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1
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Dragona E, Gagos S. Two-Replication Round, Telomere Strand-Specific, Chromosome Orientation Fluorescent In Situ Hybridization. Methods Mol Biol 2025; 2906:177-188. [PMID: 40082356 DOI: 10.1007/978-1-0716-4426-3_10] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 03/16/2025]
Abstract
Telomere strand-specific, two-replication-round, chromosome orientation fluorescence in situ hybridization (2R telomeric-CO-FISH) is a molecular cytogenetic protocol that allows simultaneous detection of genomic and telomeric sister chromatid exchanges along with telomeric intrachromosomal conservative break-induced replication. The combination of differential sister-chromatid and strand-specific telomeric labeling, provides information about intrachromosomal DNA recombinational events occurring in the last two consequent replication rounds of living cells grown in the presence of nucleic acid analogs. Despite a plethora of research applications, this methodology can be used to validate the extent of exposure to genotoxic agents and efficiency of recombinational DNA repair.
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Affiliation(s)
- Eleni Dragona
- Laboratory of Genetics, Center of Clinical Research, Experimental Surgery and Translational Research, Biomedical Research Foundation of the Academy of Athens, Greece (BRFAA), Athens, Greece
| | - Sarantis Gagos
- Laboratory of Genetics, Center of Clinical Research, Experimental Surgery and Translational Research, Biomedical Research Foundation of the Academy of Athens, Greece (BRFAA), Athens, Greece.
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2
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Hu J, Zhao F, Liu L, Huang H, Huang X. The meta-analysis of sister chromatid exchange as a biomarker in healthcare workers with occupational exposure to antineoplastic drugs. Medicine (Baltimore) 2023; 102:e34781. [PMID: 37653817 PMCID: PMC10470682 DOI: 10.1097/md.0000000000034781] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/02/2023] [Revised: 07/25/2023] [Accepted: 07/26/2023] [Indexed: 09/02/2023] Open
Abstract
BACKGROUND Sister chromatid exchange (SCE) can be used to identify early occupational health status in health care workers. Our aim is to comprehensively assess the relationship between long-term exposure to antineoplastic drugs (ADs) and SCE in health care workers via meta-analysis. METHODS Five databases were systematically searched for relevant articles published from inception to November 30, 2022. Literature data are expressed as mean difference and 95% confidence intervals (CI) or relative risk and 95% CI. For I2 > 50% trials, random effect model is used for statistical analysis, otherwise fixed effect model is used. This review was registered in the International Prospective Register of Systematic Reviews (identifier CRD42023399914). RESULTS Fourteen studies were included in this study. Results showed the level of SCE in healthcare workers exposed to ADs was significantly higher than in controls. The mean difference of the SCE trial was 0.53 (95% CI: 0.10-0.95, P = .01) under a random-effects model. CONCLUSIONS The findings suggested a significant correlation between occupational exposure to ADs in health care workers and SCE, requiring the attention of health care workers in general.
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Affiliation(s)
- Jinchen Hu
- Shenzhen Clinical Medical College, Guangzhou University of Chinese Medicine, Shenzhen, China
| | - Feifei Zhao
- Shenzhen Clinical Medical College, Guangzhou University of Chinese Medicine, Shenzhen, China
| | - Lin Liu
- Shenzhen Clinical Medical College, Guangzhou University of Chinese Medicine, Shenzhen, China
| | - Hong Huang
- Shenzhen Clinical Medical College, Guangzhou University of Chinese Medicine, Shenzhen, China
| | - Xiaohong Huang
- Department of Nursing, Longgang Central Hospital of Shenzhen, Shenzhen Guangdong & Clinical Medical College of Shenzhen, Guangzhou University of Chinese Medicine, Shenzhen, China
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3
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Kuchi Bhotla H, Balasubramanian B, Rengasamy KRR, Arumugam VA, Alagamuthu KK, Chithravel V, Chaudhary A, Alanazi AM, Pappuswamy M, Meyyazhagan A. Genotoxic repercussion of high-intensity radiation (x-rays) on hospital radiographers. ENVIRONMENTAL AND MOLECULAR MUTAGENESIS 2023; 64:123-131. [PMID: 36541415 DOI: 10.1002/em.22523] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/10/2022] [Revised: 11/29/2022] [Accepted: 12/12/2022] [Indexed: 05/24/2023]
Abstract
Recent technological advances in the medical field have increased the plausibility of exposing humans to high-intensity wavelength radiations like x-rays and gamma rays while diagnosing or treating specific medical maladies. These radiations induce nucleotide changes and chromosomal alterations in the exposed population, intentionally or accidentally. A radiological investigation is regularly used in identifying the disease, especially by the technicians working in intensive care units. The current study observes the genetic damages like chromosomal abnormalities (CA) in clinicians who are occupationally exposed to high-intensity radiations (x-rays) at their workplaces using universal cytogenetic tools like micronucleus assay (MN), sister chromatid exchange and comet assay. The study was conducted between 100 exposed practitioners from the abdominal scanning, chest scanning, cranial and orthopedic or bone scanning department and age-matched healthy controls. We observed a slightly higher rate of MN and CA (p < .05) in orthopedic and chest department practitioners than in other departments concerning increasing age and duration of exposure at work. Our results emphasize taking extra precautionary measures in clinical and hospital radiation laboratories to protect the practitioners.
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Affiliation(s)
| | | | - Kannan R R Rengasamy
- Centre of Excellence for Pharmaceutical Sciences, North-West University, Potchefstroom, South Africa
- Laboratory of Natural Products and Medicinal Chemistry (LNPMC), Department of Pharmacology, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences (SIMATS), Chennai, India
| | - Vijaya Anand Arumugam
- Medical Genetics and Epigenetics Laboratory, Department of Human Genetics and Molecular Biology, Bharathiar University, Coimbatore, Tamil Nadu, India
| | - Karthick Kumar Alagamuthu
- Department of Biotechnology, Selvamm Arts and Science College (Autonomous), Namakkal, Tamil Nadu, India
| | | | - Aditi Chaudhary
- Department of Life Sciences, CHRIST (Deemed to be University), Bangalore, India
| | - Amer M Alanazi
- Pharmaceutical Chemistry Department, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia
| | | | - Arun Meyyazhagan
- Department of Life Sciences, CHRIST (Deemed to be University), Bangalore, India
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4
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Sister chromatid exchanges induced by perturbed replication can form independently of BRCA1, BRCA2 and RAD51. Nat Commun 2022; 13:6722. [PMID: 36344511 PMCID: PMC9640580 DOI: 10.1038/s41467-022-34519-8] [Citation(s) in RCA: 22] [Impact Index Per Article: 7.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/17/2021] [Accepted: 10/27/2022] [Indexed: 11/09/2022] Open
Abstract
Sister chromatid exchanges (SCEs) are products of joint DNA molecule resolution, and are considered to form through homologous recombination (HR). Indeed, SCE induction upon irradiation requires the canonical HR factors BRCA1, BRCA2 and RAD51. In contrast, replication-blocking agents, including PARP inhibitors, induce SCEs independently of BRCA1, BRCA2 and RAD51. PARP inhibitor-induced SCEs are enriched at difficult-to-replicate genomic regions, including common fragile sites (CFSs). PARP inhibitor-induced replication lesions are transmitted into mitosis, suggesting that SCEs can originate from mitotic processing of under-replicated DNA. Proteomics analysis reveals mitotic recruitment of DNA polymerase theta (POLQ) to synthetic DNA ends. POLQ inactivation results in reduced SCE numbers and severe chromosome fragmentation upon PARP inhibition in HR-deficient cells. Accordingly, analysis of CFSs in cancer genomes reveals frequent allelic deletions, flanked by signatures of POLQ-mediated repair. Combined, we show PARP inhibition generates under-replicated DNA, which is processed into SCEs during mitosis, independently of canonical HR factors.
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5
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Akbas E, Unal F, Yuzbasioglu D. Genotoxic effects of gadobutrol and gadoversetamide active substances used in magnetic resonance imaging in human peripheral lymphocytes in vitro. Drug Chem Toxicol 2022; 45:2471-2482. [PMID: 35184618 DOI: 10.1080/01480545.2021.1957913] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/03/2022]
Abstract
Gadobutrol and gadoversetamide are gadolinium-based contrast agents (GBCAs) widely used during magnetic resonance imaging examination. In this study, the genotoxicity of two GBCAs, gadobutrol and gadoversetamide, was investigated by using different endpoints: chromosome aberration (CAs), sister chromatid exchange (SCEs), and micronucleus (MNi). Human peripheral lymphocytes (PBLs) were treated with five concentrations (7 000, 14 000, 28 000, 56 000, and 112 000 μg/mL) of both agents. While a few concentrations of gadobutrol significantly increased abnormal cell frequency and CA/Cell, nearly all the concentrations of gadoversetamide significantly elevated the same aberrations. Similarly, the effect of gadoversetamide on the formation of SCEs was higher than those of gadobutrol. Only one concentration of gadoversetamide significantly increased MN% but no gadobutrol. The comet assay was applied for the only gadobutrol which induced a significant increase in tail intensity at the highest concentration only. On the other hand, significantly decreased mitotic index (MI) was observed following both substances, again gadoversetamide was slightly higher than those of the gadobutrol. The results revealed that both the contrast agents are likely to induce genotoxic risk in PBLs. However, different concentrations and treatment periods should be examined in vitro and specifically in vivo with different test systems for the safer usage of these contrast agents.
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Affiliation(s)
- Ece Akbas
- Genetic Toxicology Laboratory, Department of Biology, Science Faculty, Gazi University, 06560, Ankara, Turkey
| | - Fatma Unal
- Genetic Toxicology Laboratory, Department of Biology, Science Faculty, Gazi University, 06560, Ankara, Turkey
| | - Deniz Yuzbasioglu
- Genetic Toxicology Laboratory, Department of Biology, Science Faculty, Gazi University, 06560, Ankara, Turkey
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6
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Biomarkers of Genotoxicity in Medical Workers Exposed to Low-Dose Ionizing Radiation: Systematic Review and Meta-Analyses. Int J Mol Sci 2021; 22:ijms22147504. [PMID: 34299125 PMCID: PMC8304237 DOI: 10.3390/ijms22147504] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/07/2021] [Revised: 06/29/2021] [Accepted: 07/08/2021] [Indexed: 12/12/2022] Open
Abstract
Medical staff represent the largest group of workers occupationally exposed to ionizing radiation (IR). Chronic exposure to low-dose IR may result in DNA damage and genotoxicity associated with increased risk of cancer. This review aims to identify the genotoxicity biomarkers that are the most elevated in IR-exposed vs. unexposed health workers. A systematic review of the literature was performed to retrieve relevant studies with various biomarkers of genotoxicity. Subsequent meta-analyses produced a pooled effect size for several endpoints. The search procedure yielded 65 studies. Chromosome aberrations (CA) and micronuclei (MN) frequencies were significantly different between IR-exposed and unexposed workers (θpooled = 3.19, 95% CI 1.46–4.93; and θpooled = 1.41, 95% CI 0.97–1.86, for total aberrant cells and MN frequencies, respectively), which was not the case for ring chromosomes and nucleoplasmic bridges. Although less frequently used, stable translocations, sister chromatid exchanges (SCE) and comet assay endpoints were also statistically different between IR-exposed and unexposed workers. This review confirms the relevance of CA and MN as genotoxicity biomarkers that are consistently elevated in IR-exposed vs. unexposed workers. Other endpoints are strong candidates but require further studies to validate their usefulness. The integration of the identified biomarkers in future prospective epidemiological studies is encouraged.
