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Sample records for cytokinesis-blocked micronucleus cytome

  1. Cytokinesis-block micronucleus assay evolves into a 'cytome' assay of chromosomal instability, mitotic dysfunction and cell death

    International Nuclear Information System (INIS)

    Fenech, Michael

    2006-01-01

    The cytokinesis-block micronucleus (CBMN) assay was originally developed as an ideal system for measuring micronuclei (MNi) however it can also be used to measure nucleoplasmic bridges (NPBs), nuclear buds (NBUDs), cell death (necrosis or apoptosis) and nuclear division rate. Current evidence suggests that (a) NPBs originate from dicentric chromosomes in which the centromeres have been pulled to the opposite poles of the cell at anaphase and are therefore indicative of DNA mis-repair, chromosome rearrangement or telomere end-fusions, (b) NPBs may break to form MNi, (c) the nuclear budding process is the mechanism by which cells remove amplified and/or excess DNA and is therefore a marker of gene amplification and/or altered gene dosage, (d) cell cycle checkpoint defects result in micronucleus formation and (e) hypomethylation of DNA, induced nutritionally or by inhibition of DNA methyl transferase can lead to micronucleus formation either via chromosome loss or chromosome breakage. The strong correlation between micronucleus formation, nuclear budding and NPBs (r = 0.75-0.77, P < 0.001) induced by either folic acid deficiency or exposure to ionising radiation is supportive of the hypothesis that folic acid deficiency and/or ionising radiation cause genomic instability and gene amplification by the initiation of breakage-fusion-bridge cycles. In its comprehensive mode, the CBMN assay measures all cells including necrotic and apoptotic cells as well as number of nuclei per cell to provide a measure of cytotoxicity and mitotic activity. The CBMN assay has in fact evolved into a 'cytome' method for measuring comprehensively chromosomal instability phenotype and altered cellular viability caused by genetic defects and/or nutrional deficiencies and/or exogenous genotoxins thus opening up an exciting future for the use of this methodology in the emerging fields of nutrigenomics and toxicogenomics and their combinations

  2. Cytokinesis-block micronucleus method in micro-blood cultures

    International Nuclear Information System (INIS)

    Liu Jinwen; Wang Lianzhi; Yang Cangzhen; Yao Yanyu

    1991-01-01

    This paper reports the cytokinesis-block micronucleus method in micro-blood cultures. The observations on detection induced micronuclei of different doses of 60 Co γ-rays irradiation and spontaneous micronucleus of different ages were performed with CB method in comporison with conventional micronucleus (CM) method. The results showed that with direct peripheral micro-blood cultures the cytoknesis-block micronuclei is also obtained. Using CB method, the micronuclei fequency of different ages was linear relationship, Y = 1.62 + 0.74 D, the spontaneous micronuclei frequency of different ages was 4.14%, the induced micronuclei also was a linear relationship, Y = 6.01 + 0.692 D. Using CM method, it showed that the induced micronuclei was a linear relationship, Y = 0.486 D - 1.968, but there is no significant difference between the micronuclei frequency of different ages. Comparison with CM and direct blood smear methods confirmed that the cytokinesis-block method of micro-blood cultures is more sensitive and precise

  3. CYTOKINESIS-BLOCK MICRONUCLEUS ASSAY IN HUMAN GLIOMA CELLS EXPOSED TO RADIATION

    Directory of Open Access Journals (Sweden)

    Jerzy Slowinski

    2011-05-01

    Full Text Available Biological tests are efficient in reflecting the biological influences of several types of generally harmful exposures. The micronucleus assay is widely used in genotoxicity studies or studies on genomic damage in general. We present methodological aspects of cytokinesis-block micronucleus assay performed in human gliomas irradiated in vitro. Eight human glioblastoma cell lines obtained from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Germany were gamma-irradiated (60Co over a dose range of 0-10 Gy. Cytokinesis-block micronucleus assay was performed to quantitate cytogenetic damage. The cells were fixed directly on dishes, stained with fluorochrome DAPI and evaluated under fluorescent and phase contrast microscope. The micronucleus frequency was expressed as a micronuclei (MN per binucleated cell (BNC ratio, calculated after scoring at least 100 BNC per dish. The frequency of spontaneous MN ranged from 0.17 to 0.613 (mean: 0.29 ± 0.14. After irradiation increase of MN frequency in the range of 0.312 - 2.241 (mean: 0.98 ± 0.68 was found at 10 Gy. Gliomas are extremely heterogenous in regard to cytogenetic effects of irradiation, as shown in this study by cytokinesis-block micronucleus assay. This test is easily performed on irradiated glioma cell lines and can assist in determining their radiosensitivity. However, in order to obtain reliable and reproducible results, precise criteria for MN scoring must be strictly followed. Simultaneous use of fluorescent and phase contrast equipment improves imaging of morphological details and can further optimize MN scoring.

  4. The cytokinesis-block micronucleus assay: a sensitive technique for measuring radiation-induced chromosome damage

    International Nuclear Information System (INIS)

    Fenech, M.; Morley, A.A.

    1987-01-01

    The sensitivity of the cytokinesis-block micronucleus assay was demonstrated by the detection in human lymphocytes of in vitro exposures of as low as 0.02 Gy of X-rays. To determine the suitability of this new method for measuring in vivo exposure to radiation the authors have performed initial longitudinal studies on (a) cancer patients undergoing partial body fractionated radiotherapy and (b) BALB-C mice following in vivo whole body irradiation with acute single doses of X-rays. The results for radiotherapy patients indicate that the dose fractions have an additive effect on the observed micronucleus frequency which appeared to decline following three months after completion of therapy. Results with irradiated mice showed a sharp increase in micronucleus frequency for splenocytes sampled immediately after treatment and the rate of decline in micronucleus frequency during the first week after treatment was dose-dependent. (author)

  5. Antioxidant evaluation of heterocyclic compounds by cytokinesis-block micronucleus assay.

    Science.gov (United States)

    Godevac, Dejan; Tesević, Vele; Vajs, Vlatka; Milosavljević, Slobodan; Stanković, Miroslava

    2013-03-01

    This article summarizes the results of using cytokinesis-block micronucleus (CBMN) assay to evaluate the antioxidant potential of heterocyclic compounds. Most studies were carried out with naturally occurring heterocyclic compounds such as plant polyphenols: flavonoids, xanthones, coumarins, and ellagitannins, or plant derived products (juices, extracts, supplements) rich in bioactive heterocyclic compounds. There are also some studies dealing with synthetic heterocyclic antioxidants. CBMN assay is an in vitro study that has been used to evaluate antioxidant and protective effects of heterocyclic compounds on induced chromosome aberration in human lymphocytes.

  6. Nucleoplasmic bridges are a sensitive measure of chromosome rearrangement in the cytokinesis-block micronucleus assay

    International Nuclear Information System (INIS)

    Fenech, M.; Umegaki, K.

    2003-01-01

    Full text: We have performed experiments using the WIL2-NS human B-lymphoblastoid cell line and primary human lymphocytes to (a) determine the importance of including measurements of nucleoplasmic bridges (NPB) in the cytokinesis-block micronucleus (CBMN) assay and (b) provide evidence that NPB originate from dicentric chromosomes and centric ring chromosomes. In addition we describe theoretical models that explain how dicentric chromosomes and centric ring chromosomes may result in the formation of NPB at anaphase. The results with WIL2-NS showed that it was possible to distinguish genotoxic effects induced by different oxidizing agents in terms of the NPB/micronucleus frequency ratio. The results with lymphocytes indicated a strong correlation (a) between NPB, centric ring chromosomes and dicentric chromosomes in metaphases (R>0.93, P 0.93, P<0.0001). The dose-response curves with gamma rays were very similar for NPB, ring chromosomes and dicentric chromosomes, as were the dose-responses for MNi, acentric rings and fragments. However, not all acentric chromosomes and dicentric chromosomes/centric rings were converted to MNi and NPB respectively, depending on the dose of radiation. Preliminary data, using FISH, suggests that NPB often represent DNA from a structural rearrangement involving only one or two homologous chromosomes. The results from this study validate the inclusion of NPB in the CBMN assay which provides a valuable measure of chromosome breakage/ rearrangement that was otherwise not available in the micronucleus assay. The CBMN assay allows NPB measurement to be achieved reliably because inhibition of cytokinesis prevents the loss of NPB that would otherwise occur if cells were allowed to divide

  7. Combined cytokinesis-block micronucleus and chromosomal aberration assay for the evaluation of radiosensitizers at low radiation doses

    International Nuclear Information System (INIS)

    Oya, Natsuo; Shibamoto, Yuta; Shibata, Toru

    1994-01-01

    Several methods have been tried for evaluating the efficacy of hypoxic cell radiosensitizers at clinically relevant low radiation doses (1-4 Gy). The cytokinesis-block micronucleus assay is known to be useful for both the in vitro and in vivo evaluation of radiosensitizers, while the chromosomal aberration assay has been commonly used to assess the mutagenicity of various agents. In the present study, the chromosomal aberration assay and the cytokinesis-block micronucleus assay were performed simultaneously to assess the radiosensitizing effect of etanidazole and KU-2285 at low radiation doses. The correlation between the two assays was also evaluated. In vitro study: EMT-6 cells were irradiated at a dose of 1-3 Gy under hypoxic conditions with or without the drugs at 1 mM. In vivo-in vitro study: EMT-6 tumor-bearing BALB/c mice received 2-4 Gy of radiation with or without administration of the drugs at 200 mg/kg. Single-cell suspensions were then obtained in both studies and were used for the cytokinesis-block micronucleus assay and the chromosomal aberration assay. The micronucleus frequency in binucleate cells was evaluated in the former assay, and the frequency of chromosomal aberrations in metaphase cells was evaluated in the latter assay. In vitro study: the sensitizer enhancement ratios of etanidazole and KU-2285 were 1.73 and 2.21, respectively, in the micronucleus assay, and 1.41 and 1.79 in the chromosomal aberration assay. In vivo-in vitro study: the sensitizer enhancement ratios of etanidazole and KU-2285 were 1.18 and 1.31, respectively, in the micronucleus assay, and 1.16 and 1.42 in the chromosomal aberration assay. In both studies, a linear correlation was observed between the micronucleus frequency and the chromosomal aberration frequency. The background (i.e., the frequency at 0 Gy) of the latter assay was considerably lower than that of the former assay, especially in the in vivo study. 31 refs., 4 figs

  8. Commentary: critical questions, misconceptions and a road map for improving the use of the lymphocyte cytokinesis-block micronucleus assay for in vivo biomonitoring of human exposure to genotoxic chemicals-a HUMN project perspective.

    Science.gov (United States)

    Kirsch-Volders, Micheline; Bonassi, Stefano; Knasmueller, Siegfried; Holland, Nina; Bolognesi, Claudia; Fenech, Michael F

    2014-01-01

    The lymphocyte cytokinesis-block micronucleus (CBMN) assay has been applied in hundreds of in vivo biomonitoring studies of humans exposed to genotoxic chemicals because it allows the measurement of both structural and numerical chromosome aberrations. The CBMN cytome assay version which, apart from measuring micronuclei (MN) already present in cells in vivo or expressed ex vivo, also includes measurement of nucleoplasmic bridges (NPB), nuclear buds (NBUD), necrosis and apoptosis, is also increasingly being used in such studies. Because of the numerous published studies there is now a need to re-evaluate the use of MN and other biomarkers within the lymphocyte CBMN cytome assay as quantitative indicators of exposure to chemical genotoxins and the genetic hazard this may cause. This review has identified some important misconceptions as well as knowledge gaps that need to be addressed to make further progress in the proper application of this promising technique and enable its full potential to be realised. The HUMN project consortium recommends a three pronged approach to further improve the knowledge base and application of the lymphocyte CBMN cytome assay to measure DNA damage in humans exposed to chemical genotoxins: (i) a series of systematic reviews, one for each class of chemical genotoxins, of studies which have investigated the association of in vivo exposure in humans with MN, NPB and NBUD induction in lymphocytes; (ii) a comprehensive analysis of the literature to obtain new insights on the potential mechanisms by which different classes of chemicals may induce MN, NPB and NBUD in vitro and in vivo and (iii) investigation of the potential advantages of using the lymphocyte CBMN cytome assay in conjunction with other promising complementary DNA damage diagnostics to obtain an even more complete assessment of the DNA damage profile induced by in vivo exposure to chemical genotoxins in humans. Copyright © 2014 The Authors. Published by Elsevier B.V. All

  9. Lack of genotoxicity of potassium iodate in the alkaline comet assay and in the cytokinesis-block micronucleus test. Comparison to potassium bromate.

    Science.gov (United States)

    Poul, J M; Huet, S; Godard, T; Sanders, P

    2004-02-01

    Iodine could be added to the diet of human population in the form of iodide or iodate but iodate had not been adequately tested for genotoxicity and carcinogenicity. In the present study, genotoxic effects of potassium iodate were evaluated in vitro using the alkaline comet assay and the cytokinesis-block micronucleus assay on CHO cells and compared to halogenate salt analogues potassium bromate and chlorate and also to their respective reduced forms (potassium iodide, bromide and chloride). The results showed that the comet assay failed to detect the presence of DNA damage after a treatment of cells by potassium iodate for concentrations up to 10 mM. This absence of primary DNA damage was confirmed in the cytokinesis-block micronucleus assay. In the same way, results showed that potassium chlorate as well as potassium iodide, bromide and chloride did not induced DNA damage in the alkaline comet assay for doses up to 10 mM. By contrast, potassium bromate exposure led to an increase in both DNA damage and frequency of micronucleated cells. The repair of bromate-induced DNA damage was incomplete 24 h after the end of treatment. These results seem to indicate that potassium bromate would induce DNA damage by several mechanisms besides oxidative stress.

  10. Test of micronucleus in lymphocytes with the cytokinesis-block like possible indicator of the answer of the patient to the radiotherapy

    International Nuclear Information System (INIS)

    Giorgio, Marina di; Nasazzi, Nora; Taja, Maria R.; Roth, Berta; Sardi, Mabel; Menendez, Pablo R.

    2001-01-01

    In order to evaluate the individual cytogenetic response to radiotherapy and its comparison with the clinical response, the cytokinesis-block micronucleus assay was applied to peripheral blood lymphocytes of patients with cervix cancer undergoing radiotherapy. The cytogenetic data were analysed using a mathematical model to evaluate the attenuation of the cytogenetic effect as a function of the time between a single exposure and blood sampling, estimating a cytogenetic recovery factor (k) that might correlate with the individual radiosensitivity, contributing with radiosensitivity tests of current use but applying a rapid methodology easy to implement in a routine clinical laboratory. Long term clinical observations could confirm the validity of k in expressing predisposition of the subject to develop delayed effects. (author)

  11. Hydrocortisone Increases the Vinblastine-Induced Chromosomal Damages in L929 Cells Investigated by the Micronucleus Assay on Cytokinesis-Blocked Binucleated Cells

    Directory of Open Access Journals (Sweden)

    Tahere Ebrahimipour

    2017-03-01

    Full Text Available Background: Stress may cause damages to DNA or/and change the ability of the cells to overcome these damages. It may also cause irregularities in the cell cycle and induce abnormal cell divisions through glucocorticoid-dependent functions. The abnormal cell divisions, in turn, lead to chromosomal mal-segregation and aneuploidy. In this study, the effects of the stress hormone, hydrocortisone (HYD, were investigated on the induced chromosomal abnormalities by vinblastine (VIN during cell cycle in L929 cells. Methods: This work was performed in winter 2013 at Department of Biology, University of Ferdowsi, Mashhad, Iran. Cultured cells were divided into different groups including control, VIN-treated, HYD treated and VIN+HYD co-treated cells. The induced chromosomal damages were investigated by micronucleus assay in cytokinesis-blocked binucleated cells. Results: Although HYD by itself did not increase the micronuclei (Mn frequency, co-treatment of cells with VIN and HYD led to significant increase (P<0.05 in the frequency of Mn in comparison to control and VIN treated groups. Conclusion: Cells treated with stress hormone are more sensitive to damages induced by VIN. Therefore, stress may not directly result in genetic instability, it can increase the harmful effects associated with other genotoxic agents.

  12. Does the recommended lymphocyte cytokinesis-block micronucleus assay for human biomonitoring actually detect DNA damage induced by occupational and environmental exposure to genotoxic chemicals?

    Science.gov (United States)

    Speit, Günter

    2013-07-01

    This commentary challenges the paradigm that the cytokinesis-block micronucleus assay (CBMN assay) with cultured human lymphocytes, as it is performed currently, is a sensitive and useful tool for detecting genotoxic effects in populations exposed occupationally or environmentally to genotoxic chemicals. Based on the principle of the assay and the available data, increased micronucleus (MN) frequencies in binucleated cells (BNC) are mainly due to MN produced in vitro during the cultivation period (i.e. MN produced in vivo do not substantially contribute to the MN frequency measured in BNC). The sensitivity of the assay for the detection of induced MN in BNC after an in vivo exposure to a genotoxic chemical is limited because cytochalasin B (Cyt-B) is added relatively late during the culture period and, therefore, the BNC that are scored do not always represent cells that have completed one cell cycle only. Furthermore, this delay means that damaged cells can be eliminated by apoptosis and/or that DNA damage induced in vivo can be repaired prior to the production of a MN in the presence of Cyt-B. A comparison with the in vitro CBMN assay used for genotoxicity testing leads to the conclusion that it is highly unlikely that DNA damage induced in vivo is the cause for increased MN frequencies in BNC after occupational or environmental exposure to genotoxic chemicals. This commentary casts doubt on the usefulness of the CBMN assay as an indicator of genotoxicity in human biomonitoring and questions the relevance of many published data for hazard identification and risk assessment. Thus, it seems worthwhile to reconsider the use of the CBMN assay as presently conducted for the detection of genotoxic exposure in human biomonitoring.

  13. Dose response curve for micronucleus of cytokinesis-block method in human lymphocytes after 60Co-gamma ray exposure

    International Nuclear Information System (INIS)

    Gao Jinsheng; Zheng Siying; Cai Feng

    1993-08-01

    The micronucleus technique of cytokines block has been proposed as a new method to measure chromosome damage in cytogenetic. The cytokines is blocked by using cytochalasin B (Cyt-B), and micronuclei are scored in cytokines-blocked (CB) cells. This can easily be done owing to the appearance of binucleate cells and large numbers accumulated by adding 3.0 μg/ml cytochalasin B at 44 hours and scoring at 72 hours. The results show that the optimum concentration of Cyt-B is 3.0 μg/ml. the Cyt-B itself can not induce the increase of micronuclei. The micronucleus frequency of normal individuals in vivo, there is an approximately linear relationship between the frequency of induced micronuclei and irradiation dose. The formula is Y 0.36 D + 2.74 (γ 2 = 0.995 P<0.01). Because the cytokines block method is simple and reliable, it is effective for assaying chromosome damage caused by genetic toxic materials

  14. The effect of an optimized imaging flow cytometry analysis template on sample throughput in the reduced culture cytokinesis-block micronucleus assay

    International Nuclear Information System (INIS)

    Rodrigues, M.A.; Beaton-Green, L.A.; Wilkins, R.C.; Probst, C.E.

    2016-01-01

    In cases of overexposure to ionizing radiation, the cytokinesis-block micronucleus (CBMN) assay can be performed in order to estimate the dose of radiation to an exposed individual. However, in the event of a large-scale radiation accident with many potentially exposed casualties, the assay must be able to generate accurate dose estimates to within ±0.5 Gy as quickly as possible. The assay has been adapted to, validated and optimized on the ImageStream"X imaging flow cyto-meter. The ease of running this automated version of the CBMN assay allowed investigation into the accuracy of dose estimates after reducing the volume of whole blood cultured to 200 μl and reducing the culture time to 48 h. The data analysis template used to identify binucleated lymphocyte cells (BNCs) and micronuclei (MN) has since been optimized to improve the sensitivity and specificity of BNC and MN detection. This paper presents a re-analysis of existing data using this optimized analysis template to demonstrate that dose estimations from blinded samples can be obtained to the same level of accuracy in a shorter data collection time. Here, we show that dose estimates from blinded samples were obtained to within ±0.5 Gy of the delivered dose when data collection time was reduced by 30 min at standard culture conditions and by 15 min at reduced culture conditions. Reducing data collection time while retaining the same level of accuracy in our imaging flow cytometry-based version of the CBMN assay results in higher throughput and further increases the relevancy of the CBMN assay as a radiation bio-dosimeter. (authors)

  15. Systematic review of the use of the lymphocyte cytokinesis-block micronucleus assay to measure DNA damage induced by exposure to polycyclic aromatic hydrocarbons

    Czech Academy of Sciences Publication Activity Database

    Šrám, Radim; Švecová, Vlasta; Rössnerová, Andrea

    2016-01-01

    Roč. 770, oct - dec (2016), s. 162-169 ISSN 1383-5742 R&D Projects: GA ČR(CZ) GA13-13458S Institutional support: RVO:68378041 Keywords : cytokinesis blocked micronuclei * peripheral blood lymphocytes * polycyclic aromatic hydrocarbons Subject RIV: DN - Health Impact of the Environment Quality Impact factor: 5.500, year: 2016

  16. The buccal cytome and micronucleus frequency is substantially altered in Down's syndrome and normal ageing compared to young healthy controls

    International Nuclear Information System (INIS)

    Thomas, Philip; Harvey, Sarah; Gruner, Tini; Fenech, Michael

    2008-01-01

    The buccal micronucleus cytome assay was used to investigate biomarkers for DNA damage, cell death and basal cell frequency in buccal cells of healthy young, healthy old and young Down's syndrome cohorts. With normal ageing a significant increase in cells with micronuclei (P < 0.05, average increase +366%), karyorrhectic cells (P < 0.001, average increase +439%), condensed chromatin cells (P < 0.01, average increase +45.8%) and basal cells (P < 0.001, average increase +233%) is reported relative to young controls. In Down's syndrome we report a significant increase in cells with micronuclei (P < 0.001, average increase +733%) and binucleated cells (P < 0.001, average increase +84.5%) and a significant decrease in condensed chromatin cells (P < 0.01, average decrease -52%), karyolytic cells (P < 0.001, average decrease -51.8%) and pyknotic cells (P < 0.001, average decrease -75.0%) relative to young controls. These changes show distinct differences between the cytome profile of normal ageing relative to that for a premature ageing syndrome, and highlight the diagnostic value of the cytome approach for measuring the profile of cells with DNA damage, cell death and proportion of cells with proliferative potential (i.e., basal cells). Significant correlations amongst cell death biomarkers observed in this study were used to propose a new model of the inter-relationship of cell types scored within the buccal micronucleus cytome assay. This study validates the use of a cytome approach to investigate DNA damage, cell death and cell proliferation in buccal cells with ageing

  17. Test of micronucleus in lymphocytes with the cytokinesis-block like possible indicator of the answer of the patient to the radiotherapy; Ensayo de micronucleos en linfocitos con bloqueo de la citocinesis como posible indicador de la respuesta del paciente a la radioterapia

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    Giorgio, Marina di; Nasazzi, Nora; Taja, Maria R. [Autoridad Regulatoria Nuclear, Buenos Aires (Argentina); Roth, Berta [Instituto de Oncologia Angel H. Roffo, Buenos Aires (Argentina); Sardi, Mabel; Menendez, Pablo R. [Hospital Italiano, Buenos Aires (Argentina)

    2001-07-01

    In order to evaluate the individual cytogenetic response to radiotherapy and its comparison with the clinical response, the cytokinesis-block micronucleus assay was applied to peripheral blood lymphocytes of patients with cervix cancer undergoing radiotherapy. The cytogenetic data were analysed using a mathematical model to evaluate the attenuation of the cytogenetic effect as a function of the time between a single exposure and blood sampling, estimating a cytogenetic recovery factor (k) that might correlate with the individual radiosensitivity, contributing with radiosensitivity tests of current use but applying a rapid methodology easy to implement in a routine clinical laboratory. Long term clinical observations could confirm the validity of k in expressing predisposition of the subject to develop delayed effects. (author)

  18. The buccal cytome and micronucleus frequency is substantially altered in Down's syndrome and normal ageing compared to young healthy controls

    Energy Technology Data Exchange (ETDEWEB)

    Thomas, Philip [CSIRO Human Nutrition, PO Box 10041, Adelaide BC, Adelaide, SA 5000 (Australia); Discipline of Physiology, School of Molecular and Biomedical Sciences, The University of Adelaide, Adelaide, SA 5005 (Australia)], E-mail: philip.thomas@csiro.au; Harvey, Sarah; Gruner, Tini [Southern Cross University, PO Box 157, Lismore, NSW 2480 (Australia); Fenech, Michael [CSIRO Human Nutrition, PO Box 10041, Adelaide BC, Adelaide, SA 5000 (Australia)], E-mail: michael.fenech@csiro.au

    2008-02-01

    The buccal micronucleus cytome assay was used to investigate biomarkers for DNA damage, cell death and basal cell frequency in buccal cells of healthy young, healthy old and young Down's syndrome cohorts. With normal ageing a significant increase in cells with micronuclei (P < 0.05, average increase +366%), karyorrhectic cells (P < 0.001, average increase +439%), condensed chromatin cells (P < 0.01, average increase +45.8%) and basal cells (P < 0.001, average increase +233%) is reported relative to young controls. In Down's syndrome we report a significant increase in cells with micronuclei (P < 0.001, average increase +733%) and binucleated cells (P < 0.001, average increase +84.5%) and a significant decrease in condensed chromatin cells (P < 0.01, average decrease -52%), karyolytic cells (P < 0.001, average decrease -51.8%) and pyknotic cells (P < 0.001, average decrease -75.0%) relative to young controls. These changes show distinct differences between the cytome profile of normal ageing relative to that for a premature ageing syndrome, and highlight the diagnostic value of the cytome approach for measuring the profile of cells with DNA damage, cell death and proportion of cells with proliferative potential (i.e., basal cells). Significant correlations amongst cell death biomarkers observed in this study were used to propose a new model of the inter-relationship of cell types scored within the buccal micronucleus cytome assay. This study validates the use of a cytome approach to investigate DNA damage, cell death and cell proliferation in buccal cells with ageing.

  19. Bio-monitoring for the genotoxic assessment in road construction workers as determined by the buccal micronucleus cytome assay.

    Science.gov (United States)

    Çelik, Ayla; Yildirim, Seda; Ekinci, Seda Yaprak; Taşdelen, Bahar

    2013-06-01

    Buccal micronucleus cytome (BMCyt) assay monitors genetic damage, cell proliferation and cell death in humans exposed to occupational and environmental agents. BMCyt is used as an indicator of genotoxic exposure, since it is associated with chromosomal instability. There is little research on the occupational exposure among road construction workers for genotoxicity testing. In the present study, we evaluated MN frequencies and other nuclear changes, karyorrhexis (KR), karyolysis (KL), broken egg (BE), binucleate (BN), condensed chromatin cell (CCC), and picnotic cell (PC) in buccal mucosa cells of 40 road construction workers (twenty smokers and twenty non-smokers) and 40 control groups consisting of healthy persons (twenty smokers and twenty non-smokers). Microscopic observation was performed of 2000 cells per individual in both road construction workers and control group. In control and worker groups, for each person repair index (RI) was calculated via formula KR+L/BE+MN. The results showed a statistically significant increase in the frequency of MN in buccal epithelial cells of exposed group compared with control group (proad construction workers, RI is lower than the control group. There is a significant difference between workers and control group (proad paving operations are absorbed by workers and that asphalt fume exposure is able to significantly induce cytogenetic damage in buccal mucosa cells of workers after controlling some possible confounding factors, such as age, sex and smoking habits. In addition to determination of nuclear changes and the micronucleus, the determination of RI value presents a new approach to genotoxic bio-monitoring assessment studies of occupationally exposed population. Copyright © 2013 Elsevier Inc. All rights reserved.

  20. Changes in buccal micronucleus cytome parameters associated with smokeless tobacco and pesticide exposure among female tea garden workers of Assam, India.

    Science.gov (United States)

    Kausar, Afifa; Giri, Sarbani; Roy, Prasenjit; Giri, Anirudha

    2014-03-01

    Assam is the highest tea producing state in India. A large number of workers are engaged in various units of tea industry. There are few reports on the health status of the tea garden workers. The present cytogenetic biomonitoring study was undertaken to investigate the genotoxic effect associated with workers in tea industries in southern Assam. Smokeless tobacco chewing along with betel nut is very common practice among the workers. Workers also get exposed periodically to mixture of pesticides. Employing buccal micronucleus cytome assay, exfoliated buccal cells were analyzed in 90 female tea garden and compared to 90 age and sex matched non-chewer control as well as 70 chewers who are not tea garden workers. Statistically significant (pworkers compared to both the control groups. The frequency of cell proliferation biomarkers was highest in the chewer controls whereas genotoxic and cell death parameters were highest in tea garden workers. Linear correlation analysis revealed strong positive correlation between the duration of occupation and the frequency of micronucleus (r=0.597; pworkers was relatively lower compared to the control group. Pesticide exposure and chewing areca nut along with smokeless tobacco use may be responsible for changes in cytome parameters in exfoliated buccal cells. Copyright © 2013 Elsevier GmbH. All rights reserved.

  1. Analysis in cytokinesis-blocked human lymphocytes

    International Nuclear Information System (INIS)

    Catena, C.; Mattoni, A.

    1987-01-01

    Biological dosimetry can be considered as an additional method to physical dosimetry for estimating dose absorption after exposure to ionizing radiation. Fully validated as well as new promising approaches in this field are reviewed. Recent experiments, carried out in our laboratory, on the analysis of micronuclei in cytokinesis-blocked human lymphocytes are presented. The possible relevance of differential human individual response to ionizing radiation, in view of the occurrence of radiosensitive syndromes, for the estimation of the absorbed dose in human is also discussed

  2. Detection of minor and major satellite DNA in cytokinesis-blocked mouse splenocytes by a PRINS tandem labelling approach.

    Science.gov (United States)

    Russo, A; Tommasi, A M; Renzi, L

    1996-11-01

    A protocol for the simultaneous visualization of minor and major satellite DNA by primed in situ DNA synthesis (PRINS) was developed in cytokinesis-blocked murine splenocytes. After individuation of optimal experimental conditions, a micronucleus (MN) test was carried out by treating splenocytes in vitro with the clastogenic agent mitomycin C and the aneugenic compound Colcemid. It was found that PRINS gives highly reproducible results, also comparable with the literature on MN results obtained by fluorescent in situ hybridization (FISH). Therefore the PRINS methodology may be proposed as a fast alternative to FISH for the characterization of induced MN.

  3. Cadmium chloride, benzo[a]pyrene and cyclophosphamide tested in the in vitro mammalian cell micronucleus test (MNvit) in the human lymphoblastoid cell line TK6 at Covance laboratories, Harrogate UK in support of OECD draft Test Guideline 487.

    Science.gov (United States)

    Fowler, Paul; Whitwell, James; Jeffrey, Laura; Young, Jamie; Smith, Katie; Kirkland, David

    2010-10-29

    The following genotoxic chemicals were tested in the in vitro micronucleus assay, at Covance Laboratories, Harrogate, UK in the human lymphoblastoid cell line TK6. Cadmium chloride (an inorganic carcinogen), benzo[a]pyrene (a polycyclic aromatic hydrocarbon requiring metabolic activation) and cyclophosphamide (an alkylating agent requiring metabolic activation) were treated with and without cytokinesis block (by addition of cytochalasin B). This work formed part of a collaborative evaluation of the toxicity measures recommended in the draft OECD Test Guideline 487 for the in vitro micronucleus test. The toxicity measures used, capable of detecting both cytostasis and cell death, were relative population doubling, relative increase in cell counts and relative cell counts for treatments in the absence of cytokinesis block, and replication index or cytokinesis blocked proliferation index in the presence of cytokinesis block. All of the chemicals tested gave significant increases in the percentage of micronucleated cells with and without cytokinesis block at concentrations giving approximately 60% toxicity (cytostasis and cell death) or less by all of the toxicity measures used. The outcomes from this series of tests support the use of relative increase in cell counts and relative population doubling, as well as relative cell counts, as appropriate measures of cytotoxicity for the non-cytokinesis blocked in the in vitro micronucleus assay. Copyright © 2010 Elsevier B.V. All rights reserved.

  4. Etoposide; colchicine; mitomycin C and cyclophosphamide tested in the in vitro mammalian cell micronucleus test (MNvit) in Chinese hamster lung (CHL) cells at Covance laboratories; Harrogate UK in support of OECD draft Test Guideline 487.

    Science.gov (United States)

    Fowler, Paul; Whitwell, James; Jeffrey, Laura; Young, Jamie; Smith, Katie; Kirkland, David

    2010-10-29

    The following genotoxic chemicals were tested in the in vitro micronucleus assay, at Covance Laboratories, Harrogate, UK in the Chinese hamster lung cell line CHL. Etoposide (a topoisomerase inhibitor), colchicine (an aneugen), mitomycin C (a DNA cross linking agent) and cyclophosphamide (an alkylating agent requiring metabolic activation) were treated with and without cytokinesis block (by addition of cytochalasin B). This work formed part of a collaborative evaluation of the toxicity measures recommended in the draft OECD Test Guideline 487 for the in vitro micronucleus test. The toxicity measures used, detecting both cytostasis and cell death, were relative population doubling, relative increase in cell counts and relative cell counts for treatments in the absence of cytokinesis block, and replication index or cytokinesis blocked proliferation index in the presence of cytokinesis block. All of the chemicals tested gave significant increases in the percentage of micronucleated cells with and without cytokinesis block at concentrations giving approximately 60% toxicity (cytostasis and cell death) or less by all of the toxicity measures used. The outcomes from this series of tests support the use of relative increase in cell counts and relative population doubling, as well as relative cell counts, as appropriate measures of cytotoxicity for the non-cytokinesis blocked in vitro micronucleus assay. Copyright © 2010 Elsevier B.V. All rights reserved.

  5. Automotive airborne brake wear debris nanoparticles and cytokinesis-block micronucleus assay in peripheral blood lymphocytes: A pilot study

    Energy Technology Data Exchange (ETDEWEB)

    Kazimirova, Alena, E-mail: alena.kazimirova@szu.sk [Department of Biology, Faculty of Medicine, Slovak Medical University, Limbova 12, 833 03 Bratislava (Slovakia); Peikertova, Pavlina [Nanotechnology Centre, VŠB – Technical University of Ostrava, 17. listopadu 15, 708 00 Ostrava (Czech Republic); IT4Innovations, VŠB – Technical University of Ostrava, 17. listopadu 15, 708 33 Ostrava-Poruba (Czech Republic); Barancokova, Magdalena; Staruchova, Marta; Tulinska, Jana [Department of Biology, Faculty of Medicine, Slovak Medical University, Limbova 12, 833 03 Bratislava (Slovakia); Vaculik, Miroslav [Nanotechnology Centre, VŠB – Technical University of Ostrava, 17. listopadu 15, 708 00 Ostrava (Czech Republic); Vavra, Ivo [Nanotechnology Centre, VŠB – Technical University of Ostrava, 17. listopadu 15, 708 00 Ostrava (Czech Republic); Institute of Electrical Engineering, Slovak Academy of Sciences, Dúbravská Cesta 9, 841 03 Bratislava (Slovakia); Kukutschova, Jana [Nanotechnology Centre, VŠB – Technical University of Ostrava, 17. listopadu 15, 708 00 Ostrava (Czech Republic); Filip, Peter [Department of Mechanical Engineering and Energy Processes, Southern Illinois University, Lincoln Drive 1263, 62901 Carbondale (United States); Dusinska, Maria [Health Effects Laboratory, Department of Environmental Chemistry, NILU-Norwegian Institute for Air Research, Instituttveien 18, 2007 Kjeller (Norway)

    2016-07-15

    Motor vehicle exhaust and non-exhaust processes play a significant role in environmental pollution, as they are a source of the finest particulate matter. Emissions from non-exhaust processes include wear-products of brakes, tires, automotive hardware, road surface, and traffic signs, but still are paid little attention to. Automotive friction composites for brake pads are composite materials which may consist of potentially hazardous materials and there is a lack of information regarding the potential influence of the brake wear debris (BWD) on the environment, especially on human health. Thus, we focused our study on the genotoxicity of the airborne fraction of BWD using a brake pad model representing an average low-metallic formulation available in the EU market. BWD was generated in the laboratory by a full-scale brake dynamometer and characterized by Raman microspectroscopy, scanning electron microscopy, and transmission electron microscopy showing that it contains nano-sized crystalline metal-based particles. Genotoxicity tested in human lymphocytes in different testing conditions showed an increase in frequencies of micronucleated binucleated cells (MNBNCs) exposed for 48 h to BWD nanoparticles (NPs) (with 10% of foetal calf serum in culture medium) compared with lymphocytes exposed to medium alone, statistically significant only at the concentration 3 µg/cm{sup 2} (p=0.032). - Highlights: • BWD was characterized by Raman microspectroscopy, scanning electron microscopy, and transmission electron microscopy. • Our result showed that BWD contains crystalline metal NPs. • Two different protocols for CBMN assay were used to study of genotoxicity of BWD. • We found significantly increased frequency of MNBNCs after 48 h exposure of BWD (with 10% of foetal calf serum in culture media) at the concentration 3 µg/cm{sup 2}.

  6. Automotive airborne brake wear debris nanoparticles and cytokinesis-block micronucleus assay in peripheral blood lymphocytes: A pilot study

    International Nuclear Information System (INIS)

    Kazimirova, Alena; Peikertova, Pavlina; Barancokova, Magdalena; Staruchova, Marta; Tulinska, Jana; Vaculik, Miroslav; Vavra, Ivo; Kukutschova, Jana; Filip, Peter; Dusinska, Maria

    2016-01-01

    Motor vehicle exhaust and non-exhaust processes play a significant role in environmental pollution, as they are a source of the finest particulate matter. Emissions from non-exhaust processes include wear-products of brakes, tires, automotive hardware, road surface, and traffic signs, but still are paid little attention to. Automotive friction composites for brake pads are composite materials which may consist of potentially hazardous materials and there is a lack of information regarding the potential influence of the brake wear debris (BWD) on the environment, especially on human health. Thus, we focused our study on the genotoxicity of the airborne fraction of BWD using a brake pad model representing an average low-metallic formulation available in the EU market. BWD was generated in the laboratory by a full-scale brake dynamometer and characterized by Raman microspectroscopy, scanning electron microscopy, and transmission electron microscopy showing that it contains nano-sized crystalline metal-based particles. Genotoxicity tested in human lymphocytes in different testing conditions showed an increase in frequencies of micronucleated binucleated cells (MNBNCs) exposed for 48 h to BWD nanoparticles (NPs) (with 10% of foetal calf serum in culture medium) compared with lymphocytes exposed to medium alone, statistically significant only at the concentration 3 µg/cm 2 (p=0.032). - Highlights: • BWD was characterized by Raman microspectroscopy, scanning electron microscopy, and transmission electron microscopy. • Our result showed that BWD contains crystalline metal NPs. • Two different protocols for CBMN assay were used to study of genotoxicity of BWD. • We found significantly increased frequency of MNBNCs after 48 h exposure of BWD (with 10% of foetal calf serum in culture media) at the concentration 3 µg/cm 2 .

