 DNA 病变的特定类型。此外，还将这些图谱与从健康对照组（HC）、治疗前乳腺癌患者（BCP）和 5 年存活者的白细胞中观察到的图谱进行了比较。结果表明，H2O2 和 MMS 都能诱导 CT-DNA 中 AP 位点的形成，且其形成与浓度和时间有关。为了确定 H2O2 或 MMS 诱导的 AP 位点的具体类型，我们使用腐胺、T7 外切酶（T7 Exo）和外切酶 III（Exo III）进行了 AP 位点裂解试验。结果表明，H2O2 在 CT-DNA 中诱导的 AP 位点主要是 5'-3'-nicked AP 位点，没有检测到完整的 AP 位点。相比之下，MMS 在 CT-DNA 中产生的 AP 位点大多不可切除，被归类为残留 AP 位点和完整 AP 位点。在人类 BEAS-2B 细胞中也采用了类似的方法，并在基于细胞的模型中证实了类似的观察结果。进一步的研究表明，在台湾 HC 中观察到的 AP 位点与用 H2O2 处理的 BEAS-2B 细胞中的 AP 位点相同，而在 BCP 中检测到的 AP 位点模式与暴露于 H2O2 的 CT-DNA 相似，这表明这些 AP 位点主要是通过活性氧（ROS）的生成而产生的。来自 BCP 的白细胞中超过 70% 的 AP 位点是 5'-nicked 和残留 AP 位点。此外，通过使用腐胺裂解检测法，在 5 年存活者中检测到的 AP 位点的特征与 HC 中的 AP 位点相似。总之，我们推测 DNA 修复级联的缺陷可能在 BCP 中检测到的特定类型 AP 位点的形成中起了介导作用。
{"title":"Investigation of the abasic sites induced by hydrogen peroxide and methyl methanesulfonate in calf thymus DNA and BEAS-2B cells","authors":"Dat Thanh Dinh , Gilang Putra Bahari , Qi Xu , Cheng-Hao Wei , Dar-Ren Chen , Wei-Chung Hsieh , Po-Hsiung Lin","doi":"10.1016/j.toxlet.2024.09.008","DOIUrl":"10.1016/j.toxlet.2024.09.008","url":null,"abstract":"<div><div>The primary goals of this study were to investigate the formation of abasic sites (AP sites) induced by methyl methanesulfonate (MMS) and hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>), and to characterize specific types of these pro-mutagenic DNA lesions in calf thymus DNA (CT-DNA), and BEAS-2B human lung normal cell line. Furthermore, these profiles were compared with those observed in leukocytes derived from healthy controls (HC), breast cancer patients (BCP) before treatment, and 5-year survivors. Results indicated that both H<sub>2</sub>O<sub>2</sub> and MMS induced the concentration- and time-dependent formation of AP sites in CT-DNA. To characterize the specific types of AP sites induced by H<sub>2</sub>O<sub>2</sub> or MMS, we performed AP site cleavage assay using putrescine, T7 exonuclease (T7 Exo), and exonuclease III (Exo III). Results showed that the AP sites induced by H<sub>2</sub>O<sub>2</sub> in CT-DNA were predominantly 5’-and 3’-nicked AP sites and no intact AP sites were detected. By contrast, the majority of AP sites generated by MMS in CT-DNA are not excisable and are classified as residual and intact AP sites. Similar approaches were performed in human BEAS-2B cells and comparable observations were confirmed in the cell-based model. Further investigation indicated that the profile of the AP sites observed in Taiwanese HC is identical to that of BEAS-2B cells treated with H<sub>2</sub>O<sub>2</sub> whereas the pattern of AP sites detected in BCP is similar to that of CT-DNA exposed to H<sub>2</sub>O<sub>2</sub>, suggesting that these AP sites were produced primarily through reactive oxygen species (ROS) generation. More than 70 % of the AP sites in leukocytes derived from BCP were 5’-nicked and residual AP sites. Furthermore, the characteristics of the AP sites detected in 5-year survivors are comparable with the ones in HC by using putrescine cleavage assay. Overall, we speculate that deficiency in the DNA repair cascade may play a role in mediating the formation of specific types of AP sites detected in BCP.</div></div>","PeriodicalId":23206,"journal":{"name":"Toxicology letters","volume":"401 ","pages":"Pages 101-107"},"PeriodicalIF":2.9,"publicationDate":"2024-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142322814","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-19DOI: 10.1016/j.toxlet.2024.09.007
Mamoon Q. Salih , Lorenz Steiner , Walter Goessler , Jawameer R. Hama , Bassam Lajin