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7
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Abstract
Homologous recombination (HR) repairs DNA double-strand breaks by using a homologous template to retrieve sequence information lost at the break site. The broken DNA molecule first engages with the homologous donor molecule and is then separated from it to complete the process. Depending on the HR subpathways used, the separation step can lead to crossovers (COs) between the participating molecules. Such events can cause genomic alterations and eventually cancer if a donor molecule other than the identical sister chromatid is used. Here, we characterize two subpathways of HR with different propensities to form COs. We show the unexpected dominance of the CO-forming subpathway and characterize the processes involved in CO formation and subpathway choice in cancer and normal, untransformed cells. Homologous recombination (HR) is an important DNA double-strand break (DSB) repair pathway that copies sequence information lost at the break site from an undamaged homologous template. This involves the formation of a recombination structure that is processed to restore the original sequence but also harbors the potential for crossover (CO) formation between the participating molecules. Synthesis-dependent strand annealing (SDSA) is an HR subpathway that prevents CO formation and is thought to predominate in mammalian cells. The chromatin remodeler ATRX promotes an alternative HR subpathway that has the potential to form COs. Here, we show that ATRX-dependent HR outcompetes RECQ5-dependent SDSA for the repair of most two-ended DSBs in human cells and leads to the frequent formation of COs, assessed by measuring sister chromatid exchanges (SCEs). We provide evidence that subpathway choice is dependent on interaction of both ATRX and RECQ5 with proliferating cell nuclear antigen. We also show that the subpathway usage varies among different cancer cell lines and compare it to untransformed cells. We further observe HR intermediates arising as ionizing radiation (IR)-induced ultra-fine bridges only in cells expressing ATRX and lacking MUS81 and GEN1. Consistently, damage-induced MUS81 recruitment is only observed in ATRX-expressing cells. Cells lacking BLM show similar MUS81 recruitment and IR-induced SCE formation as control cells. Collectively, these results suggest that the ATRX pathway involves the formation of HR intermediates whose processing is entirely dependent on MUS81 and GEN1 and independent of BLM. We propose that the predominant ATRX-dependent HR subpathway forms joint molecules distinct from classical Holliday junctions.
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8
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Abstract
The semiconservative nature of DNA replication allows the differential labeling of sister chromatids that is the fundamental requirement to perform the sister-chromatid exchange (SCE) assay. SCE assay is a powerful technique to visually detect the physical exchange of DNA between sister chromatids. SCEs could result as a consequence of DNA damage repair by homologous recombination (HR) during DNA replication. Here, we provide the detailed protocol to perform the SCE assay in cultured human cells. Cells are exposed to the thymidine analog 5-bromo-2'-deoxyuridine (BrdU) during two cell cycles, resulting in the two sister chromatids having differential incorporation of the analog. After metaphase spreads preparation and further processing, SCEs are nicely visualized under the microscope.
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Affiliation(s)
- Emanuela Tumini
- Centro Andaluz de Biología Molecular y Medicina Regenerativa, CABIMER, Universidad de Sevilla-CSIC-UPO, Seville, Spain
| | - Andrés Aguilera
- Centro Andaluz de Biología Molecular y Medicina Regenerativa, CABIMER, Universidad de Sevilla-CSIC-UPO, Seville, Spain.
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9
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Boysen G, Arora R, Degner A, Vevang KR, Chao C, Rodriguez F, Walmsley SJ, Erber L, Tretyakova NY, Peterson LA. Effects of GSTT1 Genotype on the Detoxification of 1,3-Butadiene Derived Diepoxide and Formation of Promutagenic DNA-DNA Cross-Links in Human Hapmap Cell Lines. Chem Res Toxicol 2020; 34:119-131. [PMID: 33381973 DOI: 10.1021/acs.chemrestox.0c00376] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022]
Abstract
Smoking is a leading cause of lung cancer, accounting for 81% of lung cancer cases. Tobacco smoke contains over 5000 compounds, of which more than 70 have been classified as human carcinogens. Of the many tobacco smoke constituents, 1,3-butadiene (BD) has a high cancer risk index due to its tumorigenic potency and its abundance in cigarette smoke. The carcinogenicity of BD has been attributed to the formation of several epoxide metabolites, of which 1,2,3,4-diepoxybutane (DEB) is the most toxic and mutagenic. DEB is formed by two oxidation reactions carried out by cytochrome P450 monooxygenases, mainly CYP2E1. Glutathione-S-transferase theta 1 (GSTT1) facilitates the conjugation of DEB to glutathione as the first step of its detoxification and subsequent elimination via the mercapturic acid pathway. Human biomonitoring studies have revealed a strong association between GSTT1 copy number and urinary concentrations of BD-mercapturic acids, suggesting that it plays an important role in the metabolism of BD. To determine the extent that GSTT1 genotype affects the susceptibility of individuals to the toxic and genotoxic properties of DEB, GSTT1 negative and GSTT1 positive HapMap lymphoblastoid cell lines were treated with DEB, and the extent of apoptosis and micronuclei (MN) formation was assessed. These toxicological end points were compared to the formation of DEB-GSH conjugates and 1,4-bis-(guan-7-yl)-2,3-butanediol (bis-N7G-BD) DNA-DNA cross-links. GSTT1 negative cell lines were more sensitive to DEB-induced apoptosis as compared to GSTT1 positive cell lines. Consistent with the protective effect of GSH conjugation against DEB-derived apoptosis, GSTT1 positive cell lines formed significantly more DEB-GSH conjugate than GSTT1 negative cell lines. However, GSTT1 genotype did not affect formation of MN or bis-N7G-BD cross-links. These results indicate that GSTT1 genotype significantly influences BD metabolism and acute toxicity.
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Affiliation(s)
- Gunnar Boysen
- Department of Medicinal Chemistry, University of Minnesota Minneapolis, Minnesota 55455, United States.,Department of Environmental and Occupational Health, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205, United States.,The Winthrop P Rockefeller Cancer Institute, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205, United States
| | - Rashi Arora
- University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota 55455, United States
| | - Amanda Degner
- Department of Medicinal Chemistry, University of Minnesota Minneapolis, Minnesota 55455, United States.,University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota 55455, United States
| | - Karin R Vevang
- Division of Environmental Health Sciences, University of Minnesota, Minneapolis, Minnesota 55455, United States
| | - Christopher Chao
- Department of Medicinal Chemistry, University of Minnesota Minneapolis, Minnesota 55455, United States
| | - Freddys Rodriguez
- Department of Medicinal Chemistry, University of Minnesota Minneapolis, Minnesota 55455, United States
| | - Scott J Walmsley
- University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota 55455, United States.,Institute for Health Informatics, University of Minnesota, Minneapolis, Minnesota 55455, United States
| | - Luke Erber
- Department of Medicinal Chemistry, University of Minnesota Minneapolis, Minnesota 55455, United States
| | - Natalia Y Tretyakova
- Department of Medicinal Chemistry, University of Minnesota Minneapolis, Minnesota 55455, United States.,University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota 55455, United States
| | - Lisa A Peterson
- University of Minnesota Masonic Cancer Center, Minneapolis, Minnesota 55455, United States.,Division of Environmental Health Sciences, University of Minnesota, Minneapolis, Minnesota 55455, United States
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10
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Dördü TC, Hatipoğlu R, Topaktaş M, İstifli ES. In Vitro Genotoxicity and Molecular Docking Study of Ellagic Acid. ACTA ACUST UNITED AC 2020. [DOI: 10.2174/1573407215666191102130417] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Abstract
Background:
Ellagic Acid (EA) is a polyphenolic compound that is classified in the natural
antioxidants group. Polyphenolic compounds that exert antioxidant activity possess particular importance
for scientists, food producers and consumers due to their positive effects on human health. However,
despite considerable evidence that EA shows antigenotoxic activity by binding to DNA, there is no
systematic genotoxicity study of this substance, which can covalently bind to DNA. This study aims to
reveal the possible genotoxic activity of EA using widely accepted assays for the assessment of DNA
clastogenic activity: sister chromatid exchange, chromosome aberration, micronucleus and comet assays
as well as to predict the interactions among EA and DNA through molecular docking.
Methods:
Different assays were carried out to identify the clastogenic activity of EA on human lymphocyte
DNA using Sister Chromatid Exchange (SCE), Chromosome Aberration (CA), Micronucleus (MN)
and single-cell gel electrophoresis (SCGE/comet) assays. For this aim, human peripheral blood lymphocytes
were treated with EA (60, 80 and 100 μg/ml) for 24 and 48 hrs in the SCE, CA and MN assays
and for 1 hr in the comet assay. Furthermore, molecular docking experiments were also performed to
calculate the binding energy of EA on human B-DNA structure (B-DNA dodecamer) as well as to predict
noncovalent interactions among these macromolecules.
Results:
At the concentrations and treatment times (24- or 48-hr) tested, EA did not induce either SCE or
Chromosome Aberrations (CAs) as compared to the negative and solvent controls. Although EA slightly
increased the percentage of Micronucleated Binuclear (%MNBN) cells as well as the percentage of Micronucleus
(%MN) in 24 or 48-hr treatment periods at all concentrations, this increase was not statistically
significant as compared to both controls. The effect of EA on DNA replication (nuclear division) was determined
by the Proliferation Index (PI), the Nuclear Division Index (NDI) and the Mitotic Index (MI). No
statistically significant differences were observed in the PI or NDI in 24- or 48-hr treatment periods in
human lymphocyte cultures treated with EA at various concentrations. EA generally had no significant
effect on the MI, as observed with the PI and NDI.
Discussion:
Although the concentrations of 60 and 80 μg/mL at a 24-hr treatment period and the concentrations
of 60 μg/mL and 100 μg/mL at 48-hr treatment period generally decreased the MI, those decreases
were not statistically significant when compared to negative and solvent controls. Moreover, none of the
concentrations of EA tested in this study were able to increase DNA damage determined by the tail DNA
length, %DNA in tail and tail moment parameters in the comet assay. Although the amount of DNA damage
in the comet assay decreased with increasing concentrations of EA, this decrease was not statistically
significant as compared to both controls. However, molecular docking experiments interestingly showed
that the binding free energy of EA with B-DNA was -7.84 kcal/mol-1, indicating a strong interaction between
the two molecules.
Conclusion :
Although the findings of our study show that EA does not have genotoxic potential in human
chromosomes, molecular docking experiments revealed strong hydrogen bonding between EA and
B-DNA molecules. Therefore, it has been proposed that the prevailing information suggesting that the
molecules that bind to DNA cause genotoxic effects should be reconsidered from a wider perspective.
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Affiliation(s)
- Tuba C. Dördü
- Department of Biotechnology, Institute of Basic and Applied Sciences, Cukurova University, Adana, Turkey
| | - Rüştü Hatipoğlu
- Department of Field Crops, Faculty of Agriculture, Cukurova University, Adana, Turkey
| | - Mehmet Topaktaş
- Department of Biology, Faculty of Science and Letters, Cukurova University, Adana, Turkey
| | - Erman S. İstifli
- Department of Biology, Faculty of Science and Letters, Cukurova University, Adana, Turkey
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11
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Aitken SJ, Anderson CJ, Connor F, Pich O, Sundaram V, Feig C, Rayner TF, Lukk M, Aitken S, Luft J, Kentepozidou E, Arnedo-Pac C, Beentjes SV, Davies SE, Drews RM, Ewing A, Kaiser VB, Khamseh A, López-Arribillaga E, Redmond AM, Santoyo-Lopez J, Sentís I, Talmane L, Yates AD, Semple CA, López-Bigas N, Flicek P, Odom DT, Taylor MS. Pervasive lesion segregation shapes cancer genome evolution. Nature 2020; 583:265-270. [PMID: 32581361 PMCID: PMC7116693 DOI: 10.1038/s41586-020-2435-1] [Citation(s) in RCA: 44] [Impact Index Per Article: 8.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/06/2019] [Accepted: 05/07/2020] [Indexed: 02/08/2023]
Abstract
Cancers arise through the acquisition of oncogenic mutations and grow by clonal expansion1,2. Here we reveal that most mutagenic DNA lesions are not resolved into a mutated DNA base pair within a single cell cycle. Instead, DNA lesions segregate, unrepaired, into daughter cells for multiple cell generations, resulting in the chromosome-scale phasing of subsequent mutations. We characterize this process in mutagen-induced mouse liver tumours and show that DNA replication across persisting lesions can produce multiple alternative alleles in successive cell divisions, thereby generating both multiallelic and combinatorial genetic diversity. The phasing of lesions enables accurate measurement of strand-biased repair processes, quantification of oncogenic selection and fine mapping of sister-chromatid-exchange events. Finally, we demonstrate that lesion segregation is a unifying property of exogenous mutagens, including UV light and chemotherapy agents in human cells and tumours, which has profound implications for the evolution and adaptation of cancer genomes.