  7. Automotive airborne brake wear debris nanoparticles and cytokinesis-block micronucleus assay in peripheral blood lymphocytes: A pilot study.

    Science.gov (United States)

    Kazimirova, Alena; Peikertova, Pavlina; Barancokova, Magdalena; Staruchova, Marta; Tulinska, Jana; Vaculik, Miroslav; Vavra, Ivo; Kukutschova, Jana; Filip, Peter; Dusinska, Maria

    2016-07-01

    Motor vehicle exhaust and non-exhaust processes play a significant role in environmental pollution, as they are a source of the finest particulate matter. Emissions from non-exhaust processes include wear-products of brakes, tires, automotive hardware, road surface, and traffic signs, but still are paid little attention to. Automotive friction composites for brake pads are composite materials which may consist of potentially hazardous materials and there is a lack of information regarding the potential influence of the brake wear debris (BWD) on the environment, especially on human health. Thus, we focused our study on the genotoxicity of the airborne fraction of BWD using a brake pad model representing an average low-metallic formulation available in the EU market. BWD was generated in the laboratory by a full-scale brake dynamometer and characterized by Raman microspectroscopy, scanning electron microscopy, and transmission electron microscopy showing that it contains nano-sized crystalline metal-based particles. Genotoxicity tested in human lymphocytes in different testing conditions showed an increase in frequencies of micronucleated binucleated cells (MNBNCs) exposed for 48h to BWD nanoparticles (NPs) (with 10% of foetal calf serum in culture medium) compared with lymphocytes exposed to medium alone, statistically significant only at the concentration 3µg/cm(2) (p=0.032). Copyright © 2016 Elsevier Inc. All rights reserved.

  8. Scoring of radiation-induced micronuclei in cytokinesis-blocked human lymphocytes by automated image analysis

    International Nuclear Information System (INIS)

    Verhaegen, F.; Seuntjens, J.; Thierens, H.

    1994-01-01

    The micronucleus assay in human lymphocytes is, at present, frequently used to assess chromosomal damage caused by ionizing radiation or mutagens. Manual scoring of micronuclei (MN) by trained personnel is very time-consuming, tiring work, and the results depend on subjective interpretation of scoring criteria. More objective scoring can be accomplished only if the test can be automated. Furthermore, an automated system allows scoring of large numbers of cells, thereby increasing the statistical significance of the results. This is of special importance for screening programs for low doses of chromosome-damaging agents. In this paper, the first results of our effort to automate the micronucleus assay with an image-analysis system are represented. The method we used is described in detail, and the results are compared to those of other groups. Our system is able to detect 88% of the binucleated lymphocytes on the slides. The procedure consists of a fully automated localization of binucleated cells and counting of the MN within these cells, followed by a simple and fast manual operation in which the false positives are removed. Preliminary measurements for blood samples irradiated with a dose of 1 Gy X-rays indicate that the automated system can find 89% ± 12% of the micronuclei within the binucleated cells compared to a manual screening. 18 refs., 8 figs., 1 tab

  9. Micronuclei in cytokinesis-blocked lymphocytes of medical personnel occupationally exposed to ionizing radiation

    International Nuclear Information System (INIS)

    Noditi, M.; Draghia, L.; Popescu, D.; Cincu, E.

    2006-01-01

    Bio monitoring of occupational exposures relies on surveillance of exposure and its biological consequences. The measurement of micronuclei in population of exposed cells is a cytogenetic end point used for estimation purposes. To ensure that only dividing cells are scored, cells are treated with cytochalasin B, which blocks cytokinesis and results in bi nucleated cells. Only the bi nucleated cells are evaluated for the formation of micronuclei. In 2004-2005 there have been analyzed and compared two groups of medical staff occupationally exposed to X-rays by using micronucleus test. The first group consisted in 9 doctors and nurses specialized in interventional cardiology from the Institute of Cardiology - Timisoara, Romania, males and females, smokers and nonsmokers. The mean age was 41.5 years and the mean duration of employment 10.6 years. According to personal dosimeters, some of them have had an overdose exposure. The other group consisted in 19 doctors and nurses from the radiology department of several hospitals from Timisoara with the mean age of 48.3 years and the mean duration of employment of 17.8 years. According to personal dosimeters, none of them have had an overdose exposure. The recorded frequency of micronuclei was 62.6/1000 bi nucleated cells for the interventional cardiology personnel. There have been observed cells with 2, 3 and even 5 micronuclei. For the radiology department personnel the frequency of micronuclei was 15.4/1000 bi nucleated cells and the appearance of cells with more than one micronucleus was exceptional. In general, there was a tendency of accumulation of micronuclei with age but the correlation with the age of employment was rather unclear. Due to low doses exposure confounding factors could exist. For instance, a proportion of micronuclei are formed because of other mutagen factors from the environment or smoking habit. Nevertheless the exposure of interventional cardiology personnel is more consistent and further

  10. Micronuclei in cytokinesis-blocked lymphocytes of medical personnel occupationally exposed to ionizing radiation

    Energy Technology Data Exchange (ETDEWEB)

    Noditi, M.; Draghia, L.; Popescu, D. [Institute of Public Health ' Prof. Dr. Leonida Georgescu' Timisoara (Romania); Cincu, E. [University of Agriculture Sciences of Banat, Timisoara (Romania)

    2006-07-01

    Bio monitoring of occupational exposures relies on surveillance of exposure and its biological consequences. The measurement of micronuclei in population of exposed cells is a cytogenetic end point used for estimation purposes. To ensure that only dividing cells are scored, cells are treated with cytochalasin B, which blocks cytokinesis and results in bi nucleated cells. Only the bi nucleated cells are evaluated for the formation of micronuclei. In 2004-2005 there have been analyzed and compared two groups of medical staff occupationally exposed to X-rays by using micronucleus test. The first group consisted in 9 doctors and nurses specialized in interventional cardiology from the Institute of Cardiology - Timisoara, Romania, males and females, smokers and nonsmokers. The mean age was 41.5 years and the mean duration of employment 10.6 years. According to personal dosimeters, some of them have had an overdose exposure. The other group consisted in 19 doctors and nurses from the radiology department of several hospitals from Timisoara with the mean age of 48.3 years and the mean duration of employment of 17.8 years. According to personal dosimeters, none of them have had an overdose exposure. The recorded frequency of micronuclei was 62.6/1000 bi nucleated cells for the interventional cardiology personnel. There have been observed cells with 2, 3 and even 5 micronuclei. For the radiology department personnel the frequency of micronuclei was 15.4/1000 bi nucleated cells and the appearance of cells with more than one micronucleus was exceptional. In general, there was a tendency of accumulation of micronuclei with age but the correlation with the age of employment was rather unclear. Due to low doses exposure confounding factors could exist. For instance, a proportion of micronuclei are formed because of other mutagen factors from the environment or smoking habit. Nevertheless the exposure of interventional cardiology personnel is more consistent and further

  11. Micronucleus assay for radiation workers

    International Nuclear Information System (INIS)

    Balasem, A.N.; Ali, A.S.K.

    1997-01-01

    Micronucleus assay was performed on 49 radiation workers and 22 healthy volunteers. Radiation workers were subdivided into two groups according to their employments durations in the radiation field. Group a consisted of 18 radiation workers who have been in this work between 5 and 22 years. Group b included 31 employees who have been classified as radiation workers for 1 to 4.5 years. Statistical analysis showed significant variations between the yields of micronuclei in groups A and B as well as between group A and a group of healthy controls. Meanwhile no significant difference was noticed between the yields of micronuclei in group B and the corresponding values in the healthy controls. The possible effect of age in the induction of micronuclei was discussed and a comparison with the yield of chromosomal aberrations was described. It seems that cytokinesis- blocking method may be used to detect the radiation-induced micronuclei in workers exposed occupationally to ionizing radiation in levels below the maximum permissible limit of 0.05 Sv per year

  12. HUMN project initiative and review of validation, quality control and prospects for further development of automated micronucleus assays using image cytometry systems

    Czech Academy of Sciences Publication Activity Database

    Fenech, M.; Kirsch-Volders, M.; Rössnerová, Andrea; Šrám, Radim; Romm, H.; Bolognesi, C.; Ramakumar, A.; Soussaline, F.; Schunck, CH.; Elhajouji, A.; Anwar, W.; Bonassi, S.

    2013-01-01

    Roč. 216, č. 5 (2013), s. 541-552 ISSN 1438-4639 R&D Projects: GA ČR GAP503/11/0084 Grant - others:Project NewGenesis(XE) FOOD -CT-2005-016320 Institutional support: RVO:68378041 Keywords : Micronucleus * Cytokinesis-block * Automation Subject RIV: DN - Health Impact of the Environment Quality Impact factor: 3.276, year: 2013

  13. 5-Fluorouracil, colchicine, benzo[a]pyrene and cytosine arabinoside tested in the in vitro mammalian cell micronucleus test (MNvit) in Chinese hamster V79 cells at Covance Laboratories, Harrogate, UK in support of OECD draft Test Guideline 487.

    Science.gov (United States)

    Whitwell, James; Fowler, Paul; Allars, Sarah; Jenner, Karen; Lloyd, Melvyn; Wood, Debbie; Smith, Katie; Young, Jamie; Jeffrey, Laura; Kirkland, David

    2010-10-29

    The reference genotoxic agents 5-fluorouracil (a nucleoside analogue, characterised by a steep dose response profile), colchicine (an aneugen that inhibits tubulin polymerisation), benzo[a]pyrene (a polycyclic aromatic hydrocarbon requiring metabolic activation) and cytosine arabinoside (a nucleoside analogue that inhibits the gap-filling step of excision repair) were tested in the in vitro micronucleus assay using the Chinese hamster V79 cell line at Covance Laboratories, Harrogate, UK. All chemicals were treated in the absence and presence of cytokinesis block (via addition of cytochalasin B) with this work forming part of a collaborative evaluation of the toxicity measures recommended in the draft OECD Test Guideline 487 on the In Vitro Mammalian Cell Micronucleus Test (MNvit). The toxicity measures used, detecting a possible combination of both cytostasis and cell death (though not cell death directly), were relative population doubling, relative increase in cell counts and relative cell counts for treatments in the absence of cytokinesis block, and replication index in the presence of cytokinesis block. All of the chemicals tested either gave marked increases in the percentage of micronucleated cells with and without cytokinesis block, or did not induce micronuclei at concentrations giving approximately 50-60% toxicity (cytostasis and cell death) or less by all of the toxicity measures used. The outcome from this series of tests supports the use of relative increase in cell counts and relative population doubling, as well as relative cell counts, as appropriate measures of cytotoxicity for the non-cytokinesis blocked in vitro micronucleus assay. Copyright © 2010 Elsevier B.V. All rights reserved.

  14. DNA and chromosomal damage in medical workers exposed to anaesthetic gases assessed by the lymphocyte cytokinesis-block micronucleus (CBMN) assay. A critical review

    Czech Academy of Sciences Publication Activity Database

    Vodička, Pavel; Mušák, L.; Fiorito, G.; Vymetálková, Veronika; Vodičková, Ludmila; Naccarati, Alessio

    2016-01-01

    Roč. 770, SI (2016), s. 26-34 ISSN 1383-5742 R&D Projects: GA MŠk(CZ) LD14050; GA ČR(CZ) GA15-14789S Institutional support: RVO:68378041 Keywords : micronuclei * anaesthetics gas exposure * operating room personnel Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.500, year: 2016

  15. 2-Aminoanthracene, 5-fluorouracil, colchicine, benzo[a]pyrene, cadmium chloride and cytosine arabinoside tested in the in vitro mammalian cell micronucleus test (MNvit) in Chinese hamster ovary (CHO) cells at Covance Laboratories, Harrogate UK in support of OECD draft Test Guideline 487.

    Science.gov (United States)

    Whitwell, James; Fowler, Paul; Allars, Sarah; Jenner, Karen; Lloyd, Melvyn; Wood, Debbie; Smith, Katie; Young, Jamie; Jeffrey, Laura; Kirkland, David

    2010-10-29

    The reference genotoxic agents 2-aminoanthracene (a metabolism dependent weak clastogen), 5-fluorouracil (a nucleoside analogue, characterised by a steep dose response profile), colchicine (an aneugen that inhibits tubulin polymerisation), benzo[a]pyrene (a polycyclic aromatic hydrocarbon requiring metabolic activation), cadmium chloride (an inorganic carcinogen), and cytosine arabinoside (a nucleoside analogue that inhibits the gap-filling step of excision repair) were tested in the in vitro micronucleus assay using the Chinese hamster ovary (CHO) cell line at Covance Laboratories, Harrogate, UK. All chemicals were treated in the absence and presence of cytokinesis block (via addition of cytochalasin B) with this work forming part of a collaborative evaluation of the toxicity measures recommended in the draft OECD Test Guideline 487 on the In vitro Mammalian Cell Micronucleus Test (MNvit). The toxicity measures used, detecting a possible combination of both cytostasis and cell death (though not cell death directly), were relative population doubling, relative increase in cell counts and relative cell counts for treatments in the absence of cytokinesis block, and replication index in the presence of cytokinesis block. All of the chemicals tested either gave marked positive increases in the percentage of micronucleated cells with and without cytokinesis block, or did not induce micronuclei at concentrations giving approximately 50-60% toxicity (cytostasis and cell death) or less by all of the toxicity measures used. The outcome from this series of tests supports the use of relative increase in cell counts and relative population doubling, as well as relative cell counts, as appropriate measures of cytotoxicity for the non-cytokinesis blocked in vitro micronucleus assay. Copyright © 2010 Elsevier B.V. All rights reserved.

  16. Analysis of resveratrol and radiation effects in lung cancer cells by micronucleus assay

    Energy Technology Data Exchange (ETDEWEB)

    Moreno, Carolina S.; Santos, Dymes R.A.; Vieira, Daniel P.; Rogero, Sizue O.; Rogero, Jose R., E-mail: carolina_sm@hotmail.com [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Sakuraba, Roberto K.; Weltman, Eduardo [Hospital Israelita Albert Einstein, Sao Paulo, SP (Brazil); Cruz, Aurea S.; Santos, Rezolina P. [Instituto Adolfo Lutz, Sao Paulo, SP (Brazil)

    2015-07-01

    Mucoepidermoid lung carcinoma is frequently manifested by obstructive trachea symptoms. Radiation and drugs combinations are commonly used in the lung cancer treatment. Currently there is a strong tendency to develop therapeutic strategies focused at the administration of high potential compounds to improve the ionizing radiation treatments, so as to increase the radiation effects on tumor cell while minimizing these effects to surrounding normal tissues. Resveratrol is a polyphenolic phytoalexin compound present in wines and several plants. This compound has a broad spectrum of biological activities such as antioxidant, anticarcinogenic, and induction of cell cycle arrest effects. Analysis of biological effects of ionizing radiation in the presence of resveratrol in different cell cultures has been the subject of many studies. To verify the genotoxic effects in cells exposed to ionizing radiation many methods have been proposed. The cytokinesis-block micronucleus technique is one of the preferred methods. The main of this study was to detect and quantify radioinduced DNA damage in mucoepidermoid lung carcinoma cells (NCI-H292) by cytokinesis-block micronucleus technique using cytocalasin-B. The cell culture was irradiated at a single fraction from a TrueBeam® linear accelerator (0, 0.8, 5, and 10 Gy), in the absence or presence of different resveratrol concentrations (0, 15, 30, and 60 μM). The results showed that resveratrol (15 and μM) induced significant increase frequency (p<0.05) of micronucleus formation in NCI-H292 cell culture non-irradiated and exposed at 5 Gy dose. Moreover, resveratrol (30 μM) induced micronucleus formation at 0.8 Gy dose. (author)

  17. Analysis of resveratrol and radiation effects in lung cancer cells by micronucleus assay

    International Nuclear Information System (INIS)

    Moreno, Carolina S.; Santos, Dymes R.A.; Vieira, Daniel P.; Rogero, Sizue O.; Rogero, Jose R.; Sakuraba, Roberto K.; Weltman, Eduardo; Cruz, Aurea S.; Santos, Rezolina P.

    2015-01-01

    Mucoepidermoid lung carcinoma is frequently manifested by obstructive trachea symptoms. Radiation and drugs combinations are commonly used in the lung cancer treatment. Currently there is a strong tendency to develop therapeutic strategies focused at the administration of high potential compounds to improve the ionizing radiation treatments, so as to increase the radiation effects on tumor cell while minimizing these effects to surrounding normal tissues. Resveratrol is a polyphenolic phytoalexin compound present in wines and several plants. This compound has a broad spectrum of biological activities such as antioxidant, anticarcinogenic, and induction of cell cycle arrest effects. Analysis of biological effects of ionizing radiation in the presence of resveratrol in different cell cultures has been the subject of many studies. To verify the genotoxic effects in cells exposed to ionizing radiation many methods have been proposed. The cytokinesis-block micronucleus technique is one of the preferred methods. The main of this study was to detect and quantify radioinduced DNA damage in mucoepidermoid lung carcinoma cells (NCI-H292) by cytokinesis-block micronucleus technique using cytocalasin-B. The cell culture was irradiated at a single fraction from a TrueBeam® linear accelerator (0, 0.8, 5, and 10 Gy), in the absence or presence of different resveratrol concentrations (0, 15, 30, and 60 μM). The results showed that resveratrol (15 and μM) induced significant increase frequency (p<0.05) of micronucleus formation in NCI-H292 cell culture non-irradiated and exposed at 5 Gy dose. Moreover, resveratrol (30 μM) induced micronucleus formation at 0.8 Gy dose. (author)

  18. Low dose effects detected by micronucleus assay in lymphocytes

    International Nuclear Information System (INIS)

    Koeteles, G.J.; Bojtor, I.; Kubasova, T.; Horvath, G.

    1997-01-01

    The effects of low doses of X-rays between 0.01 and 1 Gy were studied on whole blood samples of various individuals using the cytokinesis-blocked lymphocyte micronucleus assay as an endpoint. The adaptive response could be induced in G 0 cells by 0.01 Gy followed by 1 Gy challenging dose within a time period of 8 hours, in vitro. The probability distribution of micronucleus increments in those samples which had received very low doses in the range 0.01-0.05 Gy proved to be of asymmetrical type (i.e. lognormal) -very likely to the same shape which has been verified for unirradiated (control) population - while the variable turned to be normally distributed at or above 1 Gy. Profound changes have been experienced in the main characteristics of the linear dose - response relationship and in regression parameters, as well, when successively lessened dose ranges were studied toward 0.01 Gy. In the range below ∼ 0.2 Gy the response were found to be unrelated to the absorbed dose. These findings suggest that in (very) low dose range a higher attention should be needed to biological parameters like repair, protective mechanisms and antioxidant capacities, rather than to the absorbed radiation energy only. (author)

  19. Micronucleus frequency in Danish schoolchildren and their mothers from the DEMOCOPHES population

    DEFF Research Database (Denmark)

    Mørck, Thit A.; Vande Loock, Kim; Poulsen, Maria Bech

    2016-01-01

    organic pollutants and dioxin-like activity measured in the sameparticipants. The MN frequency analysis was performed with the cytokinesis-block micronucleus(CBMN) assay and included 100 children and 119 mothers. We found a significant correlationbetween mothers and children in the levels of micronuclei...... in 1000 binucleated T lymphocytes(‰MNBN) and in the proliferation index. Further the levels of ‰MNBN were significantly higherin mothers compared with their children. No significant associations were found for ‰MNBNfor traffic related exposure in neither children nor their mothers. In children, a 2.......5 times highermicronuclei in mononuclear T lymphocytes were found in children living within 50 m of a busyroad, however, this was not found in mothers or in MNBN and the effect of exposure to road trafficon MN frequency needs further investigation. No significant associations were found between...

  20. Micronucleus as biomarkers of cancer risk in anabolic androgenic steroids users.

    Science.gov (United States)

    Souza, L da Cunha Menezes; da Cruz, L A; Cerqueira, E de Moraes Marcílio; Meireles, Jrc

    2017-03-01

    The use of anabolic androgenic steroids (AAS) has grown among practitioners of recreational bodybuilding, with significant contributions of designer steroids, aiming muscle hypertrophy in healthy subjects. The abusive use of AAS in general is associated with adverse effects; one of the most worrisome is cancer development. The aim of this study was to evaluate the effectiveness of the cytokinesis block micronucleus (CBMN) test in human lymphocytes in identifying risk groups for cancer development in users of AAS. Blood was collected from 15 AAS users bodybuilders (G1), 20 non-users bodybuilders (G2) and 20 non-users sedentary (G3). MN analysis was performed on a minimum of 1000 binucleated lymphocytes. The occurrence of MN was significantly higher ( p < 0.05) in individuals of G1 compared to G2 and G3. The results indicate the sensitivity of CBMN in human lymphocytes in the identification of chromosomal damage in consequence of AAS.

  1. Micronucleus formation in cultured human keratinocytes following exposure to mitomycin C and cyclophosphamide.

    Science.gov (United States)

    van Pelt, F N; Haring, R M; Overkamp, M J; Weterings, P J

    1991-02-01

    A method is described to investigate the induction of micronuclei in cultured human keratinocytes after short-term exposure to known clastogenic agents. The cytokinesis-block method was applied to facilitate the scoring of micronucleated cells. Mitomycin C, a direct-acting compound, caused a 5-20-fold increase in micronuclei over the controls at the highest concentration tested (1 microgram/ml). Cyclophosphamide, an agent requiring metabolic activation, did not induce the formation of micronuclei in cultured keratinocytes. However, after pretreatment of the keratinocyte cultures with Aroclor 1254 for 72 h, exposure to cyclophosphamide resulted in a 3-fold increase in micronucleus frequency over the controls. No cytogenetic effect of Aroclor 1254 was observed in control experiments.

  2. Radiation risk assessment in professionals working in dental radiology area using buccal micronucleus cytome assay.

    Science.gov (United States)

    Sadatullah, Syed; Dawasaz, Ali Azhar; Luqman, Master; Assiry, Ali A; Almeshari, Ahmed A; Togoo, Rafi Ahmad

    2013-11-01

    The aim of this study was to assess the incidence of micronuclei (MN) in buccal mucosal cells of professionals working in radiology area to determine the risk of stochastic effects of radiation. All the professionals and students working in King Khalid University - College of Dentistry radiology area were included in the Risk Group (RG = 27). The Control Group (CG = 27) comprised of healthy individual matching the gender and age of the RG. Buccal mucosal scraping from all the 54 subjects of RG and CG were stained with Papanicolaou stain and observed under oil immersion lens (×100) for the presence of micronuclei (MN) in the exfoliated epithelial cells. There was no significant difference between the incidence of MN in RG and CG (p = >0.05) using t-test. Routine radiation protection protocol does minimize the risk of radiation induced cytotoxicity, however, screening of professionals should be carried out at regular intervals.

  3. Radiosensitiviness of blood lymphocytes from skin cancer patients and healthy volunteers as determined by micronucleus assay

    International Nuclear Information System (INIS)

    Lohmann, Tania Helena Ochi.

    1995-01-01

    Cancer, a major death cause in developed countries, has been related to somatic mutations that could be detected by cytogenetic analysis. Among the tools used in these tests, the micronucleus assay has been largely applied at population surveillance, biological dosimetry and early detection of groups with higher risks to developing cancers. In this study, we analysed the chromosome susceptibility of blood lymphocytes from basocellular skin cancer patients and healthy volunteers. The cytogenetic analysis was performed by a micronucleus assay, using progressive doses of ionizing radiation from a 60 Co source as mutagen. Briefly, the blood lymphocytes were irradiated in vitro, as processed by the cytokinesis-blocked method. The micronucleus frequency and distribution, cell cycle kinetics, nucleation index and dose-response relationship were determined in each patient. The results showed that the basocellular skin cancer patients lymphocytes presented higher spontaneous micronucleus frequency as compared with those from healthy young volunteers but lower than healthy now young volunteers . The radiation-induced micronucleus analysis showed that the basocellular skin cancer patients' lymphocytes presented similar proportion of damage lymphocytes as compared with those from healthy volunteers. Nevertheless, the magnitude of this damage was higher in this group with doses. Higher than 400 c Gy, which was not occurred in healthy volunteers. Cell cycle kinetics, as determined by the nucleation index, was lower in basocellular skin cancer patients as compared with healthy volunteers, indicating a more slow cell cycle. Our data showed that the lymphocytes from carcinoma basocellular patients were more radiosensitive as compared with those form healthy volunteers. (author). 159 refs., 21 figs., 16 tabs

  4. Cytogenetic Monitoring By Use Of The Micronucleus Assay Among Nuclear Malaysia Radiation Workers-A Preliminary Result

    International Nuclear Information System (INIS)

    Rahimah Abdul Rahim; Mohd Rodzi Ali; Noraisyah Mohd Yusof; Juliana Mahamad Napiah; Yahaya Talib; Rehir Dahalan

    2014-01-01

    Biological dosimetry based on the analysis of micronuclei in the cytokinesis-block micronucleus (CBMN) assay can be used as an alternative method for scoring dicentric chromosomes in the field of radiation protection. Bio dosimetry is mainly performed, in addition to physical dosimetry, with the aim of individual dose assessment. Aim of this study was to assess occupationally induced chromosomal damage in radiation workers exposed to ionizing radiation. The CBMN assay was used in the peripheral blood lymphocytes of 50 exposed workers. Number of bi-nucleated cell and micronuclei were scored and statistical analysis was done to see the effect and correlation of micronuclei with gender, age and time of worked. In conclusion, scoring of micronuclei is a useful cytogenetic monitoring for radiation workers. (author)

  5. The assessment of micronucleus frequency in lymphocytes in the cohort of coal-miners characterized by different polymorphisms of double strand break reparation genes

    Directory of Open Access Journals (Sweden)

    Maxim Yur'yevich Sinitsky

    2015-12-01

    Full Text Available Background: Coal-miners are exposed to a lot of number of harmful factors (chemical agents, ionizing radiation, heavy metals, coal dust etc.. Material and methods: Venous blood samples extracted from 129 coal-miners. Assessment of cytogenetic damage was performed using the cytokinesis-block micronucleus assay (CBMN on peripheral blood lymphocytes. PCR and gel electrophoresis were used to determine polymorphisms in the genes Lig4 (rs1805388 and XRCC4 (rs6869366. Results: We found a significant increase in the frequency of binucleated lymphocytes with micronuclei (MN and protrusions in carriers of the Ile/ Ile genotype of the Lig4 gene Thr9Ile polymorphism in comparison to Thr/Thr and Thr/Ile genotypes. Conclusions: Thr9Ile polymorphism within Lig4 gene can be used as potential molecular genetic markers of increased individual susceptibility to the complex of harmful factors in coal-mining conditions.

  6. Cytochalasin-b micronucleus test of peripheral blood lymphocytes of Kozloduy NPP workers

    International Nuclear Information System (INIS)

    Hadjidekova, V.; Hristova, R.; Atanassova, P.; Stainova, A.; Popova, L.

    2006-01-01

    Full text: The cytokinesis-block micronucleus assay in peripheral blood lymphocytes was applied to evaluate occupational radiation exposure of 65 nuclear power plant workers. Blood samples were collected from 43 workers aged between 32-54 years, mean age 41,7 years. The accumulated radiation doses for each subject varied between 7,9 - 766,4 mSv, mean level of the whole group is 237,78 mSv. Controls were 22 healthy individuals, (13 male and 9 female), aged between 27-52 years, mean age 38,8 years, selected from the administrative staff. All subjects participating in this study were interviewed concerning health status, professional history, smoking habit and other aspects relevant to the study. At least 1000 binucleated cells were analysed per person. The detected frequencies of micronuclei in the control group were ranged between 4.0 and 23.5 per 1000 binucleated cells, with the average incidence yield of 12.16 ±5.59 %. The mean group value of the frequency of micronuclei in peripheral lymphocytes of exposed workers was found to be 18.46±6.72 % in 1000 cells. The difference between the mean frequency of micronuclei in the group of exposed subjects and the control group was statistically significant (P<0,001). The correlation coefficient for duration of employment and accumulated doses is 0,30 (P<0,05). After 1,5 Gy in vitro irradiation of peripheral blood from investigated workers and controls a decreased radiosensitivity of NPP workers is detected using micronucleus assay. Decreased radiosensitivity of the professionally exposed workers could be due to the phenomenon of adaptive response. Micronucleus assay in peripheral blood lymphocytes is useful approach in cytogenetic monitoring of occupationally exposed nuclear industry workers

  7. Automation in Cytomics: A Modern RDBMS Based Platform for Image Analysis and Management in High-Throughput Screening Experiments

    NARCIS (Netherlands)

    E. Larios (Enrique); Y. Zhang (Ying); K. Yan (Kuan); Z. Di; S. LeDévédec (Sylvia); F.E. Groffen (Fabian); F.J. Verbeek

    2012-01-01

    textabstractIn cytomics bookkeeping of the data generated during lab experiments is crucial. The current approach in cytomics is to conduct High-Throughput Screening (HTS) experiments so that cells can be tested under many different experimental conditions. Given the large amount of different

  8. Cytogenetic radiosensitivity of G0-lymphocytes of breast and esophageal cancer patients as determined by micronucleus assay

    International Nuclear Information System (INIS)

    Mozdarani, H.; Mansouri, Z.; Haeri, S.A.

    2005-01-01

    Enhanced chromosomal radiosensitivity is a feature of many cancer predisposition conditions, indicative of the important role of chromosomal alterations in carcinogenesis. In this study the cytokinesis-blocked micronucleous assay was used to compare the radiosensitivity of blood lymphocytes obtained from Iranian breast or esophageal cancer patients (n=50, n=16; respectively) with that of control individuals (n=40). For each sample, one thousand binucleate lymphocytes were analyzed before and after in vitro exposure to 3 Gy of γ rays. The radiation-induced frequency of micronucleus was significantly higher in the breast cancer group (261/1,000 binucleated cells) than in esophageal cancer group (241/1,000 binucleated cells, P<0.01) or in the control group (240/1,000 binucleated cells, P<0.01). The results indicate that breast cancer patients are more radiosensitive compared to normal healthy individuals or esophageal cancer patients. Increased radiosensitivity could be due to defects in DNA repair genes involved in breast cancer formation. Since patients with esophageal cancer did not show elevated radiosensitivity, it is assumed that the contribution of radiosensitivity-related genes to the development of esophageal cancer may be smaller than the contribution of those genes to breast cancer. (author)

  9. Evaluation of apoptosis and apoptosis proteins as possible markers of radiation at doses 0.1-2 Gy, in comparison to the micronucleus assay in three cell lines

    International Nuclear Information System (INIS)

    Jaworska, A.; Angelis, P. de

    1997-01-01

    In recent years the interest in apoptosis as possible indicator of radiation damage has increased. Studies have been done to examine the induction of apoptosis after ionizing radiation using morphological criteria, characteristic DNA damage pattern(ladders), early DNA damage using flow cytometry and/or expression of the proteins involved in apoptosis. But the picture which emerges from these investigations is unclear. Some researchers suggest that apoptosis studies can be used as potential assays of biological dosimetry, others doubt if apoptosis can be used as a marker of irradiation at all. Most studies have been done using relatively high doses of radiation. In this study we focus on apoptosis induction after relatively small doses (0,1-2 Gy). We detected apoptosis with the in situ terminal deoxynucleotidyl transferase assay by flow cytometry, and measured the expression of proteins that regulate apoptosis (Bcl-2, Bax, P53) with Western blotting. As comparison we used the cytokinesis-block micronucleus assay as a reference. The studies were carried out in three lymphoid cell lines: the mouse lymphoma L5178Y resistant and sensitive cell lines widely used in radiobiological studies, and the human pre-B cell leukemia Reh cells. Our results indicate that we can not consider the examined parameters of apoptosis as markers of radiation in these cell lines. (author)

  10. Genotoxic and Antigenotoxic Assessment of Chios Mastic Oil by the In Vitro Micronucleus Test on Human Lymphocytes and the In Vivo Wing Somatic Test on Drosophila.

    Directory of Open Access Journals (Sweden)

    Dimitris Vlastos

    Full Text Available Chios mastic oil (CMO, the essential oil derived from Pistacia lentiscus (L. var. chia (Duham, has generated considerable interest because of its antimicrobial, anticancer, antioxidant and other beneficial properties. In the present study, the potential genotoxic activity of CMO as well as its antigenotoxic properties against the mutagenic agent mitomycin-C (MMC were evaluated by employing the in vitro Cytokinesis Block MicroNucleus (CBMN assay and the in vivo Somatic Mutation And Recombination Test (SMART. In the in vitro experiments, lymphocytes were treated with 0.01, 0.05 and 0.10% (v/v of CMO with or without 0.05 μg/ml MMC, while in the in vivo assay Drosophila larvae were fed with 0.05, 0.10, 0.50 and 1.00% (v/v of CMO with or without 2.50 μg/ml MMC. CMO did not significantly increase the frequency of micronuclei (MN or total wing spots, indicating lack of mutagenic or recombinogenic activity. However, the in vitro analysis suggested cytotoxic activity of CMO. The simultaneous administration of MMC with CMO did not alter considerably the frequencies of MMC-induced MN and wing spots showing that CMO doesn't exert antigenotoxic or antirecombinogenic action. Therefore, CMO could be considered as a safe product in terms of genotoxic potential. Even though it could not afford any protection against DNA damage, at least under our experimental conditions, its cytotoxic potential could be of interest.

  11. Age and smoking habit influence on the spontaneous and radiation induced frequencies of the micronucleus in human lymphocytes

    International Nuclear Information System (INIS)

    Di Giorgio, M.; Nasazzi, N.; Heredia, M.L.

    1996-01-01

    Several endpoints have been used for monitoring human population that have been exposed at work or in the environment to genotoxic agents, particularly to ionizing radiation. The cytokinesis-block micronucleus (MN) assay in peripheral lymphocytes is a reliable method for evaluating radiation induced chromosomal damage (DNA breaks and mitotic spindle disturbances) and thus, a suitable dosimeter for estimating in vivo whole body exposures. A research to determine the influence of age, sex and life style factors (smoking habits) on the MN spontaneous and radiation induced frequencies was carried out in order to define the use of this assay in Biological Dosimetry. The estimation of MN frequencies was analyzed in lymphocytes cultures from 50 health donors aged between 4 and 60 years. Based on the smoking habits, they were divided into 2 groups. A fraction of the sample was irradiated in vitro with γ-rays in the range of 0.35 Gy to 4 Gy. A statistically significant influence on the spontaneous MN frequency was observed (R 2 = 0.59) when the variables age and smoking habit were analyzed, and a statistically significant influence on the radiation induced MN frequency was also obtained (R 2 = 0.86) when dose, age and smoking habit were studied. Sex did not influence significantly MN variability, but there was a greater dispersion in the results obtained from female donors, when compared to males, possibly due to the loss of X chromosomes. The comparison of the data from smoking donors to the data from non smoking donors supports the convenience of taking into account the smoking habit for estimating in vivo whole body exposure to γ-rays for doses below 2 Gy. (authors). 8 refs., 3 figs., 2 tabs

  12. Evidence of heritable lethal mutations in progeny of X-irradiated CHO cells by micronucleus count in clon-cells

    International Nuclear Information System (INIS)

    Hagemann, G.; Kreczik, A.; Treichel, M.

    1996-01-01

    Low doses of ionizing radiation reduce the growth rates of clones following irradiation of the progenitor cells. Such reductions of clone growth have been proven by means of measurements of clone size distributions. The medians of such distributions can be used to quantify the radiation damage. Prolongations of generation times and cell death as result of heritable lethal mutations have been discussed as causes for the reduction of clone growth. The cell number of a clone of hypotetraploid CHO-cells was compared to the frequency of micronucleated binucleated cells in the same clone using the cytokinesis-block-micronucleus method. The dose dependent reduction of clone sizes is measured by the difference of the medians (after log transformation) of the clone size distributions. At cytochalasin-B concentrations of 1 μg/ml and after an incubation time of 16 h a yield of binucleated cells of about 50% was obtained. Median clone size differences as a measure of clonal radiation damage increased linearly with incubation times of 76, 100, 124, and 240 h following irradiation with 3, 5, 7, and 12 Gy. The frequency of binucleated clone cells with micronuclei strongly increased with decreasing clone size by a factor up to 20 following irradiation with 3, 5, and 7 Gy. The frequency of micronucleated binucleated clone cells was found to be independent of incubation time after irradiation. Radiation induced clone size reductions result from cell losses caused by intraclonal expression of micronuclei which have its origin in heritable lethal mutations. Measurements of clone size distributions can be done automatically. They can serve as predictive test for determination of median cell loss rates of surviving cell clones. (orig./MG) [de

  13. Increased micronucleus, nucleoplasmic bridge, and nuclear bud frequencies in the peripheral blood lymphocytes of diesel engine exhaust-exposed workers.