Hydrogen sulfide (H2S) is a toxic gas emitted through natural and anthropogenic activities. Chronic exposure to inhaled H2S at low sub-toxic levels is common among workers in oil refineries and may have important health implications. Inhaled H2S can be oxidized to thiosulfate or methylated to dimethylsulfide (DMS) which can be methylated to the novel human metabolite trimethylsulfonium (TMS) or oxidized to dimethylsulfoxide (DMSO) but the extent of methylation of inhaled H2S is currently unknown in humans. A total of 80 participants were recruited of which 40 were workers in an oil refinery in Kurdistan region, Iraq including those working in close contact with the facility area where H2S was measured at 1.5–5.0 mg m−3, and 40 controls living in a nearby city with no detectable H2S or perceptible odor (<0.1 mg m−3). A total of 240 urine samples were measured for multiple H2S-related metabolites. DMSO was consistently found in all urine samples with concentrations generally within the range of 1.0–10 µM. Although these concentrations were 10–100-fold higher than TMS urinary levels, clear correlation between DMSO and TMS was observed (rs 0.55, P < 0.0001), which supports DMS as common precursor. DMSO urinary levels were elevated in the oil refinery workers in close contact with the facilities (5.0 vs. 3.3 µM, P 0.03), but TMS was unaltered (0.13 vs. 0.14 µM, P 0.68). Overall, the results suggest that the investigated methylation metabolites are not sufficiently sensitive to low occupational exposure levels of inhaled H2S.
{"title":"Urinary excretion of H2S methylation metabolites in oil refinery workers","authors":"Mamoon Q. Salih , Lorenz Steiner , Walter Goessler , Jawameer R. Hama , Bassam Lajin","doi":"10.1016/j.toxlet.2024.09.007","DOIUrl":"10.1016/j.toxlet.2024.09.007","url":null,"abstract":"<div><div>Hydrogen sulfide (H<sub>2</sub>S) is a toxic gas emitted through natural and anthropogenic activities. Chronic exposure to inhaled H<sub>2</sub>S at low sub-toxic levels is common among workers in oil refineries and may have important health implications. Inhaled H<sub>2</sub>S can be oxidized to thiosulfate or methylated to dimethylsulfide (DMS) which can be methylated to the novel human metabolite trimethylsulfonium (TMS) or oxidized to dimethylsulfoxide (DMSO) but the extent of methylation of inhaled H<sub>2</sub>S is currently unknown in humans. A total of 80 participants were recruited of which 40 were workers in an oil refinery in Kurdistan region, Iraq including those working in close contact with the facility area where H<sub>2</sub>S was measured at 1.5–5.0 mg m<sup>−3</sup>, and 40 controls living in a nearby city with no detectable H<sub>2</sub>S or perceptible odor (<0.1 mg m<sup>−3</sup>). A total of 240 urine samples were measured for multiple H<sub>2</sub>S-related metabolites. DMSO was consistently found in all urine samples with concentrations generally within the range of 1.0–10 µM. Although these concentrations were 10–100-fold higher than TMS urinary levels, clear correlation between DMSO and TMS was observed (r<sub>s</sub> 0.55, <em>P</em> < 0.0001), which supports DMS as common precursor. DMSO urinary levels were elevated in the oil refinery workers in close contact with the facilities (5.0 vs. 3.3 µM, <em>P</em> 0.03), but TMS was unaltered (0.13 vs. 0.14 µM, <em>P</em> 0.68). Overall, the results suggest that the investigated methylation metabolites are not sufficiently sensitive to low occupational exposure levels of inhaled H<sub>2</sub>S.</div></div>","PeriodicalId":23206,"journal":{"name":"Toxicology letters","volume":"401 ","pages":"Pages 82-88"},"PeriodicalIF":2.9,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0378427424020320/pdfft?md5=2bfd8adfd903a1604913816864a52a13&pid=1-s2.0-S0378427424020320-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142296216","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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