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Affiliation(s)
- Sarah J Aitken
- Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge, UK
- Department of Pathology, University of Cambridge, Cambridge, UK
- Department of Histopathology, Cambridge University Hospitals NHS Foundation Trust, Cambridge, UK
| | - Craig J Anderson
- MRC Human Genetics Unit, MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK
| | - Frances Connor
- Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge, UK
| | - Oriol Pich
- Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, Barcelona, Spain
| | - Vasavi Sundaram
- Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge, UK
- European Molecular Biology Laboratory, European Bioinformatics Institute, Hinxton, UK
| | - Christine Feig
- Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge, UK
| | - Tim F Rayner
- Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge, UK
| | - Margus Lukk
- Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge, UK
| | - Stuart Aitken
- MRC Human Genetics Unit, MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK
| | - Juliet Luft
- MRC Human Genetics Unit, MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK
| | | | - Claudia Arnedo-Pac
- Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, Barcelona, Spain
| | - Sjoerd V Beentjes
- School of Mathematics and Maxwell Institute, University of Edinburgh, Edinburgh, UK
| | - Susan E Davies
- Department of Histopathology, Cambridge University Hospitals NHS Foundation Trust, Cambridge, UK
| | - Ruben M Drews
- Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge, UK
| | - Ailith Ewing
- MRC Human Genetics Unit, MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK
| | - Vera B Kaiser
- MRC Human Genetics Unit, MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK
| | - Ava Khamseh
- MRC Human Genetics Unit, MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK
- Higgs Centre for Theoretical Physics, University of Edinburgh, Edinburgh, UK
| | - Erika López-Arribillaga
- Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, Barcelona, Spain
| | - Aisling M Redmond
- Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge, UK
| | | | - Inés Sentís
- Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, Barcelona, Spain
| | - Lana Talmane
- MRC Human Genetics Unit, MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK
| | - Andrew D Yates
- European Molecular Biology Laboratory, European Bioinformatics Institute, Hinxton, UK
| | - Colin A Semple
- MRC Human Genetics Unit, MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK
| | - Núria López-Bigas
- Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, Barcelona, Spain
- Universitat Pompeu Fabra (UPF), Barcelona, Spain
- Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain
| | - Paul Flicek
- Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge, UK
- European Molecular Biology Laboratory, European Bioinformatics Institute, Hinxton, UK
| | - Duncan T Odom
- Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge, UK.
- German Cancer Research Center (DKFZ), Division of Regulatory Genomics and Cancer Evolution, Heidelberg, Germany.
| | - Martin S Taylor
- MRC Human Genetics Unit, MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, UK.
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12
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Cytotoxicity and Mutagenicity of Narrowband UVB to Mammalian Cells. Genes (Basel) 2020; 11:genes11060646. [PMID: 32545288 PMCID: PMC7349664 DOI: 10.3390/genes11060646] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/05/2020] [Revised: 06/04/2020] [Accepted: 06/08/2020] [Indexed: 12/15/2022] Open
Abstract
Phototherapy using narrowband ultraviolet-B (NB-UVB) has been shown to be more effective than conventional broadband UVB (BB-UVB) in treating a variety of skin diseases. To assess the difference in carcinogenic potential between NB-UVB and BB-UVB, we investigated the cytotoxicity via colony formation assay, genotoxicity via sister chromatid exchange (SCE) assay, mutagenicity via hypoxanthine phosphoribosyltransferase (HPRT) mutation assay, as well as cyclobutane pyrimidine dimer (CPD) formation and reactive oxygen species (ROS) generation in Chinese hamster ovary (CHO) and their NER mutant cells. The radiation dose required to reduce survival to 10% (D10 value) demonstrated BB-UVB was 10 times more cytotoxic than NB-UVB, and revealed that NB-UVB also induces DNA damage repaired by nucleotide excision repair. We also found that BB-UVB more efficiently induced SCEs and HPRT mutations per absorbed energy dosage (J/m2) than NB-UVB. However, SCE and HPRT mutation frequencies were observed to rise in noncytotoxic dosages of NB-UVB exposure. BB-UVB and NB-UVB both produced a significant increase in CPD formation and ROS formation (p < 0.05); however, higher dosages were required for NB-UVB. These results suggest that NB-UVB is less cytotoxic and genotoxic than BB-UVB, but can still produce genotoxic effects even at noncytotoxic doses.
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Stults DM, Killen MW, Marco-Casanova P, Pierce AJ. The Sister Chromatid Exchange (SCE) Assay. Methods Mol Biol 2020; 2102:441-457. [PMID: 31989571 DOI: 10.1007/978-1-0716-0223-2_25] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/03/2022]
Abstract
A fully optimized staining method for detecting sister chromatid exchanges in cultured cells is presented. The method gives reproducibly robust quantitative results. Sister chromatid exchange is a classic toxicology assay for genotoxicity and for detecting alterations to the biochemistry underlying cellular homologous recombination. Growth of cells in the presence of 5'-bromo-deoxyuridine for two rounds of DNA replication followed by collecting metaphase spreads on glass slides, treatment with the UV-sensitive dye Hoechst 33258, long-wave UV light exposure, and Giemsa staining gives a permanent record of the exchanges.
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Affiliation(s)
- Dawn M Stults
- Graduate Center for Toxicology, University of Kentucky, Lexington, KY, USA.,Clovis Oncology, Boulder, CO, USA
| | - Michael W Killen
- Department of Microbiology, Immunology and Molecular Genetics, University of Kentucky, Lexington, KY, USA.,Western Kentucky University, Bowling Green, KY, USA
| | | | - Andrew J Pierce
- Translational Medicine, Oncology R&D, AstraZeneca, Cambridge, United Kingdom. .,Department of Microbiology, Immunology and Molecular Genetics, Markey Cancer Center, University of Kentucky, Lexington, KY, USA.
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Nartop D, Demirel B, Güleç M, Hasanoğlu Özkan E, Kurnaz Yetim N, Sarı N, Çeker S, Öğütcü H, Ağar G. Novel polymeric microspheres: Synthesis, enzyme immobilization, antimutagenic activity, and antimicrobial evaluation against pathogenic microorganisms. J Biochem Mol Toxicol 2019; 34:e22432. [PMID: 31851403 DOI: 10.1002/jbt.22432] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/28/2019] [Revised: 09/26/2019] [Accepted: 12/02/2019] [Indexed: 11/07/2022]
Abstract
New polymeric microspheres containing azomethine (1a-1c and 2a-2c) were synthesized by condensation to compare the enzymatic properties of the enzyme glucose oxidase (GOx) and to investigate antimutagenic and antimicrobial activities. The polymeric microspheres were characterized by elemental analysis, infrared spectra (FT-IR), proton nuclear magnetic resonance spectra, thermal gravimetric analysis, and scanning electron microscopy analysis. The catalytic activity of the glucose oxidase enzyme follows Michaelis-Menten kinetics. Influence of temperature, reusability, and storage capacity of the free and immobilized glucose oxidase enzyme were investigated. It is determined that immobilized enzymes exhibit good storage stability and reusability. After immobilization of GOx in polymeric supports, the thermal stability of the enzyme increased and the maximum reaction rate (Vmax ) decreased. The activity of the immobilized enzymes was preserved even after 5 months. The antibacterial and antifungal activity of the polymeric microspheres were evaluated by well-diffusion method against some selected pathogenic microorganisms. The antimutagenic properties of all compounds were also examined against sodium azide in human lymphocyte cells by micronuclei and sister chromatid exchange tests.
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Affiliation(s)
- Dilek Nartop
- Department of Polymer Engineering, Faculty of Techonology, Düzce University, Düzce, Turkey
| | - Birtane Demirel
- Department of Chemistry, Faculty of Arts and Science, Nevşehir Hacı Bektaş Veli University, Nevşehir, Turkey
| | - Murat Güleç
- Department of Chemistry, Faculty of Arts and Science, Nevşehir Hacı Bektaş Veli University, Nevşehir, Turkey
| | | | - Nurdan Kurnaz Yetim
- Department of Chemistry, Faculty of Arts and Science, Kırklareli University, Kırklareli, Turkey
| | - Nurşen Sarı
- Department of Chemistry, Faculty of Science, Gazi University, Ankara, Turkey
| | - Selçuk Çeker
- Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Ağrı İbrahim Çeçen University, Ağrı, Turkey
| | - Hatice Öğütcü
- Department of Field Crops, Faculty of Agriculture, Ahi Evran University, Kırşehir, Turkey
| | - Güleray Ağar
- Department of Biology, Faculty of Science, Atatürk University, Erzurum, Turkey
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Nartop D, Özkan EH, Gündem M, Çeker S, Ağar G, Öğütcü H, Sarı N. Synthesis, antimicrobial and antimutagenic effects of novel polymeric-Schiff bases including indol. J Mol Struct 2019. [DOI: 10.1016/j.molstruc.2019.06.042] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/10/2023]
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Cruz-Vallejo V, Ortíz-Muñiz R, Vallarino-Kelly T, Cervantes-Ríos E, Morales-Ramírez P. In vivo Characterization of the Radiosensitizing Effect of a Very Low Dose of BrdU in Murine Cells Exposed to Low-Dose Radiation. ENVIRONMENTAL AND MOLECULAR MUTAGENESIS 2019; 60:534-545. [PMID: 30851126 DOI: 10.1002/em.22284] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/02/2019] [Revised: 02/24/2019] [Accepted: 03/02/2019] [Indexed: 06/09/2023]
Abstract
The aim of the present study was to characterize the in vivo radiosensitizing effect of a very low dose of bromodeoxyuridine (BrdU) in mice exposed to low-dose radiation by establishing the following: (1) the radiosensitizing effect during DNA synthesis using single-cell gel electrophoresis (SCGE) in murine bone marrow cells, and (2) the number and timing of the mechanisms of genotoxicity and cytotoxicity, as well as the correlation of both end points, using flow cytometry analysis of the kinetics of micronucleus induction in reticulocytes. Groups of mice received intraperitoneal injections of 0.125 mg/g of BrdU 24 h prior to irradiation with 0.5 Gy of 60 Co gamma rays. DNA breaks measured using SCGE were determined at 30 min after exposure to radiation. The kinetics of micronucleated reticulocyte (MN-RET) induction was determined every 8 h after irradiation up to 72 h. The results from both experimental models indicated that low-level BrdU incorporation into DNA increased the sensitivity to 0.5 Gy of radiation, particularly in the S phase. The formation of micronuclei by gamma rays was produced at three different times using two main mechanisms. In the BrdU-substituted cells, the second mechanism was associated with a high cytotoxic effect that was absent in the irradiated BrdU-unsubstituted cells. The third mechanism, in which micronucleus formation was increased in irradiated substituted cells compared with the irradiated nonsubstituted control cells, was also related to an increase in cytotoxicity. Environ. Mol. Mutagen. 60:534-545, 2019. © 2019 Wiley Periodicals, Inc.