    Science.gov (United States)

    Zhang, Xiao; Duan, Huawei; Gao, Feng; Li, Yuanyuan; Huang, Chuanfeng; Niu, Yong; Gao, Weimin; Yu, Shanfa; Zheng, Yuxin

    2015-02-01

    The International Agency for Research on Cancer has recently reclassified diesel engine exhaust (DEE) as a Group 1 carcinogen. Micronucleus (MN), nucleoplasmic bridge (NPB), and nuclear bud (NBUD) frequencies in peripheral blood lymphocytes (PBLs) are associated with cancer risk. However, the impact of DEE exposure on MN frequency has not been thoroughly elucidated due to mixed exposure and its impact on NPB and NBUD frequencies has never been explored in humans. We recruited 117 diesel engine testing workers with exclusive exposure to DEE and 112 non-DEE-exposed workers, and then we measured urinary levels of 4 mono-hydroxylated polycyclic aromatic hydrocarbons (OH-PAHs) using high-performance liquid chromatography-mass spectrometry as well as MN, NPB, and NBUD frequencies in PBLs using cytokinesis-block MN assay. The DEE-exposed workers exhibited significantly higher MN, NPB, and NBUD frequencies than the non-DEE-exposed workers (P < 0.05). Among all study subjects, increasing levels of all 4 urinary OH-PAHs, on both quartile and continuous scales, were associated with increased MN, NPB, and NBUD frequencies (all P < 0.05). When the associations were analyzed separately in DEE-exposed and non-DEE-exposed workers, we found that the association between increasing quartiles of urinary 9-hydroxyphenanthrene (9-OHPh) and MN frequencies persisted in DEE-exposed workers (P = 0.001). The percent of MN frequencies increased, on average, by 23.99% (95% confidential interval, 9.64-39.93) per 1-unit increase in ln-transformed 9-OHPh. Our results clearly show that exposure to DEE can induce increases in MN, NPB, and NBUD frequencies in PBLs and suggest that DEE exposure level is associated with MN frequencies. © The Author 2014. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  14. In vitro mutagenicity and genotoxicity study of 1,2-dichloroethylene, 1,1,2-trichloroethane, 1,3-dichloropropane, 1,2,3-trichloropropane and 1,1,3-trichloropropene, using the micronucleus test and the alkaline single cell gel electrophoresis technique (comet assay) in human lymphocytes.

    Science.gov (United States)

    Tafazoli, M; Kirsch-Volders, M

    1996-12-20

    The main objective of this study was to compare the cytotoxic genotoxic and mutagenic activity of a number of chlorinated aliphatic hydrocarbons, which are widely used as chemical intermediates, solvents, degreasing agents etc. in industry, and to establish the structure-toxicity relationship of the chemicals by using the most adequate determinants in estimating their toxicity. The mutagenicity and cytotoxicity of some of the candidate chemicals, namely 1,2-dichloroethylene, 1,1,2-trichloroethane, 1,3-dichloropropane, 1,2,3-trichloropropane and 1,1,3-trichloropropene were evaluated in an in vitro micronucleus assay. The cytokinesis-block methodology was applied on human lymphocytes in the presence or absence of an external metabolic activation system (S9-mix). In the micronucleus assay, all test substances, except 1,2,3-trichloropropane with and without S9-mix and 1,1,2-trichloroethane without S9-mix in the repeated experiment, exhibited a low but statistically significant mutagenic activity, compared to the concurrent control. However, none of the five chemicals was able to induce a clear and reproducible linear dose-dependent increase in micronucleus frequencies in this assay. Generally, mutagenic activity of the chemicals was found in the absence of severe cytotoxicity and/or cell cycle delay. The DNA breakage capacity and the cytotoxicity of these chemicals were also assessed in the alkaline single cell gel (SCG) electrophoresis test (comet assay) with and without S9-mix in isolated human lymphocytes. All chemical compounds induced DNA breakage, in the presence or absence of the metabolic activation system, at the doses tested. The data showed that the DNA reactivity of the chemicals increased with increasing degree of halogenation. The results of the present work suggested that the comet assay might be a more suitable and sensitive screening method than the micronucleus test for this particular class of compound. However, both assays do detect different

  15. Genotoxic evaluation of [DOTA,Tyr3]octreotate labeled with 131I and 177Lu in human peripheral lymphocytes in vitro by micronucleus assay

    International Nuclear Information System (INIS)

    Suzauki, Miriam Fussae; Silva, Marcia Augusta da; Caldeira Filho, Jose de Souza; Colturato, Maria Tereza; Araujo, Elaine Bortoleti de; Bartolini, Paolo; Okazaki, Kayo

    2005-01-01

    The radiolabeled receptor-binding peptides have being used for cancer diagnosis and therapy. The octreotate, a somatostatin analogue peptide, bound to various tumors expressing sst receptors (thyroid, pancreas, prostrate, melanoma and lymphomas). The amount and the type of receptors for somatostatin influence the tissue uptake. The [DOTA, Tyr 3 ]octreotate has been used because of its high affinity to somatostatin subtype receptors sstr 2 and sstr 5 . The pharmacokinetic study showed that the blood clearance is rapid and only 9% of the intravenous injected activity remains in human blood after one hour. The aim of this study was to evaluate the cytogenetic effect of radiolabeled [DOTA, Tyr 3 ]octreotate in blood cells in vitro, using the cytokinesis-block micronucleus (MN) assay. This technique allows evaluating the mutagenic effects of both endogenous and exogenous agents at chromosome level. Blood samples of healthy donors were collected in heparinized syringes and exposed to different activities of [DOTA, Tyr 3 ]octreotate labeled with with 131 I (n=3) and 177 Lu (n=3), where radioactive concentration ranged from 600 to 5600 kBq/mL, corresponding to an injected activity of 3.1 to 28.9 GBq in a reference man of 70 kg weight. 131 I and 177 Lu are beta- and gamma-emitters. After one-hour exposition to radiopharmaceuticals at 37 deg C, the cells were washed with culture medium for removing the non internalised octreotate and cultivated for 72 hours, according to criteria adopted by the IAEA. The results showed a positive correlation between radioactive concentrations (X) and the frequency of binucleated cells with micronuclei (Y) (P 131 I-DOTA, Tyr 3 ]octreotate was Y = (1.634 ± 0.236) + (0.912 ± 0.137) 10 -3 X and for [ 177 Lu-DOTA, Tyr 3 ]octreotate was Y = (1.715 ± 0.342) + (0.743 ± 0.135) 10 -3 X. The non labeled molecule, [DOTA, Tyr 3 ]octreotate, has no influence in the induction of cytogenetic damage. The micronucleus assay with rat pancreatic tumor cells

  16. Flow cytometric determination of micronucleus frequency.

    Science.gov (United States)

    Elhajouji, Azeddine; Lukamowicz-Rajska, Magdalena

    2013-01-01

    During the last two decades the micronucleus (MN) test has been extensively used as a genotoxicity screening tool of chemicals and in a variety of exploratory and mechanistic investigations. The MN is a biomarker for chromosomal damage or mitotic abnormalities, since it can originate from chromosome fragments or whole chromosomes that fail to be incorporated into daughter nuclei during mitosis (Fenech et al., Mutagenesis 26:125-132, 2011; Kirsch-Volders et al., Arch Toxicol 85:873-899, 2011). The simplicity of scoring, accuracy, amenability to automation by image analysis or flow cytometry, and readiness to be applied to a variety of cell types either in vitro or in vivo have made it a versatile tool that has contributed to a large extent in our understanding of key toxicological issues related to genotoxins and their effects at the cellular and organism levels. Recently, the final acceptance of the in vitro MN test guideline 487 (OECD Guideline for Testing of Chemicals, In vitro mammalian cell micronucleus test 487. In vitro mammalian cell micronucleus test (MNVIT). Organization for Economic Cooperation and Development, Paris, 2010) together with the standard in vivo MN test OECD guideline 474 (OECD Guideline for The Testing of Chemicals, Mammalian erythrocyte micronucleus test no. 474. Organization for Economic Cooperation and Development, Paris, 1997) will further position the assay as a key driver in the determination of the genotoxicity potential in exploratory research as well as in the regulatory environment. This chapter covers to some extent the protocol designs and experimental steps necessary for a successful performance of the MN test and an accurate analysis of the MN by the flow cytometry technique.

  17. 40 CFR 79.64 - In vivo micronucleus assay.

    Science.gov (United States)

    2010-07-01

    ... micronucleus assay. (a) Purpose. The micronucleus assay is an in vivo cytogenetic test which uses erythrocytes... that, because it contains RNA, can be differentiated by appropriate staining techniques from a normochromatic erythrocyte (NCE), which lacks RNA. In one to two days, a PCE matures into a NCE. (c) Test method...

  18. Micronucleus frequency and content in healthy relatives of cancer patients

    Czech Academy of Sciences Publication Activity Database

    Zölzer, F.; Křížová, M.; Skalická, Z.F.; Rössnerová, Andrea; Šrám, Radim

    2017-01-01

    Roč. 22, č. 7 (2017), s. 667-673 ISSN 1354-750X Institutional support: RVO:68378041 Keywords : micronucleus centromere test * chromosomal damage * inheritable trait Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Human genetics Impact factor: 2.006, year: 2016

  19. What Can a Micronucleus Teach? Learning about Environmental Mutagenesis

    Science.gov (United States)

    Linde, Ana R.; Garcia-Vazquez, Eva

    2009-01-01

    The micronucleus test is widely employed in environmental health research. It can also be an excellent tool for learning important concepts in environmental health. In this article we present an inquiry-based laboratory exercise where students explore several theoretical and practical aspects of environmental mutagenesis employing the micronucleus…

  20. Micronucleus induction as a measure of I-131 exposure

    Energy Technology Data Exchange (ETDEWEB)

    Kasuba, V; Horvat, D [Inst. for Medical Recearch and Occupational Health, Zagreb (Croatia). Laboratory for Mutagenesis; Kusic, Z [Clinical Hospital Sestre Milosrdnice, Zagreb (Croatia). Dept. of Oncology and Nuclear Medicine; Vlatkovic, M [Clinical Hospital Centre, Zagreb (Croatia). Dept. of Nuclear Medicine and Radiation Protection

    1994-10-01

    The change of cell numbers in the peripheral blood following irradiation has been studied for many years, particularly in patients undergoing radiotherapy. Recently, attention is directed towards the use of cytogenetic-mutagenetic methods to estimate the biological effects of received radiation dose. The aim of our study was to identify the difference in number and distribution of micronucleus, depending of applied therapeutic dose of iodine-131. According to their diagnosis, six patients have received iodine-131 in range from 80 to 140 mCi, while in the other group of patients the dose values varied from 7 to 32 mCi. On in vitro peripheral blood lymphocyte cultures micronucleus test was applied. Micronucleus analyses were carried out before the treatment, 24, 48 and 96 hours after the oral application of radiopharmaceutical. The number of micronucleus is showing increase, depending on applied radioactivity of iodine-131 and duration of exposition. The clear dose response relationship was never found. These results illustrate the problem associated with the inhomogeneous distribution of dose which results from the concentration of incorporated radionuclide into thyroid or other tissues. (author).

  1. The role of the micronucleus in stomatogenesis in sexual reproduction of Paramecium tetraurelia: laser microbeam irradiation of the micronucleus

    International Nuclear Information System (INIS)

    Tam Laiwa; Ng, S.F.

    1986-01-01

    Fifteen amicronucleate cell lines and 22 cell lines with defective micronuclei were obtained following selective laser microbeam irradiation of the micronucleus. The amicronucleate cell lines showed reduced growth rate and formed abnormal oral apparatuses in asexual reproduction, and failed to produce any oral apparatus in autogamy. The 22 cell lines with defective micronucleus exhibited various abnormalities of the oral apparatus newly formed during autogamy. These abnormalities included the arrest of membranelle assembly, reduction in the length of the buccal cavity and oral membranelles, disruption of the organization of the membranelles, quadrulation of the dorsal peniculus, and failure of addition of membranellar basal body rows. Hence the micronucleus plays multiple roles in sexual stomatogenesis. Our results agree with the notion that the micronucleus acts during a critical period between the second meiotic division and up to the formation of the zygotic nucleus to control the early stage of oral membranelle assembly. Laser microbeam irradiation might have created recessive mutations and/or chromosomal aberrations, which were expressed during this critical period with the formation of abnormal postmeiotic nuclei. (author)

  2. Role of the micronucleus in stomatogenesis in sexual reproduction of Paramecium tetraurelia: laser microbeam irradiation of the micronucleus

    Energy Technology Data Exchange (ETDEWEB)

    Tam Laiwa; Ng, S.F.

    1986-12-01

    Fifteen amicronucleate cell lines and 22 cell lines with defective micronuclei were obtained following selective laser microbeam irradiation of the micronucleus. The amicronucleate cell lines showed reduced growth rate and formed abnormal oral apparatuses in asexual reproduction, and failed to produce any oral apparatus in autogamy. The 22 cell lines with defective micronucleus exhibited various abnormalities of the oral apparatus newly formed during autogamy. These abnormalities included the arrest of membranelle assembly, reduction in the length of the buccal cavity and oral membranelles, disruption of the organization of the membranelles, quadrulation of the dorsal peniculus, and failure of addition of membranellar basal body rows. Hence the micronucleus plays multiple roles in sexual stomatogenesis. Our results agree with the notion that the micronucleus acts during a critical period between the second meiotic division and up to the formation of the zygotic nucleus to control the early stage of oral membranelle assembly. Laser microbeam irradiation might have created recessive mutations and/or chromosomal aberrations, which were expressed during this critical period with the formation of abnormal postmeiotic nuclei.

  3. Micronucleus formation in cultured human keratinocytes: Involvement of intercellular bioactivation.

    Science.gov (United States)

    van Pelt, F N; Haring, R M; Weterings, P J

    1991-01-01

    Micronucleus formation in cultured human keratinocytes was studied after exposure to benzo[a]pyrene, cyclophosphamide and 12-O-tetradecanoylphorbol-13-acetate without the addition of an exogenous metabolizing system. The first two agents need bioactivation by specific isoenzymes of cytochrome P-450 to form genotoxic intermediates. Benzo[a]pyrene induced the micronucleus formation in both uninduced and Aroclor 1254-pretreated cultures. Clastogenic effects of cyclophosphamide were observed only in Aroclor 1254-pretreated cells. The tumour promotor 12-O-tetradecanoylphorbol-13-acetate did not affect the frequency of micronuclei in human keratinocytes. The data indicate that cultured human keratinocytes can be used to study the tissue-specific response to genotoxic agents as well as interindividual variation in biotransformation capacity.

  4. Increased Chromosomal and Oxidative DNA Damage in Patients with Multinodular Goiter and Their Association with Cancer

    Directory of Open Access Journals (Sweden)

    Hamiyet Donmez-Altuntas

    2017-01-01

    Full Text Available Thyroid nodules are a common clinical problem worldwide. Although thyroid cancer accounts for a small percentage of thyroid nodules, the majority are benign. 8-Hydroxy-2′-deoxyguanosine (8-OHdG levels are a marker of oxidative stress and play a key role in the initiation and development of a range of diseases and cancer types. This study evaluates cytokinesis-block micronucleus cytome (CBMN-cyt assay parameters and plasma 8-OHdG levels and their association with thyroid nodule size and thyroid hormones in patients with multinodular goiter. The study included 32 patients with multinodular goiter and 18 age- and sex-matched healthy controls. CBMN-cyt assay parameters in peripheral blood lymphocytes of patients with multinodular goiter and controls were evaluated, and plasma 8-OHdG levels were measured. The micronucleus (MN frequency (chromosomal DNA damage, apoptotic and necrotic cells (cytotoxicity, and plasma 8-OHdG levels (oxidative DNA damage were significantly higher among patients with multinodular goiter. Our study is the first report of increased chromosomal and oxidative DNA damage in patients with multinodular goiter, which may predict an increased risk of thyroid cancer in these patients. MN frequency and plasma 8-OHdG levels may be markers of the carcinogenic potential of multinodular goiters and could be used for early detection of different cancer types, including thyroid cancer.

  5. Evaluation of genetic damage in tobacco and arsenic exposed population of Southern Assam, India using buccal cytome assay and comet assay.

    Science.gov (United States)

    Roy, Prasenjit; Mukherjee, Anita; Giri, Sarbani

    2016-02-01

    Ground water is the principal source of drinking water in Assam. Ground water contamination of arsenic in drinking water is a great concern for human health and considered as a human carcinogen. The present cytogenetic biomonitoring study was undertaken to investigate the genotoxic effects associated with people of southern Assam consuming arsenic contaminated water and chewing tobacco. Employing the buccal cytome assay, exfoliated cells were analyzed in 138 individuals of age range 22-42 years and divided into four groups. Group I (n=54) are participants residing in localities where ground water contains arsenic concentration below the permissible limit (comet assay, percent of tail DNA gradually increases among the groups and has statistical significance. Spearman correlation revealed strong positive correlation between the arsenic exposed peoples and the binucleated cells (r=0.4763; Pcomet assay. Copyright © 2015 Elsevier Inc. All rights reserved.

  6. Micronucleus frequency in exfoliated buccal cells from hairdresser who expose to hair products

    Directory of Open Access Journals (Sweden)

    Koh Hui Yee

    2015-06-01

    Full Text Available Background: Hairdresser is one of the fastest growing occupations in today’s society. Hairdresser help styling, cutting, colouring, perming, curling, straightening hair and various treatment to customer. Somehow, hairdresser are constantly exposed to chemical substances such as aromatic amines, hydrogen peroxide, thioglycolic acid, formaldehyde in hair products which can cause damage to human’s genome. Micronucleus is one of the effective biomarker for processes associated with the induction of DNA damage. Purpose: The aim of this study was to determine the micronucleus frequencies in buccal mucosa epithelial cells of hairdresser who were exposed to chemical of hair products. Method: This study was conducted on twenty female subjects, who were divided into 2 groups: exposed and non-exposed (control group. All subjects recruited were working in the same beauty salon. Buccal cells were obtained from each individual by using cytobrush. The cells were stained with modified Feulgen-Ronssenback method and counting of micronucleus per 1000 cell was done under light microscope. The data were analyzed using independent t-test and one-way Anova (p<0.05. Result: The result showed a significant difference in micronucleus frequency between 2 groups. There were a significantly increase of micronucleus frequency in hairdressers and increase of  micronucleus frequency with the longer duration of exposure. Conclusion: It concluded that the chemical substances of hair products had affected the micronucleus frequency ofthe epithelial cells in buccal mucosa of hairdressers.

  7. Cytological assay of micronucleus induction by radiation in oral cancer

    International Nuclear Information System (INIS)

    Bhattathiri, V.N.; Bindu, L.; Remani, P.; Chandralekha, B.; Davis, C.A.; Krishnan Nair, M.

    1996-01-01

    A study was conducted to analyze the dose-response relationship of micronucleus (MN) induction by radiation in eighty-three patients with oral cancers. Serial scrape smears were taken before treatment and after delivery of various fractions of a course of radical radiotherapy. The smears were stained with Giemsa's stain and frequency of micronucleated cells (MNC) evaluated. Before treatment 70.5% of tumours showed MNC, the mean value being 4.18 MNC/1000 cells. The frequency of MNC, increased with increasing dose of radiation. As regards relation to treatment duration, there was initially a slight increase, followed by rapid increase and then a plateauing. Radiosensitive and resistant tumours showed differing pattern of change. The MN test by serial cytological assay has potential as a tool to understand the dynamics of radiation induced cell death and predict radiosensitivity. (author). 4 refs., 5 figs

  8. An improved in vitro micronucleus assay to biological dosimetry

    International Nuclear Information System (INIS)

    Ocampo, Ivette Z.; Okazaki, Kayo; Vieira, Daniel P.

    2013-01-01

    The biological dosimetry is widely used to estimate the absorbed dose in people occupationally or accidentally exposed to the radiation for a better medical treatment, minimizing the harmful effects. Many techniques and methods have been proposed to detect and quantify the radioinduced lesions in genetic material, among them, the micronucleus (MN) assay. In the present study, we proposed an improved in vitro micronucleus technique that is rapid, sensitive and with minor cell manipulations. Assays were carried out with human tumor cells (MCF-7) seeded (3x10 4 cells) in slides placed into Petri dishes. Adherent cells were maintained with RPMI medium, supplemented with fetal calf serum, 1 % antibiotics, cytochalasin B (2 μg/mL), and incubated at 37 deg C in the presence of 5% CO2 for 72h. Cells were pre-treated for 24h with aminoguanidine, a nitric oxide synthase inhibitor. Nitric oxide is an intracellular free-radical, involved in DNA double-strand break repair mechanisms. After incubation, adherent cells on slides were briefly fixed with paraformaldehyde and stained with acridine orange (100 μg/mL) for analysis through fluorescence microscopy. Dye fluorescence permitted accurate discrimination between nuclei and micronuclei (bright green) and cytoplasm (red), and made possible a faster counting of binucleated cells. Aminoguanidine (2 mM) induced significant increase (p< 0.05) in frequencies of binucleated cells with micronuclei and in the number of micronuclei per binucleated cell. Data showed that proposed modifications permit to understand an early aspect of NO inhibition and suggested an improved protocol to MN assays. (author)

  9. Evaluation of river water genotoxicity using the piscine micronucleus test.

    Science.gov (United States)

    Ergene, Serap; Cavaş, Tolga; Celik, Ayla; Köleli, Nurcan; Aymak, Cemil

    2007-07-01

    The Berdan River, which empties into the Mediterranean Sea on the east coast of Turkey, receives discharges of industrial and municipal waste. In the present study, the in vivo piscine micronucleus (MN) test was used to evaluate the genotoxicity of water samples collected from different locations along the Berdan River. Nile tilapia (Oreochromis niloticus) were exposed in the laboratory for 2, 4, and 6 days, and micronuclei were evaluated in peripheral blood erythrocytes, gill cells, and caudal fin epithelial cells. A single dose of 5 mg/L cyclophosphamide was used as a positive control. In addition to micronuclei, nuclear abnormalities (NAs), such as binucleated cells and blebbed, notched, and lobed nuclei, were assessed in the erythrocytes, and chemical analyses were carried out to determine the amount of heavy metals in the water samples. MN and NA frequencies were significantly elevated (up to 2- to 3-fold) in fish exposed to river water samples taken downstream of potential discharges, and the elevated responses in gill and fin cells were related to the concentration of heavy metals in the water. MN frequencies (expressed as micronucleated cells/1,000 cells), in both treated and untreated fish, were greatest in gill cells (range: 0.80-3.70), and generally lower in erythrocytes (range: 0.50-2.80), and fin cells (range: 0.45-1.70). The results of this study indicate that the Berdan River is contaminated with genotoxic pollutants and that the genotoxicity is related to the discharge of wastes into the river water.

  10. In vitro safety assessment of the strawberry tree (Arbutus unedo L.) water leaf extract and arbutin in human peripheral blood lymphocytes.

    Science.gov (United States)

    Jurica, K; Brčić Karačonji, I; Mikolić, A; Milojković-Opsenica, D; Benković, V; Kopjar, N

    2018-04-25

    Strawberry tree (Arbutus unedo L.) leaves have long been used in the traditional medicine of the Mediterranean region. One of their most bioactive constituents is the glycoside arbutin, whose presence makes A. unedo suitable as a potential substitute for bearberry [Arctostaphylos uva ursi (L.) Spreng] leaves, an herbal preparation widely used for treating urinary tract infections. The safety and biocompatibility of strawberry tree water leaf extract have not yet been documented well. This study estimated arbutin content in strawberry tree water leaf extract (STE) using high performance liquid chromatography. Furthermore, we performed an in vitro safety assessment of the 24 h exposure to three presumably non-toxic concentrations of standardized STE and arbutin in human peripheral blood lymphocytes using the apoptosis/necrosis assay, the alkaline comet assay, and the cytokinesis-block micronucleus cytome assay. The STE was also tested for total antioxidant capacity and lipid peroxidation. At a concentration corresponding to the maximum allowable daily intake of arbutin, the tested extract was not cytotoxic, had a negligible potential for causing primary DNA damage and even hindered micronuclei formation in lymphocytes. It also showed a valuable antioxidant capacity, and did not exert marked lipid peroxidation. These promising results represent a solid frame for further development of STE-based herbal preparations. Although arbutin generally had a low DNA damaging potential, the slowing down of lymphocyte proliferation observed after 24 h of exposure points to a cytostatic effect, which merits further research.

  11. Cyto- and genotoxicity assessment of Gold nanoparticles obtained by laser ablation in A549 lung adenocarcinoma cells

    International Nuclear Information System (INIS)

    Bucchianico, Sebastiano Di; Migliore, Lucia; Marsili, Paolo; Vergari, Chiara; Giammanco, Francesco; Giorgetti, Emilia

    2015-01-01

    Gold nanoparticles have attracted enormous interest in biomedical applications, based on their unique optical properties. However, their toxicity on human tissues is still an open issue. Beyond the potential intrinsic toxicity of nanostructured gold, a non-negligible contribution of stabilizers or reaction by-products related to current wet chemical synthesis procedures can be expected. Aimed at isolating gold contribution from that of any other contaminant, we produced colloidal suspensions of Gold nanoparticles having average size <10 nm in deionized water or acetone by pulsed laser ablation, that permits preparation of uncoated and highly stable Gold nanoparticles in pure solvents. Subsequently, we investigated the role of surface chemistry, size, and dispersivity of synthesized Gold nanoparticles in exerting toxicity in a cell model system of deep respiratory tract, representing the main route of exposure to NPs, namely adenocarcinoma epithelial A549 cells. Gold nanoparticles prepared in water showed no particular signs of cytotoxicity, cytostasis, and/or genotoxicity as assessed by MTT colorimetric viability test and Cytokinesis-block micronucleus cytome assay up to concentrations of the order of 5 μg/mL. In contrast, Gold nanoparticles produced in pure acetone and then transferred into deionized water showed impaired cell viability, apoptosis responses, micronuclei, and dicentric chromosomes induction as well as nuclear budding, as a function of the amount of surface contaminants like amorphous carbon and enolate ions

  12. Cyto- and genotoxicity assessment of Gold nanoparticles obtained by laser ablation in A549 lung adenocarcinoma cells

    Science.gov (United States)

    Di Bucchianico, Sebastiano; Migliore, Lucia; Marsili, Paolo; Vergari, Chiara; Giammanco, Francesco; Giorgetti, Emilia

    2015-05-01

    Gold nanoparticles have attracted enormous interest in biomedical applications, based on their unique optical properties. However, their toxicity on human tissues is still an open issue. Beyond the potential intrinsic toxicity of nanostructured gold, a non-negligible contribution of stabilizers or reaction by-products related to current wet chemical synthesis procedures can be expected. Aimed at isolating gold contribution from that of any other contaminant, we produced colloidal suspensions of Gold nanoparticles having average size exposure to NPs, namely adenocarcinoma epithelial A549 cells. Gold nanoparticles prepared in water showed no particular signs of cytotoxicity, cytostasis, and/or genotoxicity as assessed by MTT colorimetric viability test and Cytokinesis-block micronucleus cytome assay up to concentrations of the order of 5 μg/mL. In contrast, Gold nanoparticles produced in pure acetone and then transferred into deionized water showed impaired cell viability, apoptosis responses, micronuclei, and dicentric chromosomes induction as well as nuclear budding, as a function of the amount of surface contaminants like amorphous carbon and enolate ions.

  13. Cyto- and genotoxicity assessment of Gold nanoparticles obtained by laser ablation in A549 lung adenocarcinoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Bucchianico, Sebastiano Di [Karolinska Institutet, Institute of Environmental Medicine (Sweden); Migliore, Lucia [University of Pisa, Department of Translational Research and New Technologies in Medicine and Surgery, Division of Medical Genetics (Italy); Marsili, Paolo [Institute of Complex Systems (ISC-CNR) (Italy); Vergari, Chiara [Plasma Diagnostics and Technologies s.r.l. (Italy); Giammanco, Francesco [University of Pisa, Department of Physics “E. Fermi” (Italy); Giorgetti, Emilia, E-mail: emilia.giorgetti@fi.isc.cnr.it [Institute of Complex Systems (ISC-CNR) (Italy)

    2015-05-15

    Gold nanoparticles have attracted enormous interest in biomedical applications, based on their unique optical properties. However, their toxicity on human tissues is still an open issue. Beyond the potential intrinsic toxicity of nanostructured gold, a non-negligible contribution of stabilizers or reaction by-products related to current wet chemical synthesis procedures can be expected. Aimed at isolating gold contribution from that of any other contaminant, we produced colloidal suspensions of Gold nanoparticles having average size <10 nm in deionized water or acetone by pulsed laser ablation, that permits preparation of uncoated and highly stable Gold nanoparticles in pure solvents. Subsequently, we investigated the role of surface chemistry, size, and dispersivity of synthesized Gold nanoparticles in exerting toxicity in a cell model system of deep respiratory tract, representing the main route of exposure to NPs, namely adenocarcinoma epithelial A549 cells. Gold nanoparticles prepared in water showed no particular signs of cytotoxicity, cytostasis, and/or genotoxicity as assessed by MTT colorimetric viability test and Cytokinesis-block micronucleus cytome assay up to concentrations of the order of 5 μg/mL. In contrast, Gold nanoparticles produced in pure acetone and then transferred into deionized water showed impaired cell viability, apoptosis responses, micronuclei, and dicentric chromosomes induction as well as nuclear budding, as a function of the amount of surface contaminants like amorphous carbon and enolate ions.

  14. Genotoxic potential of montmorillonite clay mineral and alteration in the expression of genes involved in toxicity mechanisms in the human hepatoma cell line HepG2.

    Science.gov (United States)

    Maisanaba, Sara; Hercog, Klara; Filipic, Metka; Jos, Ángeles; Zegura, Bojana

    2016-03-05

    Montmorillonite, also known as Cloisite(®)Na(+) (CNa(+)), is a natural clay with a wide range of well-documented and novel applications, such as pharmaceutical products or food packaging. Although considered a low toxic product, the expected increased exposure to CNa(+) arises concern on the potential consequences on human and environmental health especially as its genotoxicity has scarcely been investigated so far. Thus, we investigated, for the first time, the influence of non-cytotoxic concentrations of CNa(+) (15.65, 31.25 and 62.5 μg/mL) on genomic instability of human hepatoma cell line (HepG2) by determining the formation of micronuclei (MNi), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) with the Cytokinesis block micronucleus cytome assay. Further on we studied the influence of CNa(+) on the expression of several genes involved in toxicity mechanisms using the real-time quantitative PCR. The results showed that CNa(+) increased the number of MNi, while the numbers of NBUDs and NPBs were not affected. In addition it deregulated genes in all the groups studied, mainly after longer time of exposure. These findings provide the evidence that CNa(+) is potentially genotoxic. Therefore further studies that will elucidate the molecular mechanisms involved in toxic activity of CNa(+) are needed for hazard identification and human safety assessment. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. A novel comprehensive evaluation platform to assess nanoparticle toxicity in vitro

    Science.gov (United States)

    Hirsch, C.; Kaiser, J.-P.; Wessling, F.; Fischer, K.; Roesslein, M.; Wick, P.; Krug, H. F.

    2011-07-01

    The amount of engineered nanomaterials (ENM) is constantly increasing. Their unique properties, compared to their bulk counterparts, render them suitable for various applications in many areas of life. Hence, nanomaterials appear in a variety of different consumer products leading to the exposure of human beings and the environment during their lifecycle. Even though results on biological effects of ENM are available, harmonized and validated test systems are still missing. One major problem concerning the reliable and robust toxicity testing arises from interactions of ENM with different assay systems. Modifications or damage to DNA can have fatal consequences, such as the formation of tumor cells and hence carcinogenesis. Therefore we focused on the re-evaluation of two genotoxicity assays concerning their nanomaterial compatibility; namely the cytokinesis-block micronucleus cytome assay (MN-assay) and the alkaline single cell gel electorphoresis assay (comet assay). We demonstrate the interference of ENM agglomerates with the read-out of both assays and discuss possibilities how to acquire relevant genotoxicity data.

  16. A novel comprehensive evaluation platform to assess nanoparticle toxicity in vitro

    International Nuclear Information System (INIS)

    Hirsch, C; Kaiser, J-P; Wessling, F; Fischer, K; Roesslein, M; Wick, P; Krug, H F

    2011-01-01

    The amount of engineered nanomaterials (ENM) is constantly increasing. Their unique properties, compared to their bulk counterparts, render them suitable for various applications in many areas of life. Hence, nanomaterials appear in a variety of different consumer products leading to the exposure of human beings and the environment during their lifecycle. Even though results on biological effects of ENM are available, harmonized and validated test systems are still missing. One major problem concerning the reliable and robust toxicity testing arises from interactions of ENM with different assay systems. Modifications or damage to DNA can have fatal consequences, such as the formation of tumor cells and hence carcinogenesis. Therefore we focused on the re-evaluation of two genotoxicity assays concerning their nanomaterial compatibility; namely the cytokinesis-block micronucleus cytome assay (MN-assay) and the alkaline single cell gel electorphoresis assay (comet assay). We demonstrate the interference of ENM agglomerates with the read-out of both assays and discuss possibilities how to acquire relevant genotoxicity data.

  17. Screening potential genotoxic effect of aquatic plant extracts using the mussel micronucleus test

    Directory of Open Access Journals (Sweden)

    Bettina Eck-Varanka

    2016-01-01

    Full Text Available Objective: To assess the genotoxic potential of selected aquatic macrophytes: Ceratophyllum demersum L. (hornwort, family Ceratophyllaceae, Typha angustifolia L. (narrowleaf cattail, family Typhaceae, Stratiotes aloides L. (water soldier, family Butomaceae, and Oenanthe aquatica (L. Poir. (water dropwort, family Umbelliferae. Methods: For genotoxicity assessment, the mussel micronucleus test was applied. Micronucleus frequency was determined from the haemolymph of Unio pictorum L. (painter’s mussel. In parallel, total and hydrolisable tannin contents were determined. Results: All plant extracts elucidated significant mutagenic effect. Significant correlation was determined between tannin content and mutagenic capacity. Conclusions: The significant correlation between genotoxicity as expressed by micronucleus frequency and tannin content (both total and hydrolisable tannins indicate that tannin is amongst the main compounds being responsible for the genotoxic potential. It might be suggested that genotoxic capacity of these plants elucidate a real ecological effect in the ecosystem.

  18. Radiation dose-response relationship of micronucleus occurrence in pollen mother cells of tradescantia

    International Nuclear Information System (INIS)

    Kim, Jin Kyu; Kim, Yeon Ku; Song, Hi Sup

    1999-01-01

    This study was carried out to investigate the radiation dose-response of micronucleus frequencies in Tradescantia pollen mother cells. The number of micronuclei increased in the tetrads as a result of chromosome deletion after irradiation. The maximal frequency of micronucleus showed a good dose-response relationship in the range of dose 0∼50 cGy. On the basis of the relationship, a dose of 1 cGy resulted in two additional micronuclei in 100 tetrads. The radiation dose-response relationship of micronucleus occurrence is prerequisite to biological monitoring of radiation and can be modified for biological risk assessment of toxicants, and to safety test of water or soil integrity

  19. Micronucleus induction by repeated exposure of diagnostic X-ray on oral buccal mucosa

    International Nuclear Information System (INIS)

    Lohith Tejashvi, K.; Suchetha Kumari, N.; Shetty, Shishir Ram

    2012-01-01

    Radiography is the important diagnostic tools essential for diagnosis and planning of orthodontic treatment. X-ray is ionizing radiation which showed various effects include breaking the bond of biological molecules, inducing loss of ability of cell death, increases nuclear alterations. Micronuclei - x000D - (MN) are small chromatin bodies that appear in the cytoplasm by the - x000D - condensation of acrocentric chromosomal fragments or by whole chromosomes. This - x000D - is a sensitive indicator of genetic damage. - x000D - x000D - . To evaluate micronucleus induction by repeated exposure of diagnostic X-ray on human buccal cell. Methods: 25 patients who visiting to ABSMIDS, Department of Oral medicine and Radiology for dental checkup exposed to diagnostic X-ray more than 4 times have been selected for this study. The buccal cell for analysis was collected from the cheek mucosa by means of gentle scraping of epithelial using ice-cream sticks and placed in Buffer saline. This sample was smeared on glass slide and then fixed in methanol:glacial acetic acid (3:1). Air dried and stained with Giemsa for 15-25 minutes. Then 250 cells in each slides were analyzed under microscope and frequency of micronucleus was scored (n=4). Repeated X-ray exposed cells showed micronucleus (1.25%) and nuclear alteration (2.3%) compare to the control. Repeated X-ray exposure leads to induces detectable number of micronucleus and nuclear alterations. (author)

  20. Absence of micronucleus formation in CHO-K1 cells cultivated in platelet lysate enriched medium.

    Science.gov (United States)

    Bernardi, Martina; Adami, Valentina; Albiero, Elena; Madeo, Domenico; Rodeghiero, Francesco; Astori, Giuseppe

    2014-03-01

    Human platelet lysate (PL) represents an effective substitute of fetal bovine serum (FBS) for mesenchymal stromal cell (MSC) cultivation. Compared to FBS, PL favors MSC proliferation significantly shortening the population doubling time and avoiding the risks related to the use of animal derivatives. Growth factors contained in the platelets are released upon platelet disruption following freezing/thawing cycles or as we have recently described by using ultrasound. We have investigated whether the increased cell proliferation achieved by using PL could induce mitotic stress and whether the potential formation of free radicals during PL production by ultrasound could cause chromosomal instability in mammalian cells. We have applied an image analysis assisted high content screening (HCS) in vitro micronucleus assay in the Chinese Hamster Ovarian K1 (CHO-K1) rodent mammalian cell line. PL was produced by sonication; for the micronucleus assay, CHO-K1 cells were exposed to increasing concentrations of PL. Cytokinesis was blocked by cytochalasin B, nuclei were stained with bisbenzimide and images were acquired and analyzed automatically using an HCS system, both with a 20× and a 10× objective. Our results suggest that growth stimulus induced by the use of PL did not significantly increase micronucleus formation in CHO-K1 cells compared to negative control. Micronucleus testing in conjunction with HCS could represent a valid tool to evaluate the safety of ancillary materials used in the production of cell-based medicinal products. Copyright © 2013 Elsevier GmbH. All rights reserved.

  1. 40 CFR 798.5395 - In vivo mammalian bone marrow cytogenetics tests: Micronucleus assay.

    Science.gov (United States)

    2010-07-01

    ... Genetic Toxicity § 798.5395 In vivo mammalian bone marrow cytogenetics tests: Micronucleus assay. (a... and documented with data, only this one time point need be sampled. (ii) If a repeated treatment... slides, spread as a smear and stained. (2) Analysis. Slides shall be coded before microscopic analysis...

  2. Association between micronucleus frequency and cervical intraepithelial neoplasia grade in Thinprep cytological test and its significance.