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Affiliation(s)
- Virginia Cruz-Vallejo
- Instituto Nacional de Investigaciones Nucleares, Carretera México-Toluca s/n, La Marquesa, Ocoyoacac, Estado de México C. P., 52750, Mexico
- Doctorado en Biología Experimental, Universidad Autónoma Metropolitana, Avenida San Rafael Atlixco 186 CP, 09340, Ciudad de México, Mexico
| | - Rocío Ortíz-Muñiz
- Departamento de Ciencias de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Avenida San Rafael Atlixco 186 CP, 09340, Ciudad de México, Mexico
| | - Teresita Vallarino-Kelly
- Instituto Nacional de Investigaciones Nucleares, Carretera México-Toluca s/n, La Marquesa, Ocoyoacac, Estado de México C. P., 52750, Mexico
| | - Elsa Cervantes-Ríos
- Departamento de Ciencias de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Avenida San Rafael Atlixco 186 CP, 09340, Ciudad de México, Mexico
| | - Pedro Morales-Ramírez
- Instituto Nacional de Investigaciones Nucleares, Carretera México-Toluca s/n, La Marquesa, Ocoyoacac, Estado de México C. P., 52750, Mexico
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Yun JW, Kim SH, Kim YS, Choi EJ, You JR, Cho EY, Yoon JH, Kwon E, Kim HC, Jang JJ, Park JS, Che JH, Kang BC. Preclinical study of safety of Dendropanax morbifera Leveille leaf extract: General and genetic toxicology. JOURNAL OF ETHNOPHARMACOLOGY 2019; 238:111874. [PMID: 30986520 DOI: 10.1016/j.jep.2019.111874] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 09/05/2018] [Accepted: 04/09/2019] [Indexed: 06/09/2023]
Abstract
ETHNOPHARMACOLOGY RELEVANCE Dendropanax morbifera Leveille (DM) has been used in traditional medicines for infectious and skin diseases, and dysmenorrhea. It exhibits a diverse therapeutic potential including anti-cancer, anti-thrombotic, anti-diabetic, anti-oxidant, and anti-inflammatory activities. AIM OF THE STUDY Despite promising health benefits of DM, knowledge of its potential adverse effects is very limited. The current study focused on the investigation of subchronic toxicity and genotoxicity of extract obtained from DM according to the test guidelines published by the Organization for Economic Cooperation and Development. MATERIALS AND METHODS We conducted a toxicological evaluation of DM extracts using 14-day repeated-dose toxicity study and 13-week repeated-dose toxicity study in Sprague-Dawley rats administered orally at doses of 500, 1000, or 2000 mg/kg/day. The clastogenicity of DM extract was also evaluated by in vitro chromosome aberration assay and in vivo micronucleus assay. RESULTS Assessment of subchronic toxicity of DM extract by oral administration in rats revealed unremarkable treatment-related findings with respect to food/water consumption, body weight, mortality, urinalysis, hematology, serum biochemistry, necropsy, organ weight and histopathology at doses of 500, 1000, and 2000 mg/kg. Accordingly, the level of no-observed-adverse-effect for DM extract in 13-week subchronic toxicity study was considered to be 2000 mg/kg/day in rats. The data observed from in vitro chromosome aberration assay and in vivo micronucleus assay exclude any clastogenicity of DM extract. CONCLUSION The results suggest that the oral consumption of DM extract has no adverse effects in humans and represents a safe traditional medicine.
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Affiliation(s)
- Jun-Won Yun
- Department of Biotechnology, The Catholic University of Korea, Bucheon, Republic of Korea
| | - Seung-Hyun Kim
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Yun-Soon Kim
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Eun Jin Choi
- Jeonnam Bioindustry Foundation, Jeonnam Institute of Natural Resources Research (JINR), Jangheung-gun, Jeollanam-do, Republic of Korea
| | - Ji-Ran You
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Eun-Young Cho
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Jung-Hee Yoon
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Euna Kwon
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Hyoung-Chin Kim
- Laboratory Animal Resource Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju, Republic of Korea
| | - Ja-June Jang
- Department of Pathology, Seoul National University College of Medicine, Seoul, Republic of Korea
| | - Jin-Sung Park
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Jeong-Hwan Che
- Biomedical Center for Animal Resource and Development, Seoul National University College of Medicine, Seoul, Republic of Korea.
| | - Byeong-Cheol Kang
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea; Biomedical Center for Animal Resource and Development, Seoul National University College of Medicine, Seoul, Republic of Korea; Graduate School of Translational Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea; Designed Animal and Transplantation Research Institute, Institute of GreenBio Science Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, Republic of Korea.
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Investigation of Safety Profile of Four Copaifera Species and of Kaurenoic Acid by Salmonella/Microsome Test. EVIDENCE-BASED COMPLEMENTARY AND ALTERNATIVE MEDICINE 2019; 2019:7631531. [PMID: 30733813 PMCID: PMC6348810 DOI: 10.1155/2019/7631531] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 07/18/2018] [Revised: 12/12/2018] [Accepted: 12/31/2018] [Indexed: 12/19/2022]
Abstract
Trees of the Copaifera genus are native to the tropical regions of Latin America and Western Africa. Copaifera sp is widely used as a popular medicine and it has various ethnopharmacological indications, including gonorrhea, bronchitis, asthma, skin ulcers, ulcers, sore throat, uterine infections, general inflammations, cancer, and leishmanioses. Kaurenoic acid is a naturally occurring diterpene found in Copaifera and has been used as an anti-inflammatory, treatment of ulcer, leishmaniasis, and cancer. Bearing in mind the fact that the Ames test is an excellent tool to assess the safety of extracts, oils, and phytochemicals isolated from medicinal plants, from it, we evaluate the mutagenic potential of four species, between oleoresins (C. oblongifolia; C. langsdorffii) and leaves extracts (C. lucens; C. multijuga), of the Copaifera genus and also of kaurenoic acid, which is one of its major compounds. The results showed that the Copaifera spp. and kaurenoic acid did not induce an increase in the number of revertant colonies, without mutagenic effect in experiments, in the all concentrations evaluated by Ames test. The results obtained in our study support the safe use of the Copaifera genus medicinal plants selected and of kaurenoic acid.
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Morales-Ramírez P, Vallarino-Kelly T, Cruz-Vallejo VL. The OECD's micronucleus test guideline for single exposure to an agent and the genotox-kinetic alternative. Mutagenesis 2018; 32:411-415. [PMID: 28472308 DOI: 10.1093/mutage/gex010] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/08/2016] [Accepted: 04/05/2017] [Indexed: 11/13/2022] Open
Abstract
The 'Organization for Economic Co-operation and Development (OECD) guidelines for the Testing of Chemicals' aims to identify whether a chemical is a genotoxic hazard, and these guidelines 'are periodically reviewed in the light of scientific progress, changing regulatory needs and animal welfare considerations'. OECD published a mammalian erythrocyte micronucleus test guideline for testing chemicals (1) that proposes: 'Animals are treated with the test chemical once…Samples of peripheral blood are taken at least twice (from the same group of animals), starting not earlier than 36 h after treatment, with appropriate intervals following the first sample, but not extending beyond 72 h'. This guidelines are base on the report by the Collaborative Study Group for the Micronucleus Test (CSGMT), which was based on the sampling of mice peripheral blood every 24 h We investigated the kinetics of micronucleus induction by taking samples prior to administration and every 8 or 10 h after single treatment. The comparisons suggest that 24-h sampling could cause not only an underestimation of the responses to various agents but also a misestimation of the time of maximal induction. We proposed that samples of peripheral blood must be collected at two different times during an optimal 25-h sampling range (from 25 to 50 h). Besides, we hypothesize that the time of maximal EPC-MN induction depends on the time required for the mechanisms involved in micronucleus production; and we suggest that a kinetic analysis of MN-PCE induction by several agents with well-known mechanisms of micronuclei induction would allow derivation of a specific relationship between the kinetics of MN-PCE induction and some process of DNA breaks and/or micronuclei induction.
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Affiliation(s)
- Pedro Morales-Ramírez
- Instituto Nacional de Investigaciones Nucleares, Apartado Postal 18-1027 México, D.F., México
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Rencüzoğulları E, Aydın M. Genotoxic and mutagenic studies of teratogens in developing rat and mouse. Drug Chem Toxicol 2018; 42:409-429. [PMID: 29745766 DOI: 10.1080/01480545.2018.1465950] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/25/2022]
Abstract
In this review, genotoxic and mutagenic effects of teratogenic chemical agents in both rat and mouse have been reviewed. Of these chemicals, 97 are drugs and 33 are pesticides or belong to other groups. Large literature searches were conducted to determine the effects of chemicals on chromosome abnormalities, sister chromatid exchanges, and micronucleus formation in experimental animals such as rats and mice. In addition, studies that include unscheduled DNA synthesis, DNA adduct formations, and gene mutations, which help to determine the genotoxicity or mutagenicity of chemicals, have been reviewed. It has been estimated that 46.87% of teratogenic drugs and 48.48% of teratogenic pesticides are positive in all tests. So, all of the teratogens involved in this group have genotoxic and mutagenic effects. On the other hand, 36.45% of the drugs and 21.21% of the pesticides have been found to give negative results in at least one test, with the majority of the tests giving positive results. However, only 4.16% of the drugs and 18.18% of the pesticides were determined to give negative results in the majority of the tests. Among tests with major negative results, 12.50% of the teratogenic drugs and 12.12% of the teratogenic pesticides were negative in all conducted tests.
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Affiliation(s)
- Eyyüp Rencüzoğulları
- a Department of Biology, Faculty of Science and Letters , Adiyaman University , Adiyaman , Turkey
| | - Muhsin Aydın
- a Department of Biology, Faculty of Science and Letters , Adiyaman University , Adiyaman , Turkey
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21
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Rahmouni F, Saoudi M, Amri N, El-Feki A, Rebai T, Badraoui R. Protective effect of Teucrium polium on carbon tetrachloride induced genotoxicity and oxidative stress in rats. Arch Physiol Biochem 2018; 124:1-9. [PMID: 28714319 DOI: 10.1080/13813455.2017.1347795] [Citation(s) in RCA: 18] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 10/19/2022]
Abstract
This study aimed to investigate the protective effects of Teucrium polium (TP) on carbon tetrachloride (CCl4) induced spleen, erythrocyte's oxidative stress, and genotoxicity in rats. TP was found to contain large amounts of polyphenols (150 mg GAE/G of dry plant) and flavonoids (60 mg QE/g of quercetin dry plant). The CCl4 (0.5 ml/kg) treated rats exhibited significant reductions in serum vitamin A (VA), vitamin E (VE) and total antioxidant status (TAS). Thiobarbituric acid reactive substances (TBARS) and conjugated dienes (CD) were significantly high in the CCl4 group compared to controls. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were significantly decreased in CCl4 rats. Cytogenetic trials revealed remarkable increases in the frequency of chromosomal aberrations (CAs) and sister chromatid exchange (SCE) following CCl4 administration. Pretreatment with TP prevented damages caused by CCl4. Spleen characterised by necrosis was detected in CCl4 as compared to controls. Pretreatment with TP considerably decreased the perturbation.
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Affiliation(s)
- Fatma Rahmouni
- a Laboratory of Histology , Medicine Faculty of Sfax University , Sfax , Tunisia
| | - Mongi Saoudi
- b Laboratory of Animal Physiology , Sciences Faculty of Sfax University , Sfax , Tunisia
| | - Nahed Amri
- a Laboratory of Histology , Medicine Faculty of Sfax University , Sfax , Tunisia
| | - Abdelfattah El-Feki
- b Laboratory of Animal Physiology , Sciences Faculty of Sfax University , Sfax , Tunisia
| | - Tarek Rebai
- a Laboratory of Histology , Medicine Faculty of Sfax University , Sfax , Tunisia
| | - Riadh Badraoui
- a Laboratory of Histology , Medicine Faculty of Sfax University , Sfax , Tunisia
- c Laboratory of Histology , Medicine College of Tunis El-Manar University , Tunis , Tunisia
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Yun JW, Kim SH, Kim YS, You JR, Cho EY, Yoon JH, Kwon E, Yun IJ, Oh JH, Jang JJ, Park JS, Che JH, Kang BC. Enzymatic extract from Ecklonia cava: Acute and subchronic oral toxicity and genotoxicity studies. Regul Toxicol Pharmacol 2018; 92:46-54. [PMID: 29108849 DOI: 10.1016/j.yrtph.2017.10.034] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/08/2017] [Revised: 10/30/2017] [Accepted: 10/31/2017] [Indexed: 11/20/2022]
Abstract
Ecklonia cava (EC) is known to have antioxidant, anti-inflammatory, antidiabetic, and anticancer properties. Despite its wide use and beneficial properties, comprehensive toxicological information regarding EC extract is currently limited. Therefore, the purpose of this study was to investigate acute toxicity, subchronic toxicity, and genotoxicity of enzymatic EC extract according to test guidelines published by Organization for Economic Cooperation and Development. The acute oral LD50 values of this EC extract administered to rats and dogs were estimated to be more than 3000 mg/kg BW. In an oral 13-week toxicity study, changes in body weights of rats exposed to the EC extract up to 3000 mg/kg BW were found to be normal. In addition, repeated doses of EC extract failed to influence any systematic parameters of treatment-related toxic symptoms such as food/water consumption, mortality, urinalysis, hematology, serum biochemistry, organ weight, or histopathology. These results indicated that the no-observed-adverse-effect level for the EC extract was 3000 mg/kg/day for male and female rats. Data obtained from Ames test, chromosome aberration assay, and micronucleus assay indicated that EC extract was not mutagenic or clastogenic. Taken together, these results support the safety of enzymatic EC extract as a potential therapeutic for human consumption against various diseases.
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Affiliation(s)
- Jun-Won Yun
- Department of Biotechnology, The Catholic University of Korea, Bucheon, Republic of Korea
| | - Seung-Hyun Kim
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Yun-Soon Kim
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Ji-Ran You
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Eun-Young Cho
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Jung-Hee Yoon
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Euna Kwon
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - In-Jue Yun
- Ju Yeong NS Co., Ltd., Chuncheon, Kangwon-do, Republic of Korea
| | - Je-Hun Oh
- Ju Yeong NS Co., Ltd., Chuncheon, Kangwon-do, Republic of Korea
| | - Ja-June Jang
- Department of Pathology, Seoul National University College of Medicine, Seoul, Republic of Korea
| | - Jin-Sung Park
- Department of Neurogenetics, Kolling Institute, Royal North Shore Hospital and University of Sydney, St. Leonards, Australia
| | - Jeong-Hwan Che
- Biomedical Center for Animal Resource and Development, Seoul National University College of Medicine, Seoul, Republic of Korea.
| | - Byeong-Cheol Kang
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea; Biomedical Center for Animal Resource and Development, Seoul National University College of Medicine, Seoul, Republic of Korea; Graduate School of Translational Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea; Designed Animal and Transplantation Research Institute, Institute of GreenBio Science Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, Republic of Korea.
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23
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Nagatomo A, Oguri M, Nishida N, Ogawa M, Ichikawa A, Tanaka-Azuma Y. Evaluation of genotoxicity and subchronic toxicity of standardized rose hip extract. Hum Exp Toxicol 2017; 37:725-741. [DOI: 10.1177/0960327117730881] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/17/2022]
Abstract
Rose hip is the fruit of the rose plant, which is widely used in food, cosmetics and as a traditional medicine. Therefore, rose hip is considered safe and has a sufficient history of consumption as food. However, few studies have reported on the safety of rose hip extracts in toxicological analyses. Thus, to evaluate the safety of rosehip polyphenol MJ (RHPMJ), an aqueous ethanol extract standardized with the trans-tiliroside content, we performed genotoxicity and 90-day repeated oral dose toxicity studies in compliance with the Organisation for Economic Co-operation and Development-Good Laboratory Practice. RHPMJ did not induce gene mutations in reverse mutation tests of Salmonella typhimurium TA98, TA100, TA1535, TA1537 and Escherichia coli WP2 uvrA strains and did not induce chromosomal aberrations in cultured Chinese hamster lung (CHL/IU) cells. Moreover, micronucleus tests using rat bone marrow showed RHPMJ had no micronucleus-inducing potential. Finally, 90-day repeated oral dose toxicity studies (100–1000 mg/kg) in male and female rats showed no treatment-related toxicity in rats. These data indicate that the RHPMJ had no genotoxicity and a no-observed-adverse-effect level greater than 1000 mg/kg in rats.
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Affiliation(s)
| | - M Oguri
- Morishita Jintan Co., Ltd, Osaka, Japan
| | - N Nishida
- Morishita Jintan Co., Ltd, Osaka, Japan
| | - M Ogawa
- Bioresearch Center, CMIC Pharma Science Co., Ltd, Yamanashi, Japan
| | - A Ichikawa
- Bioresearch Center, CMIC Pharma Science Co., Ltd, Yamanashi, Japan
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Yun JW, Kim YS, Kwon E, Kim SH, You JR, Kim HH, Che JH, Kang BC. Evaluation of in vitro and in vivo genotoxicity of Angelica acutiloba in a standard battery of assays. Lab Anim Res 2017; 33:231-236. [PMID: 29046698 PMCID: PMC5645601 DOI: 10.5625/lar.2017.33.3.231] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/06/2017] [Revised: 09/02/2017] [Accepted: 09/03/2017] [Indexed: 12/21/2022] Open
Abstract
Among three representative species of Angelica found in Asian countries, including Korea, China, and Japan, Angelica acutiloba (AA) has been used as traditional herbal medicine with antitumor, anti-inflammatory, anti-obesity, and anti-diabetes activities. In this study, the potential genotoxicity and mutagenicity of the AA extract were examined in a battery of in vitro and in vivo tests (bacterial reverse mutation assay, in vitro chromosomal aberrations assay, and in vivo micronucleus assay) in accordance with the test guidelines for toxicity testing developed by the Organization for Economic Cooperation and Development. Upon testing in the bacterial mutation assay (Ames test) using five Salmonella typhimurium TA98, TA100, TA102, TA1535 and TA1537, no significant increase the number of revertant colonies in the metabolic activation system and non-activation system was noted in the AA extract groups. Also, in the chromosome aberration test, the AA extract did not cause chromosomal aberration with or without metabolic activation by S9 mix. A bone marrow micronucleus test of mice demonstrated that the incidence of micronucleated polychromatic erythrocytes in the AA extract groups (500, 1000 and 2000 mg/kg BW) was equivalent to that of the negative control group. Based on these results from a standard battery of assays, the AA extract was concluded to have no genotoxic at the proper dose.
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Affiliation(s)
- Jun-Won Yun
- Department of Biotechnology, The Catholic University of Korea, Bucheon, Gyeonggi-do, Korea
| | - Yun-Soon Kim
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Korea
| | - Euna Kwon
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Korea
| | - Seung-Hyun Kim
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Korea
| | - Ji-Ran You
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Korea
| | - Hyeon Hoe Kim
- Department of Urology, Seoul National University College of Medicine, Seoul, Korea
| | - Jeong-Hwan Che
- Biomedical Center for Animal Resource and Development, Seoul National University College of Medicine, Seoul, Korea
| | - Byeong-Cheol Kang
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Korea
- Biomedical Center for Animal Resource and Development, Seoul National University College of Medicine, Seoul, Korea
- Graduate School of Translational Medicine, Seoul National University College of Medicine, Seoul, Korea
- Designed Animal and Transplantation Research Institute, Institute of GreenBio Science Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, Korea
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Yun JW, Kim SH, Kim YS, You JR, Cho EY, Yoon JH, Kwon E, Lee SJ, Kim SP, Seo JH, In JP, Ahn JH, Jang JJ, Park JS, Che JH, Kang BC. Absence of subchronic oral toxicity and genotoxicity of rice koji with Aspergillus terreus. Regul Toxicol Pharmacol 2017; 89:244-252. [PMID: 28802559 DOI: 10.1016/j.yrtph.2017.08.004] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/15/2017] [Revised: 08/04/2017] [Accepted: 08/07/2017] [Indexed: 01/08/2023]
Abstract
Koji products have been considered as an effective fermented food consumed in East Asia with many health benefits. Particularly, rice koji with Aspergillus terreus (RAT) has been reported to be able to prevent hyperlipidemia and hepatic steatosis through regulating cholesterol synthesis. Despite its biological activities, there is a lack of comprehensive information to give an assurance of its safety. Therefore, the objective of this study was to perform a series of toxicological studies (repeated dose oral toxicity and genotoxicity) according to test guidelines published by the Organization for Economic Cooperation and Development. Along with acute toxicity study using rats and beagle dogs, a 13-week toxicity study revealed no clear RAT-related toxic changes, including body weight, mortality, hematology, serum biochemistry, organ weight, and histopathology after oral administration at doses of 500, 1000, and 2000 mg/kg BW. The no-observed-adverse-effect level of RAT was considered to be more than 2000 mg/kg BW/day in rats of both genders. In addition, potential genotoxicity was evaluated using a standard battery of tests (Ames test, chromosome aberration assay, and micronucleus assay) which revealed that RAT showed no genotoxicity. Accordingly, these results suggest that RAT is a safe and non-toxic functional food for human consumption at proper dose.
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Affiliation(s)
- Jun-Won Yun
- Department of Biotechnology, The Catholic University of Korea, Bucheon, Republic of Korea
| | - Seung-Hyun Kim
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Yun-Soon Kim
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Ji-Ran You
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Eun-Young Cho
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Jung-Hee Yoon
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Euna Kwon
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | | | | | | | | | - Jae Hun Ahn
- Graduate School of Translational Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea
| | - Ja-June Jang
- Department of Pathology, Seoul National University College of Medicine, Seoul, Republic of Korea
| | - Jin-Sung Park
- Department of Biotechnology, The Catholic University of Korea, Bucheon, Republic of Korea; Department of Neurogenetics, Kolling Institute, Royal North Shore Hospital and University of Sydney, St. Leonards, Australia
| | - Jeong-Hwan Che
- Biomedical Center for Animal Resource and Development, Seoul National University College of Medicine, Seoul, Republic of Korea.
| | - Byeong-Cheol Kang
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea; Graduate School of Translational Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea; Biomedical Center for Animal Resource and Development, Seoul National University College of Medicine, Seoul, Republic of Korea; Designed Animal and Transplantation Research Institute, Institute of GreenBio Science Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, Republic of Korea.
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26
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Yun JW, Kim SH, Kim YS, You JR, Cho EY, Yoon JH, Kwon E, Ahn JH, Jang JJ, Che JH, Kang BC. A comprehensive study on in vitro and in vivo toxicological evaluation of Artemisia capillaris. Regul Toxicol Pharmacol 2017; 88:87-95. [PMID: 28487065 DOI: 10.1016/j.yrtph.2017.05.010] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/22/2016] [Revised: 05/04/2017] [Accepted: 05/05/2017] [Indexed: 01/08/2023]
Abstract
Artemisia capillaris (AC) has been used as an alternative therapy in obesity, atopic dermatitis, and liver diseases through several biological activity including anti-steatotic, antioxidant, and anti-inflammatory activities. Despite its ethnomedicinal benefits, no sufficient background information is available about the long-term safety and genotoxicity of the AC extract. Therefore, the present study was carried out to investigate the 13-week subchronic toxicity and genotoxicity of the AC extract according to the test guidelines published by the Organization for Economic Cooperation and Development. In the 13-week toxicity study using doses of 25, 74, 222, 667, and 2000 mg/kg body weight, oral administration of the AC extract in male and female rats did not result in any significant adverse effects in food/water consumption, body weight, mortality, hematology, serum biochemistry, organ weight and histopathology. Accordingly, the no-observed-adverse-effect level in rats of both genders was established for the AC extract at 2000 mg/kg/day, the highest dose level tested. In addition, the AC extract was not genotoxic in a battery of tests including Ames test, in vitro chromosome aberration assay and in vivo micronucleus assay. In conclusion, we demonstrated that the AC extract is considered as a safe traditional medicine for human consumption.
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Affiliation(s)
- Jun-Won Yun
- Department of Biotechnology, The Catholic University of Korea, Bucheon, Republic of Korea
| | - Seung-Hyun Kim
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Yun-Soon Kim
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Ji-Ran You
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Eun-Young Cho
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Jung-Hee Yoon
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Euna Kwon
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Jae Hun Ahn
- Graduate School of Translational Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea
| | - Ja-June Jang
- Department of Pathology, Seoul National University College of Medicine, Seoul, Republic of Korea
| | - Jeong-Hwan Che
- Biomedical Center for Animal Resource and Development, Seoul National University College of Medicine, Seoul, Republic of Korea.
| | - Byeong-Cheol Kang
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea; Graduate School of Translational Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea; Biomedical Center for Animal Resource and Development, Seoul National University College of Medicine, Seoul, Republic of Korea; Designed Animal and Transplantation Research Institute, Institute of GreenBio Science Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, Republic of Korea.