    Science.gov (United States)

    Shi, Yong-Hua; Wang, Bo-Wei; Tuokan, Talaf; Li, Qiao-Zhi; Zhang, Ya-Jing

    2015-01-01

    A micronucleus is an additional small nucleus formed due to chromosomes or chromosomal fragments fail to be incorporated into the nucleus during cell division. In this study, we assessed the utility of micronucleus counting as a screening tool in cervical precancerous lesions in Thinprep cytological test smears under oil immersion. High risk HPV was also detected by hybrid capture-2 in Thinprep cytological test smears. Our results showed that micronucleus counting was significantly higher in high-grade squamous intraepithelial lesion (HSIL) and invasive carcinoma cases compared to low-grade squamous intraepithelial lesion (LSIL) and non-neoplastic cases. Receiver operating characteristic (ROC) curve analysis revealed that micronucleus counting possessed a high degree of sensitivity and specificity for identifying HSIL and invasive carcinoma. Cut-off of 7.5 for MN counting gave a sensitivity of 89.6% and a specificity of 66.7% (P = 0.024 and AUC = 0.892) for detecting HSIL and invasive carcinoma lesions. Multiple linear regression analysis showed that only HSIL and invasive cancer lesions not age, duration of marital life and number of pregnancy are significantly associated with MN counting. The positive rate of high risk HPV was distinctly higher in LSIL, HSIL and invasive cancer than that in non-neoplstic categories. In conclusions, MN evaluation may be viewed as an effective biomarker for cervical cancer screening. The combination of MN count with HPV DNA detection and TCT may serve as an effective means to screen precancerous cervical lesions in most developing nations.

  3. Dose-rate effects for apoptosis and micronucleus formation in gamma-irradiated human lymphocytes

    International Nuclear Information System (INIS)

    Boreham, D.R.; Dolling, J.-A.; Maves, S.R.; Siwarungsun, N.; Mitchel, R.E.J.

    2000-01-01

    We have compared dose-rate effects for γ-radiation-induced apoptosis and micronucleus formation in human lymphocytes. Long-term assessment of individual radiation-induced apoptosis showed little intraindividual variation but significant interindividual variation. The effectiveness of radiation exposure to cause apoptosis or micronucleus formation was reduced by low-dose-rate exposures, but the reduction was apparent at different dose rates for these two end points. Micronucleus formation showed a dose-rate effect when the dose rate was lowered to 0.29 cGy/min, but there was no accompanying cell cycle delay. A further increase in the dose-rate effect was seen at 0.15 cGy/min, but was now accompanied by cell cycle delay. There was no dose-rate effect for the induction of apoptosis until the dose rate was reduced to 0.15 cGy/min, indicating that the mechanisms or signals for processing radiation-induced lesions for these two end points must be different at least in part. There appear to be two mechanisms that contribute to the dose-rate effect for micronucleus formation. One of these does not affect binucleate cell frequency and occurs at dose rates higher than that required to produce a dose-rate effect for apoptosis, and one affects binucleate cell frequency, induced only at the very low dose rate which coincidentally produces a dose-rate effect for apoptosis. Since the dose rate at which cells showed reduced apoptosis as well as a further reduction in micronucleus formation was very low, we conclude that the processing of the radiation-induced lesions that induce apoptosis, and some micronuclei, is very slow in quiescent and PHA-stimulated lymphocytes, respectively. (author)

  4. Dose-rate effects for apoptosis and micronucleus formation in gamma-irradiated human lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Boreham, D.R.; Dolling, J.-A.; Maves, S.R. [Atomic Energy of Canada Limited, Chalk River, Ontario (Canada); Siwarungsun, N. [Chulalongkorn Univ., Bangkok (Thailand); Mitchel, R.E.J. [Atomic Energy of Canada Limited, Chalk River, Ontario (Canada)

    2000-07-01

    We have compared dose-rate effects for {gamma}-radiation-induced apoptosis and micronucleus formation in human lymphocytes. Long-term assessment of individual radiation-induced apoptosis showed little intraindividual variation but significant interindividual variation. The effectiveness of radiation exposure to cause apoptosis or micronucleus formation was reduced by low-dose-rate exposures, but the reduction was apparent at different dose rates for these two end points. Micronucleus formation showed a dose-rate effect when the dose rate was lowered to 0.29 cGy/min, but there was no accompanying cell cycle delay. A further increase in the dose-rate effect was seen at 0.15 cGy/min, but was now accompanied by cell cycle delay. There was no dose-rate effect for the induction of apoptosis until the dose rate was reduced to 0.15 cGy/min, indicating that the mechanisms or signals for processing radiation-induced lesions for these two end points must be different at least in part. There appear to be two mechanisms that contribute to the dose-rate effect for micronucleus formation. One of these does not affect binucleate cell frequency and occurs at dose rates higher than that required to produce a dose-rate effect for apoptosis, and one affects binucleate cell frequency, induced only at the very low dose rate which coincidentally produces a dose-rate effect for apoptosis. Since the dose rate at which cells showed reduced apoptosis as well as a further reduction in micronucleus formation was very low, we conclude that the processing of the radiation-induced lesions that induce apoptosis, and some micronuclei, is very slow in quiescent and PHA-stimulated lymphocytes, respectively. (author)

  5. Automatic analysis of the micronucleus test in primary human lymphocytes using image analysis.

    Science.gov (United States)

    Frieauff, W; Martus, H J; Suter, W; Elhajouji, A

    2013-01-01

    The in vitro micronucleus test (MNT) is a well-established test for early screening of new chemical entities in industrial toxicology. For assessing the clastogenic or aneugenic potential of a test compound, micronucleus induction in cells has been shown repeatedly to be a sensitive and a specific parameter. Various automated systems to replace the tedious and time-consuming visual slide analysis procedure as well as flow cytometric approaches have been discussed. The ROBIAS (Robotic Image Analysis System) for both automatic cytotoxicity assessment and micronucleus detection in human lymphocytes was developed at Novartis where the assay has been used to validate positive results obtained in the MNT in TK6 cells, which serves as the primary screening system for genotoxicity profiling in early drug development. In addition, the in vitro MNT has become an accepted alternative to support clinical studies and will be used for regulatory purposes as well. The comparison of visual with automatic analysis results showed a high degree of concordance for 25 independent experiments conducted for the profiling of 12 compounds. For concentration series of cyclophosphamide and carbendazim, a very good correlation between automatic and visual analysis by two examiners could be established, both for the relative division index used as cytotoxicity parameter, as well as for micronuclei scoring in mono- and binucleated cells. Generally, false-positive micronucleus decisions could be controlled by fast and simple relocation of the automatically detected patterns. The possibility to analyse 24 slides within 65h by automatic analysis over the weekend and the high reproducibility of the results make automatic image processing a powerful tool for the micronucleus analysis in primary human lymphocytes. The automated slide analysis for the MNT in human lymphocytes complements the portfolio of image analysis applications on ROBIAS which is supporting various assays at Novartis.

  6. Necrosis is increased in lymphoblastoid cell lines from children with autism compared with their non-autistic siblings under conditions of oxidative and nitrosative stress

    Science.gov (United States)

    Fenech, Michael F.

    2013-01-01

    Autism spectrum disorders are a heterogeneous group of neurodevelopmental conditions characterised by impairments in reciprocal social interaction, communication and stereotyped behaviours. As increased DNA damage events have been observed in a range of other neurological disorders, it was hypothesised that they would be elevated in lymphoblastoid cell lines (LCLs) obtained from children with autism compared with their non-autistic siblings. Six case–sibling pairs of LCLs from children with autistic disorder and their non-autistic siblings were obtained from the Autism Genetic Resource Exchange (AGRE) and cultured in standard RPMI-1640 tissue culture medium. Cells were exposed to medium containing either 0, 25, 50, 100 and 200 µM hydrogen peroxide (an oxidative stressor) or 0, 5, 10, 20 and 40 µM s-nitroprusside (a nitric oxide producer) for 1h. Following exposure, the cells were microscopically scored for DNA damage, cytostasis and cytotoxicity biomarkers as measured using the cytokinesis-block micronucleus cytome assay. Necrosis was significantly increased in cases relative to controls when exposed to oxidative and nitrosative stress (P = 0.001 and 0.01, respectively). Nuclear division index was significantly lower in LCLs from children with autistic disorder than their non-autistic siblings when exposed to hydrogen peroxide (P = 0.016), but there was no difference in apoptosis, micronucleus frequency, nucleoplasmic bridges or nuclear buds. Exposure to s-nitroprusside significantly increased the number of micronuclei in non-autistic siblings compared with cases (P = 0.003); however, other DNA damage biomarkers, apoptosis and nuclear division did not differ significantly between groups. The findings of this study show (i) that LCLs from children with autism are more sensitive to necrosis under conditions of oxidative and nitrosative stress than their non-autistic siblings and (ii) refutes the hypothesis that children with autistic disorder are abnormally

  7. Necrosis is increased in lymphoblastoid cell lines from children with autism compared with their non-autistic siblings under conditions of oxidative and nitrosative stress.

    Science.gov (United States)

    Main, Penelope A E; Thomas, Philip; Esterman, Adrian; Fenech, Michael F

    2013-07-01

    Autism spectrum disorders are a heterogeneous group of neurodevelopmental conditions characterised by impairments in reciprocal social interaction, communication and stereotyped behaviours. As increased DNA damage events have been observed in a range of other neurological disorders, it was hypothesised that they would be elevated in lymphoblastoid cell lines (LCLs) obtained from children with autism compared with their non-autistic siblings. Six case-sibling pairs of LCLs from children with autistic disorder and their non-autistic siblings were obtained from the Autism Genetic Resource Exchange (AGRE) and cultured in standard RPMI-1640 tissue culture medium. Cells were exposed to medium containing either 0, 25, 50, 100 and 200 µM hydrogen peroxide (an oxidative stressor) or 0, 5, 10, 20 and 40 µM s-nitroprusside (a nitric oxide producer) for 1h. Following exposure, the cells were microscopically scored for DNA damage, cytostasis and cytotoxicity biomarkers as measured using the cytokinesis-block micronucleus cytome assay. Necrosis was significantly increased in cases relative to controls when exposed to oxidative and nitrosative stress (P = 0.001 and 0.01, respectively). Nuclear division index was significantly lower in LCLs from children with autistic disorder than their non-autistic siblings when exposed to hydrogen peroxide (P = 0.016), but there was no difference in apoptosis, micronucleus frequency, nucleoplasmic bridges or nuclear buds. Exposure to s-nitroprusside significantly increased the number of micronuclei in non-autistic siblings compared with cases (P = 0.003); however, other DNA damage biomarkers, apoptosis and nuclear division did not differ significantly between groups. The findings of this study show (i) that LCLs from children with autism are more sensitive to necrosis under conditions of oxidative and nitrosative stress than their non-autistic siblings and (ii) refutes the hypothesis that children with autistic disorder are abnormally

  8. Genotoxic potential of montmorillonite clay mineral and alteration in the expression of genes involved in toxicity mechanisms in the human hepatoma cell line HepG2

    Energy Technology Data Exchange (ETDEWEB)

    Maisanaba, Sara, E-mail: saramh@us.es [Area of Toxicology, Faculty of Pharmacy, University of Sevilla, Profesor García González no. 2, 41012 Seville (Spain); Hercog, Klara; Filipic, Metka [National Institute of Biology, Department for Genetic Toxicology and Cancer Biology, Vecna pot 111, 1000 Ljubljana (Slovenia); Jos, Ángeles [Area of Toxicology, Faculty of Pharmacy, University of Sevilla, Profesor García González no. 2, 41012 Seville (Spain); Zegura, Bojana [National Institute of Biology, Department for Genetic Toxicology and Cancer Biology, Vecna pot 111, 1000 Ljubljana (Slovenia)

    2016-03-05

    Highlights: • Cloisite{sup ®}Na{sup +} has a wide range of well-documented and novel applications. • Cloisite{sup ®}Na{sup +} induces micronucleus, but not nuclear bridges or nuclear buds in HepG2 cells. • Cloisite{sup ®}Na{sup +} induces changes in the gene expression. • Gene alteration is presented mainly after 24 h of exposure to Cloisite{sup ®}Na{sup +}. - Abstract: Montmorillonite, also known as Cloisite{sup ®}Na{sup +} (CNa{sup +}), is a natural clay with a wide range of well-documented and novel applications, such as pharmaceutical products or food packaging. Although considered a low toxic product, the expected increased exposure to CNa{sup +} arises concern on the potential consequences on human and environmental health especially as its genotoxicity has scarcely been investigated so far. Thus, we investigated, for the first time, the influence of non-cytotoxic concentrations of CNa{sup +} (15.65, 31.25 and 62.5 μg/mL) on genomic instability of human hepatoma cell line (HepG2) by determining the formation of micronuclei (MNi), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) with the Cytokinesis block micronucleus cytome assay. Further on we studied the influence of CNa{sup +} on the expression of several genes involved in toxicity mechanisms using the real-time quantitative PCR. The results showed that CNa{sup +} increased the number of MNi, while the numbers of NBUDs and NPBs were not affected. In addition it deregulated genes in all the groups studied, mainly after longer time of exposure. These findings provide the evidence that CNa{sup +} is potentially genotoxic. Therefore further studies that will elucidate the molecular mechanisms involved in toxic activity of CNa{sup +} are needed for hazard identification and human safety assessment.

  9. A cytogenetic study of Korean native goat bred in the nuclear power plant using the micronucleus assay

    International Nuclear Information System (INIS)

    Kang, Chang-Mo; Ji, Young-Hoon; Lee, Hae-June; Kim, Se-Ra; Kim, Jong-Choon; Kim, Sung-Ho

    2005-01-01

    Cytogenetic and hematological analysis was performed on the peripheral blood lymphocytes (PBLs) obtained from Korean native goats bred in two nuclear power plants (Wolsong and Uljin) and a control area. The frequencies of gamma-ray-induced micronuclei (MN) in the cytokinesis-blocked (CB) lymphocytes at several doses were measured in three Korean native goats. The measurements performed after irradiation showed dose-related increases in the MN frequency in each of the donors. The results were analyzed using a linear-quadratic model with a line of best fit of y=0.1019D+0.0045D 2 +0.0093 (y=number of MN/GB cells and D=irradiation dose in Gy). The MN rates in the goats from the Wolsong and Uljin nuclear power plant, and the control area were 9.60±2.88, 6.83±1.47 and 9.88±4.32 per 1,000 CB lymphocytes, respectively. The apparent difference is not statistically significant. The MN frequencies of PBLs from goats bred in three areas means that the values are within the background variation in this experiment. The MN frequencies and hematological values were similar regardless of whether the goats were bred in the nuclear power plant or the control area. (author)

  10. State of the Art High-Throughput Approaches to Genotoxicity: Flow Micronucleus, Ames II, GreenScreen and Comet

    Science.gov (United States)

    State of the Art High-Throughput Approaches to Genotoxicity: Flow Micronucleus, Ames II, GreenScreen and Comet (Presented by Dr. Marilyn J. Aardema, Chief Scientific Advisor, Toxicology, Dr. Leon Stankowski, et. al. (6/28/2012)

  11. Assessment of the genotoxicity of 137Cs radiation using Vicia-micronucleus, Tradescantia-micronucleus and Tradescantia-stamen-hair mutation bioassays.

    Science.gov (United States)

    Minouflet, Marion; Ayrault, Sophie; Badot, Pierre-Marie; Cotelle, Sylvie; Ferard, Jean-François

    2005-01-01

    Since the middle of the 20th century, ionizing radiations from radioactive isotopes including 137Cs have been investigated to determine their genotoxic impact on living organisms. The present study was designed to compare the effectiveness of three plant bioassays to assess DNA damage induced by low doses of 137Cs: Vicia-micronucleus test (Vicia-MCN), Tradescantia-micronucleus test (Trad-MCN) and Tradescantia-stamen-hair mutation test (Trad-SH) were used. Vicia faba (broad bean) and Tradescantia clone 4430 (spiderwort) were exposed to 137Cs according to different scenarios: external and internal (contamination) irradiations. Experiments were conducted with various levels of radioactivity in solution or in soil, using solid or liquid 137Cs sources. The three bioassays showed different sensitivities to the treatments. Trad-MCN appeared to be the most sensitive test (significative response from 1.5 kBq/200 ml after 30 h of contamination). Moreover, at comparable doses, internal irradiations led to larger effects for the three bioassays. These bioassays are effective tests for assessing the genotoxic effects of radioactive 137Cs pollution.

  12. Assessment of the genotoxicity of 137Cs radiation using Vicia-micronucleus, Tradescantia-micronucleus and Tradescantia-stamen-hair mutation bioassays

    International Nuclear Information System (INIS)

    Minouflet, Marion; Ayrault, Sophie; Badot, Pierre-Marie; Cotelle, Sylvie; Ferard, Jean-Francois

    2005-01-01

    Since the middle of the 20th century, ionizing radiations from radioactive isotopes including 137 Cs have been investigated to determine their genotoxic impact on living organisms. The present study was designed to compare the effectiveness of three plant bioassays to assess DNA damage induced by low doses of 137 Cs: Vicia-micronucleus test (Vicia-MCN), Tradescantia-micronucleus test (Trad-MCN) and Tradescantia-stamen-hair mutation test (Trad-SH) were used. Vicia faba (broad bean) and Tradescantia clone 4430 (spiderwort) were exposed to 137 Cs according to different scenarios: external and internal (contamination) irradiations. Experiments were conducted with various levels of radioactivity in solution or in soil, using solid or liquid 137 Cs sources. The three bioassays showed different sensitivities to the treatments. Trad-MCN appeared to be the most sensitive test (significative response from 1.5 kBq/200 ml after 30 h of contamination). Moreover, at comparable doses, internal irradiations led to larger effects for the three bioassays. These bioassays are effective tests for assessing the genotoxic effects of radioactive 137 Cs pollution

  13. Evaluation of 10 aliphatic halogenated hydrocarbons in the mouse bone marrow micronucleus test.

    Science.gov (United States)

    Crebelli, R; Carere, A; Leopardi, P; Conti, L; Fassio, F; Raiteri, F; Barone, D; Ciliutti, P; Cinelli, S; Vericat, J A

    1999-03-01

    Ten halogenated aliphatic hydrocarbons (carbon tetrachloride, 1-chlorohexane, 2,3-dichlorobutane, 1,2-dichloroethane, 1,2-dichloroethylene, 1,3-dichloropropane, hexachloroethane, 1,1,2-trichloroethane, 1,2,3-trichloropropane and 1,1,3-trichloropropene), previously assayed in genetic assays in fungi, were evaluated in the mouse bone marrow micronucleus test in order to assess their genotoxicity in vivo. All chemicals were administered once i.p. at 40 and 70-80% of their respective LD50 to male and female CD-1 mice, 24 and 48 h before killing. All treatments produced evident clinical symptoms, but no marked depression of bone marrow proliferation. No statistically significant increases in the incidence of micronucleated polychromatic erythrocytes over the control values were observed at any sampling time with any of the 10 halogenated hydrocarbons assayed. The comparison of the results obtained in this study with the findings provided by in vitro micronucleus assays on the same chemicals, reported by other authors, indicate that mouse bone marrow is weakly sensitive to the genotoxic effects induced by halogenated hydrocarbons in other test systems. This suggests that the role of such an assay in carcinogen screening may be questionable for this chemical class. An examination of mouse bone marrow micronucleus test results with the halogenated aliphatic hydrocarbons classified as carcinogens by IARC supports this conclusion.

  14. Variability in micronucleus induction with different mutagens applied to several species of fish

    Directory of Open Access Journals (Sweden)

    Cesar Koppe Grisolia

    2000-03-01

    Full Text Available Fish are often used for screening genotoxicity of water. For such programs, a knowledge of the sensitivity to clastogens, spontaneous micronucleus frequency and cell cycle kinetics of the target tissue is necessary. To investigate the pattern of inter-specific sensitivity to micronucleus induction three species of fish, Tilapia rendalli, Oreochromis niloticus and Cyprinus carpio, were exposed to the clastogens bleomycin (BLM, cyclophosphamide (CP, 5-fluorouracil (5-FU, and mitomycin C (MMC. The binucleate/mononucleate ratio in peripheral erythrocytes exposed to cytochalasin B was also used to evaluate the time-dependent response of micronucleus formation during hematopoesis in the kidney and the micronucleus peak in peripheral erythrocytes. Micronucleus frequencies induced by CP were significantly greater than their respective controls for the three fish species throughout all treatment periods. During the whole evaluation period (30 days CP was also the most effective clastogen. In general, until the 14th day of evaluation period T. rendalii was the most sensitive species to clastogens. No difference in micronucleus frequencies among species was observed in the 4th evaluation (at the 30th day. A micronucleus peak was observed at the 7th day after treatment. After the 14th day the frequencies were stabilized. The cytochalasin B experiment was carried out to demonstrate that micronuclei induced in the young kidney erythrocyte cells were detected in the circulating blood 2-4 days later.Este estudo fez uma avaliação da indução de micronúcleos em eritrócitos de sangue periférico de peixes Tilapia rendalli, Oreochromis niloticus e Cyprinus carpio após o tratamento com mitomicina C, ciclofosfamida, 5-fluorouracil e bleomicina. Foram colhidas amostras periódicas de sangue com 2, 7, 14 e 30 dias após o tratamento único. Os tratamentos com citocalasina B tiveram como objetivo analisar as proporções entre células binucleadas

  15. Health assessment of gasoline and fuel oxygenate vapors: micronucleus and sister chromatid exchange evaluations.

    Science.gov (United States)

    Schreiner, Ceinwen A; Hoffman, Gary M; Gudi, Ramadevi; Clark, Charles R

    2014-11-01

    Micronucleus and sister chromatid exchange (SCE) tests were performed for vapor condensate of baseline gasoline (BGVC), or gasoline with oxygenates, methyl tert-butyl ether (G/MTBE), ethyl tert butyl ether (G/ETBE), t-amyl methyl ether (G/TAME), diisopropyl ether (G/DIPE), t-butyl alcohol (TBA), or ethanol (G/EtOH). Sprague Dawley rats (the same 5/sex/group for both endpoints) were exposed to 0, 2000, 10,000, or 20,000mg/m(3) of each condensate, 6h/day, 5days/week over 4weeks. Positive controls (5/sex/test) were given cyclophosphamide IP, 24h prior to sacrifice at 5mg/kg (SCE test) and 40mg/kg (micronucleus test). Blood was collected from the abdominal aorta for the SCE test and femurs removed for the micronucleus test. Blood cell cultures were treated with 5μg/ml bromodeoxyuridine (BrdU) for SCE evaluation. No significant increases in micronucleated immature erythrocytes were observed for any test material. Statistically significant increases in SCE were observed in rats given BGVC alone or in female rats given G/MTBE. G/TAME induced increased SCE in both sexes at the highest dose only. Although DNA perturbation was observed for several samples, DNA damage was not expressed as increased micronuclei in bone marrow cells. Inclusion of oxygenates in gasoline did not increase the effects of gasoline alone or produce a cytogenetic hazard. Copyright © 2014 Elsevier Inc. All rights reserved.

  16. Study on ionizing radiation to the workers' lymphocyte micronucleus rate and chromosome aberrations

    International Nuclear Information System (INIS)

    Li Jianhua; Wang Linchao; He Wei

    2007-01-01

    Objective: To study lymphocyte genetic material of an iron and steel enterprise workers exposed to the ionizing radiation, find out measures to protect their health and reduce ionizing radiation occupation harm. Methods: 342 workers were choseh as the exposed group who worked in an iron and steel enterprise in the beam installment operation, to examine their circumference blood lymphocyte micronucleus rate and the chromosome aberrations, simultaneously select 280 chefs as the control group, The irradiation dosage was determined and statistical analysis was carded out wich the consideration of their length of work and differences in work post. Results: Exposed group: the micronucleus rate masculine gender (MNR), 4 people, the masculine gender pick out rate is 12.87%. The chromosome aberration factor masculine gender (CAF), 12 people, the masculine rate is 3.51%. Control group: MNR 3 people, the asculine gender pick out rate is 1.07%; CAF 2 people, masculine gender rate is 0.72%. Comparing the two groups, every item has the significant difference. Workers in is the exposed group workers have the average exposure dose of 6.73mSv/a, MNR,CAF are illuminated to the dosage have a positive line correlation. They become increased as the job lenght prolongs. The nucleon name, the material calculation and the medical X-radial are responsible for the highest ratio. Conclusion: In iron and steel enterprises, long-time ionizing radiation can cause the workers' circumference blood lymphocyte micronucleus rate and the chromosome aberrations obvious to rise. The beam protection measures strengthened so as to reduce the harms to workers. (authors)

  17. p53 protein expression versus micronucleus induction as an indicator of DNA damage

    International Nuclear Information System (INIS)

    Hickman, A.W.; Carpenter, T.R.; Johnson, N.F.

    1994-01-01

    In vitro assays for detecting DNA damage play an important role in evaluating the possible adverse health effects of chemical compounds. Exposure to many DNA-damaging agents in vitro has been shown to cause elevated levels of the tumor-suppressor protein p53. Work in our laboratory has shown that induction of the p53 protein is useful as a biodosimeter for determining the radiation dose to cells. The purpose of this investigation was to compare the sensitivity of this assay to that of micronucleus induction, which is commonly used as a marker of radiation-induced damage

  18. Patient exposure and micronucleus assay during extracorporeal shock wave lithotripsy (ESWL) for kidney stones

    International Nuclear Information System (INIS)

    Shao, M.

    1992-01-01

    This study reports results of radiation exposure to 20 patients with kidney stones during Extracorporeal Shock Wave Lithotripsy (ESWL). A domestically made JT-ESWL-I Lithotripter was used. Data indicated that the amount of radiation exposure is related to the numbers, size, location and radiodensity of the stone, and also the number of shock wave and the time of fluoroscopy exposure given to the patients. The results of the micronucleus frequency of lymphocytes assay in the human peripheral blood are reported. This effect increased with increasing radiation exposure doses. (author). 4 refs., 2 tabs

  19. Tradescantia micronucleus test indicates genotoxic potential of traffic emissions in European cities

    International Nuclear Information System (INIS)

    Klumpp, Andreas; Ansel, Wolfgang; Klumpp, Gabriele; Calatayud, Vicent; Garrec, Jean Pierre; He Shang; Penuelas, Josep; Ribas, Angela; Ro-Poulsen, Helge; Rasmussen, Stine; Sanz, Maria Jose; Vergne, Phillippe

    2006-01-01

    Urban atmospheres contain complex mixtures of air pollutants including mutagenic and carcinogenic substances such as benzene, diesel soot, heavy metals and polycyclic aromatic hydrocarbons. In the frame of a European network for the assessment of air quality by the use of bioindicator plants, the Tradescantia micronucleus (Trad-MCN) test was applied to examine the genotoxicity of urban air pollution. Cuttings of Tradescantia clone no. 4430 were exposed to ambient air at 65 monitoring sites in 10 conurbations employing a standardised methodology. The tests revealed an elevated genotoxic potential mainly at those urban sites which were exposed to severe car traffic emissions. This bioassay proved to be a suitable tool to detect local 'hot spots' of mutagenic air pollution in urban areas. For its use in routine monitoring programmes, however, further standardisation of cultivation and exposure techniques is recommended in order to reduce the variability of results due to varying environmental conditions. - The Tradescantia micronucleus test can be used to assess genotoxic potential at urban sites

  20. Tradescantia micronucleus test indicates genotoxic potential of traffic emissions in European cities

    Energy Technology Data Exchange (ETDEWEB)

    Klumpp, Andreas [Institute for Landscape and Plant Ecology (320), University of Hohenheim, 70593 Stuttgart (Germany)]. E-mail: aklumpp@uni-hohenheim.de; Ansel, Wolfgang [Institute for Landscape and Plant Ecology (320), University of Hohenheim, 70593 Stuttgart (Germany); Klumpp, Gabriele [Institute for Landscape and Plant Ecology (320), University of Hohenheim, 70593 Stuttgart (Germany); Calatayud, Vicent [Fundacion CEAM, Parque Tecnologico, c/Charles Darwin 14, 46980 Paterna, Valencia (Spain); Garrec, Jean Pierre [INRA Nancy, Laboratoire Pollution Atmospherique, 54280 Champenoux (France); He Shang [INRA Nancy, Laboratoire Pollution Atmospherique, 54280 Champenoux (France); Penuelas, Josep [Unitat Ecofisiologia CSIC-CEAB-CREAF, Universitat Autonoma de Barcelona, Ed. C, 08193 Bellaterra, Barcelona (Spain); Ribas, Angela [Unitat Ecofisiologia CSIC-CEAB-CREAF, Universitat Autonoma de Barcelona, Ed. C, 08193 Bellaterra, Barcelona (Spain); Ro-Poulsen, Helge [Botanical Institute, University of Copenhagen, Oster Farimagsgade 2D, 1353 Copenhagen K (Denmark); Rasmussen, Stine [Botanical Institute, University of Copenhagen, Oster Farimagsgade 2D, 1353 Copenhagen K (Denmark); Sanz, Maria Jose [Fundacion CEAM, Parque Tecnologico, c/Charles Darwin 14, 46980 Paterna, Valencia (Spain); Vergne, Phillippe [ENS Lyon and Lyon Botanical Garden, 46 Allee d' Italie, 69364 Lyon Cedex 07 (France)

    2006-02-15

    Urban atmospheres contain complex mixtures of air pollutants including mutagenic and carcinogenic substances such as benzene, diesel soot, heavy metals and polycyclic aromatic hydrocarbons. In the frame of a European network for the assessment of air quality by the use of bioindicator plants, the Tradescantia micronucleus (Trad-MCN) test was applied to examine the genotoxicity of urban air pollution. Cuttings of Tradescantia clone no. 4430 were exposed to ambient air at 65 monitoring sites in 10 conurbations employing a standardised methodology. The tests revealed an elevated genotoxic potential mainly at those urban sites which were exposed to severe car traffic emissions. This bioassay proved to be a suitable tool to detect local 'hot spots' of mutagenic air pollution in urban areas. For its use in routine monitoring programmes, however, further standardisation of cultivation and exposure techniques is recommended in order to reduce the variability of results due to varying environmental conditions. - The Tradescantia micronucleus test can be used to assess genotoxic potential at urban sites.

  1. Examination of Mutagenic Effects of GAL-57 Herbicide (Bentazone+Dicamba Using Mouse Micronucleus Test

    Directory of Open Access Journals (Sweden)

    Vesela Karan

    2007-01-01

    Full Text Available A micronucleus test was run to investigate mutagenic potential of the herbicide GAL-57, a formulated mixture of bentazone and dicamba.The test was applied to mice of both sexes (strain: CRL: NMRI BR and the herbicide (product was administered by gavage at 2000 mg/kg rate, twice within 24 hs. Cyclophosphamide (positive control was administered at 60 mg/kg, while distilled water as a solvent was negative control. The animals were sacrificed 24 hs after second treatment, their bone marrow cells isolated from femur, and effects evaluated.The data acquired showed that repeated treatment of mice with GAL-57 caused neither biological nor significant statistical increase in the number of micronuclei in treated animals. At the same time, the number of micronucleated polychromatic erythrocytes in the bone marrow of animals treated with cyclophosphamide (positive control showed a significant statistical increase. The results suggest that the herbicide product tested did not show any mutagenic activity under the conditions of mouse micronucleus test.

  2. Comparison of micronucleus frequencies and proliferation kinetics in three X-irradiated cell lines

    International Nuclear Information System (INIS)

    Kaffenberger, W.; Becker, K.; Beuningen, D. van

    1990-01-01

    The kinetics of the occurrence of micronuclei was correlated with the survival of three mammalian cell lines of human, monkey, and mouse origin after irradiation with 240 kV X-rays. Particular attention was paid to the evaluation of the individual proliferation kinetics of the cell lines as well as to the characterization of micronuclei subpopulation with respect to size and possible biological importance using DNA and BUdR labelling techniques, fluorescence microscopy, and image analysis. The results demonstrate very characteristic size distributions of micronuclei for the three cell lines independent of radiation dose and time after irradiation. A close correlation between cell death and the occurrence of micronuclei (expressed as a calculated 'MN index') after irradiation could be established only when the kinetics of progression of cells through the cell cycle (e.g. the doubling time) and the biological characteristics of micronuclei (e.g. BUdR positivity, the micronucleus frequencies, and the number of micronuclei per main nucleus) were taken into account. Therefore, the micronucleus assay might not be useful as a quantitative perdictive assay in vivo but may allow qualitative estimations of radiation damage only because the necessary proliferation parameters of the cells might not be possible to establish in vivo. (orig.) [de

  3. Micronucleus assay as a biomarker of genotoxicity in the occupational exposure to agrochemicals in rural workers.

    Science.gov (United States)

    Gentile, N; Mañas, F; Bosch, B; Peralta, L; Gorla, N; Aiassa, D

    2012-06-01

    This paper aims to evaluate the genotoxic effect of agrochemicals in rural workers occupationally exposed by the micronucleus assay in peripheral blood lymphocytes and to promote the development of health and environmental preventive and protective practices. A total of 30 blood samples from 20 individuals occupationally exposed to different agrochemicals and 10 unexposed persons, who formed the reference group, were analyzed. We found statistically significant differences (p < 0.0005, Student's t Test) in the frequency of micronuclei between the two groups (7.20 ± 1.55 and 15.15 ± 5.10 CBMN for reference and exposed groups respectively). The analysis of age showed a positive correlation (Pearson Correlation Test) with the frequency of micronuclei in exposed population (p < 0.05; r(2) = 0.47), in contrast with smoking habits and years of exposure. Micronucleus assay allows an early detection of populations at higher risk of having genetic damage, allowing us to implement strategies of intervention for the purpose of contributing to reduce that risk.

  4. Hyperthermia-induced micronucleus formation in a human keratinocyte cell line

    International Nuclear Information System (INIS)

    Hintzsche, Henning; Riese, Thorsten; Stopper, Helga

    2012-01-01

    Elevated temperature can cause biological effects in vitro and in vivo. Many studies on effects of hypo- and hyperthermia have been conducted, but only few studies systematically investigated the formation of genomic damage in the micronucleus test in human cells in vitro as a consequence of different temperatures. In the present study, HaCaT human keratinocytes were exposed to different temperatures from 37 °C to 42 °C for 24 h in a regular cell culture incubator. Micronucleus frequency as a marker of genomic damage was elevated in a temperature-dependent and statistically significant manner. Apoptosis occurred at temperatures of 39 °C or higher. Cell proliferation was unaffected up to 40 °C and decreased at 41 °C and 42 °C. Expression of the heat shock protein Hsp70 was elevated, particularly at temperatures of 40 °C and higher. These findings are in agreement with several in vivo studies and some in vitro studies looking at single, specific temperatures, but a systematically investigated temperature-dependent increase of genomic damage in human keratinocytes in vitro is demonstrated for the first time here.

  5. Effects of estradiol and progesterone on the variability of the micronucleus assay

    International Nuclear Information System (INIS)

    Baeyens, Ans; Vandersickel, Veerle; Thierens, Hubert; Ridder, Leo De; Vral, Anne

    2005-01-01

    To investigate chromosomal radiosensitivity of lymphocytes the micronucleus (MN) assay has been used for many years. The results of these studies suggest the use of the MN assay as a biomarker for cancer predisposition. However, the MN assay has still some limitations associated with the reproducibility and sensitivity. Especially a high intra-individual variability has been observed. An explanation for this high intra-individual variability is not yet available. In literature it is suggested that the high variability among females is attributable to hormonal status. In this study we investigated if the high intra-individual variability in micronucleus formation in lymphocytes of females after in vitro exposure to ionising radiation is caused by variations in hormone levels of estradiol (E2) and progesterone (PROG). For this, the MN assay was performed on blood samples of 18 healthy women during 7 consecutive weeks while the estradiol and progesterone levels were determined at the same time. The MN assay was also examined in cultures of isolated blood lymphocytes with estradiol or progesterone levels added in vitro. The results demonstrated that estradiol and progesterone levels have no influence on the variations in radiation-induced MN yields observed in blood samples of healthy women. These conclusions were confirmed by the 'in vitro' experiments as no correlation between the MN yields and the concentrations of hormones (estradiol or progesterone) added in vitro to isolated lymphocytes cultures was observed

  6. Cytogenetic instability in populations with residential proximity to open-pit coal mine in Northern Colombia in relation to PM10 and PM2.5 levels.

    Science.gov (United States)

    Espitia-Pérez, Lyda; da Silva, Juliana; Espitia-Pérez, Pedro; Brango, Hugo; Salcedo-Arteaga, Shirley; Hoyos-Giraldo, Luz Stella; de Souza, Claudia T; Dias, Johnny F; Agudelo-Castañeda, Dayana; Valdés Toscano, Ana; Gómez-Pérez, Miguel; Henriques, João A P

    2018-02-01

    Epidemiological studies indicate that living in proximity to coal mines is correlated with numerous diseases including cancer, and that exposure to PM 10 and PM 2.5 components could be associated with this phenomenon. However, the understanding of the mechanisms by which PM exerts its adverse effects is still incomplete and comes mainly from studies in occupationally exposed populations. The aims of this study were to: (1) evaluate DNA damage in lymphocytes assessing the cytokinesis-block micronucleus cytome assay (CBMN-cyt) parameters; (2) identify aneugenic or clastogenic effects in lymphocytes of exposed populations using CREST immunostaining for micronuclei; (3) evaluate multi-elemental composition of atmospheric particulate matter; and (4) verify relation between the DNA damage and PM 2.5 and PM 10 levels around the mining area. Analysis revealed a significant increase in micronuclei frequency in binucleated (MNBN) and mononucleated (MNMONO) cells of individuals with residential proximity to open-pit coal mines compared to residents from non-mining areas. Correlation analysis demonstrated a highly significant association between PM 2.5 levels, MNBN frequencies and CREST+ micronuclei induction in exposed residents. These results suggest that PM 2.5 fraction generated in coal mining activities may induce whole chromosome loss (aneuploidy) preferentially, although there are also chromosome breaks. Analysis of the chemical composition of PM 2.5 by PIXE demonstrated that Si, S, K and Cr concentrations varied significantly between coal mining and reference areas. Enrichment factor values (EF) showed that S, Cr and Cu were highly enriched in the coal mining areas. Compared to reference area, mining regions had also higher concentrations of extractable organic matter (EOM) related to nonpolar and polar compounds. Our results demonstrate that PM 2.5 fraction represents the most important health risk for residents living near open-pit mines, underscoring the need for

  7. In vitro evaluation of the cyto-genotoxic potential of Ruthenium(II) SCAR complexes: a promising class of antituberculosis agents.

    Science.gov (United States)

    De Grandis, Rone Aparecido; Resende, Flávia Aparecida; da Silva, Monize Martins; Pavan, Fernando Rogério; Batista, Alzir Azevedo; Varanda, Eliana Aparecida

    2016-03-01

    Tuberculosis is a top infectious disease killer worldwide, caused by the bacteria Mycobacterium tuberculosis. Increasing incidences of multiple drug-resistance (MDR) strains are emerging as one of the major public health threats. However, the drugs in use are still incapable of controlling the appalling upsurge of MDR. In recent years a marked number of research groups have devoted their attention toward the development of specific and cost-effective antimicrobial agents against targeted MDR-Tuberculosis. In previous studies, ruthenium(II) complexes (SCAR) have shown a promising activity against MDR-Tuberculosis although few studies have indeed considered ruthenium toxicity. Therefore, within the preclinical requirements, we have sought to determine the cyto-genotoxicity of three SCAR complexes in this present study. The treatment with the SCARs induced a concentration-dependent decrease in cell viability in CHO-K1 and HepG2 cells. Based on the clonogenic survival, SCAR 5 was found to be more cytotoxic while SCAR 6 exhibited selectivity action on tumor cells. Although SCAR 4 and 5 did not indicate any mutagenic activity as evidenced by the Ames and Cytokinesis block micronucleus cytome assays, the complex SCAR 6 was found to engender a frameshift mutation detected by Salmonella typhimurium in the presence of S9. Similarly, we observed a chromosomal damage in HepG2 cells with significant increases of micronuclei and nucleoplasmic bridges. These data indicate that SCAR 4 and 5 complexes did not show genotoxicity in our models while SCAR 6 was considered mutagenic. This study presented a comprehensive genotoxic evaluation of SCAR complexes were shown to be genotoxic in vitro. All in all, further studies are required to fully elucidate how the properties can affect human health. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Evaluation of a liver micronucleus assay in young rats (III): a study using nine hepatotoxicants by the Collaborative Study Group for the Micronucleus Test (CSGMT)/Japanese Environmental Mutagen Society (JEMS)-Mammalian Mutagenicity Study Group (MMS).

    Science.gov (United States)

    Takasawa, Hironao; Suzuki, Hiroshi; Ogawa, Izumi; Shimada, Yasushi; Kobayashi, Kazuo; Terashima, Yukari; Matsumoto, Hirotaka; Aruga, Chinami; Oshida, Keiyu; Ohta, Ryo; Imamura, Tadashi; Miyazaki, Atsushi; Kawabata, Masayoshi; Minowa, Shigenori; Hayashi, Makoto

    2010-04-30

    We have been investigating a liver micronucleus assay to detect genotoxic chemicals using young rats for several years, and had established its advantages with respect to using autonomous proliferation of young rat hepatocytes. Nine chemicals known to induce hepatotoxic effects such as necrosis (2,6-dinitrotolune, bromobenzene, isoniazid, phenacetin, allyl alcohol and thioacetamide), cholestasis (chlorpromazine hydrochloride and alpha-naphthyl isothiocyanate) and oxidative stress (clofibrate) were selected for this study. A liver micronucleus assay was conducted in 4-week-old male F344 rats using two or three dose levels of test chemicals given orally by gavage to evaluate the compound's ability to induce micronucleated hepatocytes. Several of these test chemicals were additionally examined in a peripheral blood micronucleus assay conducted concurrently and in the same animals. The genotoxic rodent hepatocarcinogen, 2,6-dinitrotoluene showed a positive result in the liver micronucleus assay, but the nongenotoxic hepatocarcinogens, clofibrate and thioacetamide gave negative responses. Bromobenzene, known to produce DNA adducts but is noncarcinogenic in rodent liver, was judged equivocal in this assay. alpha-Naphthyl isothiocyanate is noncarcinogenic and showed negative response in the liver. The other four chemicals, known to be either noncarcinogenic or carcinogenic in other non-liver target organs, showed negative results in the liver micronucleus assay. Based on the results in the present study and previous report described above, it was concluded that this technique is able to effectively predict genotoxic rodent hepatocarcinogenicity, and does not give false positives due to hepatotoxicity. Copyright 2010 Elsevier B.V. All rights reserved.

  9. Dependence of the bystander effect for micronucleus formation on dose of heavy-ion radiation in normal human fibroblasts

    International Nuclear Information System (INIS)

    Matsumoto, Yoshitaka; Hamada, Nobuyuki; Aoki-Nakano, Mizuho; Furusawa, Yoshiya; Funayama, Tomoo; Sakashita, Tetsuya; Kobayashi, Yasuhiko; Wada, Seiichi; Kakizaki, Takehiko

    2015-01-01

    Ionising radiation-induced bystander effects are well recognised, but its dependence on dose or linear energy transfer (LET) is still a matter of debate. To test this, 49 sites in confluent cultures of AG01522D normal human fibroblasts were targeted with microbeams of carbon (103 keV μm -1 ), neon (375 keV μm -1 ) and argon ions (1260 keV μm -1 ) and evaluated for the bystander-induced formation of micronucleus that is a kind of a chromosome aberration. Targeted exposure to neon and argon ions significantly increased the micronucleus frequency in bystander cells to the similar extent irrespective of the particle numbers per site of 1- 6. In contrast, the bystander micronucleus frequency increased with increasing the number of carbon-ion particles in a range between 1 and 3 particles per site and was similar in a range between 3 and 8 particles per site. These results suggest that the bystander effect of heavy ions for micronucleus formation depends on dose. (authors)

  10. Significance of the proportion of binucleate cells in the micronucleus assay; A methodological study

    Energy Technology Data Exchange (ETDEWEB)

    Imamura, Masahiro; Edgren, M.R. (Karolinska Inst., Stockholm (Sweden). Dept. of Radiation Physics)

    1994-03-01

    Using treatment with cytochalasin-B (Cyt-B) for the induction of a cytokinetic block, the significance of the proportion of binucleate cells (BNC) in the micronucleus (MN) assay was investigated in a methodological study. A Chinese hamster cell line V79 was used in which MN were induced by radiation. In complementary tests the radiation effect in inducing MN was enhanced by depletion of the cellular glutathione content with buthionine sulfoximine (BSO). The data indicated that the concentration of Cyt-B is the major factor which determines the proportion of BNC. This proportion was shown to be independent of radiation dose and of BSO. Furthermore, the MN frequency was not related to the percentage of BNC. Therefore, a high proportion of BNC may be practical for the MN assay, but may not make the technique more accurate. (author).

  11. Significance of the proportion of binucleate cells in the micronucleus assay

    International Nuclear Information System (INIS)

    Imamura, Masahiro; Edgren, M.R.

    1994-01-01

    Using treatment with cytochalasin-B (Cyt-B) for the induction of a cytokinetic block, the significance of the proportion of binucleate cells (BNC) in the micronucleus (MN) assay was investigated in a methodological study. A Chinese hamster cell line V79 was used in which MN were induced by radiation. In complementary tests the radiation effect in inducing MN was enhanced by depletion of the cellular glutathione content with buthionine sulfoximine (BSO). The data indicated that the concentration of Cyt-B is the major factor which determines the proportion of BNC. This proportion was shown to be independent of radiation dose and of BSO. Furthermore, the MN frequency was not related to the percentage of BNC. Therefore, a high proportion of BNC may be practical for the MN assay, but may not make the technique more accurate. (author)

  12. Genotoxicity and ELF-magnetic fields: a review through the micronucleus assay

    International Nuclear Information System (INIS)

    Alcaraz, M.; Andreu-Galvez, M.; Sanchez-Villalobos, J. M.; Achel, D. G.; Olmos, E.; Martinez-Hernandez, C. M.

    2012-01-01

    Thirty for (34) published studies, conducted from 1994 to the present to evaluate the genotoxic effect of magnetic fields using ELF-EMF and diagnostic resonance on humans by the micronucleus assay have been reviewed. some characteristics of the assay methods, their significance to genotoxicity and basic interpretations of the results of these assays are discussed. of the studies analysed 70.5% implicated genotoxic effects induced by these magnetic fields: 52.9% were due to exposure to magnetic fields only and 17,6% by exposure to magnetic fields in combination with some treatment types, resulting in additive or synergistic effect. Evidence exist to support the notion that exposure of humans to magnetic fields stimulates genotoxic effects, although the actual mechanisms of action or even the true human health consequences resulting from these exposure still remain unclear. (Author) 80 refs.

  13. Genotoxicity assessment of cobalt chloride in Eisenia hortensis earthworms coelomocytes by comet assay and micronucleus test.

    Science.gov (United States)

    Ciğerci, İbrahim Hakkı; Ali, Muhammad Muddassir; Kaygısız, Şöhret Yüksek; Liman, Recep

    2016-02-01

    Cobalt and its different compounds are extensively used worldwide and considered as possible environmental pollutant. Earthworms are useful model organism and its different species are used to monitor soil pollution. No study has been found to detect cobalt chloride (CoCl2) genotoxicity in earthworms. So, current study aimed to evaluate CoCl2 induced genotoxicity in Eisenia hortensis earthworms coelomocytes by alkaline comet assay (CA) and micronucleus (MN) test. The earthworms (n = 10 for each group) were exposed to different series of CoCl2 concentrations (100 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm, 600 ppm) to find LD50. The LD50 for CoCl2 was found at 226 ppm. Then, doses of LD50/2, LD50 and 2XLD50 for 48 h were used. CA and MN demonstrated the significant increase (P earthworms. Copyright © 2015 Elsevier Ltd. All rights reserved.

  14. Structural and numerical chromosome aberration inducers in liver micronucleus test in rats with partial hepatectomy.

    Science.gov (United States)

    Itoh, Satoru; Hattori, Chiharu; Nagata, Mayumi; Sanbuissho, Atsushi

    2012-08-30

    The liver micronucleus test is an important method to detect pro-mutagens such as active metabolites not reaching bone marrow due to their short lifespan. We have already reported that dosing of the test compound after partial hepatectomy (PH) is essential to detect genotoxicity of numerical chromosome aberration inducers in mice [Mutat. Res. 632 (2007) 89-98]. In naive animals, the proportion of binucleated cells in rats is less than half of that in mice, which suggests a species difference in the response to chromosome aberration inducers. In the present study, we investigated the responses to structural and numerical chromosome aberration inducers in the rat liver micronucleus test. Two structural chromosome aberretion inducers (diethylnitrosamine and 1,2-dimethylhydrazine) and two numerical chromosome aberration inducers (colchicine and carbendazim) were used in the present study. PH was performed a day before or after the dosing of the test compound in 8-week old male F344 rats and hepatocytes were isolated 4 days after the PH. As a result, diethylnitrosamine and 1,2-dimethylhydrazine, structural chromosome aberration inducers, exhibited significant increase in the incidence of micronucleated hepatocyte (MNH) when given either before and after PH. Colchicine and carbendazim, numerical chromosome aberration inducers, did not result in any toxicologically significant increase in MNH frequency when given before PH, while they exhibited MNH induction when given after PH. It is confirmed that dosing after PH is essential in order to detect genotoxicity of numerical chromosome aberration inducers in rats as well as in mice. Regarding the species difference, a different temporal response to colchicine was identified. Colchicine increased the incidence of MNH 4 days after PH in rats, although such induction in mice was observed 8-10 days after PH. Copyright © 2012 Elsevier B.V. All rights reserved.

  15. Analysis of the Genotoxic Effects of Mobile Phone Radiation using Buccal Micronucleus Assay: A Comparative Evaluation.

    Science.gov (United States)

    Banerjee, Sumita; Singh, Narendra Nath; Sreedhar, Gadiputi; Mukherjee, Saikat

    2016-03-01

    Micronucleus (MN) is considered to be a reliable marker for genotoxic damage and it determines the presence and the extent of the chromosomal damage. The MN is formed due to DNA damage or chromosomal disarrangements. The MN has a close association with cancer incidences. In the new era, mobile phones are constantly gaining popularity specifically in the young generation, but this device uses radiofrequency radiation that may have a possible carcinogenic effect. The available reports related to the carcinogenic effect of mobile radiation on oral mucosa are contradictory. To explore the effects of mobile phone radiation on the MN frequency in oral mucosal cells. The subjects were divided into two major groups: low mobile phone users and high mobile phone users. Subjects who used their mobile phone since less than five years and less than three hours a week comprised of the first group and those who used their mobile since more than five years and more than 10 hours a week comprised of the second group. Net surfing and text messaging was not considered in this study. Exfoliated buccal mucosal cells were collected from both the groups and the cells were stained with DNA-specific stain acridine orange. Thousand exfoliated buccal mucosal cells were screened and the cells which were positive for micronuclei were counted. The micronucleus frequency was represented as mean±SD, and unpaired Student t-test was used for intergroup comparisons. The number of micronucleated cells/ 1000 exfoliated buccal mucosal cells was found to be significantly increased in high mobile phone users group than the low mobile phone users group. The use of mobile phone with the associated complaint of warmth around the ear showed a maximum increase in the number of micronucleated cells /1000 exfoliated buccal mucosal cells. Mobile phone radiation even in the permissible range when used for longer duration causes significant genotoxicity. The genotoxicity can be avoided to some extent by the

  16. [Influence of Four Kinds of PPCPs on Micronucleus Rate of the Root-Tip Cells of Vicia-faba and Garlic].

    Science.gov (United States)

    Wang, Lan-jun; Wang, Jin-hua; Zhu, Lu-sheng; Wang, Jun; Zhao, Xiang

    2016-04-15

    In order to determine the degree of biological genetic injury induced by PPCPs, the genotoxic effects of the doxycycline (DOX), ciprofloxacin (CIP), triclocarban (TCC) and carbamazepine (CBZ) in the concentration range of 12.5-100 mg · L⁻¹ were studied using micronucleus rate and micronucleus index of Vicia-fabe and garlic. The results showed that: (1) When the Vicia-faba root- tip cells were exposed to DOX, CIP, TCC and CBZ, micronucleus rates were higher than 1.67 ‰ (CK₁), it was significantly different from that of the control group (P garlic root tip cells were exposed to DOX, CIP, TCC and CBZ respectively, the micronucleus rates were less than those of the Vicia-faba, while in most treatments significantly higher than that of the control group (0.67‰). The micronucleus index was higher than 3.5 in the groups exposed to CIP with concentrations of 25, 50, 100 mg · L⁻¹ and TCC and CBZ with concentrations of 25 mg · L⁻¹; With the increase of exposure concentrations, the micronucleus rate showed a trend of first increasing and then decreasing as well. (3) Under the same experimental conditions, the cells micronucleus rates of the garlic cells caused by the four tested compounds were significantly lower than those of Vicia-faba. (4) The micronucleus index of the root tip cells of Vicia-faba and garlic treated with the four kinds of compounds followed the order of CIP > CBZ > TCC > DOX. These results demonstrated that the four compounds caused biological genetic injury to root-tip cells of Vicia-faba and garlic, and the genetic damage caused to garlic was significantly lower than that to Vicia-faba. The damages caused by the four kinds of different compounds were also different.

  17. Spontaneous micronucleus frequencies in human peripheral blood lymphocytes as a screening test for an individual variation in a different population and radiation-induced micronucleus induction

    International Nuclear Information System (INIS)

    Kang, Chang-Mo; Jeon, Hye-Jeong; Cho, Chul-Koo

    2004-01-01

    Our studies were to evaluate the role of epigenetic factors in the variation of radiosensitivity on human peripheral blood lymphocytes by measuring the frequencies of micronucleus (MN) from 293 healthy subjects of different population for assessing the radiation health risk in Korea. We analyzed the frequencies of both spontaneous and in vitro 60 Co γ-rays or 50MeV neutron-induced MNs. The frequencies of spontaneous NMs not only vary greatly between individuals, but also working or living areas. The increased levels of cells with spontaneous MNs were observed with an increasing age. The frequencies of spontaneous MNs were significantly higher in females than in males. For both sexes, MN frequency was significantly and positively correlated with age. Age and gender are the most important demographic variables impacting on the MN index. Donors who had ever smoked showed significantly increased frequencies of MNs compared to nonsmokers. The main lifestyle factors influencing the MN index in the subjects are correlated significantly and positively with smoke while measuring the spontaneous frequencies of micronuclei. Therefore, it is evident that with regard to the application of MN assay all future studies to evaluate the association between radiosensitivity and susceptibility for radiation health risks in different populations should take into account the effect of age, gender and lifestyle. For the dose-response study, the induced MNs were observed at all doses, and the numerical changes according to doses. The dose-response curves were fitted with a linear-quadratic forms of the dose, and the results were different for γ-rays and neutrons significantly. Neutrons were more effective than γ-rays in producing MN with a dose-dependent manner. The frequency of MN varies with dose. The RBE for a micronuclei was 2.37 ± 0.17. The results suggested that the MN assay have a high potential to ensure appropriate quality control and a standard documentation protocol, which

  18. Evaluation of genetic alteration induced by radon gas using the micronucleus test (Tradescantia sp. clone KU-20)

    International Nuclear Information System (INIS)

    Bruschi, Armando L.; Azevedo, Heliana de; Macacini, Jose F.; Roque, Claudio V.

    2011-01-01

    The first observations over the existence of radon gas (Rn), initially known as 'thorium emanation', were carried out between the end of 19 th and beginning of 20 th centuries. A result of uranium-238 (U 238 ) radioactive decay, radon is a tasteless, odorless and colorless gas under room temperature, with a 3.825-day half life and particle α emission in its decay, and as final product of its disintegration, the stable lead-206 isotope (Pb 206 ). Being it is the gas with the highest density known, closed and poor ventilated environments are favorable to its accumulation, with its inhalation being the highest health risk. The use of vegetal bioindicators has shown to be excellent on the monitoring of air quality and on mutagenic potential of various pollutants contained in the atmosphere. Within this context, the objective of this study was to evaluate the micronucleus test application potential utilizing the Tradescantia sp. clone KU-20, in order to evaluate genetic alterations induced by radon gas. Stems of Tradescantia sp. clone KU-20, previously immerse in Hoagland solution, were introduced in a radon detection equipment's calibration chamber (Alphaguard), containing radium salt. Afterwards, the accommodated stems were exposed to radon gas (the average radon concentration was 7.639 KBq/m3) for 24 hours. The results demonstrated an increase on micronucleus formation (39.23 + 2.143 MCN/100 tetrads) in stems exposed in relation to the negative control (18.00 + 1.396 MCN/100 tetrads). The difference between the values indicated a significant increase on micronucleus frequency in the inflorescences subjected to radon gas. The presented results demonstrated the micronucleus test application potential using Tradescantia clone KU-20 to evaluate genetic effects induced by radon gas. (author)

  19. Assessment of the genotoxicity of Cu and Zn in raw and anaerobically digested slurry with the Vicia faba micronucleus test

    OpenAIRE

    Marcato, Claire-Emmanuelle; Pinelli, Eric; Pourrut, Bertrand; Silvestre, Jérôme; Guiresse, Agnès Maritchù

    2009-01-01

    Genotoxicity of Cu and Zn was assessed by use of the micronucleus (MN) test on Vicia faba roots. Plants were exposed to various leachates of rawand anaerobically digested pig slurry, with maximum total concentrations of 200MCu and 600MZn. The results indicated stabilisation of the organic matter during anaerobic digestion of the slurry and bioconversion of some phytotoxic organic compounds (e.g. phenols or p-cresol), but did not showa relationship between Cu and Zn concentrations and MN fr...

  20. In vivo micronucleus test as a biomarker of genotoxicity in free-range goats from suspected contaminated environment

    Directory of Open Access Journals (Sweden)

    Afusat Jagun Jubril

    2017-09-01

    Conclusion: The finding indicates the prevalence and frequency of micronucleus as a biomarker of genotoxicity and an indicator of exposure to environmental genotoxic subtances. Hence, this highlights the relevance of these goats as important sentinel animal model. These findings, therefore, serve as a preliminary data for further studies on the latent genotoxic environmental contaminants and their potential deleterious impact. [J Adv Vet Anim Res 2017; 4(3.000: 281-287

  1. Kinetics of micronucleus induction and cytotoxicity caused by distinct antineoplastics and alkylating agents in vivo.

    Science.gov (United States)

    Morales-Ramírez, Pedro; Vallarino-Kelly, Teresita; Cruz-Vallejo, Virginia

    2014-01-30

    This mini-review aims to compare the differences in the kinetics of the induction of micronucleated polychromatic erythrocytes (MN-PCE) and cytotoxicity by distinct antineoplastic and genotoxic agents in murine peripheral blood in vivo and to correlate these kinetics with the underlying processes. Comparisons were carried out using our previously obtained data with nominal doses causing similar levels of cytotoxicity, as measured in terms reduction of PCE. The aneuploidogens caused the most rapid induction of MN-PCEs and had the highest rates of cytotoxicity and genotoxicity. The promutagens cyclophosphamide and dimethylnitrosamine showed the most delayed responses and had the lowest genotoxic and cytotoxic efficiencies. DNA crosslinking agents had a similar delay of 4-5 h, greater than those of aneuploidogens, but differed in their cytotoxic and genotoxic efficiencies. Methylnitrosourea and 5-aza-cytidine caused greater delays than crosslinking agents. These delays can be due to the methylnitrosourea-mediated induction of formation of mono alkyl adducts which are interpreted as mismatches during DNA duplication, whereas 5-aza-cytidine requires incorporation into the DNA to induce breakage. This review allows us to conclude that the requirement for metabolic activation and the mechanisms of DNA breakage and of micronucleus induction are the main factors that affect the time of maximal MN-PCE induction. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  2. Inhibitory effects of Enteromorpha linza polysaccharide on micronucleus of Allium sativum root cells.

    Science.gov (United States)

    Zhang, Zhongshan; Wang, Xiaomei; Li, Jingfen; Liu, Chongbin; Zhang, Quanbin

    2016-06-01

    In this study, the antimutagenic function of the polysaccharide from Enteromorpha linza with the micronucleus test of Allium sativum root cells induced by sulfur dioxide and ultraviolet was studied. The concentration-effect relation of the two inducers was firstly evaluated. The results showed that an increase of genotoxicity damage was demonstrated and micronuclei frequency induced by sulfur dioxide and ultraviolet displayed dose dependent increases. All the doses of polysaccharide did affect the micronuclei frequency formation compared with the negative control. And also, the significant increase in inhibition rate of micronuclei frequency was observed with the increase of the dose of polysaccharide. It was showed maximum inhibition of micronuclei frequency cells (71.74% and 66.70%) at a concentration of 200g/mL in three experiments. The low molecular weight polysaccharide showed higher inhibition rate than raw polysaccharide at the higher concentration (50g/mL) in the absence of sulfur dioxide and ultraviolet. It was confirmed to be a good mutant inhibitor. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Effect of mobile phone station on micronucleus frequency and chromosomal aberrations in human blood cells.

    Science.gov (United States)

    Yildirim, M S; Yildirim, A; Zamani, A G; Okudan, N

    2010-01-01

    The use of mobile telephones has rapidly increased worldwide as well as the number of mobile phone base stations that lead to rise low level radiofrequency emissions which may in turn have possible harm for human health. The national radiation protection board has published the known effects of radio waves exposure on humans living close to mobile phone base stations. However, several studies have claimed that the base station has detrimental effects on different tissues. In this study, we aimed to evaluate the effects of mobile phone base stations on the micronucleus (MN) frequency and chromosomal aberrations on blood in people who were living around mobile phone base stations and healthy controls. Frequency of MN and chromosomal aberrations in study and control groups was 8.96 +/- 3.51 and 6.97 +/- 1.52 (p: 0.16); 0.36 +/- 0.31 and 0.75 +/- 0.61 (p: 0.07), respectively. Our results show that there was not a significant difference of MN frequency and chromosomal aberrations between the two study groups. The results claim that cellular phones and their base stations do not produce important carcinogenic changes.

  4. Use of micronucleus test in the assessment of radiation effects in aquatic environments

    International Nuclear Information System (INIS)

    Araujo, Edvaldo F. de; Silva, Luanna R.S.; Lima, Pedro A. de S.; Amancio, Francisco F.; Melo, Ana Maria M. de A.; Silva, Edvane B. da; Silva, Ronaldo C. da

    2011-01-01

    The study of the effects of radioactive substances on the environment is accomplished by radioecology. This science has played an important role in combating all forms of pollution. The uncontrolled use of physical and chemical agents has been a concern for environmental regulatory agencies, due to the serious damage to ecosystems. Aquatic organisms are exposed to a variety of pollutants harmful to aquatic systems. The mollusks Biomphalaria glabrata has been featured as a bioindicator to possess characteristics such as short reproductive cycle ease of maintenance in the laboratory and low maintenance cost. The micronucleus assay has been shown to be a great test to identify mutagenic effects caused by physical and chemical agents. In this study the frequency of micronuclei in haemocytes of Biomphalaria glabrata exposed to high doses of 60 Co gamma radiation contributing to a further standardization of this test as an indicator of the presence of radioactive contamination in aquatic environments. The young adult snails of Biomphalaria glabrata were divided into groups and subjected to a dose of 0 (control), 40 and 60 Gy of gamma radiation. The results showed that snails irradiated with 40 Gy showed a smaller number of haemocytes, whereas those exposed to 60 Gy had a greater quantity of these cells compared to control group. It can be concluded that the morphological analysis and the frequency of micronuclei in haemocytes of Biomphalaria glabrata exposed to 60 Co gamma radiation may be used in studies of the action of high doses of radiation in aquatic environments (author)

  5. Effects of heavy metals/metalloids contamination of soils on micronucleus induction in Tradescantia pallida

    Directory of Open Access Journals (Sweden)

    Neelima Meravi

    2013-06-01

    Full Text Available The present study was conducted in GGV campus, Bilaspur in which heavy metals/metalloids speciation of soil (for Cr, Fe, Ni, Cd and Pb was performed for assessing the genotoxicity of these metals. The metals concentrations were measured with the help of AAS 7000 (Shimadzu and the standard solution was prepared using standard metal solution of Inorganic Ventures. The concentrations of Cr, Fe, Ni, Cd and Pb (in ug/100 g soil were 12.4, 33.9, 3.1, 0.07 and 2.4 respectively. The flowers of Tradescantia pallida plants growing in this soil were taken and their micronucleus (Trad-MCN bioassay was performed. Trad-MCN bioassay was performed using the protocols established by Ma (1981. The study revealed that at these concentrations of metals micronuclei (stained objects that were smaller than the nuclei and not connected to the nuclei are classified as MCN were formed. Therefore it can be inferred from the present study that soil of GGV campus is genotoxic for the Tradescantia pallida.

  6. Micronucleus Assay in Exfoliated Buccal Epithelial Cells Using Liquid Based Cytology Preparations in Building Construction Workers.

    Science.gov (United States)

    Arul, P; Smitha, Shetty; Masilamani, Suresh; Akshatha, C

    2018-01-01

    Cytogenetic damage in exfoliated buccal epithelial cells due to environmental and occupational exposure is often monitored by micronucleus (MN) assay using liquid based cytology (LBC) preparations. This study was performed to evaluate MN in exfoliated buccal epithelial cells of building construction workers using LBC preparations. LBC preparations of exfoliated buccal epithelial cells from 100 subjects [50 building construction workers (cases) and 50 administrative staffs (controls)] was evaluated by May-Grunwald Giemsa, Hematoxylin and Eosin and Papanicolaou stains. Student's t test was used for statistical analysis and a P value of 5 years) and smokers and non-smokers of cases (P=0.001). However, there were meaningful differences regarding mean frequencies of MN between smokers, non-smokers, those with alcohol consumption or not in cases and controls using various stains (P=0.001). There was an increased risk of cytogenetic damage in building construction workers. However, evaluation of MN of exfoliated buccal epithelial cells in building construction workers serve as a minimally invasive biomarker for cytogenetic damage. LBC preparations can be applied for MN assay as it improves the quality of smears and cell morphology, decreases the confounding factors and reduces false positive results.

  7. Synergistic interaction of radiation and octylphenol evaluated by tradescantia-micronucleus assay

    International Nuclear Information System (INIS)

    Shin, H. S.; Lee, J. H.; Lee, B. H.; Kim, J. K.

    2003-01-01

    Many kinds of synthetic chemicals have been being used for various purposes. Some of them are called 'endocrine disruptors' because they can disturb the endocrine system of organisms. Presently no technique is established for the quantitative assessment of biological risk of the environmental hormones. The pollen mother cells (PMC) of tradescantia are very sensitive to chemical toxicants or ionizing radiation, and thus can be used as a biological end- point for assessing their effects. Micronucleus frequencies in PMC showed a good dose- and concentration-response relationship for radiation and bisphenol A. The MCN frequencies in the pollen mother cells treated with octylphenol were 4.20, 7.27, 4.93 MCN/100 tetrads for 1, 5 and 10 μM, respectively. On the other hand, the frequencies were 10.13, 19.27, 24.47 MCN/100 tetrads for the octylphenol treatments (1, 5, and 10 μM) combined with 30 cGy irradiation. The MCN frequency of 30 cGy control was 8.00 MCN/100 tetrads for the octylphenol treatments (1, 5, and 10μM) combined with 30 cGy irradiation. The MCN frequency of 30 cGy control was 8.00 MCN/100 tetrads. It is known from the result that the Trad-MCN assay can be an excellent tool for the detection of biologically harmful effects of environmental toxicants or synthetic chemicals

  8. Biomonitoring of Serbian population revealed by CB micronucleus test before and after the bombing of Yugoslavia

    International Nuclear Information System (INIS)

    Joksic, G.; Stankovic, M.; Guc-Scekic, M.; Vranjes, A.

    2002-01-01

    The induction of micronuclei in mitotically active cells has been widely used and promoted as a biological marker of exposure to environmental toxins. Biomonitoring of population using cytochalasin block micronucleus test (CBMN) has been performed for 11 years in our country; the incidence of micronuclei was evaluated in many groups of occupationally exposed persons as well as healthy unexposed controls. The spontaneous frequency of MN per 1000 binucleated cells was 9±3 (mean±SD) for woman, 7±2 for men. The average incidence of micronuclei in lymphocytes of newborns was 5.3±0.6, in their mothers 15±3 per 1000 binucleated cells, respectively. After the bombing of Yugoslavia significantly higher incidence of micronuclei was found in all groups of examines: health adults and newborns. In healthy adults, the average incidence of micronuclei was 28.16±14.63; in young pregnant woman 25.3±5.02 and their foetuses 20.14±9.6 respectively. One year latter (2001) the incidence of MN declined in all adults but enhance in foetal blood lymphocytes. (author)

  9. Micronucleus test for radiation biodosimetry in mass casualty events: Evaluation of visual and automated scoring

    Energy Technology Data Exchange (ETDEWEB)

    Bolognesi, Claudia, E-mail: claudia.bolognesi@istge.i [Environmental Carcinogenesis Unit, National Cancer Research Institute, Largo R. Benzi 10, 16132 Genoa (Italy); Balia, Cristina; Roggieri, Paola [Environmental Carcinogenesis Unit, National Cancer Research Institute, Largo R. Benzi 10, 16132 Genoa (Italy); Cardinale, Francesco [Clinical Epidemiology Unit, National Cancer Research Institute, Largo R. Benzi 10, 16132 Genoa (Italy); Department of Health Sciences, University of Genoa, Genoa (Italy); Bruzzi, Paolo [Clinical Epidemiology Unit, National Cancer Research Institute, Largo R. Benzi 10, 16132 Genoa (Italy); Sorcinelli, Francesca [Environmental Carcinogenesis Unit, National Cancer Research Institute, Largo R. Benzi 10, 16132 Genoa (Italy); Laboratory of Genetics, Histology and Molecular Biology Section, Army Medical and Veterinary, Research Center, Via Santo Stefano Rotondo 4, 00184 Roma (Italy); Lista, Florigio [Laboratory of Genetics, Histology and Molecular Biology Section, Army Medical and Veterinary, Research Center, Via Santo Stefano Rotondo 4, 00184 Roma (Italy); D' Amelio, Raffaele [Sapienza, Universita di Roma II Facolta di Medicina e Chirurgia and Ministero della Difesa, Direzione Generale Sanita Militare (Italy); Righi, Enzo [Frascati National Laboratories, National Institute of Nuclear Physics, Via Enrico Fermi 40, 00044 Frascati, Rome (Italy)

    2011-02-15

    In the case of a large-scale nuclear or radiological incidents a reliable estimate of dose is an essential tool for providing timely assessment of radiation exposure and for making life-saving medical decisions. Cytogenetics is considered as the 'gold standard' for biodosimetry. The dicentric analysis (DA) represents the most specific cytogenetic bioassay. The micronucleus test (MN) applied in interphase in peripheral lymphocytes is an alternative and simpler approach. A dose-effect calibration curve for the MN frequency in peripheral lymphocytes from 27 adult donors was established after in vitro irradiation at a dose range 0.15-8 Gy of {sup 137}Cs gamma rays (dose rate 6 Gy min{sup -1}). Dose prediction by visual scoring in a dose-blinded study (0.15-4.0 Gy) revealed a high level of accuracy (R = 0.89). The scoring of MN is time consuming and requires adequate skills and expertise. Automated image analysis is a feasible approach allowing to reduce the time and to increase the accuracy of the dose estimation decreasing the variability due to subjective evaluation. A good correlation (R = 0.705) between visual and automated scoring with visual correction was observed over the dose range 0-2 Gy. Almost perfect discrimination power for exposure to 1-2 Gy, and a satisfactory power for 0.6 Gy were detected. This threshold level can be considered sufficient for identification of sub lethally exposed individuals by automated CBMN assay.

  10. Nuclear localization signal targeting to macronucleus and micronucleus in binucleated ciliate Tetrahymena thermophila.

    Science.gov (United States)

    Iwamoto, Masaaki; Mori, Chie; Osakada, Hiroko; Koujin, Takako; Hiraoka, Yasushi; Haraguchi, Tokuko

    2018-06-08

    Ciliated protozoa possess two morphologically and functionally distinct nuclei: a macronucleus (MAC) and a micronucleus (MIC). The MAC is transcriptionally active and functions in all cellular events. The MIC is transcriptionally inactive during cell growth, but functions in meiotic events to produce progeny nuclei. Thus, these two nuclei must be distinguished by the nuclear proteins required for their distinct functions during cellular events such as cell proliferation and meiosis. To understand the mechanism of the nuclear transport specific to either MAC or MIC, we identified specific nuclear localization signals (NLSs) in two MAC- and MIC-specific nuclear proteins, macronuclear histone H1 and micronuclear linker histone-like protein (Mlh1), respectively. By expressing GFP-fused fragments of these proteins in Tetrahymena thermophila cells, two distinct regions in macronuclear histone H1 protein were assigned as independent MAC-specific NLSs and two distinct regions in Mlh1 protein were assigned as independent MIC-specific NLSs. These NLSs contain several essential lysine residues responsible for the MAC- and MIC-specific nuclear transport, but neither contains any consensus sequence with known monopartite or bipartite NLSs in other model organisms. Our findings contribute to understanding how specific nuclear targeting is achieved to perform distinct nuclear functions in binucleated ciliates. © 2018 The Authors. Genes to Cells published by Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd.

  11. Fluorescence in situ hybridization in combination with the comet assay and micronucleus test in genetic toxicology

    Directory of Open Access Journals (Sweden)

    Hovhannisyan Galina G

    2010-09-01

    Full Text Available Abstract Comet assay and micronucleus (MN test are widely applied in genotoxicity testing and biomonitoring. While comet assay permits to measure direct DNA-strand breaking capacity of a tested agent MN test allows estimating the induced amount of chromosome and/or genome mutations. The potential of these two methods can be enhanced by the combination with fluorescence in situ hybridization (FISH techniques. FISH plus comet assay allows the recognition of targets of DNA damage and repairing directly. FISH combined with MN test is able to characterize the occurrence of different chromosomes in MN and to identify potential chromosomal targets of mutagenic substances. Thus, combination of FISH with the comet assay or MN test proved to be promising techniques for evaluation of the distribution of DNA and chromosome damage in the entire genome of individual cells. FISH technique also permits to study comet and MN formation, necessary for correct application of these methods. This paper reviews the relevant literature on advantages and limitations of Comet-FISH and MN-FISH assays application in genetic toxicology.

  12. Study design and statistical analysis of data in human population studies with the micronucleus assay.

    Science.gov (United States)

    Ceppi, Marcello; Gallo, Fabio; Bonassi, Stefano

    2011-01-01

    The most common study design performed in population studies based on the micronucleus (MN) assay, is the cross-sectional study, which is largely performed to evaluate the DNA damaging effects of exposure to genotoxic agents in the workplace, in the environment, as well as from diet or lifestyle factors. Sample size is still a critical issue in the design of MN studies since most recent studies considering gene-environment interaction, often require a sample size of several hundred subjects, which is in many cases difficult to achieve. The control of confounding is another major threat to the validity of causal inference. The most popular confounders considered in population studies using MN are age, gender and smoking habit. Extensive attention is given to the assessment of effect modification, given the increasing inclusion of biomarkers of genetic susceptibility in the study design. Selected issues concerning the statistical treatment of data have been addressed in this mini-review, starting from data description, which is a critical step of statistical analysis, since it allows to detect possible errors in the dataset to be analysed and to check the validity of assumptions required for more complex analyses. Basic issues dealing with statistical analysis of biomarkers are extensively evaluated, including methods to explore the dose-response relationship among two continuous variables and inferential analysis. A critical approach to the use of parametric and non-parametric methods is presented, before addressing the issue of most suitable multivariate models to fit MN data. In the last decade, the quality of statistical analysis of MN data has certainly evolved, although even nowadays only a small number of studies apply the Poisson model, which is the most suitable method for the analysis of MN data.