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Genotoxicity kinetics in murine normoblasts as an approach for the in vivo action of difluorodeoxycytidine. Cancer Chemother Pharmacol 2017; 79:843-853. [DOI: 10.1007/s00280-017-3290-0] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/12/2016] [Accepted: 03/14/2017] [Indexed: 12/30/2022]
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Pepelko WE, Peirano WB. Health Effects of Exposure to Diesel Engine Emissions: A Summary of Animal Studies Conducted by the U.S. Environmental Protection Agency's Health Effects Research Laboratories at Cincinnati, Ohio. ACTA ACUST UNITED AC 2016. [DOI: 10.3109/10915818309141011] [Citation(s) in RCA: 27] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022]
Abstract
In order to evaluate the potentially harmful effects of diesel engine emissions, inhalation exposure studies were carried out using a variety of animal species and strains, and measuring a wide range of toxicological parameters. Exhaust was provided by a 6 cylinder Nissan diesel engine operated 20 hours/day, 7 days/week, during a 2 month preliminary trial and 8 hours/day, 7 days/week during a 2 year, 4 month long-term study. The exhaust was diluted to produce a concentration of 6 mg/m3 particulate matter during the preliminary study and during the first year of the long term study and 12 mg/m3 thereafter. Exposure to diesel emissions did not result in detectable genotoxic effects as measured by increases in sister chromatid exchange (SCE), micronucleus testing and metaphase analysis in Chinese hamster and mouse bone marrow cells or in the morphology of cat sperm. SCE in lung cells from Syrian hamsters, however, was increased at the higher exposure level. The incidence of heritable mutations was not increased in mice. An increase in lung tumor incidence was detected in female Senear mice exposed from conception, but not in males, or in either male or female strain A mice exposed from 6 weeks of age. Voluntary activity was decreased in rats during exposure and learning ability was impaired in adult animals exposed during the early postnatal period. Susceptibility to an infectious challenge was increased in diesel exposed mice. Clearance of particles from the lung was found to be minimal even after 90 days recovery in clean air. Pulmonary function alterations in diesel exposed cats suggested the presence of a lesion restricting the air flow. The integrity of the lung as measured by leakage of 131I labeled protein into the alveoli did not seem to be impaired, but increased collagen synthesis suggested that fibrosis may develop with long-term exposure. The preliminary inhalation study resulted in the induction of xenobiotic metabolizing enzymes in the liver. In later studies, however, enzyme induction was found only in male mice injected with large doses of diesel particulate extract.
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Affiliation(s)
- W. E. Pepelko
- U.S. Environmental Protection Agency 26 West St. Clair Street Cincinnati, Ohio 45268
| | - W. B. Peirano
- U.S. Environmental Protection Agency 26 West St. Clair Street Cincinnati, Ohio 45268
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29
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Sanchez-Vicente L, Herraez E, Briz O, Nogales R, Molina-Alcaide E, Marin JJG. Biodetection of potential genotoxic pollutants entering the human food chain through ashes used in livestock diets. Food Chem 2016; 205:81-8. [PMID: 27006217 DOI: 10.1016/j.foodchem.2016.02.160] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/17/2015] [Revised: 02/21/2016] [Accepted: 02/28/2016] [Indexed: 10/22/2022]
Abstract
Ash derived from energy generation is used as a source of minerals in livestock feeds. The microbial biosensor recApr-Luc2 was built to detect genotoxic hazard in recycled ash. Escherichia coli SOS gene (recA, lexA, dinI and umuC) expression in response to cisplatin-induced DNA damage led to the selection of the recA promoter. The biosensor required functional RecA expression to respond to genotoxic heavy metals (Cr>Cd≈Pb), and polluted ash induced a strong recApr-Luc2 response. In human liver and intestinal cells, heavy metals induced acute toxicity (Cr>Cd>Pb) at concentrations sufficient to activate recApr-Luc2. Cytostatic effects, including genotoxicity, were cell- and metal-dependent, apart from Cr. In agreement with the recApr-Luc2 bioassay, Cr had the strongest effect in all cells. In conclusion, recApr-Luc2 could be useful for evaluating the genotoxic risk of pollutants present in ash that might be concentrated in animal products and, thus, entering the human food chain.
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Affiliation(s)
- Laura Sanchez-Vicente
- Laboratory of Experimental Hepatology and Drug Targeting (HEVEFARM), IBSAL, University of Salamanca, Salamanca, Spain
| | - Elisa Herraez
- Laboratory of Experimental Hepatology and Drug Targeting (HEVEFARM), IBSAL, University of Salamanca, Salamanca, Spain; Centre for the Study of Liver and Gastrointestinal Diseases (CIBERehd), Carlos III National Institute of Health, Madrid, Spain
| | - Oscar Briz
- Laboratory of Experimental Hepatology and Drug Targeting (HEVEFARM), IBSAL, University of Salamanca, Salamanca, Spain; Centre for the Study of Liver and Gastrointestinal Diseases (CIBERehd), Carlos III National Institute of Health, Madrid, Spain
| | | | | | - Jose J G Marin
- Laboratory of Experimental Hepatology and Drug Targeting (HEVEFARM), IBSAL, University of Salamanca, Salamanca, Spain; Centre for the Study of Liver and Gastrointestinal Diseases (CIBERehd), Carlos III National Institute of Health, Madrid, Spain.
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30
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Peschke M, Nagel S, Haamann F, Melzer S, Meier K. Cytogenetic monitoring of pharmaceutical staff working with cytostatic drug preparations: A 5-year follow-up. J Oncol Pharm Pract 2016. [DOI: 10.1177/107815529500100104] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/17/2022]
Abstract
Introduction. Cytostatic drug preparation in hos pital pharmacies leads to a concentration of expo sure on a few people and requires considerable protective measures. Methods. To prove the efficiency of those standards, 45 people who routinely prepare cyto static drugs were monitored with cytogenetic tests, Micronucleus-rate (MN), and Sister Chroma tid Exchange-rate (SCE), once a year over a period up to 5 years. All participants of the study received a physical examination before they began prepar ing cytostatic drugs. Baseline MN and SCE were performed at the time. Individual comparisons for the tests between pre- and post-exposure, and a comparison between an exposed and an unex posed group were then performed. Results. During the 5-year follow-up there were no hints of genotoxic effects as a result of handling cytostatic drugs. Conclusion. Thus, it appeared that the safety precautions employed at this Institute were ade quate.
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Affiliation(s)
- Michael Peschke
- Dienst der Freien und Hansestadt Hamburg, Alter Steinweg 4, D-20459 Hamburg
| | - Sibylle Nagel
- Dienst der Freien und Hansestadt Hamburg, Alter Steinweg 4, D-20459 Hamburg
| | - Frank Haamann
- Dienst der Freien und Hansestadt Hamburg, Alter Steinweg 4, D-20459 Hamburg
| | - Simone Melzer
- Allg. Krankenhauses Harburg, Eissendorfer Pferdeweg 52, D-21075 Hamburg, Germany
| | - Klaus Meier
- Allg. Krankenhauses Harburg, Eissendorfer Pferdeweg 52, D-21075 Hamburg, Germany
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Maeda J, Yurkon CR, Fujii Y, Fujisawa H, Kato S, Brents CA, Uesaka M, Fujimori A, Kitamura H, Kato TA. Solution Radioactivated by Hadron Radiation Can Increase Sister Chromatid Exchanges. PLoS One 2015; 10:e0144619. [PMID: 26657140 PMCID: PMC4682810 DOI: 10.1371/journal.pone.0144619] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/28/2015] [Accepted: 10/21/2015] [Indexed: 11/20/2022] Open
Abstract
When energetic particles irradiate matter, it becomes activated by nuclear reactions. Radioactivation induced cellular effects are not clearly understood, but it could be a part of bystander effects. This investigation is aimed at understanding the biological effects from radioactivation in solution induced by hadron radiation. Water or phosphate buffered saline was activated by being exposed to hadron radiation including protons, carbon- and iron-ions. 1 mL of radioactivated solution was transferred to flasks with Chinese hamster ovary (CHO) cells cultured in 5 mL of complete media. The induction of sister chromatid exchanges (SCE) was used to observe any increase in DNA damage responses. The energy spectrum and the half-lives of the radioactivation were analyzed by NaI scintillation detector in order to identify generated radionuclides. In the radioactivated solution, 511 keV gamma-rays were observed, and their half-lives were approximately 2 min, 10 min, and 20 min. They respectively correspond to the beta+ decay of 15O, 13N, and 11C. The SCE frequencies in CHO cells increased depending on the amount of radioactivation in the solution. These were suppressed with a 2-hour delayed solution transfer or pretreatment with dimethyl sulfoxide (DMSO). Our results suggest that the SCE induction by radioactivated solution was mediated by free radicals produced by the annihilated gamma-rays. Since the SCE induction and DMSO modulation are also reported in radiation-induced bystander effects, our results imply that radioactivation of the solution may have some contribution to the bystander effects from hadron radiation. Further investigations are required to assess if radioactivation effects would attribute an additional level of cancer risk of the hadron radiation therapy itself.
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Affiliation(s)
- Junko Maeda
- Department of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, Colorado, United States of America
| | - Charles R. Yurkon
- Department of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, Colorado, United States of America
| | - Yoshihiro Fujii
- Department of Radiological Sciences, Ibaraki Prefectural University of Health Sciences, Inashiki, Ibaraki, Japan
| | - Hiroshi Fujisawa
- Graduate School of Engineering, The University of Tokyo, Tokyo, Japan
| | - Sayaka Kato
- Department of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, Colorado, United States of America
| | - Colleen A. Brents
- Department of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, Colorado, United States of America
| | - Mitsuru Uesaka
- Graduate School of Engineering, The University of Tokyo, Tokyo, Japan
| | - Akira Fujimori
- Research Center for Charged Particle Therapy, International Open Laboratory, National Institute of Radiological Sciences, Chiba, Chiba, Japan
| | - Hisashi Kitamura
- Research Development and Support Center, National Institute of Radiological Sciences, Chiba, Chiba, Japan
| | - Takamitsu A. Kato
- Department of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, Colorado, United States of America
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32
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Yun JW, You JR, Kim YS, Cho EY, Kim SH, Yoon JH, Kwon E, Chung DH, Kim YT, Jang JJ, Che JH, Kang BC. Pre-clinical in vitro and in vivo safety evaluation of Cimicifuga heracleifolia. Regul Toxicol Pharmacol 2015; 73:303-10. [DOI: 10.1016/j.yrtph.2015.07.006] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/28/2015] [Revised: 07/08/2015] [Accepted: 07/09/2015] [Indexed: 01/14/2023]
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Akyıl D, Konuk M. Detection of genotoxicity and mutagenicity of chlorthiophos using micronucleus, chromosome aberration, sister chromatid exchange, and Ames tests. ENVIRONMENTAL TOXICOLOGY 2015; 30:937-945. [PMID: 24515492 DOI: 10.1002/tox.21968] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 12/06/2013] [Revised: 01/27/2014] [Accepted: 01/28/2014] [Indexed: 06/03/2023]
Abstract
Potential mutagenic and genotoxic effects of Chlorthiophos, an organophosphate pesticide, were evaluated using four standard assays. Five different concentrations of the pesticide were tested by an Ames test using Salmonella typhimurium strains TA97, TA98, TA100, and TA102, with and without S9 metabolic activation. No concentrations of Chlorthiophos showed mutagenic activity on the TA97, TA100, and TA102 strains, with and without S9 fraction, but were all mutagenic to the TA98 strain without S9. Sister chromatid exchange (SCE), chromosome aberration (CA), and micronucleus (MN) tests were used to investigate the genotoxic effects of Chlorthiophos in human peripheral lymphocytes treated with 25, 50, 100, and 200 µg/mL concentrations of Chlorthiophos for 24 and 48 h. The nuclear division index (NDI), replication index (RI), and mitotic index (MI) were also calculated to determine the cytotoxicity of Chlorthiophos. No increase in SCE frequency was seen for any treatment period or concentration, but Chlorthiophos at 200 µg/mL increased the frequency of CAs. Increases in MN formation were only observed at Chlorthiophos concentrations of 200 µg/mL following 24 and 48 h treatments. Chlorthiophos treatment reduced the MI and NDI significantly, but had no effect on the RI.