  13. Effects of electromagnetic fields induced from the visual display terminal on the micronucleus frequencies in tradescantia

    International Nuclear Information System (INIS)

    Shin, Hae Shik; Kim, Jin Kyu; Lee, Jin Hong

    2002-01-01

    Humans are exposed daily to electromagnetic fields (EMFs) originating from a variety of devices and VDT (Visual Display Terminal) workstations. This research was designed to examine the biological effects of electromagnetic fields from VDT using the Tradescantia- micronuclesus (Trad-MCN) bioassay. Tradescantia BNL 4430 clone was used to evaluate the influence of EMFs radiated from the VDT workstation. Trad-MCN assay is a cytogenetic test based on the formation of micronuclei that result from chromosome breakage in the meiotic pollen mother cells. No study has established unequivocally a causal relationship between EMFs and animals or plants. Fresh cuttings bearing young flower buds were exposed for 24hours in front of the VDT workstation. The cuttings were placed at 30, 50, 70, and 90cm distances from the workstations. The micronucleus were scored under a light microscope (400 x magnification). Three hundreds of tetrads were scored from each of the slides in every the experimental group. The frequencies expressed in terms of MCN / 100 tetrads. Trad-MCN frequencies were 8.93 ± 0.32, 11.2 ± 0.50, 7.67 ± 0.61, and 6.22 ± 1.78/ 100 tetrads at 30, 50, 70, and 90cm, respectively. In conclusion, EMFs from VDT give rise to damage the chromosome this plant. In addition, The results of the study indicate that Trad-MCN assay can detect chromosome damage due to EMFs from the electrical device workstation. In conclusion, the Trad-MCN assay is sensitive, reproducible, easy to perform, well standardized, inexpensive and undemanding in equipment

  14. Dose-response relationship of octylphenol and radiation evaluated by tradescantia-micronucleus assay

    International Nuclear Information System (INIS)

    Kim, J. K.; Cheon, K. J.; Lee, B. H.; Shin, H. S.; Lee, J. H.

    2002-01-01

    Many kinds of synthetic chemicals have been being used for various purposes. Some of them are called 'Endocrine Disruptor's because they can disturb the endocrine system of organisms. Presently no technique is established for the quantitative assessment of biological risk of the environmental hormones. The pollen mother cells (PMC) of Tradescantia are very sensitive to chemical toxicants or ionizing radiation, and thus can be used as a biological end-point assessing their effect. Micronucleus frequencies in PMC showed a good dose- and concentration-response relationship for radiation, bisphenol A and octylphenol. A parallel series of experiment using five increasing doses of gamma-ray at 10, 20, 30, 40 and 50 cGy was conducted. The MCN frequencies of 12.0, 25.2, 41.7, 76 and 83 MCN/100 tetrads were observed from each of the increasing gamma-ray dosage groups, respectively. Lenear regression analysis of the gamma-ray data MCN frequencies yielded a correlation coefficient of 0.95. the MCN frequencies in pollen mother cells treated with bisphenol a and octylphenol showed dose-response relationship in a concentration of 0, 1, 2, 4 μM and 0, 4, 10, 20 μM. the MCN frequency for the bisphenol a and octylphenol group yields 2.33, 8.06, 12.7 and 19.6 MCN/100 tetrads for the bisphenol a and 2.33, 2.33, 11.47, 17.6 MCN/100 tetrads for the octylphenol. The MCN frequency of the control was 2.33 MCN/100 tetrads. It is known from the result that Trad-MCN assay can be an excellent tool for detection of biological risk due to environmental toxicants or synthetic chemicals

  15. Micronucleus evaluation for determining the chromosomal breakages after radionuclide synovectomy in patients with hemophilia

    International Nuclear Information System (INIS)

    Kavakli, K.; Cogulu, O.; Karaca, E.

    2012-01-01

    To investigate the genotoxic effects of 90 Y and 186 Re in patients with hemophilia who were undergoing radionuclide synovectomy (RS) procedure in the last 3 years. Nineteen patients were enrolled in the study. Most of the patients (n=17) were hemophilia-A (mean age 20.6±10.5 years) and 18 patients (mean age 22.6±10.6 years) with hemophilia who were not exposed to RS procedure were included in the study as control group. Most cases in the control group (n=13) were hemophilia-A. 90 Y for knee joints and 186 Re for elbow or ankle joints were used to perform RS in hemophilic patients. We studied the micronucleus (MN) test on peripheral blood lymphocytes as an indicator of radiation-induced cytogenetic damage and calculated nuclear division index. There was no significant difference between the patients with and without RS with respect to MN values. However, both values obtained in RS-exposed patients and control group were much elevated than values reported in literature from healthy controls. The mean MN values of patients below 20 years old were much lower but not significant than those above 20 years old. MN frequencies between 186 Re and 90 Y groups were also analyzed, and no significant difference was observed. Hemophilia patients who were treated with 186 Re showed higher levels of MN compared to patients treated with 90 Y although the difference was not significant. Radioisotope synovectomy (RS) seems to be a safe procedure not causing a significant genotoxic effect on hemophilic patients, however, further studies including larger series of patients are needed to better understand the effects of RS on patients' health. (author)

  16. Influence of GSTM1 and GSTT1 genotypes and confounding factors on the frequency of sister chromatid exchange and micronucleus among road construction workers.

    Science.gov (United States)

    Kumar, Anil; Yadav, Anita; Giri, Shiv Kumar; Dev, Kapil; Gautam, Sanjeev Kumar; Gupta, Ranjan; Aggarwal, Neeraj

    2011-07-01

    In the present study, we have investigated the influence of polymorphism of GSTM1 and GSTT1 genes and confounding factors such as age, sex, exposure duration and consumption habits on cytogenetic biomarkers. Frequency of sister chromatid exchanges (SCEs), high frequency cell (HFC) and cytokinesis blocked micronuclei (CBMN) were evaluated in peripheral blood lymphocytes of 115 occupationally exposed road construction workers and 105 unexposed individuals. The distribution of null and positive genotypes of glutathione-S transferase gene was evaluated by multiplex PCR among control and exposed subjects. An increased frequency of CBMN (7.03±2.08); SCE (6.95±1.76) and HFC (6.28±1.69) were found in exposed subjects when compared to referent (CBMN - 3.35±1.10; SCE - 4.13±1.30 and HFC - 3.98±1.56). These results were found statistically significant at p<0.05. When the effect of confounding factors on the frequency of studied biomarkers was evaluated, a strong positive interaction was found. The individuals having GSTM1 and GSTT1 null genotypes had higher frequency of CBMN, SCE and HFC. The association between GSTM1 and GSTT1 genotypes and studied biomarkers was found statistically significant at p<0.05. Our findings suggest that individuals having null type of GST are more susceptible to cytogenetic damage by occupational exposure regardless of confounding factors. There is a significant effect of polymorphism of these genes on cytogenetic biomarkers which are considered as early effects of genotoxic carcinogens. Copyright © 2011 Elsevier Ltd. All rights reserved.

  17. Biological Dosimetry Using Micronucleus Assay in Simulated Partial-Body Exposure to Ionizing Radiation

    Directory of Open Access Journals (Sweden)

    S. Purnami

    2017-08-01

    Full Text Available In radiation accidents, it is common that only several parts of the body are exposed to radiation. As a consequence there is a mixture of exposed and unexposed lymphocytes in peripheral blood cells of the samples. This phenomenon will cause the dose value estimated using the exposed lymphocytes to be lower than the actual dose. In this study, an assessment of partial body exposures using micronucleus assay by estimating the partial body dose and fraction of irradiated blood was conducted. An optimal D0 value also has been determined in this study to estimate the fraction of irradiated cells. Peripheral blood lymphocytes (PBLs from three healthy donors were irradiated in vitro with 2 Gy of X-rays. Partial radiation exposure was simulated by mixing the irradiated and non-irradiated blood in different proportions. The proportions of mixtures of blood samples irradiated in vitro were 5, 10, 15, 20, and 30 %. Blood samples were then cultured and harvested based on micronuclei assay protocol. At least 2000 binucleated cells with well-preserved cytoplasm were scored for the MN frequency. Dose Estimate 5.1 software was used to calculate the dispersion index (σ2/y and normalized unit of this index (U in each proportion of bloods. The fractions of irradiated cells were calculated with CABAS (Chromosomal Aberration Calculation Software for several different D0 values (2.7; 3.8; 5.4. The results showed that D0 value at 5.4 gave the closest results to the actual proportion of irradiated bloods, while for the dose estimation the estimated doses value from all proportions in all donors were higher than the actual dose. The factor that may cause this phenomenon was that the dose response calibration curve used to predict the radiation dose was not constructed in the laboratory used. Overall it can be concluded that a biodosimetry using MN assay can be used to estimate the radiation dose in partial body exposure. In order to establish a biodosimetry using MN

  18. Alkaline Comet Assay and Micronucleus Test Parameters in Children Exposed to Diagnostic X-Ray Examination

    International Nuclear Information System (INIS)

    Gajski, G.; Geric, M.; Garaj-Vrhovac, V.; Milkovic, Dj.; Beck, N.; Ranogajec-Komor, M.; Miljanic, S.; Knezevic, Z.

    2011-01-01

    Chest radiograms represent the basic radiological examination of thorax and are the most frequently performed radiological diagnostic procedure in the child population. Understanding the risks of low doses of radiation is an important aspect in the risk benefit analysis in paediatric populations. To provide the best care for the young patients the effects of radiation should be minimized thus chest X-rays must be performed by highest standards to ensure that the young patient has the lowest risk possible. Since children are the most sensitive to radiation, there is a need for follow up of the young populations that receive these X-ray diagnostic examinations. Follow up would be especially advisable for children that are at higher risk of radiation induced damage, for example children with a predisposition to DNA damage, or for children that are constantly exposed to numerous radiological examinations due to their illness. In that manner, present study was undertaken to evaluate application of different dosimetry systems in conjunction with alkaline comet assay and micronucleus test for the assessment of different types of DNA and chromosomal alterations in child population exposed to acute diagnostic X-rays examination. For that purpose doses were measured using thermoluminescence (TL) and radiophotoluminescent (RPL) dosimetry systems. The study demonstrated that immediately after exposure to diagnostic X-irradiation, mean percentage of DNA in tail of the comets, which is indirect measures of DNA damage, was significantly changed. The same was noticed for mean total number of micronuclei as well. It was shown that children with pulmonary diseases subjected to diagnostic procedure develop a significant increase in mean total number of each measured parameter which are the biomarkers of genetic damage for carcinogenesis, than prior to diagnostic procedure and that interindividual differences exist for each monitored child. Our results show that genetic damage arises

  19. Genotoxic potential generated by biomass burning in the Brazilian Legal Amazon by Tradescantia micronucleus bioassay: a toxicity assessment study

    Directory of Open Access Journals (Sweden)

    Artaxo Paulo

    2011-05-01

    Full Text Available Abstract Background The Brazilian Amazon has suffered impacts from non-sustainable economic development, especially owing to the expansion of agricultural commodities into forest areas. The Tangará da Serra region, located in the southern of the Legal Amazon, is characterized by non-mechanized sugar cane production. In addition, it lies on the dispersion path of the pollution plume generated by biomass burning. The aim of this study was to assess the genotoxic potential of the atmosphere in the Tangará da Serra region, using Tradescantia pallida as in situ bioindicator. Methods The study was conducted during the dry and rainy seasons, where the plants were exposed to two types of exposure, active and passive. Results The results showed that in all the sampling seasons, irrespective of exposure type, there was an increase in micronucleus frequency, compared to control and that it was statistically significant in the dry season. A strong and significant relationship was also observed between the increase in micronucleus incidence and the rise in fine particulate matter, and hospital morbidity from respiratory diseases in children. Conclusions Based on the results, we demonstrated that pollutants generated by biomass burning in the Brazilian Amazon can induce genetic damage in test plants that was more prominent during dry season, and correlated with the level of particulates and elevated respiratory morbidity.

  20. Biomonitoring of genotoxic risk in radar facility workers: comparison of the comet assay with micronucleus assay and chromatid breakage assay

    International Nuclear Information System (INIS)

    Garaj-Vrhovac, V.; Kopjar, N.

    2003-01-01

    Genotoxic risks of occupational exposure in a radar facility were evaluated by using alkaline comet assay, micronucleus assay and chromatid breakage assay on peripheral blood leukocytes in exposed subjects and corresponding controls. Results show that occupational exposure to microwave radiation correlates with an increase of genome damage in somatic cells. The levels of DNA damage in exposed subjects determined by using alkaline comet assay were increased compared to control and showed interindividual variations. Incidence of micronuclei was also significantly increased compared to baseline control values. After short exposure of cultured lymphocytes to bleomycin, cells of occupationally exposed subjects responded with high numbers of chromatid breaks. Although the level of chromosome damage generated by bleomycin varied greatly between individuals, in exposed subjects a significantly elevated number of chromatid breaks was observed. Our results support data reported in literature indicating that microwave radiation represents a potential DNA-damaging hazard. Alkaline comet assay is confirmed as a sensitive and highly reproducible technique for detection of primary DNA damage inflicted in somatic cells. Micronucleus assay was confirmed as reliable bio-markers of effect and chromatid breakage assay as sensitive bio-marker of individual cancer susceptibility. The results obtained also confirm the necessity to improve measures and to perform accurate health surveillance of individuals occupationally exposed to microwave radiation

  1. 3-Nitrobenzanthrone (3-NBA) induced micronucleus formation and DNA damage in human hepatoma (HepG2) cells.

    Science.gov (United States)

    Lamy, Evelyn; Kassie, Fekadu; Gminski, Richard; Schmeiser, Heinz H; Mersch-Sundermann, Volker

    2004-01-15

    3-Nitrobenzanthrone (3-NBA), identified in diesel exhaust and in airborne particulate matter, is a potent mutagen in Salmonella, induces micronuclei formation in mice and in human cells and DNA adducts in rats. In the present study, we investigated the genotoxic potency of 3-NBA in human HepG2 cells using the micronucleus (MN) assay and the single cell gel electrophoresis (SCGE). 3-NBA caused a genotoxic effect at concentrations > or =12 nM in both assays. In the micronucleus assay, we found 98.7+/-10.3 MN/1000 BNC at a concentration of 100 nM 3-NBA in comparison to 27.3+/-0.6 MN/1000 BNC with the negative control. At the same concentration, the DNA-migration (SCGE) showed an Olive tail moment (OTM) of 2.7+/-0.45 and %DNA in the tail of 8.28+/-0.76; OTM and %DNA in the tail of cells treated with the negative control were 0.73+/-0.08 and 2.81+/-0.30, respectively. The results are discussed under consideration of former studies.

  2. Biomonitoring of agricultural workers exposed to pesticide mixtures in Guerrero state, Mexico, with comet assay and micronucleus test.

    Science.gov (United States)

    Carbajal-López, Yolanda; Gómez-Arroyo, Sandra; Villalobos-Pietrini, Rafael; Calderón-Segura, María Elena; Martínez-Arroyo, Amparo

    2016-02-01

    The aim of this study was to evaluate the genotoxic effect of pesticides in exfoliated buccal cells of workers occupationally exposed in Guerrero, Mexico, using the comet assay and the micronucleus test. The study compared 111 agricultural workers in three rural communities (Arcelia 62, Ajuchitlan 13, and Tlapehuala 36), with 60 non-exposed individuals. All the participants were males. The presence of DNA damage was investigated in the exfoliated buccal cells of study participants with the comet assay and the micronucleus (MN) test; comet tail length was evaluated in 100 nuclei and 3000 epithelial cells of each individual, respectively; other nuclear anomalies such as nuclear buds, karyolysis, karyorrhexis, and binucleate cells were also evaluated. Study results revealed that the tail migration of DNA and the frequency of MN increased significantly in the exposed group, which also showed nuclear anomalies associated with cytotoxic or genotoxic effect. No positive correlation was noted between exposure time and tail length and micronuclei frequencies. No significant effect on genetic damage was observed as a result of age, smoking, and alcohol consumption. The MN and comet assay in exfoliated buccal cells are useful and minimally invasive methods for monitoring genetic damage in individuals exposed to pesticides. This study provided valuable data for establishing the possible risk to human health associated with pesticide exposure.

  3. Influence of Magnolol on the bystander effect induced by alpha-particle irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Wong, T.P.W.; Law, Y.L. [Department of Physics and Materials Science, City University of Hong Kong, Tat Chee Avenue, Kowloon Tong (Hong Kong); Tse, A.K.W.; Fong, W.F. [Research and Development Division, School of Chinese Medicine, Hong Kong Baptist University, Baptist University Road, Kowloon Tong (Hong Kong); Yu, K.N. [Department of Physics and Materials Science, City University of Hong Kong, Tat Chee Avenue, Kowloon Tong (Hong Kong)], E-mail: peter.yu@cityu.edu.hk

    2010-04-15

    In this work, the influence of Magnolol on the bystander effect in alpha-particle irradiated Chinese hamster ovary (CHO) cells was examined. The bystander effect was studied through medium transfer experiments. Cytokinesis-block micronucleus (CBMN) assay was performed to quantify the chromosome damage induced by alpha-particle irradiation. Our results showed that the alpha-particle induced micronuclei (MN) frequencies were suppressed with the presence of Magnolol.

  4. Application of liquid-based cytology preparation in micronucleus assay of exfoliated buccal epithelial cells in road construction workers.

    Science.gov (United States)

    Arul, P

    2017-01-01

    Asphalts are bitumens that consist of complex of hydrocarbon mixtures and it is used mainly in road construction and maintenance. This study was undertaken to evaluate the micronucleus (MN) assay of exfoliated buccal epithelial cells in road construction workers using liquid-based cytology (LBC) preparation. Three different stains (May-Grunwald Giemsa, hematoxylin and eosin, and Papanicolaou) were used to evaluate the frequency of MN in exfoliated buccal epithelial of 100 participants (fifty road construction workers and fifty administrative staff) using LBC preparation. Statistical analysis was performed with Student's t-test, and Proad construction exhibit a higher frequency of MN in exfoliated buccal epithelial cells and they are under the significant risk of cytogenetic damage. LBC preparation has potential application for the evaluation of frequency of MN. This technique may be advocated in those who are occupationally exposed to potentially carcinogenic agents in view of improvement in the smear quality and visualization of cell morphology.

  5. Micronucleus formation compared to the survival rate of human melanoma cells after X-ray and neutron irradiation and hyperthermia

    Energy Technology Data Exchange (ETDEWEB)

    van Beuningen, D.; Streffer, C.; Bertholdt, G.

    1981-09-01

    After neutron and X-ray irradiation and combined X-ray irradiation and hyperthermia (3 hours, 42/sup 0/C), the survival rate of human melanoma cells was measured by means of the colony formation test and compared to the formation of micronuclei. Neutrons had a stronger effect on the formation of micronuclei than the combination of X-rays and hyperthermia. X-rays had the lowest effect. The dose effect curve showed a break at that dose level at which a reduction of cells was observed in the cultures. A good relation between survival rate and formation of micronuclei was found for the X-ray irradiation, but not for the neutron irradiation and the combined treatment. These observations are discussed. At least for X-rays, the micronucleus test has turned out to be a good screening method for the radiosensitivity of a biologic system.

  6. Genotoxic, Cytotoxic, Antigenotoxic, and Anticytotoxic Effects of Sulfonamide Chalcone Using the Ames Test and the Mouse Bone Marrow Micronucleus Test.

    Directory of Open Access Journals (Sweden)

    Carolina Ribeiro E Silva

    Full Text Available Chalcones present several biological activities and sulfonamide chalcone derivatives have shown important biological applications, including antitumor activity. In this study, genotoxic, cytotoxic, antigenotoxic, and anticytotoxic activities of the sulfonamide chalcone N-{4-[3-(4-nitrophenylprop-2-enoyl]phenyl} benzenesulfonamide (CPN were assessed using the Salmonella typhimurium reverse mutation test (Ames test and the mouse bone marrow micronucleus test. The results showed that CPN caused a small increase in the number of histidine revertant colonies in S. typhimurium strains TA98 and TA100, but not statistically significant (p > 0.05. The antimutagenicity test showed that CPN significantly decreased the number of His+ revertants in strain TA98 at all doses tested (p 0.05. Additionally, CPN co-administered with MMC significantly increased PCE/NCE ratio at all doses tested, demonstrating its anticytotoxic effect. In summary, CPN presented genotoxic, cytotoxic, antigenotoxic, and anticytotoxic properties.

  7. Evaluation of genotoxicity after application of Listerine(R) on human lymphocytes by micronucleus and single cell gel electrophoresis assays.

    Science.gov (United States)

    Türkez, Hasan; Togar, Basak; Arabaci, Taner

    2012-04-01

    Listerine (LN) is one of the most commonly used mouth rinses worldwide although very limited information is available concerning its genotoxicity. In another view, the biological safety profile of oral care products is frequently assumed on the basis of simplistic test models. Therefore, the present study was undertaken to investigate the in vitro genotoxic potential of LN using micronucleus and single cell gel electrophoresis tests as genetic endpoints. Different concentrations of LN (0-100% of ml/culture, v/v) were applied to whole human blood cultures (n = 5). The result of the present study showed that there were no statistically significant differences (p > 0.05) between the control group and the groups treated with LN alone in both analysed endpoints. In conclusion, our result first demonstrated the absence of genotoxicity of LN on human lymphocytes.

  8. Fish eco-genotoxicology: Comet and micronucleus assay in fish erythrocytes as in situ biomarker of freshwater pollution

    Directory of Open Access Journals (Sweden)

    Bilal Hussain

    2018-02-01

    Full Text Available Owing to white meat production Labeo rohita have vast economic importance, but its population has been reduced drastically in River Chenab due to pollution. Atomic absorption spectrophotometry showed a merciless toxicity level of Cd, Cu, Mn, Zn, Pb, Cr, Sn and Hg. Comet assay results indicated significant (p < .05 DNA fragmentation in Labeo rohita as 42.21 ± 2.06%, 31.26 ± 2.41% and 21.84 ± 2.21% DNA in comet tail, tail moment as 17.71 ± 1.79, 10.30 ± 1.78 and 7.81 ± 1.56, olive moment as 13.58 ± 1.306, 8.10 ± 1.04 and 5.88 ± 0.06, respectively, from three different polluted sites on the river. Micronucleus assay showed similar findings of single micronucleus induction (MN as 50.00 ± 6.30‰, double MN 14.40 ± 2.56‰, while nuclear abnormalities (NA were found as 150.00 ± 2.92‰. These higher frequencies of MN induction and NA were found to be the cause of reduction of 96% of the population of this fish species in an experimental area of the River Chenab. This fish species has been found near extinction through the length of the river Chenab and few specimens in rainy seasons if restored by flood, may die in sugarcane mill season. Due to sweeping extinction Labeo rohita showed the highest sensitivity for pollution and could be used as bioindicator and DNA fragmentation in this column feeder fish species as a biomarker of the pollution load in freshwater bodies.

  9. Sacha Inchi (Plukenetia volubilis L. powder: acute toxicity, 90 days oral toxicity study and micronucleus assay in rodents

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    Idania Rodeiro

    2018-02-01

    Full Text Available Context: Sacha Inchi has been consumed for years by indigenous peoples. Meanwhile, its toxicological potential has not been sufficiently studied. Aims: To assess the acute, sub-chronic toxicity and genotoxicity evaluation of Sacha Inchi powder obtained from Plukenetia volubilis L. Methods: A dose of 2000 mg/kg was orally administered to rats and mice and toxicity symptoms for 14 days were observed. In repeated dose study, the product was orally administered to Sprague Dawley rats of both sexes. Animals received 50, 250 and 500 mg/kg/day of the product for 90 days. At the end, animals were sacrificed and samples were done for hematological and biochemical analysis, organ weighs and histopathological examination. Genotoxicity potential of Sacha Inchi powder was evaluated through micronucleus test in mice. Negative controls received the vehicle (carboxymethyl cellulose, 0.5% used. Results: No morbidity or mortality at 2000 mg/kg of the product were found. Sacha Inchi powder oral administration during 90 days to rats did not lead to death, body weight gain, food consumption, or adverse events. No significant changes on hematological or biochemical parameters, organ weights or histopathological findings were observed. Induction of micronucleus formation attributable to the product was not found in mice. Conclusions: No toxicity effects after oral acute exposure of Sacha Inchi power to rats and mice were observed. Neither toxicity attributable to oral doses of the product up to 500 mg/kg during 90 days to rats were found. Results suggested Sacha Inchi powder does not have genotoxicity potential under our experimental conditions.

  10. Role of Macronutrients and Micronutrients in DNA Damage: Results From a Food Frequency Questionnaire.

    Science.gov (United States)

    Ladeira, Carina; Carolino, Elisabete; Gomes, Manuel C; Brito, Miguel

    2017-01-01

    The links between diet and genomic instability have been under investigation for several decades, and evidence suggests a significant causal or preventive role for various dietary factors. This study investigates the influence of macronutrients (calories, protein, and glucides) and micronutrients, such as vitamins and minerals, as assessed by a food frequency questionnaire, on genotoxicity biomarkers measured by cytokinesis-blocked micronucleus assay and comet assay. The results found significant positive and negative correlations. Micronucleus frequency tends to increase with higher intake of caffeine, calcium, magnesium, zinc, and protein ( P macronutrients and micronutrients.

  11. Changes of chromosome aberration rate and micronucleus frequency along with accumulated dose in continuously irradiated mice with a low dose rate of γ-rays

    International Nuclear Information System (INIS)

    Tanaka, Kimio; Izumi, Jun; Yanai, Takanori; Ichinohe, Kazuaki; Matsumoto, Tsuneya

    2003-01-01

    Chromosome aberrations in chronically exposed workers in nuclear facilities and medical radiologists have been reported. However chronological change of chromosome aberration rates along with accumulated dose has not been well studied. Chromosome aberrations and micronuclei in spleen lymphocytes were observed serially in mice continuously irradiated with a low dose rate of 20 mGy/day up to 400 days. Chromosome aberration rates were rapidly increased to 11.1% at 1 Gy, while micronucleus incidence increased at 5 Gy. After these doses their increase rates were saturated. Micronucleus incidence in bone marrow erythroblasts was higher than in spleen cells. These chronological changes of cytogenetic aberrations seem to be induced through a balance between developments of chromosome aberrations and micronuclei, and life span of spleen lymphocytes. These results will be helpful for risk assessment in low dose rate radiation exposure. (author)

  12. Use of the micronucleus assay for the selective detection of radiosensitivity in BUdR-unincorporated cells after pulse-labelling of exponentially growing tumour cells

    Energy Technology Data Exchange (ETDEWEB)

    Masunaga, S.; Ono, K.; Fushiki, M.; Abe, M. (Kyoto Univ. (Japan). Faculty of Medicine); Wandl, E.O. (Vienna Univ. (Austria). Klinik fuer Strahlentherapie und Strahlenbiologie)

    1990-08-01

    To determine the radiosensitivity of non S-phase tumour cells in vitro, survival curves of SCC VII tumour cells were obtained after a short block with hydroxyurea. Dose-response curves of micronucleus (MN) frequency appearing in non-S-phase cells were also determined by excluding S-phase cells with immunofluorescence staining to 5-bromo-2'-deoxyuridine (BUdR). Both the dose response curves of MN frequency and survival curves were analysed by a linear-quadratic model (surviving fraction =exp (-{alpha}D-{beta}D{sup 2}), MN frequency =aD+bD{sup 2}+c). A good correlation between the {alpha}/{beta} and a/b ratios was observed. In both BUdR-unincorporated and asynchronous cell cultures, the regression lines between the surviving fraction and micronucleus frequency were statistically identical. (author).

  13. Aneuploidogenic effects and DNA oxidation induced in vitro by differently sized gold nanoparticles

    Directory of Open Access Journals (Sweden)

    Di Bucchianico S

    2014-05-01

    Full Text Available Sebastiano Di Bucchianico,1 Maria Rita Fabbrizi,1 Silvia Cirillo,1 Chiara Uboldi,1 Douglas Gilliland,2 Eugenia Valsami-Jones,3,4 Lucia Migliore11Department of Translational Research and New Technologies in Medicine and Surgery, Medical Genetics Unit, University of Pisa, Pisa, Italy; 2European Commission-Joint Research Centre, Institute for Health and Consumer Protection, NanoBioSciences Unit, Ispra, Italy; 3School of Geography, Earth and Environmental Sciences, University of Birmingham, Edgbaston, Birmingham, UK; 4Earth Sciences, Natural History Museum, Cromwell Road, London, UKAbstract: Gold nanoparticles (Au NPs are used in many fields, including biomedical applications; however, no conclusive information on their potential cytotoxicity and genotoxicity mechanisms is available. For this reason, experiments in human primary lymphocytes and murine macrophages (Raw264.7 were performed exposing cells to spherical citrate-capped Au NPs with two different nominal diameters (5 nm and 15 nm. The proliferative activity, mitotic, apoptotic, and necrotic markers, as well as chromosomal damage were assessed by the cytokinesis-block micronucleus cytome assay. Fluorescence in situ hybridization with human and murine pancentromeric probes was applied to distinguish between clastogenic and aneuploidogenic effects. Our results indicate that 5 nm and 15 nm Au NPs are able to inhibit cell proliferation by apoptosis and to induce chromosomal damage, in particular chromosome mis-segregation. DNA strand breaks were detected by comet assay, and the modified protocol using endonuclease-III and formamidopyrimidine-DNA glycosylase restriction enzymes showed that pyrimidines and purines were oxidatively damaged by Au NPs. Moreover, we show a size-independent correlation between the cytotoxicity of Au NPs and their tested mass concentration or absolute number, and genotoxic effects which were more severe for Au NP 15 nm compared to Au NP 5 nm. Results indicate that

  14. Genomic instability related to zinc deficiency and excess in an in vitro model: is the upper estimate of the physiological requirements recommended for children safe?

    Science.gov (United States)

    Padula, Gisel; Ponzinibbio, María Virginia; Gambaro, Rocío Celeste; Seoane, Analía Isabel

    2017-08-01

    Micronutrients are important for the prevention of degenerative diseases due to their role in maintaining genomic stability. Therefore, there is international concern about the need to redefine the optimal mineral and vitamin requirements to prevent DNA damage. We analyzed the cytostatic, cytotoxic, and genotoxic effect of in vitro zinc supplementation to determine the effects of zinc deficiency and excess and whether the upper estimate of the physiological requirement recommended for children is safe. To achieve zinc deficiency, DMEM/Ham's F12 medium (HF12) was chelated (HF12Q). Lymphocytes were isolated from healthy female donors (age range, 5-10 yr) and cultured for 7 d as follows: negative control (HF12, 60 μg/dl ZnSO 4 ); deficient (HF12Q, 12 μg/dl ZnSO 4 ); lower level (HF12Q + 80 μg/dl ZnSO 4 ); average level (HF12Q + 180 μg/dl ZnSO 4 ); upper limit (HF12Q + 280 μg/dl ZnSO 4 ); and excess (HF12Q + 380 μg/dl ZnSO 4 ). The comet (quantitative analysis) and cytokinesis-block micronucleus cytome assays were used. Differences were evaluated with Kruskal-Wallis and ANOVA (p < 0.05). Olive tail moment, tail length, micronuclei frequency, and apoptotic and necrotic percentages were significantly higher in the deficient, upper limit, and excess cultures compared with the negative control, lower, and average limit ones. In vitro zinc supplementation at the lower and average limit (80 and 180 μg/dl ZnSO 4 ) of the physiological requirement recommended for children proved to be the most beneficial in avoiding genomic instability, whereas the deficient, upper limit, and excess (12, 280, and 380 μg/dl) cultures increased DNA and chromosomal damage and apoptotic and necrotic frequencies.

  15. Genotoxicity of freshwater ecosystem shows DNA damage in preponderant fish as validated by in vivo micronucleus induction in gill and kidney erythrocytes.

    Science.gov (United States)

    Obiakor, M O; Okonkwo, J C; Ezeonyejiaku, C D

    2014-12-01

    Genotoxicity of Anambra River was studied by micronucleus (MN) assay of preponderant fish species in the river. The micronucleus indices obtained were used as biomarker to estimate and predict pollution profile and possible danger of feeding on the aquatic species. Micronuclei profile of the fish was measured from gill and kidney erythrocytes using microscopic technique. Season, species and location effects on micronuclei, together with their interactions were also determined. Two major seasons (rainy and dry) and preponderant fish species in the river (Synodontis clarias, Linnaeus, 1758 and Tilapia nilotica, Linnaeus, 1757) were studied at five distinct locations that displayed differential environmental stresses. The study showed that the micronucleus index of fish is an excellent biomarker for measuring pollution level and genotoxicity of freshwater habitat. Season, species of fish and location affect micronuclei profile of the fish species sampled in the river. Disease outbreak among rural dwellers depending on the river for domestic and other uses is imminent and they lack knowledge on its health implication. Moreover, the study maintained that the micronuclei in fish could be measured from either the gill or kidney; however, gill is more efficient as it enables collection of several samples from the same individuals without sacrificing it, and Synodontis clarias fish species appeared to be more vulnerable to the genotoxic damage than Tilapia nilotica. Consequently, the study recommended regular monitoring (micronucleus tests) of edible aquatic life such as Synodontis clarias in order to eliminate the danger of people feeding on toxic metals, some of which are carcinogenic. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Mutagenic effects of tributyltin and inorganic lead (Pb II on the fish H. malabaricus as evaluated using the comet assay and the piscine micronucleus and chromosome aberration tests

    Directory of Open Access Journals (Sweden)

    Ferraro Marcos Vinícius M.

    2004-01-01

    Full Text Available Genotoxicity studies on toxic metals and their organic compounds are very important, especially so in the investigation of the effects of these compounds on the aquatic environments where they tend to accumulate. The use of endemic aquatic organisms as biological sentinels has proved useful to environmental monitoring. We assessed the mutagenic potential of tributyltin (TBT and inorganic lead (PbII using samples of the fish Hoplias malabaricus (commonly called traíra using the comet assay and the piscine micronucleus and chromosome aberration tests. Eighteen H. malabaricus were acclimatized in three individual aquariums, each containing six fish, six fish being exposed to 0.3 mg/g of body weight (bw of TBT, six to 21 mg/g bw of PbII and six being used as controls. Exposure to TBT and PbII was achieved by feeding the fish every five days with Astyanax (a small fish that is part of the normal diet of H. malabaricus which had been injected with solutions of TBT, PbII or with water (the control group. After two months the H. malabaricus were sacrificed and their peripheral blood collected and subjected to the comet and micronucleus assays, the chromosome aberration assay being conducted using kidney-tissue. Although the comet assay showed now mutagenic effects at the lead concentrations used but encountered results with TBT, the micronucleus and chromosome aberrations assays both indicated that TBT and PbII are potentially mutagenic (p < 0.01, the micronucleus assay showing morphological alterations of the nucleus.

  17. Evaluation of the repeated-dose liver micronucleus assay using 2,4-dinitrotoluene: a report of a collaborative study by CSGMT/JEMS.MMS.

    Science.gov (United States)

    Maeda, Akihisa; Tsuchiyama, Hiromi; Asaoka, Yoshiji; Hirakata, Mikito; Miyoshi, Tomoya; Oshida, Keiyu; Miyamoto, Yohei

    2015-03-01

    The liver micronucleus assay using young adult rats has the potential to detect liver carcinogens by repeated dosing, and could be expected to be integrated into repeated-dose toxicity studies using a hepatocyte isolation method without the traditional in situ collagenase perfusion. In this study, to assess the performance of the repeated-dose liver micronucleus assay, 2,4-dinitrotoluene (DNT), which is a rodent liver carcinogen, was administered orally to male rats at doses of 50, 100 and 200 mg/kg/day once daily for 14 or 28 consecutive days, and the frequencies of micronucleated hepatocytes (MNHEPs) and micronucleated immature erythrocytes (MNIMEs) were examined. Significant increases in the MNHEPs were observed at 50 mg/kg/day or more in the 14-day treatment, and 50 and 100 mg/kg/day in the 28-day treatment. These increases were dependent on both the dose and the number of administrations, which indicates the possibility that the MNHEPs accumulate as a result of repeated dosing. In contrast, no increase in the MNIMEs was observed. In conclusion, the repeated-dose liver micronucleus assay using young adult rats is sufficiently sensitive to detect the genotoxicity of 2,4-DNT at a low dose.

  18. The effect of gamma radiation on the Common carp (Cyprinus carpio): In vivo genotoxicity assessment with the micronucleus and comet assays.

    Science.gov (United States)

    M K, Praveen Kumar; Soorambail K, Shyama; Bhagatsingh Harisingh, Sonaye; D'costa, Avelyno; Ramesh Chandra, Chaubey

    2015-10-01

    Radioactive wastes may be leached into freshwater, either accidentally or in industrial effluents. We have studied gamma radiation-induced DNA damage in the freshwater fish Cyprinus carpio. Fish were irradiated with 2-10Gy gamma radiation and genotoxic effects in blood cells were studied with the micronucleus (MN) and comet assays. Micronuclei and a dose-dependent increase in comet-tail DNA were seen in dose- and time-dependent studies. The highest % tail DNA was observed at 24h, declining until 72h, which may indicate the repair of radiation-induced DNA single-strand breaks after gamma radiation. However, double-stranded DNA damage may not have been repaired, as indicated by increased micronuclei at later periods. A positive correlation was observed between the comet and micronucleus assay results. This study confirms the mutagenic/genotoxic potential of gamma radiation in the Common carp, as well as the possible combined use of the micronucleus and comet assays for in vivo laboratory studies with fresh-water fish for screening the genotoxic potential of radioactive pollution. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Ultrastructural changes of the UV-irradiated micronucleus in vegetative cells of paramecium bursaria and its functional importance

    International Nuclear Information System (INIS)

    Borkhsenius, O.N.; Fokin, S.I.

    1982-01-01

    Ultrastructural micronucleus (MI) changes of vegetative cells in Paramecium bursaria in 0.5-7 h after MI ultraviolet irradiation and cell posterity with irradiated MI after different periods (2.6 and 30 days, three years) after ultraviolet irradiation have been studied. It is established that MI irradiation at a dose of 306J/m 2 doesn't result in its loss in postradiation generations however in posterity MI considerable ultrastructural changes occur. In two days after operation the shell of descendant MI of irradiated Paramecium bursaria forms multiple blades; small chromatin blocks considerably increase in sizes and, as a rule, occupy the central position in a nucleus. During later periods afeter irradiation (6 days) density of chromatin elements in MI begins to change. By the 30-st day in MI fine-fibrillar karyoplasm detected are only not numerous chromatin structures scattered in disorder. The results obtained point to the existence of the ''cryptic'' type MIs are not revealed at the light-optical level but preserved in a series of postradiation generations. The presence of such MIs in viable cultures of P. bursaria confirms indirectly MI significance in vegetatic life of P. bursaria

  20. The Tradescantia micronucleus assay is a highly sensitive tool for the detection of low levels of radioactivity in environmental samples.