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Affiliation(s)
- Dilek Akyıl
- Biology Department, Faculty of Sciences and Literatures, Afyon Kocatepe University, 03200, Afyonkarahisar, Turkey
| | - Muhsin Konuk
- Molecular Biology and Genetics Department, Faculty of Engineering and Natural Sciences, Üsküdar University, Altunizade, 34662, Istanbul, Turkey
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Yun JW, Che JH, Kwon E, Kim YS, Kim SH, You JR, Kim WH, Kim HH, Kang BC. Safety evaluation of Angelica gigas: Genotoxicity and 13-weeks oral subchronic toxicity in rats. Regul Toxicol Pharmacol 2015; 72:473-80. [PMID: 26032491 DOI: 10.1016/j.yrtph.2015.05.025] [Citation(s) in RCA: 23] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/30/2015] [Revised: 05/18/2015] [Accepted: 05/25/2015] [Indexed: 10/23/2022]
Abstract
As a well-known traditional medicine, Angelica gigas (AG) and its active constituents, including decursin and decursinol, have been shown to possess several health beneficial properties such as anti-bacterial, immunostimulating, anti-tumor, neuroprotective, anti-nociceptive and anti-amnestic activities. However, there is lack of toxicity studies to assess potential toxicological concerns, especially long-term toxicity and genotoxicity, regarding the AG extract. Therefore, the safety of AG extract was assessed in subchronic toxicity and genotoxicity assays in accordance with the test guidelines published by the Organization for Economic Cooperation and Development. In a subchronic toxicity study for 13 weeks (125, 250, 500, 1000 and 2000 mg/kg body weight, delivered by gavage), data revealed no significant adverse effects of the AG extract in food consumption, body weight, mortality, hematology, biochemistry, necropsy, organ weight and histopathology throughout the study in male and female rats. These results suggest that no observed adverse effect level of the AG extract administered orally was determined to be greater than 2000 mg/kg/day, the highest dose tested. In addition, a battery of tests including Ames test, in vitro chromosome aberration assay and in vivo micronucleus assay suggested that the AG extract was not genotoxic. In conclusion, the AG extract appears to be safe as a traditional medicine for oral consumption.
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Affiliation(s)
- Jun-Won Yun
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Jeong-Hwan Che
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea; Biomedical Center for Animal Resource and Development, N-BIO, Seoul National University, Seoul, Republic of Korea
| | - Euna Kwon
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Yun-Soon Kim
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Seung-Hyun Kim
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Ji-Ran You
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea
| | - Woo Ho Kim
- Department of Pathology, Seoul National University College of Medicine, Seoul, Republic of Korea
| | - Hyeon Hoe Kim
- Department of Urology, Seoul National University College of Medicine, Seoul, Republic of Korea
| | - Byeong-Cheol Kang
- Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea; Biomedical Center for Animal Resource and Development, N-BIO, Seoul National University, Seoul, Republic of Korea; Graduate School of Translational Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea; Designed Animal and Transplantation Research Institute, Institute of GreenBio Science Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, Republic of Korea.
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35
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Ceker S, Agar G, Alpsoy L, Nardemir G, Kizil HE. Antagonistic effects of Satureja hortensis essential oil against AFB, on human lymphocytes in vitro. CYTOL GENET+ 2014. [DOI: 10.3103/s0095452714050028] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/30/2022]
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36
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Norizadeh Tazehkand M, Topaktas M. Thein vitrogenotoxic and cytotoxic effects of remeron on human peripheral blood lymphocytes. Drug Chem Toxicol 2014; 38:266-71. [DOI: 10.3109/01480545.2014.947425] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022]
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37
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The relationship between dioxin congeners in the breast milk of Vietnamese women and sister chromatid exchange. Int J Mol Sci 2014; 15:7485-99. [PMID: 24786289 PMCID: PMC4057685 DOI: 10.3390/ijms15057485] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/14/2014] [Revised: 04/11/2014] [Accepted: 04/14/2014] [Indexed: 11/16/2022] Open
Abstract
The aim of this study was to clarify the relationship between dioxin concentrations in breast milk and the sister chromatid exchange (SCE) frequency in women from herbicide-sprayed and non sprayed areas. Blood samples were taken from 21 women with high TCDD (tetrachlorodibenzo-p-dioxin) levels from sprayed areas, 23 women with moderate TCDD levels from sprayed areas, and 19 women from non sprayed areas to determine their SCE frequency. The SCE frequencies for the high and moderate TCDD groups from the sprayed area and for the non sprayed area group were 2.40, 2.19, and 1.48 per cell, respectively. Multiple regression analysis showed that the standardized β values for 1,2,3,6,7,8-hexaCDD (β = 0.60), 1,2,3,4,6,7,8-heptaCDD (β = 0.64), and octaCDD (β = 0.65) were higher than those for TCDD (β = 0.34) and 1,2,3,7,8-pentaCDD (β = 0.42). The adjusted R² value for polyCDDs (R² = 0.38) was higher than that for polyCDD toxic equivalents (TEQ (toxic equivalents); R² = 0.23). This study therefore shows that levels of hexa-, hepta-, and octaCDD, which were previously regarded as being less toxic than TCDD, are closely related to SCE frequency and that the level of dioxin (pg/g lipid) is potentially more useful as an indicator than TEQ value for explaining SCE frequency.
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Multiplexed DNA repair assays for multiple lesions and multiple doses via transcription inhibition and transcriptional mutagenesis. Proc Natl Acad Sci U S A 2014; 111:E1823-32. [PMID: 24757057 DOI: 10.1073/pnas.1401182111] [Citation(s) in RCA: 107] [Impact Index Per Article: 9.7] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/23/2022] Open
Abstract
The capacity to repair different types of DNA damage varies among individuals, making them more or less susceptible to the detrimental health consequences of damage exposures. Current methods for measuring DNA repair capacity (DRC) are relatively labor intensive, often indirect, and usually limited to a single repair pathway. Here, we describe a fluorescence-based multiplex flow-cytometric host cell reactivation assay (FM-HCR) that measures the ability of human cells to repair plasmid reporters, each bearing a different type of DNA damage or different doses of the same type of DNA damage. FM-HCR simultaneously measures repair capacity in any four of the following pathways: nucleotide excision repair, mismatch repair, base excision repair, nonhomologous end joining, homologous recombination, and methylguanine methyltransferase. We show that FM-HCR can measure interindividual DRC differences in a panel of 24 cell lines derived from genetically diverse, apparently healthy individuals, and we show that FM-HCR may be used to identify inhibitors or enhancers of DRC. We further develop a next-generation sequencing-based HCR assay (HCR-Seq) that detects rare transcriptional mutagenesis events due to lesion bypass by RNA polymerase, providing an added dimension to DRC measurements. FM-HCR and HCR-Seq provide powerful tools for exploring relationships among global DRC, disease susceptibility, and optimal treatment.
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Abstract
A fully optimized staining method for detecting sister chromatid exchanges in cultured cells is presented. The method gives reproducibly robust quantitative results. Sister chromatid exchange is a classic toxicology assay for genotoxicity and for detecting alterations to the biochemistry underlying cellular homologous recombination. Growth of cells in the presence of 5'-bromo-deoxyuridine for two rounds of DNA replication followed by collecting metaphase spreads on glass slides, treatment with the UV-sensitive dye Hoechst 33258, long-wave UV light exposure, and Giemsa staining gives a permanent record of the exchanges.
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Affiliation(s)
- Dawn M Stults
- Markey Cancer Center, University of Kentucky, 222 Combs Research Building, 800 Rose Street, Lexington, KY, 40536-0096, USA
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Sevindik N, Rencuzogullari E. The genotoxic and antigenotoxic effects of Salvia fruticosa leaf extract in human blood lymphocytes. Drug Chem Toxicol 2013; 37:295-302. [PMID: 24224704 DOI: 10.3109/01480545.2013.851689] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/17/2023]
Abstract
The aim of this study was to investigate the genotoxic and antigenotoxic effects of Salvia fruticosa (Sf) leaf extract with the absence and presence of S9 mix using sister chromatid exchange (SCE), chromosome aberration (CA) and micronucleus (MN) formation test systems in human peripheral blood lymphocytes (HPBLs) that were treated with 1.5-, 3.0- and 6.0-µL/mL concentrations for 24- and 48-hour treatment periods. The cytotoxicity of Sf leaf extract was also investigated by calculating the mitotic index (MI), proliferation index (PI) and nuclear division index (NDI). In the absence of S9 mix, Sf leaf extract alone increased SCE frequency at the 48-hour treatment period; however, it induced the CA and MN at all concentrations and at all treatment periods. Sf plus MMC (mitomycin C) synergically induced SCE and CA, except the highest concentration of Sf leaf extract and MMC on induction of SCE. In addition, Sf leaf extract induced the effect of MMC on MN frequency for 24 hours, but it significantly decreased the effect of MMC on MN frequency for the 48-hour treatment period. Sf leaf extract showed a cytotoxic effect by decreasing the MI; however, it did not decrease the PI and NDI. In the presence of S9 mix, Sf leaf extract did not increase the SCE, when compared to solvent control, whereas it reduced the effect of cyclophosphamide (Cyp). Sf leaf extract induced the CA and MN, but could not increase the effect of Cyp on CA and MN formation. Sf leaf extract had no cytotoxic effect; however, it induced the cytotoxicity of Cyp.
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Affiliation(s)
- Nadire Sevindik
- Deparment of Biology, Natural and Applied Science Institute, Çukurova University , Adana , Turkey and
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Tug E, Kayhan G, Kan D, Guntekin S, Ergun MA. The evaluation of long-term effects of ionizing radiation through measurement of current sister chromatid exchange (SCE) rates in radiology technologists, compared with previous SCE values. Mutat Res 2013; 757:28-30. [PMID: 23867852 DOI: 10.1016/j.mrgentox.2013.04.025] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/31/2012] [Revised: 02/12/2013] [Accepted: 04/05/2013] [Indexed: 11/29/2022]
Abstract
Ionizing radiation is a strong physical mutagen, causing breakage of phosphodiester bonds in DNA at any stage of the mitotic cycle. Analysis of sister chromatid exchange (SCE) has come into use as a sensitive DNA-damage indicator. We investigated the SCE rates in radiology technologists who are occupationally and chronically exposed to ionizing radiation. The study included 39 radiology technologists and 35 sex- and age-matched healthy controls. There was a statistically significant difference in the SCE frequency between radiology technologists and controls (p<0.0001). Additionally, previous SCE data of 10 radiology technologists were compared with current results regarding radiation exposure time. There was statistically significant difference between previous and current SCE values (p=0.005). The significant increase in the frequency of SCE in radiology technologists emphasizes the importance of radiation-protection procedures in order to minimize radiation exposure and avoid possible genotoxic effects. Comparison of two studies that measured SCE values of radiology technologists after 8 years also suggests that the genotoxic effect is reversible. In conclusion, radiation is still an important mutagenic agent despite improvements in daily working hours and conditions.
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Affiliation(s)
- E Tug
- Gazi University, Faculty of Medicine, Department of Medical Genetics, Ankara, Turkey.