    Science.gov (United States)

    Mišík, Miroslav; Krupitza, Georg; Mišíková, Katarina; Mičieta, Karol; Nersesyan, Armen; Kundi, Michael; Knasmueller, Siegfried

    2016-12-01

    Environmental contamination with radioactive materials of geogenic and anthropogenic origin is a global problem. A variety of mutagenicity test procedures has been developed which enable the detection of DNA damage caused by ionizing radiation which plays a key role in the adverse effects caused by radioisotopes. In the present study, we investigated the usefulness of the Tradescantia micronucleus test (the most widely used plant based genotoxicity bioassay) for the detection of genetic damage caused by environmental samples and a human artifact (ceramic plate) which contained radioactive elements. We compared the results obtained with different exposure protocols and found that direct exposure of the inflorescences is more sensitive and that the number of micronuclei can be further increased under "wet" conditions. The lowest dose rate which caused a significant effect was 1.2 μGy/h (10 h). Comparisons with the results obtained with other systems (i.e. with mitotic cells of higher plants, molluscs, insects, fish and human lymphocytes) show that the Tradescantia MN assay is one to three orders of magnitude more sensitive as other models, which are currently available. Taken together, our findings indicate that this method is due to its high sensitivity a unique tool, which can be used for environmental biomonitoring in radiation polluted areas. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Ultrastructural changes of the UV-irradiated micronucleus in vegetative cells of Paramecium bursaria and its functional importance

    Energy Technology Data Exchange (ETDEWEB)

    Borkhsenius, O.N.; Fokin, S.I. (Leningradskij Gosudarstvennyj Univ. (USSR). Biologicheskij Nauchno-Issledovatel' skij Inst.)

    1982-01-01

    Ultrastructural micronucleus (MI) changes of vegetative cells in Paramecium bursaria in 0.5-7 h after MI ultraviolet irradiation and cell posterity with irradiated MI after different periods (2.6 and 30 days, three years) after ultraviolet irradiation have been studied. It is established that MI irradiation at a dose of 306J/m/sup 2/ doesn't result in its loss in postradiation generations however in posterity MI considerable ultrastructural changes occur. In two days after operation the shell of descendant MI of irradiated Paramecium bursaria forms multiple blades; small chromatin blocks considerably increase in sizes and, as a rule, occupy the central position in a nucleus. During later periods after irradiation (6 days) density of chromatin elements in MI begins to change. By the 30th day in MI fine-fibrillar karyoplasm detected are only not numerous chromatin structures scattered in disorder. The results obtained point to the existence of the ''cryptic'' type MIs are not revealed at the light-optical level but preserved in a series of postradiation generations. The presence of such MIs in viable cultures of P. bursaria confirms indirectly MI significance in vegetatic life of P. bursaria.

  2. Micronucleus frequency and hematologic index in Colossoma macropomum (Pisces, Ariidae) for environmental impact assessment at a protected area in Brazil

    International Nuclear Information System (INIS)

    Sousa, Debora Batista Pinheiro; Neta, Raimunda Nonata Fortes Carvalho

    2014-01-01

    This study used micronucleus assays and erythrocyte indices in the freshwater fish tambaqui, Colossoma macropomum, to assess environmental impacts in the Environmental Protection Area at Maracanã, São Luis, Brazil. Fish were sampled from two locations within the protected area, Serena Lagoon and Ambude River, on four occasions. Biometric data (length and weight) and an aliquot of blood were collected from each fish for analysis. Erythrocyte indices including: mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration were calculated, and blood samples were examined for micronuclei and nuclear morphological changes. Micronuclei were found in fish from both locations, although the frequency was higher in fish from Ambude River. Nuclear morphological changes were identified only in fish collected from Ambude River. Several nuclear morphological changes were found in erythrocytes stained with Giemsa, including: micronuclei and binucleate nuclei. On average, erythrocyte indices were lower in fish collected from Ambude River than in those from Serena Lagoon. Our results indicate that micronuclei and erythrocyte indices can be used in C. macropomum as indicators of environmental health

  3. Evaluating the genotoxic effects of workers exposed to lead using micronucleus assay, comet assay and TCR gene mutation test

    International Nuclear Information System (INIS)

    Chen Zhijian; Lou Jianlin; Chen Shijie; Zheng Wei; Wu Wei; Jin Lifen; Deng Hongping; He Jiliang

    2006-01-01

    To evaluate the genotoxic effects of lead (Pb) exposure, 25 workers in a workplace producing storage battery were monitored for three genetic end-points using micronucleus (MN) assay, comet assay and TCR gene mutation test. Twenty-five controls were matched with workers according to age, gender and smoking. The air Pb concentration in the workplace was 1.26 mg/m 3 . All subjects were measured for Pb concentration of blood by atom absorption spectrophotometry. The mean Pb concentration of blood in workers (0.32 mg/l) was significantly higher than that in controls (0.02 mg/l). The results of MN test showed that the mean micronuclei rate (MNR) and mean micronucleated cells rate (MCR) in workers were 9.04 ± 1.51 per mille and 7.76 ± 1.23 per mille , respectively, which were significantly higher than those (2.36 ± 0.42 per mille and 1.92 ± 0.31 per mille ) in controls (P -4 and 1.74 ± 0.17 x 10 -4 , respectively, there was no significant difference between workers and controls (P > 0.05). The results of our study indicated that the genetic damage was detectable in 25 workers occupationally exposed to lead

  4. Micronucleus frequency and hematologic index in Colossoma macropomum (Pisces, Ariidae) for environmental impact assessment at a protected area in Brazil

    Energy Technology Data Exchange (ETDEWEB)

    Sousa, Debora Batista Pinheiro, E-mail: deborabpsousa@gmail.com [Postgraduate Program of Aquatic Resources and Fishery (PPGRAP/UEMA), State University of Maranhão (Brazil); Neta, Raimunda Nonata Fortes Carvalho [Department of Chemistry and Biology, State University of Maranhão (Brazil)

    2014-10-06

    This study used micronucleus assays and erythrocyte indices in the freshwater fish tambaqui, Colossoma macropomum, to assess environmental impacts in the Environmental Protection Area at Maracanã, São Luis, Brazil. Fish were sampled from two locations within the protected area, Serena Lagoon and Ambude River, on four occasions. Biometric data (length and weight) and an aliquot of blood were collected from each fish for analysis. Erythrocyte indices including: mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration were calculated, and blood samples were examined for micronuclei and nuclear morphological changes. Micronuclei were found in fish from both locations, although the frequency was higher in fish from Ambude River. Nuclear morphological changes were identified only in fish collected from Ambude River. Several nuclear morphological changes were found in erythrocytes stained with Giemsa, including: micronuclei and binucleate nuclei. On average, erythrocyte indices were lower in fish collected from Ambude River than in those from Serena Lagoon. Our results indicate that micronuclei and erythrocyte indices can be used in C. macropomum as indicators of environmental health.

  5. Techniques for induction of premature chromosome condensation (PCC) by Calyculin-A and micronucleus assay for biodosimetry in Vietnam

    International Nuclear Information System (INIS)

    Pham Ngoc Duy; Tran Que; Hoang Hung Tien; Bui Thi Kim Luyen; Nguyen Thi Kim Anh; Ha Thi Ngoc Lien

    2014-01-01

    The International Atomic Energy Agency (IAEA) and World Health Organization are interested in biological dosimetry method for radiation emergency medicine currently. Some cytogenetic techniques such as premature chromosome condensation (PCC) induced by Calyculin-A and micronucleus (MN) assay are necessary to develop biodosimetry in Vietnam. In this study, we optimized the condition for MN assay with 6 µg/ml Cytochalasin-B concentration and 72.5 hours for peripheral lymphocyte blood culture. The optimization for PCC method is 50 nM Calyculin-A concentration for 45 minutes peripheral lymphocyte blood treatment. For samples exposed to 3.0 Gy gamma 60 Co (dose rate 0.0916 Gy/s), the frequency of MN is 19.02 ± 0.38%, NBP is 1.95 ± 0.28%, dicentric and ring is 41.43 ± 8.12% and frag and min is 63.33 ± 5.16%. For samples exposed to 6.0 Gy gamma 60 Co (dose rate 0.0916 Gy/s), the frequency of ring-PCC is 17.73 ± 2.46%, extra unite is 218.91± 7.58%, dicentric is 83.81 ± 1.09%, ring is 10.75 ± 1.74%, fragment and minute is 193.17 ± 13.10%. MN and ring-PCC are specific marker applying for biodosimetry. (author)

  6. Improvement of techniques for the detection of radio-induced micronuclei in human blood lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Voisin, P.; Paillole, N.

    1995-12-31

    Scoring of micronuclei in cytokinesis-blocked peripheral human lymphocytes, after an accidental overexposure, seems an easier and faster alternative for biological dosimetry than conventional cytogenetics (dicentric chromosomes). Several variations of the cytokinesis-block micronucleus assay have been tested, in order to obtain a sufficient number of micronuclei in bi-nucleated cells by the shortest time possible for operational purposes. The methods differ in the use of hypotonic treatment as well as culture time (48 to 72 h), amount of blood and materials used. We have compared frequencies of bi-nucleated cells and micronuclei in normal lymphocytes and after {gamma}-({sup 60}Co) irradiation in vitro with {sup 60}Co for doses up to 6 Gy. Main results and the final choice of the technique are presented. (authors). 3 refs., 3 figs.

  7. Improvement of techniques for the detection of radio-induced micronuclei in human blood lymphocytes

    International Nuclear Information System (INIS)

    Voisin, P.; Paillole, N.

    1995-01-01

    Scoring of micronuclei in cytokinesis-blocked peripheral human lymphocytes, after an accidental overexposure, seems an easier and faster alternative for biological dosimetry than conventional cytogenetics (dicentric chromosomes). Several variations of the cytokinesis-block micronucleus assay have been tested, in order to obtain a sufficient number of micronuclei in bi-nucleated cells by the shortest time possible for operational purposes. The methods differ in the use of hypotonic treatment as well as culture time (48 to 72 h), amount of blood and materials used. We have compared frequencies of bi-nucleated cells and micronuclei in normal lymphocytes and after γ-( 60 Co) irradiation in vitro with 60 Co for doses up to 6 Gy. Main results and the final choice of the technique are presented. (authors). 3 refs., 3 figs

  8. Flow cytometry based micronucleus assay for evaluation of genotoxic potential of 2-ACBs in hepatic cells HepG2

    International Nuclear Information System (INIS)

    Barbezan, Angélica B.; Santos, Carla J.B.; Carvalho, Luma R.; Vieira, Daniel P.; Villavicêncio, Anna L.C.H.; Santelli, Glaucia M.M.

    2017-01-01

    Food irradiation is approved for use in more than 60 countries for applications and purposes in a wide variety of foods, being an effective and safe method for preservation and long-term storage. 2-Alkylcyclobutanones (2-ACBs) are the only known radiolytic products generated from foods that contain fatty acids (Triglycerides) when irradiated. The acids analyzed in this study are palmitic and stearic, which when irradiated form 2-Dodecylcyclobutanones (2-dDCB) and 2-Tetradecylcyclobutanone (2-tDCB). Part of the 2-ACBs ingested is excreted through feces and part is deposited in adipose tissues. In vitro studies so far have been only in colon cells. The work used a human hepatoma cell line (HepG2) since the accumulation of fat in this organ is quite common. Micronucleus test was selected to evaluate possible genotoxic effects of 2-dDCB and 2-tDCB compounds when exposed to high concentrations (447, 1422 and 2235 μM) for 4 and 24 hours. Tests were performed in quadriplicates using flow cytometric analysis. None detectable genotoxic damage was observed after 4 hours of exposure to the compounds, and cytotoxic effects were only significant at the highest concentration (2235 μM) of 2-dDCB. After 24 hours of exposure, slight genotoxic damage was observed at all concentrations evaluated, and cytotoxic effects were only present when exposed to compound 2-tDCB. Although there is a genotoxic and cytotoxic effect in some of the situations tested, the two compounds predominantly induced proliferation reduction effects of this hepatic tumor cell line. (author)

  9. Dose-Response Assessment of Four Genotoxic Chemicals in a Combined Mouse and Rat Micronucleus and Comet Assay Protocol

    Science.gov (United States)

    Recio, Leslie; Hobbs, Cheryl; Caspary, William; Witt, Kristine L.

    2012-01-01

    The in vivo micronucleus (MN) assay has proven to be an effective measure of genotoxicity potential. However, sampling a single tissue (bone marrow) for a single indicator of genetic damage using the MN assay provides a limited genotoxicity profile. The in vivo alkaline (pH>13) Comet assay, which detects a broad spectrum of DNA damage, can be applied to a variety of rodent tissues following administration of test agents. To determine if the Comet assay is a useful supplement to the in vivo MN assay, a combined test protocol (MN/Comet assay) was conducted in male B6C3F1 mice and F344/N rats using four model genotoxicants: ethyl methanesulfonate (EMS), acrylamide (ACM), cyclophosphamide (CP), and vincristine sulfate (VS). Test compounds were administered on 4 consecutive days at 24-hour intervals (VS was administered to rats for 3 days); animals were euthanized 4 hours after the last administration. All compounds induced significant increases in micronucleated reticulocytes (MN-RET) in the peripheral blood of mice, and all but ACM induced MN-RET in rats. EMS and ACM induced significant increases in DNA damage, measured by the Comet assay, in multiple tissues of mice and rats. CP-induced DNA damage was detected in leukocytes and duodenum cells. VS, a spindle fiber disrupting agent, was negative in the Comet assay. Based on these results, the MN/Comet assay holds promise for providing more comprehensive assessments of potential genotoxicants, and the National Toxicology Program is presently using this combined protocol in its overall evaluation of the genotoxicity of substances of public health concern. PMID:20371966

  10. Evaluation of spontaneous and radiation-induced micronucleus frequency in cultured human peripheral blood lymphocytes depending on age and sex

    Energy Technology Data Exchange (ETDEWEB)

    Jeon, H. J.; Kang, C. M.; Chung, H. C. [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)] (and others)

    2002-12-15

    The goal of this study was to provide data on the dose-dependent production of MicroNucleus (MN) in human lymphocytes irradiated with {sup 60}Co {gamma}-rays and 50MeV neutron, and to evaluate predictive markers of intrinsic radiosensitivity in individuals for monitoring occupational or environmental radiation exposure. For the dose-response study, heparinized whole blood of 10 healthy volunteers was irradiated with {sup 60}Co {gamma}-rays employing of 0.25-8Gy. The MNs were observed all doses, and the numerical changes according to doses. In dose-response curves fit linear-quadratic form (alpha =0.31{+-}0.049, beta =0.0022{+-}0.0022) for {gamma}-rays, but (alpha=0.99{+-}0.528, beta =0.0093{+-}0.0047) for neutron. Neutrons were than {gamma}-rays effective in producing MN with dose-dependent manner. The frequency of MN varies with dose. The RBE (Relative Biological Effectiveness) for micronuclei was 2.370.17. Further studies were carried out to provide evidence for the existence of individual variations in age-dependent responses to radiation. Spontaneous and radiation-induced MN varies greatly among individuals, and little is known about the molecular mechanisms of this variability. It was shown that the increased level of spontaneous cell with MN was observed with increasing age. The relationship between radiosensitivity and the increased spontaneous level of MN may be in inverse proportion. These studies indicated that the MN assay have a high potential as a rapid, sensitive and accurate method which can be used to monitor a large population exposed to radiation for rapid triage in the case of a large-scale accident.

  11. Evaluation of spontaneous and radiation-induced micronucleus frequency in cultrued human peripheral blood lymphocytes depending on age and sex

    Energy Technology Data Exchange (ETDEWEB)

    Jeon, H. J.; Kang, C. M.; Chung, H. C. [Korea Cancer Center Hospital, Seoul (Korea, Republic of)] [and others

    2002-07-01

    The goal of this study was to provide data on the dose-dependent production of micronucleus (MN) in human lymphocytes irradiated with {sub 60} Co {gamma} -rays and 50MeV neutron, and to evaluate predictive markers of intrinsic radiosensitivity in individuals for monitoring occupational or environmental radiation exposure. For the dose-response study, heparinized whole blood of 10 healthy volunteers was irradiated with {sub 60} Co {gamma} -rays employing of 0.25-8Gy. The MNs were observed all doses, and the numerical changes according to doses. In dose-response curves fit linear- quadratic form (alpha =0.31{+-}0.049, beta =0.0022{+-}0.0022) for {gamma} -rays, but (alpha =0.99{+-}0.528, beta =0.0093{+-}0.0047) for neutron. Neutrons were than {gamma} -rays effective in producing MN with dose-dependent manner. The frequency of MN varies with dose. The RBE for micronuclei was 2.37{+-}0.17. Further studied are carried out to provide evidence for the existence of individual variations in age-dependent responses to radiation. Spontaneous and radiation-induced MN varies greatly between individuals, and little is known about the molecular mechanisms of this variability. It was shown that the increased level of spontaneous cell with MN was observed with increasing age. The relationship between radiosensitivity and the increased spontaneous level of MN may be in inverse proportion. These studies indicates that the MN assay have a high potential as a rapid, sensitive and accurate method which can be used to monitor a large population exposed to radiation for rapid triage in the case of a large-scale accident.

  12. Micronucleus induction and reproductive death in a human cell line exposed to low-energy argon beam

    International Nuclear Information System (INIS)

    Courdi, A.; Mari, D.; Herault, J.; Chauvel, P.

    1995-01-01

    The aim of this study was to measure the biological efficiency of a low-energy argon beam (E=7.1 MeV/nucleon, LET=1590 keV/μm) on a human melanoma cell line (CAL4) established in our Institute. Two different methods were used: the micronucleus (MN) test and the colony-forming assay. MN are scored in binucleate cells (BNC) and are formed from acentric fragments or whole chromosomes that have not been incorporated into daughter nuclei at mitosis. The colony-forming assay quantifies reproductive death. Parallel experiments were run with cobalt gamma-rays for comparison. After Co irradiation, the MN-free BNC dose-response curve coincided with that of the loss of colony-forming ability, suggesting the potential of the former as a predictive test of cell killing. After Ar irradiation, there was a dissociation between the two effects, especially at high doses: cell death was greater than the frequency of BNC with MN. The inactivation cross-section was 74 μm 2 ; it was 39 μm 2 for MN yield. Therefore, the relative biological effectiveness (RBE) was higher for cell killing than for MN yield (0.8 and 0.5, respectively, at a Co dose of 3 Gy). The total MN count in BNC followed the same pattern of response as the fraction of BNC with MN. However, multiple (>2) MN in BNC were more frequently observed after low-dose Ar irradiation than after gamma-ray exposure (RBE > 1). Moreover, the frequency of multiple MN induction exceeded that expected from a Poisson distribution at all dose levels of Ar irradiation. (orig.)

  13. Genotoxicity of Heterocyclic PAHs in the Micronucleus Assay with the Fish Liver Cell Line RTL-W1

    Science.gov (United States)

    Brinkmann, Markus; Blenkle, Henning; Salowsky, Helena; Bluhm, Kerstin; Schiwy, Sabrina; Tiehm, Andreas; Hollert, Henner

    2014-01-01

    Heterocyclic aromatic hydrocarbons are, together with their un-substituted analogues, widely distributed throughout all environmental compartments. While fate and effects of homocyclic PAHs are well-understood, there are still data gaps concerning the ecotoxicology of heterocyclic PAHs: Only few publications are available investigating these substances using in vitro bioassays. Here, we present a study focusing on the identification and quantification of clastogenic and aneugenic effects in the micronucleus assay with the fish liver cell line RTL-W1 that was originally derived from rainbow trout (Oncorhynchus mykiss). Real concentrations of the test items after incubation without cells were determined to assess chemical losses due to, e.g., sorption or volatilization, by means of gas chromatography-mass spectrometry. We were able to show genotoxic effects for six compounds that have not been reported in vertebrate systems before. Out of the tested substances, 2,3-dimethylbenzofuran, benzothiophene, quinoline and 6-methylquinoline did not cause substantial induction of micronuclei in the cell line. Acridine caused the highest absolute induction. Carbazole, acridine and dibenzothiophene were the most potent substances compared with 4-nitroquinoline oxide, a well characterized genotoxicant with high potency used as standard. Dibenzofuran was positive in our investigation and tested negative before in a mammalian system. Chemical losses during incubation ranged from 29.3% (acridine) to 91.7% (benzofuran) and may be a confounding factor in studies without chemical analyses, leading to an underestimation of the real potency. The relative potency of the investigated substances was high compared with their un-substituted PAH analogues, only the latter being typically monitored as priority or indicator pollutants. Hetero-PAHs are widely distributed in the environment and even more mobile, e.g. in ground water, than homocyclic PAHs due to the higher water solubility. We

  14. Correlation of In Vivo Versus In Vitro Benchmark Doses (BMDs) Derived From Micronucleus Test Data: A Proof of Concept Study.

    Science.gov (United States)

    Soeteman-Hernández, Lya G; Fellows, Mick D; Johnson, George E; Slob, Wout

    2015-12-01

    In this study, we explored the applicability of using in vitro micronucleus (MN) data from human lymphoblastoid TK6 cells to derive in vivo genotoxicity potency information. Nineteen chemicals covering a broad spectrum of genotoxic modes of action were tested in an in vitro MN test using TK6 cells using the same study protocol. Several of these chemicals were considered to need metabolic activation, and these were administered in the presence of S9. The Benchmark dose (BMD) approach was applied using the dose-response modeling program PROAST to estimate the genotoxic potency from the in vitro data. The resulting in vitro BMDs were compared with previously derived BMDs from in vivo MN and carcinogenicity studies. A proportional correlation was observed between the BMDs from the in vitro MN and the BMDs from the in vivo MN assays. Further, a clear correlation was found between the BMDs from in vitro MN and the associated BMDs for malignant tumors. Although these results are based on only 19 compounds, they show that genotoxicity potencies estimated from in vitro tests may result in useful information regarding in vivo genotoxic potency, as well as expected cancer potency. Extension of the number of compounds and further investigation of metabolic activation (S9) and of other toxicokinetic factors would be needed to validate our initial conclusions. However, this initial work suggests that this approach could be used for in vitro to in vivo extrapolations which would support the reduction of animals used in research (3Rs: replacement, reduction, and refinement). © The Author 2015. Published by Oxford University Press on behalf of the Society of Toxicology.

  15. In situ monitoring of urban air in Córdoba, Argentina using the Tradescantia-micronucleus (Trad-MCN) bioassay

    Science.gov (United States)

    Carreras, H. A.; Pignata, M. L.; Saldiva, P. H. N.

    During the last decades, a significant deterioration of ambient air quality has been observed in Argentina. However, the availability of air pollution monitoring stations is still limited to only few cities. In this study, we investigated the genotoxicity of ambient levels of air pollution in Córdoba using the Tradescantia micronucleus assay. The experiment was performed from October, 2004 to April 2005. Pots with Tradescantia pallida were placed in three sites: Córdoba city center, characterized by important avenues with high traffic activity (cars, taxis, and public transport vehicles); the university campus, along a side road with heavy traffic of gasoline and diesel powered vehicles, buses and trucks; and a residential area, with no significant local sources of air pollution. Twenty young T. pallida inflorescences were collected from each sampling site in November, February and April. Micronuclei frequencies were determined in early tetrads of pollen mother cells and expressed as MCN/100 tetrads. Simultaneously, the environmental levels of total suspended particles (24 h mean) were determined for each site. A significant difference in micronuclei frequency was observed among sites ( p=0.036). Post-hoc analysis revealed that the residential area exhibited a lower micronuclei frequency than the university and city center areas. In conclusion, we found that the gradients of ambient air pollution of Córdoba are associated with changes in the spontaneous micronuclei frequency of Tradescantia pollen mother cells. These results indicate that in situ biomonitoring with higher plants may be useful for characterizing air pollution in areas without instrumental monitoring techniques, or for exploring the distribution of air contaminants at a microscale.

  16. Flow cytometry based micronucleus assay for evaluation of genotoxic potential of 2-ACBs in hepatic cells HepG2

    Energy Technology Data Exchange (ETDEWEB)

    Barbezan, Angélica B.; Santos, Carla J.B.; Carvalho, Luma R.; Vieira, Daniel P.; Villavicêncio, Anna L.C.H., E-mail: abarbezan@ipen.br [Instituto de Pesquisas Energéticas e Nucleares (IPEN/CNEN-SP), São Paulo, SP (Brazil); Santelli, Glaucia M.M. [Universidade de São Paulo (USP), SP (Brazil). Departamento de Biologia Celular e do Desenvolvimento

    2017-07-01

    Food irradiation is approved for use in more than 60 countries for applications and purposes in a wide variety of foods, being an effective and safe method for preservation and long-term storage. 2-Alkylcyclobutanones (2-ACBs) are the only known radiolytic products generated from foods that contain fatty acids (Triglycerides) when irradiated. The acids analyzed in this study are palmitic and stearic, which when irradiated form 2-Dodecylcyclobutanones (2-dDCB) and 2-Tetradecylcyclobutanone (2-tDCB). Part of the 2-ACBs ingested is excreted through feces and part is deposited in adipose tissues. In vitro studies so far have been only in colon cells. The work used a human hepatoma cell line (HepG2) since the accumulation of fat in this organ is quite common. Micronucleus test was selected to evaluate possible genotoxic effects of 2-dDCB and 2-tDCB compounds when exposed to high concentrations (447, 1422 and 2235 μM) for 4 and 24 hours. Tests were performed in quadriplicates using flow cytometric analysis. None detectable genotoxic damage was observed after 4 hours of exposure to the compounds, and cytotoxic effects were only significant at the highest concentration (2235 μM) of 2-dDCB. After 24 hours of exposure, slight genotoxic damage was observed at all concentrations evaluated, and cytotoxic effects were only present when exposed to compound 2-tDCB. Although there is a genotoxic and cytotoxic effect in some of the situations tested, the two compounds predominantly induced proliferation reduction effects of this hepatic tumor cell line. (author)

  17. Genotoxicity of heterocyclic PAHs in the micronucleus assay with the fish liver cell line RTL-W1.

    Directory of Open Access Journals (Sweden)

    Markus Brinkmann

    Full Text Available Heterocyclic aromatic hydrocarbons are, together with their un-substituted analogues, widely distributed throughout all environmental compartments. While fate and effects of homocyclic PAHs are well-understood, there are still data gaps concerning the ecotoxicology of heterocyclic PAHs: Only few publications are available investigating these substances using in vitro bioassays. Here, we present a study focusing on the identification and quantification of clastogenic and aneugenic effects in the micronucleus assay with the fish liver cell line RTL-W1 that was originally derived from rainbow trout (Oncorhynchus mykiss. Real concentrations of the test items after incubation without cells were determined to assess chemical losses due to, e.g., sorption or volatilization, by means of gas chromatography-mass spectrometry. We were able to show genotoxic effects for six compounds that have not been reported in vertebrate systems before. Out of the tested substances, 2,3-dimethylbenzofuran, benzothiophene, quinoline and 6-methylquinoline did not cause substantial induction of micronuclei in the cell line. Acridine caused the highest absolute induction. Carbazole, acridine and dibenzothiophene were the most potent substances compared with 4-nitroquinoline oxide, a well characterized genotoxicant with high potency used as standard. Dibenzofuran was positive in our investigation and tested negative before in a mammalian system. Chemical losses during incubation ranged from 29.3% (acridine to 91.7% (benzofuran and may be a confounding factor in studies without chemical analyses, leading to an underestimation of the real potency. The relative potency of the investigated substances was high compared with their un-substituted PAH analogues, only the latter being typically monitored as priority or indicator pollutants. Hetero-PAHs are widely distributed in the environment and even more mobile, e.g. in ground water, than homocyclic PAHs due to the higher water

  18. Evaluation of a liver micronucleus assay in young rats (IV): a study using a double-dosing/single-sampling method by the Collaborative Study Group for the Micronucleus Test (CSGMT)/Japanese Environmental Mutagen Society (JEMS)-Mammalian Mutagenicity Study Group (MMS).

    Science.gov (United States)

    Takasawa, Hironao; Suzuki, Hiroshi; Ogawa, Izumi; Shimada, Yasushi; Kobayashi, Kazuo; Terashima, Yukari; Matsumoto, Hirotaka; Oshida, Keiyu; Ohta, Ryo; Imamura, Tadashi; Miyazaki, Atsushi; Kawabata, Masayoshi; Minowa, Shigenori; Maeda, Akihisa; Hayashi, Makoto

    2010-04-30

    A collaborative study was conducted to evaluate whether a liver micronucleus assay using four-week-old male F344 rats can be used to detect genotoxic rat hepatocarcinogens using double-dosing with a single-sampling 4 days after the second dose. The assay methods were thoroughly validated by the seven laboratories involved in the study. Seven chemicals, 2,4-diaminotoluene, diethyl nitrosamine, p-dimethylaminoazobenzene, 1,2-dimethylhydrazine dihydrochloride, 2,4-dinitrotolunene, 2,6-dinitrotoluene and mitomycin C, known to produce positive responses in the single-dosing/triple-sampling method were selected for use in the present study, and each chemical was examined in two laboratories with the exception of 2,4-dinitrotolunene. Although several of the compounds were examined at lower doses for reasons of toxicity than in the single-dosing/triple-sampling method, all chemicals tested in the present study induced micronuclei in liver cells indicating a positive result. These findings suggest that the liver micronucleus assay can be used in young rats to detect genotoxic rat hepatocarcinogens using a double-dosing/single-sampling procedure. Further, the number of animals used in the liver micronucleus assay can be reduced by one-third to a half by using the double-dosing/single-sampling method. This reduction in animal numbers also has significant savings in time and resource for liver perfusion and hepatocyte isolation. Copyright 2010 Elsevier B.V. All rights reserved.

  19. Effect of smoking habit on the frequency of micronuclei in human lymphocytes: results from the human micronucleus project

    International Nuclear Information System (INIS)

    Bonassi, Stefano; Neri, M.; Lando, Cecilia

    2004-01-01

    Full text: The effect of tobacco smoking on the frequency of micronuclei (MN) in human lymphocytes has been the object of many population studies. In most reports, the results were unexpectedly negative, and in many instances smokers had lower frequencies of MN than non-smokers. A pooled re-analysis of 24 databases from the HUMN international collaborative project has been performed with the aim of understanding the impact of smoking habits on MN frequency. The complete database included 5710 subjects, with 3501 non-smokers, 1409 current smokers, and 800 former smokers, among subjects in occupational and environmental surveys. The overall result of the re-analysis confirmed the small decrease of MN frequencies in current smokers (frequency ratio (FR =0.97, 95% confidence interval (CI =0.93-1.01) and in former smokers (FR =0.96, 95% CI =0.91-1.01), when compared to non-smokers. MN frequency was not influenced by the number of cigarettes smoked per day among subjects occupationally exposed to genotoxic agents, whereas a typical U-shaped curve is observed for non-exposed smokers, showing a significant increase of MN frequency in individuals smoking 30 cigarettes or more per day (FR =1.59, 95% CI =1.35-1.88). This analysis confirmed that smokers do not experience an overall increase in MN frequency, although when the interaction with occupational exposure is taken into account, heavy smokers were the only group showing a significant increase in genotoxic damage as measured by the micronucleus assay in lymphocytes. From these results some general recommendations for the design of bio monitoring studies involving smokers can be formulated. Quantitative data about smoking habit should always be collected because, in the absence of such data, the simple comparison of smokers versus non-smokers could be misleading. The sub-group of heavy smokers (≥30 cigarettes per day) should be specifically evaluated whenever it is large enough to satisfy statistical requirements. The

  20. Application of the micronucleus test and comet assay in Trachemys callirostris erythrocytes as a model for in situ genotoxic monitoring.

    Science.gov (United States)

    Zapata, Lina M; Bock, Brian C; Orozco, Luz Yaneth; Palacio, Jaime A

    2016-05-01

    Trachemys callirostris is a turtle species endemic to northern South America. In northern Colombia it occurs in the middle and lower Magdalena River drainage and its principal tributaries (lower Cauca and San Jorge rivers) and in other minor drainages such as the lower Sinú River. In recent years, industrial, agricultural, and mining activities have altered natural habitats in Colombia where this species occurs, and many of the pollutants released there are known to induce genetic alterations in wildlife species. The micronucleus test and comet assay are two of the most widely used methods to characterize DNA damage induced by physical and chemical agents in wildlife species, but have not been employed previously for genotoxic evaluations in T. callirostris. The goal of this study was to optimize these genotoxic biomarkers for T. callirostris erythrocytes in order to establish levels of DNA damage in this species and thereby evaluate its potential as a sentinel species for monitoring genotoxic effects in freshwater environments in northern Colombia. Both genotoxic techniques were applied on peripheral blood erythrocytes from 20 captive-reared T. callirostris individuals as a negative control, as well as from samples obtained from 49 individuals collected in Magangué (Magdalena River drainage) and 24 individuals collected in Lorica (Sinú River drainage) in northern Colombia. Negative control individuals exhibited a baseline frequency of micronuclei of 0.78±0.58 and baseline values for comet tail length and tail moment of 3.34±0.24µm and 10.70±5.5, respectively. In contrast, samples from both field sites exhibited significantly greater evidence of genotoxic effects for both tests. The mean MN frequencies in the samples from Magangué and Lorica were 8.04±7.08 and 12.19±12.94, respectively. The mean tail length for samples from Magangué and Lorica were 5.78±3.18 and 15.46±7.39, respectively. Finally, the mean tail moment for samples from Magangué and

  1. Effects of β-glucan polysaccharide revealed by the dominant lethal assay and micronucleus assays, and reproductive performance of male mice exposed to cyclophosphamide

    Directory of Open Access Journals (Sweden)

    Rodrigo Juliano Oliveira

    2014-01-01

    Full Text Available β-glucan is a well-known polysaccharide for its chemopreventive effect. This study aimed to evaluate the chemopreventive ability of β-glucan in somatic and germ cells through the dominant lethal and micronucleus assays, and its influence on the reproductive performance of male mice exposed to cyclophosphamide. The results indicate that β-glucan is capable of preventing changes in DNA in both germ cells and somatic ones. Changes in germ cells were evaluated by the dominant lethal assay and showed damage reduction percentages of 46.46% and 43.79% for the doses of 100 and 150 mg/kg. For the somatic changes, evaluated by micronucleus assay in peripheral blood cells in the first week of treatment, damage reduction percentages from 80.63-116.32% were found. In the fifth and sixth weeks, the percentage ranged from 10.20-52.54% and -0.95-62.35%, respectively. Besides the chemopreventive efficiency it appears that the β-glucan, when combined with cyclophosphamide, is able to improve the reproductive performance of males verified by the significant reduction in rates of post-implantation losses and reabsorption in the mating of nulliparous females with males treated with cyclophosphamide.

  2. Evaluation of repeated dose micronucleus assays of the liver using N-nitrosopyrrolidine: a report of the collaborative study by CSGMT/JEMS.MMS.

    Science.gov (United States)

    Ogawa, Izumi; Hagioa, Soichiro; Furukawa, Satoshi; Abe, Masayoshi; Kuroda, Yusuke; Hayashi, Seigo; Wako, Yumi; Kawasako, Kazufumi

    2015-03-01

    The repeated dose liver micronucleus (RDLMN) assay has the potential to detect liver carcinogens, and can be integrated into a general toxicological study. To assess the performance of the assay, N-nitrosopyrrolidine (NPYR), a genotoxic hepatocarcinogen, was tested in 14- or 28-day RDLMN assays. NPYR was orally administered to rats at a daily dose of 25, 50 or 100 mg/kg. One day after the last administration, a portion of the liver was removed and hepatocyte micronucleus (MN) specimens were prepared by the new method recently established by Narumi et al. In addition, a bone marrow MN assay and a histopathological examination of the liver were conducted. The detection of Phospho-Histone H3 was performed by immunohistochemistry to evaluate the proliferation rate of hepatocytes. The results showed significant increase in the number of micronucleated hepatocytes and Phospho-Histone H3-positive cells from the lowest dose in both 14- and 28-day RDLMN assays. On the other hand, the bone marrow MN assay yielded a negative result, which was in accordance with the existing report of the bone marrow MN assay using mice. Upon histopathological examination, inflammatory lesions and hypertrophy were noted, which may explain the increase in the hepatocyte proliferation and the enhancement of MN induction by NPYR. Our findings indicate that the RDLMN assay could be a useful tool for comprehensive risk assessment of carcinogenicity by providing information on both genotoxicity and histopathology when integrated into a general repeat dosing toxicity assay.

  3. Lack of genotoxic effect of food dyes amaranth, sunset yellow and tartrazine and their metabolites in the gut micronucleus assay in mice.

    Science.gov (United States)

    Poul, Martine; Jarry, Gérard; Elhkim, Mostafa Ould; Poul, Jean-Michel

    2009-02-01

    The food dyes amaranth, sunset yellow and tartrazine were administered twice, at 24h intervals, by oral gavage to mice and assessed in the in vivo gut micronucleus test for genotoxic effects (frequency of micronucleated cells) and toxicity (apoptotic and mitotic cells). The concentrations of each compound and their main metabolites (sulfanilic acid and naphthionic acid) were measured in faeces during a 24-h period after single oral administrations of the food dyes to mice. Parent dye compounds and their main aromatic amine metabolites were detected in significant amounts in the environment of colonic cells. Acute oral exposure to food dye additives amaranth, sunset yellow and tartrazine did not induce genotoxic effect in the micronucleus gut assay in mice at doses up to 2000 mg/kg b.w. Food dyes administration increased the mitotic cells at all dose levels when compared to controls. These results suggest that the transient DNA damages previously observed in the colon of mice treated by amaranth and tartrazine by the in vivo comet assay [Sasaki, Y.F., Kawaguchi, S., Kamaya, A., Ohshita, M., Kabasawa, K., Iwama, K., Taniguchi, K., Tsuda, S., 2002. The comet assay with 8 mouse organs: results with 39 currently used food additives. Mutat. Res. 519, 103-119] are unable to be fixed in stable genotoxic lesions and might be partly explained by local cytotoxicity of the dyes.