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Ceker S, Orhan F, Kizil HE, Alpsoy L, Gulluce M, Aslan A, Agar G. Genotoxic and antigenotoxic potentials of two Usnea species. Toxicol Ind Health 2013; 31:990-9. [DOI: 10.1177/0748233713485889] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/18/2022]
Abstract
For ages, lichens have long been investigated popularly for biological roles, mainly antitumor, antimicrobial and antioxidant activities. Many positive results were obtained in these previous research. Thus, in this study, we aimed to determine whether extracts of Usnea articulata (UAE) and Usnea filipendula (UFE) possessing a protection against aflatoxin B1 (AFB1)-induced genotoxic and oxidative damage. The results of our studies showed that 5 μM concentrations of AFB1 increased the frequencies of sister chromatid exchange (SCE) and the level of malondialdehyde (MDA) and decreased the activities of superoxide dismutase (SOD), glutathione (GSH) and glutathione peroxidase (GPx). However, when 5, 10 and 20 µg/mL concentrations of UAE and UFE was added to AFB1, the frequencies of SCE and MDA level were decreased and SOD, GSH and GPx level were increased. The Ames ( Salmonella typhimurium TA1535, TA1537) and WP2 ( Escherichia coli) test systems carried out evinced that UAE and UFE possess any mutagenicity, but have antimutagenic effects. Consequently, the results of this experiment have clearly shown that UAE and UFE have strong antioxidative and antigenotoxic effects that are associated with its antioxidant nature. A detailed study can be performed to determine the antioxidant properties of each compound that will extend the use of lichen extracts in food and pharmacy industries.
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Affiliation(s)
- Selcuk Ceker
- Central Research and Application Laboratories, Agri Ibrahim Cecen University, Agri, Turkey
- Department of Biology, Faculty of Science, Ataturk University, Erzurum, Turkey
| | - Furkan Orhan
- Central Research and Application Laboratories, Agri Ibrahim Cecen University, Agri, Turkey
- Department of Biology, Faculty of Science, Ataturk University, Erzurum, Turkey
| | | | - Lokman Alpsoy
- Department of Biology, Faculty of Science, Fatih University, Istanbul, Turkey
| | - Medine Gulluce
- Department of Biology, Faculty of Science, Ataturk University, Erzurum, Turkey
| | - Ali Aslan
- Department of Biology Education, Kazım Karabekir Faculty of Education, Ataturk University, Erzurum, Turkey
| | - Guleray Agar
- Department of Biology, Faculty of Science, Ataturk University, Erzurum, Turkey
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Bajpayee M, Pandey AK, Parmar D, Dhawan A. Current Status of Short-Term Tests for Evaluation of Genotoxicity, Mutagenicity, and Carcinogenicity of Environmental Chemicals and NCEs. Toxicol Mech Methods 2012; 15:155-80. [PMID: 20021080 DOI: 10.1080/15376520590945667] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/23/2022]
Abstract
The advent of the industrial revolution has seen a significant increase in the number of new chemical entities (NCEs) released in the environment. It becomes imperative to check the toxic potential of NCEs to nontarget species before they are released for commercial purposes because some of these may exert genotoxicity, mutagenicity, or carcinogenicity. Exposure to such compounds produces chemical changes in DNA, which are generally repaired by the DNA repair enzymes. However, DNA damage and its fixation may occur in the form of gene mutations, chromosomal damage, and numerical chromosomal changes and recombination. This may affect the incidence of heritable mutations in man and may be transferred to the progeny or lead to the development of cancer. Hence, adequate tests on NCEs have to be undertaken for the risk assessment and hazard prediction. Compounds that are positive in tests that detect such damages have the potential to be human mutagens/carcinogens. Only long-term animal bioassays, involving lifetime studies on animals, were used earlier to classify substances as mutagens/carcinogens. These tests were cumbersome and time consuming and required a lot of facilities and personnel. Short-term tests, therefore, were brought into practice. A "battery" of three to four of these short-term tests has been proposed now by a number of regulatory authorities for the classification of compounds as mutagenic or carcinogenic. This review deals with the current status of these short-term tests.
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Affiliation(s)
- Mahima Bajpayee
- Developmental Toxicology Division, Industrial Toxicology Research Center, M.G. Marg, LucknowIndia
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Medley CD, Smith JE, Wigman LS, Chetwyn NP. A DNA-conjugated magnetic nanoparticle assay for assessing genotoxicity. Anal Bioanal Chem 2012; 404:2233-9. [DOI: 10.1007/s00216-012-6350-7] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/10/2012] [Revised: 08/03/2012] [Accepted: 08/09/2012] [Indexed: 11/29/2022]
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Thompson LH. Losing and finding myself in DNA repair. DNA Repair (Amst) 2012; 11:637-48. [PMID: 23012750 DOI: 10.1016/j.dnarep.2011.10.005] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/17/2022]
Affiliation(s)
- Larry H Thompson
- Biology & Biotechnology Division, L452, Lawrence Livermore National Laboratory, Livermore, CA 94551-0808, USA.
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Ila HB, Ilhan A. In vitro genotoxic perspective of Tamiflu. Cytotechnology 2012; 64:443-9. [PMID: 22252233 DOI: 10.1007/s10616-011-9422-1] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/15/2011] [Accepted: 12/20/2011] [Indexed: 11/25/2022] Open
Abstract
The aim of this study was to investigate the genotoxic and/or cytotoxic effects of Tamiflu, commercial form of the oseltamivir antiviral and most frequently prescribed for the treatment of influenza infections, on cultured human peripheral lymphocytes by using sister chromatid exchange (SCE), chromosomal aberration (CA), and cytokinesis-blocked micronucleus (CBMN) assays. Cells were treated with 0.5, 1, 2 μg/mL oseltamivir, the Tamiflu capsule ingredient, for 24 or 48 h in the absence or presence of an exogenous metabolic activation system (S9 mix). The test chemical did not demonstrate any genotoxic effect dose-dependently but it showed a weak cytotoxicity on cells in this study. On the other hand, some concentrations of Tamiflu (2 μg/mL without S9 mix for 48 h and 1 μg/mL with S9 mix) induced SCE and also decreased significantly the proliferation index (PI) (48 h period) and the nuclear division index (NDI) (24 h period) (P < 0.05) in the absence of S9 mix. Considering the results, Tamiflu did not induce significant increases of CA or micronucleated cells in vitro in cultured peripheral blood lymphocytes under the treatment conditions used but weak SCE induction was observed. On the other hand, the weak cytotoxic effects observed disappeared in the cultures treated in presence of the S9 mix.
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Affiliation(s)
- Hasan Basri Ila
- Department of Biology, Faculty of Science and Letters, Cukurova University, 01330, Adana, Turkey,
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Karadeniz B, Ulker Z, Alpsoy L. Genotoxic and cytotoxic effects of storax in vitro. Toxicol Ind Health 2011; 29:181-6. [PMID: 22155886 DOI: 10.1177/0748233711428642] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/17/2022]
Abstract
The aim of this study is to investigate the effects of the storax balsam, which is a kind of sweet gum obtained from the Liquidambar orientalis Mill trees, on cell viability, cytotoxicity and genotoxicity in human lymphocyte in vitro. We studied the genotoxic effects of the extract of storax balsam (SE) using sister chromatid exchange (SCE) test system. Also the cytotoxic and inhibitory effects on cell proliferation of SE were evaluated using lactate dehydrogenase (LDH) assay and cell proliferation (WST-1) assay. The SCE frequency was increased when the cells were treated with 1.6 and 4.0 µg/mL SE concentrations (p < 0.05). Moreover, treatment of the cells with the same concentrations significantly depleted the cell number at 24th and 48th hours and elevated the LDH levels (p < 0.05) at 48th hour. These results suggest that SE can be used as an alternative antibacterial and antipathogenic agent due to its cytotoxic and genotoxic effects.
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Aslan A, Agar G, Alpsoy L, Kotan E, Ceker S. Protective role of methanol extracts of two lichens on oxidative and genotoxic damage caused by AFB1 in human lymphocytes in vitro. Toxicol Ind Health 2011; 28:505-12. [PMID: 21986884 DOI: 10.1177/0748233711416944] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022]
Abstract
In this study, the antigenotoxic and antioxidant effects of Umbilicaria vellea (UME) and Xantho somloensis (XME) extracts were determined using sister chromatid exchange (SCE), micronuclei (MN) assays, and superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities and malondialdehyde (MDA) levels against the effects of aflatoxin B(1) (AFB(1))-induced oxidative stress and genotoxicity in human lymphocytes in vitro. The results showed that the frequencies of SCE, MN, and MDA level decreased, but the activities of SOD and GPx increased when 5 μg/mL and 10 μg/mL doses of UME and XME were added to AFB(1)-treated cultures. Also the present results indicate that strong antioxidative and the antigenotoxicity mechanisms of UME and XME are associated with its antioxidant nature.
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Affiliation(s)
- Ali Aslan
- Department of Biology Teacher Training, Ataturk University, Erzurum, Turkey
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Liao PH, Lin RH, Yang ML, Li YC, Kuan YH. Evaluation of differential representative values between Chinese hamster cells and human lymphocytes in mitomycin C-induced cytogenetic assays and caspase-3 activity. Toxicol Ind Health 2011; 28:174-80. [PMID: 21768208 DOI: 10.1177/0748233711409974] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022]
Abstract
Chinese hamster ovary (CHO) cells, its lung fibroblasts (V79), and human lymphocytes are routinely used in in vitro cytogenetic assays, which include micronuclei (MN), sister chromatid exchange (SCE), and chromosome aberration (CA) assays. Mitomycin C (MMC), a DNA cross-link alkylating agent, is both an anticancer medicine and a carcinogen. To study the differential representative values of cell types in MMC-treated cytogenetic assays and its upstream factor, cysteine aspartic acid-specific protease (caspase)-3. Among the chosen cell types, lymphocytes expressed the highest sensitivity in all three MMC-induced assays, whereas CHO and V79 showed varied sensitivity in different assays. In MN assay, the sensitivity of CHO is higher than or equal to V79; in SCE assay, the sensitivity of CHO is the same as V79; and in CA assay, the sensitivity of CHO is higher than V79. In-depth analysis of CA revealed that in chromatid breaks and dicentrics formation, lymphocyte was the most sensitive of all and CHO was more sensitive than V79; and in acentrics and interchanges formation, lymphocyte was much more sensitive than the others. Furthermore, we found caspase-3 activity plays an important role in MMC-induced cytogenetic assays, with MMC-induced caspase-3 activity resulting in more sensitivity in lymphocytes than in CHO and V79. Based on these findings, lymphocyte will make a suitable predictive or representative control reference in cytogenetic assays and caspase-3 activity with its high specificity, positive predictive value, and sensitivity.
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Affiliation(s)
- Pei-Hu Liao
- 1Department of Pharmacology, School of Medicine, Chung Shan Medical University and Chung Shan Medical University Hospital, Taichung, Taiwan, Republic of China
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Stults DM, Killen MW, Shelton BJ, Pierce AJ. Recombination phenotypes of the NCI-60 collection of human cancer cells. BMC Mol Biol 2011; 12:23. [PMID: 21586152 PMCID: PMC3112106 DOI: 10.1186/1471-2199-12-23] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/01/2011] [Accepted: 05/17/2011] [Indexed: 11/10/2022] Open
Abstract
Background The NCI-60 is a collection of tumor cell lines derived from a variety of human adult cancer tissue types and is commonly used for genetic analysis and screening of potential chemotherapeutic agents. We wanted to understand the contributions of specific mechanisms of genomic instability to the etiology of cancers represented by the NCI-60. Results We screened the NCI-60 for dysregulated homologous recombination by using the gene cluster instability (GCI) assay we pioneered, and for defects in base excision repair by sensitivity to 5-hydroxymethyl-2'-deoxyuridine (hmdUrd). We identified subsets of the NCI-60 lines that either displayed the characteristic molecular signature of GCI or were sensitive to hmdUrd. With the exception of the NCI-H23 lung cancer line, these phenotypes were not found to overlap. None of the lines examined in either subset exhibited significant changes in the frequency of sister chromatid exchanges (SCE), neither did any of the lines in either subset exhibit microsatellite instability (MSI) indicative of defects in DNA mismatch repair. Conclusions Gene cluster instability, sensitivity to hmdUrd and sister chromatid exchange are mechanistically distinct phenomena. Genomic instability in the NCI-60 appears to involve only one mechanism of instability for each individual cell line.
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Affiliation(s)
- Dawn M Stults
- Department of Toxicology, University of Kentucky, Lexington, KY, USA
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