  4. A follow-up survey of the lymphocyte micronucleus for three victims seventheen years after Shanghai '6.25' 60Co radiation accident

    International Nuclear Information System (INIS)

    Du Jie; Chen Ying; Zhang Xueqing; Liu Xiulin; Yan Xuekun

    2008-01-01

    Objective: To study the long-term effect on lymphocyte micronucleus and its clinical significance for the three victims seventheen years after Shanghai '6.25' 60 Co radiation accident. Method: The MN and MNC frequency were observed by CBMN method. Results: The MN and MNC frequency were still higher than normal even 17 years after the expo- sure. The MN frequency is related with the former radiation dose. The MNC consisting of ≥2MN is the hightest for 'Long', but for 'Jun' it consisted of 1 MN. The MN frequency is lower 17 years after the exposure than that of 5 years, but higher than 10 years. Conclusions: The MN is one of important signs for long-term radiation effect. Special attention should be payed to the high level of MN remaining in the body for so long time. (authors)

  5. Protection of nucleated bone marrow cells of mice against effect of radiation-induced micronucleus formation by using polysaccharides extracted from 'Zi Zhi' (a ganoderma)

    International Nuclear Information System (INIS)

    Chu Fang; Luo Houliang; Luo Gui; Chen Shunle; Liu Zhifang

    1988-01-01

    This paper describes the influence of polysaccharides extracted from 'Zi Zhi' (Ganoderma Sinese Zhao, Xu et Zhang) on the frequency of micronucleated cells induced by 60 Co gamma irradiation at different doses in bone marrow of mice. These polysaccharides of 'Zi Zhi' were shown to be of ability to protect nucleated bone marrow cells from micronucleus formation in irradiated mice. For Swiss mice, a dose reduction factor (DRF) was found to be 1.72 in the range of 0 to 4.728 Gy and for LACA mice, to be 1.73 in the range of 0 to 3.152 Gy. Such findings indicate that these polysaccharides are comparable to L-cysteine in their effeciency of protection

  6. Development of a data-processing method based on Bayesian k-means clustering to discriminate aneugens and clastogens in a high-content micronucleus assay.

    Science.gov (United States)

    Huang, Z H; Li, N; Rao, K F; Liu, C T; Huang, Y; Ma, M; Wang, Z J

    2018-03-01

    Genotoxicants can be identified as aneugens and clastogens through a micronucleus (MN) assay. The current high-content screening-based MN assays usually discriminate an aneugen from a clastogen based on only one parameter, such as the MN size, intensity, or morphology, which yields low accuracies (70-84%) because each of these parameters may contribute to the results. Therefore, the development of an algorithm that can synthesize high-dimensionality data to attain comparative results is important. To improve the automation and accuracy of detection using the current parameter-based mode of action (MoA), the MN MoA signatures of 20 chemicals were systematically recruited in this study to develop an algorithm. The results of the algorithm showed very good agreement (93.58%) between the prediction and reality, indicating that the proposed algorithm is a validated analytical platform for the rapid and objective acquisition of genotoxic MoA messages.

  7. Biomonitoring air quality during and after a public transportation strike in the center of Uberlândia, Minas Gerais, Brazil by Tradescantia micronucleus bioassay.

    Science.gov (United States)

    Pereira, Boscolli Barbosa; de Campos, Edimar Olegário; de Lima, Euclides Antônio Pereira; Barrozo, Marcos Antonio Souza; Morelli, Sandra

    2014-03-01

    The aim of this study was to address the lack of information concerning the air quality in the city of Uberlândia, Minas Gerais, Brazil. In this study, we conducted an unprecedented experiment involving the in situ biomonitoring of air genotoxicity in the city center during and after a public transportation strike using the Tradescantia micronucleus test. The frequency of micronuclei was significantly higher in the city center compared with the reference site (Mann-Whitney test, p transport stoppage (Kruskal-Wallis, Dunn p transport stoppage and the increase in the concentration of particulate matter from the increased flow of vehicles in the city center during the strike positively influenced the MN frequency. The climatic factors did not change during the biomonitoring period, reflecting the fact that climatic factors did not influence the MN frequency.

  8. Alkaline comet assay in liver and stomach, and micronucleus assay in bone marrow, from rats treated with 2-acetylaminofluorene, azidothymidine, cisplatin, or isobutyraldehyde.

    Science.gov (United States)

    Kraynak, A R; Barnum, J E; Cunningham, C L; Ng, A; Ykoruk, B A; Bennet, B; Stoffregen, D; Merschman, M; Freeland, E; Galloway, S M

    2015-07-01

    As part of the Japanese Center for the Validation of Alternative Methods (JaCVAM) initiative international validation study of the in vivo rat alkaline comet assay (comet assay), we examined the ability of the assay to determine the genotoxicity of 2-acetylaminofluorene (AAF), azidothymidine (AZT), cisplatin (CPN), and isobutyraldehyde (IBA) in liver and glandular stomach of male Sprague-Dawley rats. Rats were given oral doses of test compound or control once daily for three days. High dose levels were approximately maximum tolerated doses and were based on preliminary range-finding studies. Tissues were harvested 3h after the final dose (48h after the initial dose). A bone marrow micronucleus assay (MN) was also conducted on the rats treated with AZT, CPN, and IBA. Acute toxic effects of treatment were determined primarily through histomorphologic analysis of liver and stomach but also by body weight and serum liver enzyme changes. The comet assay was conducted on fresh tissue preparations but frozen samples from two studies were also assayed. Statistically significant dose-related differences in comet % DNA in tail were found in liver and stomach for the genotoxin AZT and in liver for the genotoxin CPN, but not in liver or stomach for the non-genotoxin IBA. Statistically significant differences in % DNA in tail were measured in liver for the low and mid dose of the genotoxin AAF, but not the high dose. The comet assays of frozen liver suspensions from CPN- and AAF-treated rats yielded comparable results to the assays of fresh preparations. There were no indications of significant toxicity induced by any treatment. The micronucleus assay was positive for CPN and AZT and negative for IBA. In conclusion, the in vivo comet assay is capable of detecting genotoxic effects of a variety of chemicals and may fill an important role in the genotoxicity test battery. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Genotoxic and Cytotoxic Effects in Exfoliated Buccal and Nasal Cells of Chromium and Cobalt Exposed Electroplaters

    Czech Academy of Sciences Publication Activity Database

    Wultsch, G.; Nersesyan, A.; Kundi, M.; Mišík, M.; Setayesh, T.; Waldherr, M.; Vodička, Pavel; Vodičková, Ludmila; Knasmüller, S.

    2017-01-01

    Roč. 80, 13-15 (2017), s. 651-660 ISSN 1528-7394 Institutional support: RVO:68378041 Keywords : micronucleus cytome assay * hexavalent chromium * nuclear anomalies Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Biochemistry and molecular biology Impact factor: 2.731, year: 2016

  10. Evaluation of the repeated-dose liver and gastrointestinal tract micronucleus assays with 22 chemicals using young adult rats: summary of the collaborative study by the Collaborative Study Group for the Micronucleus Test (CSGMT)/The Japanese Environmental Mutagen Society (JEMS) - Mammalian Mutagenicity Study Group (MMS).

    Science.gov (United States)

    Hamada, Shuichi; Ohyama, Wakako; Takashima, Rie; Shimada, Keisuke; Matsumoto, Kazumi; Kawakami, Satoru; Uno, Fuyumi; Sui, Hajime; Shimada, Yasushi; Imamura, Tadashi; Matsumura, Shoji; Sanada, Hisakazu; Inoue, Kenji; Muto, Shigeharu; Ogawa, Izumi; Hayashi, Aya; Takayanagi, Tomomi; Ogiwara, Yosuke; Maeda, Akihisa; Okada, Emiko; Terashima, Yukari; Takasawa, Hironao; Narumi, Kazunori; Wako, Yumi; Kawasako, Kazufumi; Sano, Masaki; Ohashi, Nobuyuki; Morita, Takeshi; Kojima, Hajime; Honma, Masamitsu; Hayashi, Makoto

    2015-03-01

    The repeated-dose liver micronucleus (RDLMN) assay using young adult rats has the potential to detect hepatocarcinogens. We conducted a collaborative study to assess the performance of this assay and to evaluate the possibility of integrating it into general toxicological studies. Twenty-four testing laboratories belonging to the Mammalian Mutagenicity Study Group, a subgroup of the Japanese Environmental Mutagen Society, participated in this trial. Twenty-two model chemicals, including some hepatocarcinogens, were tested in 14- and/or 28-day RDLMN assays. As a result, 14 out of the 16 hepatocarcinogens were positive, including 9 genotoxic hepatocarcinogens, which were reported negative in the bone marrow/peripheral blood micronucleus (MN) assay by a single treatment. These outcomes show the high sensitivity of the RDLMN assay to hepatocarcinogens. Regarding the specificity, 4 out of the 6 non-liver targeted genotoxic carcinogens gave negative responses. This shows the high organ specificity of the RDLMN assay. In addition to the RDLMN assay, we simultaneously conducted gastrointestinal tract MN assays using 6 of the above carcinogens as an optional trial of the collaborative study. The MN assay using the glandular stomach, which is the first contact site of the test chemical when administered by oral gavage, was able to detect chromosomal aberrations with 3 test chemicals including a stomach-targeted carcinogen. The treatment regime was the 14- and/or 28-day repeated-dose, and the regime is sufficiently promising to incorporate these methods into repeated-dose toxicological studies. The outcomes of our collaborative study indicated that the new techniques to detect chromosomal aberrations in vivo in several tissues worked successfully. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  11. Evaluation of the repeated-dose liver micronucleus assay using N-nitrosomorpholine in young adult rats: report on collaborative study by the Collaborative Study Group for the Micronucleus Test (CSGMT)/Japanese Environmental Mutagen Society (JEMS)-Mammalian Mutagenicity Study (MMS) Group.

    Science.gov (United States)

    Hayashi, Aya; Kosaka, Mizuki; Kimura, Aoi; Wako, Yumi; Kawasako, Kazufumi; Hamada, Shuichi

    2015-03-01

    The present study was conducted to evaluate the suitability of a repeated-dose liver micronucleus (LMN) assay in young adult rats as a collaborative study by the Mammalian mutagenicity study (MMS) group. All procedures were performed in accordance with the standard protocols of the MMS Group. Six-week-old male Crl:CD(SD) rats (5 animals/group) received oral doses of the hepatocarcinogen N-nitrosomorpholine (NMOR) at 0 (control), 5, 10, and 30mg/kg/day (10mL/kg) for 14 days. Control animals received vehicle (water). Hepatocytes were collected from the liver 24h after the last dose, and the number of micronucleated hepatocytes (MNHEPs) was determined by microscopy. The number of micronucleated immature erythrocytes (MNIMEs) in the femoral bone marrow was also determined. The liver was examined using histopathologic methods after formalin fixation. The results showed statistically significant and dose-dependent increases in the number of MNHEPs in the liver at doses of 10mg/kg and greater when compared with the vehicle control. However, no significant increase was noted in the number of MNIMEs in the bone marrow at doses of up to 30mg/kg. Histopathology of the liver revealed hypertrophy and single cell necrosis of hepatocytes at doses of 5mg/kg and above. These results showed that the induction of micronuclei by NMOR was detected by the repeated-dose LMN assay, but not by the repeated-dose bone marrow micronucleus assay. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Genotoxic and antigenotoxic properties of selenium compounds in the in vitro micronucleus assay with human whole blood lymphocytes and tk6 lymphoblastoid cells

    Directory of Open Access Journals (Sweden)

    Eduard Cemeli

    2006-01-01

    Full Text Available Selenium is known to possess both genotoxic and antigenotoxic properties. In the present study, we have evaluated the genotoxicity and antigenotoxicity of three selenium compounds (sodium selenate, sodium selenite and selenous acid by measuring in vitro micronucleus induction. Assays were conducted in whole blood lymphocytes and in the TK6 lymphoblastoid cell line, with and without co-treatment with potassium dichromate, a known genotoxic compound. In general, the compounds were more active in TK6 cells than they were in blood lymphocytes. Only 1 μM selenous acid increased the frequency of binucleated cells containing micronuclei (BNMN in blood lymphocytes, while all three selenium compounds increased BNMN in TK6 cells. In addition, combinations of selenous acid and potassium dichromate resulted in lower frequencies of BNMN than potassium dichromate alone in blood lymphocytes, while combinations of sodium selenate and potassium dichromate produced lower frequencies of BNMN than potassium dichromate alone in TK6 cells. The concentrations of selenium compounds that were used, in combination with the medium components and the biological physiology of the whole blood lymphocytes and TK6 cells, could have affected the redox potential of the compounds, switching the chemicals from a pro-oxidant to antioxidant status and vice-versa. The lower activities of the compounds in blood lymphocytes may be due to the protective effects of blood components. The results indicate that the genotoxic and antigenotoxic properties of selenium compounds are highly dependent upon the conditions under which they are evaluated.

  13. Enhanced micronucleus formation in the descendants of {gamma}-ray-irradiated tobacco cells: Evidence for radiation-induced genomic instability in plant cells

    Energy Technology Data Exchange (ETDEWEB)

    Yokota, Yuichiro, E-mail: yokota.yuichiro@jaea.go.jp [Life Science and Biotechnology Division, Quantum Beam Science Directorate, Japan Atomic Energy Agency, 1233 Watanuki-machi, Takasaki, Gunma 370-1292 (Japan); Funayama, Tomoo; Hase, Yoshihiro [Life Science and Biotechnology Division, Quantum Beam Science Directorate, Japan Atomic Energy Agency, 1233 Watanuki-machi, Takasaki, Gunma 370-1292 (Japan); Hamada, Nobuyuki [Radiation Safety Research Center, Nuclear Technology Research Laboratory, Central Research Institute of Electric Power Industry, 2-11-1 Iwado-kita, Komae, Tokyo 201-8511 (Japan); Kobayashi, Yasuhiko; Tanaka, Atsushi; Narumi, Issay [Life Science and Biotechnology Division, Quantum Beam Science Directorate, Japan Atomic Energy Agency, 1233 Watanuki-machi, Takasaki, Gunma 370-1292 (Japan)

    2010-09-10

    Ionizing radiation-induced genomic instability has been documented in various end points such as chromosomal aberrations and mutations, which arises in the descendants of irradiated mammalian or yeast cells many generations after the initial insult. This study aimed at addressing radiation-induced genomic instability in higher plant tobacco cells. We thus investigated micronucleus (MN) formation and cell proliferation in tobacco cells irradiated with {gamma}-rays and their descendants. In {gamma}-irradiated cells, cell cycle was arrested at G{sub 2}/M phase at around 24 h post-irradiation but released afterward. In contrast, MN frequency peaked at 48 h post-irradiation. Almost half of 40 Gy-irradiated cells had MN at 48 h post-irradiation, but proliferated as actively as sham-irradiated cells up to 120 h post-irradiation. Moreover, the descendants that have undergone at least 22 generations after irradiation still showed a two-fold MN frequency compared to sham-irradiated cells. This is the direct evidence for radiation-induced genomic instability in tobacco cells.

  14. Vinclozolin, a widely used fungizide, enhanced BaP-induced micronucleus formation in human derived hepatoma cells by increasing CYP1A1 expression.

    Science.gov (United States)

    Wu, Xin-Jiang; Lu, Wen-qing; Roos, Peter H; Mersch-Sundermann, Volker

    2005-10-15

    Vinclozolin, a widely used fungicide, can be identified as a residue in numerous vegetable and fruit samples. To get insight in its genetic toxicity, we investigated the genotoxic effect of vinclozolin in the human derived hepatoma cell line HepG2 using the micronucleus (MN) assay. Additionally, to evaluate the co- or anti-mutagenic potency of vinclozolin, we treated HepG2 cells with different concentrations of vinclozolin for 24 h. Subsequently, the cells were exposed to benzo[a]pyrene (BaP) for 1h. Exposure of HepG2 cells to 50-400 microM vinclozolin alone did not cause any induction of micronuclei. However, a pronounced co-mutagenic effect was observed. MN frequencies caused by BaP increased by 30.6%, 52.8% and 65.3% after pretreatment of the cell cultures with 50, 100 and 200 microM vinclozolin, respectively. The highest concentration (400 microM) of vinclozolin tested caused cytotoxicity. Therefore, micronuclei were not considered for that concentration. To clarify the mechanism of cogenotoxicity, we assayed cytochrome P450 1A1 (CYP1A1), which plays a pivotal role in activation of BaP. Cells exposed to vinclozolin led to significant increase of CYP1A1 expression in Western blot. The result suggested that induction of CYP1A1 by vinclozolin account for its enhancing effect on genotoxicity caused by BaP.

  15. Evaluation of repeated dose micronucleus assays of the liver and gastrointestinal tract using potassium bromate: a report of the collaborative study by CSGMT/JEMS.MMS.

    Science.gov (United States)

    Okada, Emiko; Fujiishi, Yohei; Narumi, Kazunori; Kado, Shoichi; Wako, Yumi; Kawasako, Kazufumi; Kaneko, Kimiyuki; Ohyama, Wakako

    2015-03-01

    The food additive potassium bromate (KBrO3) is known as a renal carcinogen and causes chromosomal aberrations in vitro without metabolic activation and in vivo in hematopoietic and renal cells. As a part of a collaborative study by the Mammalian Mutagenicity Study group, which is a subgroup of the Japanese Environmental Mutagen Society, we administered KBrO3 to rats orally for 4, 14, and 28 days and examined the micronucleated (MNed) cell frequency in the liver, glandular stomach, colon, and bone marrow to confirm whether the genotoxic carcinogen targeting other than liver and gastrointestinal (GI) tract was detected by the repeated dose liver and GI tract micronucleus (MN) assays. In our study, animals treated with KBrO3 showed some signs of toxicity in the kidney and/or stomach. KBrO3 did not increase the frequency of MNed cells in the liver and colon in any of the repeated dose studies. However, KBrO3 increased the frequency of MNed cells in the glandular stomach and bone marrow. Additionally, the MNed cell frequency in the glandular stomach was not significantly affected by the difference in the length of the administration period. These results suggest that performing the MN assay using the glandular stomach, which is the first tissue to contact agents after oral ingestion, is useful for evaluating the genotoxic potential of chemicals and that the glandular stomach MN assay could be integrated into general toxicity studies. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Evaluation of the repeated dose liver micronucleus assay using young adult rats with cyclophosphamide monohydrate: a report of a collaborative study by CSGMT/JEMS.MMS.

    Science.gov (United States)

    Matsumoto, Kazumi; Zaizen, Kazuyo; Miyamoto, Atsushi; Wako, Yumi; Kawasako, Kazufumi; Ishida, Hisao

    2015-03-01

    The repeated dose liver micronucleus (RDLMN) assay using young adult rats has the potential to detect liver carcinogens, and this assay could be integrated into general toxicological studies. In this study, in order to assess the performance of the assay, cyclophosphamide monohydrate (CP) was tested in a 14-day RDLMN assay. Based on the results of the 4-day repeated dose-finding study, 10 mg/kg/day of CP was selected as the highest dose and the lower doses were set at 5, 2.5, 1.25, and 0.625 mg/kg/day for the 14-day RDLMN assay. On the day after the completion of the dosing period, specimens of hepatocytes and bone marrow cells were prepared and the induction of micronuclei was assessed. No changes were observed in the incidences of micronucleated hepatocytes. Nevertheless, the incidences of micronucleated immature erythrocytes in the bone marrow were increased significantly at CP doses of 1.25 mg/kg/day or more. These findings are consistent with reports that CP induces tumors in various tissues but it does not induce liver tumors.

  17. Micronucleus assay for human peripheral blood lymphocytes as a biomarker of individual sensitivity to assessing radiation health risk in different population

    International Nuclear Information System (INIS)

    Kang, C.-M.; Jeon, H.-J.; Lee, Y.-S.; Lee, S.-J.; Jin, Y.-H.; Kim, Y.-H.; Kim, T.-W.; Cho, C.-K.

    2003-01-01

    Full text: Our studies were to evaluate micronucleus (MN) assay for human peripheral blood lymphocytes (HPBL) as a biomarker of individual sensitivity to assessing radiation health risk in different population in Korea. Further studied are carried out to provide evidence for the existence of individual variations in age-dependent responses. For the MN assay, HPBLs were irradiated with doses of 0, 1, 2, 4, 8Gy 60 Co γ-rays. Spontaneous frequencies not only vary greatly between individuals, but also working or living areas because of the groups with different lifestyle living in different ecological situation and the reaction to radiation exposure. It was shown that the increased level of spontaneous cell with MN was observed with increased age. The relationship between radiosensitivity and the increased spontaneous level of MN may be in inverse proportion. Age and gender are the most important demographic variables impact on MN index with MN frequencies in female being greater than those in male by a factor of depending on the age group. For both sexes, MN frequency was significantly and positively correlated with age. The main lifestyle factors influencing the MN index in subjects are significantly and positively correlated with smoking in measuring the spontaneous frequencies of micronuclei. The described results show that the genetic damaged rate like MN index in human populations is correlated significantly with age, sex and lifestyle factors. So far, it is evident that with regard to the application of MN assay all future studies to evaluate radiation health risks in different population have to take into account the influence of age, gender, and lifestyle. The results suggested that the MN assay have a high potential to ensure appropriate quality control and standard documentation protocol which can be used to monitor a large population exposed to radiation epidemiologically. We conclude that the determination of individual radiosensitivity with MN assay is

  18. Differences in micronucleus induction in peripheral blood reticulocytes of mice exposed to N-ethyl-N-nitrosourea at light and dark dosing times.

    Science.gov (United States)

    Itoh, Keiichi; Masumori, Shoji; Nakajima, Madoka; Hayashi, Makoto; Sakakibara, Hiroyuki; Shimoi, Kayoko

    2012-01-01

    Mammals, including human beings, have a circadian clock system to regulate behavioral and physiological processes. In this study, we investigated the effect of dosing time on micronucleus induction in the bone marrow by evaluating the frequencies of micronucleated peripheral reticulocytes (MNRETs) in mice exposed to N-ethyl-N-nitrosourea (ENU) to assess any difference in genotoxic sensitivity to chemicals between light and dark periods (inactive phase for rodents and active phase for rodents). Male C3H/He mice were treated intraperitoneally with ENU (12.5 or 25 mg/kg body weight) at zeitgeber time (ZT) 3 in the light period or ZT15 in the dark period, and then the time courses of the frequencies of the MNRETs were determined. The frequencies of the MNRETs induced by ENU increased time-dependently and peaked at 48 hr after treatment for ZT3 and ZT15, and were obviously higher in the ZT15 treatment group than the ZT3 treatment group. The MNRETs were measured at 48 hr after treatment with ENU (25 mg/kg body weight) at various dosing times (ZT0, 3, 6, 12, 15 and 18). The frequencies of the MNRETs in mice treated at ZT0, 15 and 18 were significantly higher than those in mice treated at ZT3, 6 and 12. These results suggest that genotoxic sensitivity to chemicals in mouse bone marrow is different between light and dark periods maybe due to different biological responses (detoxification, cell cycle, DNA repair, etc.) related to circadian rhythms.

  19. Duodenal crypt health following exposure to Cr(VI): Micronucleus scoring, γ-H2AX immunostaining, and synchrotron X-ray fluorescence microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, Chad M.; Wolf, Jeffrey C.; Elbekai, Reem H.; Paranjpe, Madhav G.; Seiter, Jennifer M.; Chappell, Mark A.; Tappero, Ryan V.; Suh, Mina; Proctor, Deborah M.; Bichteler, Anne; Haws, Laurie C.; Harris, Mark A.

    2015-08-01

    Lifetime exposure to high concentrations of hexavalent chromium [Cr(VI)] in drinking water results in intestinal damage and an increase in duodenal tumors in B6C3F1 mice. To assess whether these tumors could be the result of a direct mutagenic or genotoxic mode of action, we conducted a GLP-compliant 7-day drinking water study to assess crypt health along the entire length of the duodenum. Mice were exposed to water (vehicle control), 1.4, 21, or 180 ppm Cr(VI) via drinking water for 7 consecutive days. Crypt enterocytes in Swiss roll sections were scored as normal, mitotic, apoptotic, karyorrhectic, or as having micronuclei. A single oral gavage of 50 mg/kg cyclophosphamide served as a positive control for micronucleus induction. Exposure to 21 and 180 ppm Cr(VI) significantly increased the number of crypt enterocytes. Micronuclei and γ-H2AX immunostaining were not elevated in the crypts of Cr(VI)-treated mice. In contrast, treatment with cyclophosphamide significantly increased numbers of crypt micronuclei and qualitatively increased γ-H2AX immunostaining. Synchrotron-based X-ray fluorescence (XRF) microscopy revealed the presence of strong Cr fluorescence in duodenal villi, but negligible Cr fluorescence in the crypt compartment. Together, these data indicate that Cr(VI) does not adversely effect the crypt compartment where intestinal stem cells reside, and provide additional evidence that the mode of action for Cr(VI)-induced intestinal cancer in B6C3F1 mice involves chronic villous wounding resulting in compensatory crypt enterocyte hyperplasia.

  20. Use of the fluorescent micronucleus assay to detect the genotoxic effects of radiation and arsenic exposure in exfoliated human epithelial cells

    International Nuclear Information System (INIS)

    Moore, L.E.; Warner, M.L.; Smith, A.H.

    1996-01-01

    The exfoliated cell micronucleus (MN) assay using fluorescent in situ hybridization (FISH) with a centromeric probe is a rapid method for determining the mechanism of MN formation in epithelial tissues exposed to carcinogenic agents. Here, we describe the use of this assay to detect the presence or absence of centromeric DNA in MN induced in vivo by radiation therapy and chronic arsenic (As) ingestion. We examined the buccal cells of an individual receiving 6,500 rads of photon radiation to the head and neck. Exfoliated cells were collected before, during, and after treatment. After radiation exposure a 16.6-fold increase in buccal cell MN frequency was seen. All induced MN were centromere negative (MN-) resulting from chromosome breakage. This finding is consistent with the clastogenic action of radiation and confirmed the reliability of the method. Three weeks post-therapy, MN frequencies returned to baseline. The assay was used on 18 people chronically exposed to high levels of inorganic arsenic (In-As) in drinking water (average level, 1,312 μg As/L) and 18 matched controls (average level, 16 μg As/L). The combined increase in MN frequency was 1.8-fold (P = 0.001, Fisher's exact test). Frequencies of micronuclei containing acentric fragments (MN-) and those containing whole chromosomes (MN+) both increased, suggesting that arsenic may have both clastogenic and weak aneuploidogenic properties in vivo. After stratification on sex, the effect was stronger in male than in female bladder cells. In males the MN-frequency increased 2.06-fold (P =0.07) while the frequency of MN+ increased 1.86-fold (P = 0.08). In addition, the frequencies of MN and MN+ were positively associated with urinary arsenic and its metabolites. The association was stronger for micronuclei containing acentric fragments. By using FISH with centromeric probes, the mechanism of chemically induced genotoxicity can not be determined in epithelial tissues. 35 refs., 4 tabs

  1. Nongenotoxic effects and a reduction of the DXR-induced genotoxic effects of Helianthus annuus Linné (sunflower) seeds revealed by micronucleus assays in mouse bone marrow.

    Science.gov (United States)

    Boriollo, Marcelo Fabiano Gomes; Souza, Luiz Silva; Resende, Marielly Reis; Silva, Thaísla Andrielle da; Oliveira, Nelma de Mello Silva; Resck, Maria Cristina Costa; Dias, Carlos Tadeu dos Santos; Fiorini, João Evangelista

    2014-04-02

    This research evaluated the genotoxicity of oil and tincture of H. annuus L. seeds using the micronucleus assay in bone marrow of mice. The interaction between these preparations and the genotoxic effects of doxorubicin (DXR) was also analysed (antigenotoxicity test). Experimental groups were evaluated at 24-48 h post treatment with N-Nitroso-N-ethylurea (positive control - NEU), DXR (chemotherapeutic), NaCl (negative control), a sunflower tincture (THALS) and two sources of sunflower oils (POHALS and FOHALS). Antigenotoxic assays were carried out using the sunflower tincture and oils separately and in combination with NUE or DXR. For THALS, analysis of the MNPCEs showed no significant differences between treatment doses (250-2,000 mg.Kg-1) and NaCl. A significant reduction in MNPCE was observed when THALS (2,000 mg.Kg-1) was administered in combination with DXR (5 mg.Kg-1). For POHALS or FOHALS, analysis of the MNPCEs also showed no significant differences between treatment doses (250-2,000 mg.Kg-1) and NaCl. However, the combination DXR + POHALS (2,000 mg.Kg-1) or DXR + FOHALS (2,000 mg.Kg-1) not contributed to the MNPCEs reduction. This research suggests absence of genotoxicity of THALS, dose-, time- and sex-independent, and its combination with DXR can reduce the genotoxic effects of DXR. POHALS and FOHALS also showed absence of genotoxicity, but their association with DXR showed no antigenotoxic effects.

  2. Micronucleus test of varying amounts of potassium bromate (KBrO3) on the meristematic cells of Allium cepa var. aggregatum root tips

    International Nuclear Information System (INIS)

    Cajigal Romnick, M.; Somera, Leomerto A.

    1999-03-01

    Four hundred twenty onion bulbs of the multiplier variety Allium cepa var. aggregatum were used as test materials to assay the micronucleus induction capacity of potassium bromate doses of 0, 5, 10, 25, 50, 75, and 100 parts per million. Microscopic analyses were done using onion root tips prepared according to a modified technique of Medina (1994). These analyses were done on root tips taken from onions grown in KBrO 3 for three days and for five days. The study was conducted following a completely randomized design and the data were statistically analyzed using a non-parametric equivalent of the analysis of variance. A significant amount of micronucleated cells (MCN) were found among treated onions compared with the almost non-occurrence in the control groups (0 ppm). The Kruskal-Wallis H-test and the Wilcoxon two-samples tests revealed significant differences among treatment means and that a significant increase in the number of MCN occurs as the dose of KBr0 3 increased in both day experiments. Results from the higher doses of 50, 75, and 100 ppm were found to be significantly the same for the day 3 experiments while those of the day 5 higher doses are characterized by lack of clear cellular and nuclear outline such that scoring is difficult. Differences in MCN averages for the day 3 and 5 experiments appear to be insignificant. However, day 3 results show averages that are more significantly different from each other. These prove that the MCN can be used as an efficient and time-saving parameter for the allium test of chemicals with chromosome breaking capacities. (Author)

  3. No evidence of chromosome damage in children and adolescents with differentiated thyroid carcinoma after receiving {sup 131}I radiometabolic therapy, as evaluated by micronucleus assay and microarray analysis

    Energy Technology Data Exchange (ETDEWEB)

    Federico, Giovanni; Fiore, Lisa; Massart, Francesco; Saggese, Giuseppe [Azienda Ospedaliero-Universitaria Pisana, Department of Pediatrics, Unit of Pediatric Endocrinology and Diabetes, Pisa (Italy); Boni, Giuseppe; Lazzeri, Patrizia; Mariani, Giuliano [Azienda Ospedaliero-Universitaria Pisana, Unit of Nuclear Medicine, Pisa (Italy); Fabiani, Barbara; Verola, Carmela; Scarpato, Roberto [University of Pisa, Department of Biology, Unit of Genetics, Mutagenesis and Environmental Epidemiology, Pisa (Italy); Traino, Claudio [Azienda Ospedaliero-Universitaria Pisana, Health Physics Service, Pisa (Italy)

    2008-11-15

    As {sup 131}I therapy, used to achieve ablation of thyroid gland remnant, can cause chromosome damage in cultured peripheral lymphocytes especially, we investigated whether administration of radioiodine may induce early genome damage in peripheral T lymphocytes of adolescents with differentiated thyroid carcinoma (DTC). We studied 11 patients, aged 14.8 {+-} 3.1 years, who assumed {sup 131}I (range: 1.11-4.44 GBq) to ablate thyroid remnant. A blood sample for micronucleus assay and for evaluating expression of some genes involved in the DNA repair or the apoptosis pathways was obtained from each patient 1 h before (T{sub 0}) and 24 (T{sub 1}) and 48 h (T{sub 2}) post-radioiodine administration. Compared to T{sub 0}, we did not find any difference in the number of micronucleated cells at both T{sub 1} and T{sub 2} in any subject. Nine out of 11 patients had altered expression levels in a majority of the DNA repair and apoptosis genes at T{sub 1}, which decreased at T{sub 2}. We demonstrated for the first time that peripheral cells of DTC children and adolescents who received {sup 131}I at a mean dosage of 3.50 {+-} 0.37 GBq did not show chromosome damage within 48 h from the end of radiometabolic therapy. This may be due to a prompt activation of the cell machinery that maintains the integrity of the genome to prevent harmful double-strand breaks from progressing to chromosome mutations, either by repairing the lesions or by eliminating the most seriously damaged cells via apoptosis. (orig.)

  4. Micronucleus test of varying amounts of potassium bromate (KBrO{sub 3}) on the meristematic cells of Allium cepa var. aggregatum root tips

    Energy Technology Data Exchange (ETDEWEB)

    Cajigal Romnick, M; Somera, Leomerto A

    1999-03-01

    Four hundred twenty onion bulbs of the multiplier variety Allium cepa var. aggregatum were used as test materials to assay the micronucleus induction capacity of potassium bromate doses of 0, 5, 10, 25, 50, 75, and 100 parts per million. Microscopic analyses were done using onion root tips prepared according to a modified technique of Medina (1994). These analyses were done on root tips taken from onions grown in KBrO{sub 3} for three days and for five days. The study was conducted following a completely randomized design and the data were statistically analyzed using a non-parametric equivalent of the analysis of variance. A significant amount of micronucleated cells (MCN) were found among treated onions compared with the almost non-occurrence in the control groups (0 ppm). The Kruskal-Wallis H-test and the Wilcoxon two-samples tests revealed significant differences among treatment means and that a significant increase in the number of MCN occurs as the dose of KBr0{sub 3} increased in both day experiments. Results from the higher doses of 50, 75, and 100 ppm were found to be significantly the same for the day 3 experiments while those of the day 5 higher doses are characterized by lack of clear cellular and nuclear outline such that scoring is difficult. Differences in MCN averages for the day 3 and 5 experiments appear to be insignificant. However, day 3 results show averages that are more significantly different from each other. These prove that the MCN can be used as an efficient and time-saving parameter for the allium test of chemicals with chromosome breaking capacities. (Author)

  5. In vitro genotoxicity of neutral red after photo-activation and metabolic activation in the Ames test, the micronucleus test and the comet assay.

    Science.gov (United States)

    Guérard, Melanie; Zeller, Andreas; Singer, Thomas; Gocke, Elmar

    2012-07-04

    Neutral red (Nr) is relatively non-toxic and is widely used as indicator dye in many biological test systems. It absorbs visible light and is known to act as a photosensitizer, involving the generation of reactive oxygen species (type-I reaction) and singlet oxygen (type-II reaction). The mutagenicity of Nr was determined in the Ames test (with Salmonella typhimurium strains TA1535, TA97, TA98, TA98NR, TA100, and TA102) with and without metabolic activation, and with and without photo-activation on agar plates. Similarly to the situation following metabolic activation, photo-mutagenicity of Nr was seen with all Salmonella strains tested, albeit with different effects between these strains. To our knowledge, Nr is the only photo-mutagen showing such a broad action. Since the effects are also observed in strains not known to be responsive to ROS, this indicates that ROS production is not the sole mode of action that leads to photo-genotoxicity. The reactive species produced by irradiation are short-lived as pre-irradiation of an Nr solution did not produce mutagenic effects when added to the bacteria. In addition, mutagenicity in TA98 following irradiation was stronger than in the nitroreductase-deficient strain TA98NR, indicating that nitro derivatives that are transformed by bacterial nitroreductase to hydroxylamines appear to play a role in the photo-mutagenicity of Nr. Photo-genotoxicity of Nr was further investigated in the comet assay and micronucleus test in L5178Y cells. Concentration-dependent increases in primary DNA damage and in the frequency of micronuclei were observed after irradiation. Copyright © 2012 Elsevier B.V. All rights reserved.

  6. Role of Macronutrients and Micronutrients in DNA Damage: Results From a Food Frequency Questionnaire

    Directory of Open Access Journals (Sweden)

    Carina Ladeira

    2017-02-01

    Full Text Available The links between diet and genomic instability have been under investigation for several decades, and evidence suggests a significant causal or preventive role for various dietary factors. This study investigates the influence of macronutrients (calories, protein, and glucides and micronutrients, such as vitamins and minerals, as assessed by a food frequency questionnaire, on genotoxicity biomarkers measured by cytokinesis-blocked micronucleus assay and comet assay. The results found significant positive and negative correlations. Micronucleus frequency tends to increase with higher intake of caffeine, calcium, magnesium, zinc, and protein ( P  < .05, Spearman correlation. Calorie and omega-6 intakes are negatively correlated with DNA damage measured by the comet assay. These results are somewhat controversial because some of the correlations found are contrary to dominant views in the literature; however, we suggest that unraveling the association between diet and genetic instability requires a much better understanding of the modulating role of macronutrients and micronutrients.

  7. Dose dependency of the frequency of micronucleated binucleated clone cells and of division related median clone sizes difference. Pt. 2

    International Nuclear Information System (INIS)

    Hagemann, G,; Kreczik, A.; Treichel, M.

    1996-01-01

    Following irradiation of the progenitor cells the clone growth of CHO cells decreases as a result of cell losses. Lethally acting expressions of micronuclei are produced by heritable lethal mutations. The dependency of the frequency of micronucleated binucleated clone cells and of the median clone sizes difference on the radiation dose was measured and compared to non-irradiated controls. Using the cytokinesis-block-micronucleus-method binucleated cells with micronuclei were counted as ratio of all binucleated cells within a clone size distribution. This ratio (shortened: micronucleus yield) was determined for all clone size distributions, which had been exposed to different irradiation doses and incubation times. The micronucleus yields were compared to the corresponding median clone sizes differences. The micronucleus yield is linearly dependent on the dose and is independent of the incubation time. The same holds true for the division related median clone sizes difference, which as a result is also linearly dependent on the micronucleus yield. Due to the inevitably errors of the cell count of micronucleated binucleated cells, an automatic measurement of the median clone sizes differences is the preferred method for evaluation of cellular radiation sensitivity for heritable lethal mutations. This value should always be determined in addition, if clone survival fractions are used as predictive test because it allows for an estimation of the remission probability of surviving cells. (orig.) [de

  8. Response of thyroid follicular cells to gamma irradiation compared to proton irradiation. I. Initial characterization of DNA damage, micronucleus formation, apoptosis, cell survival, and cell cycle phase redistribution

    Science.gov (United States)

    Green, L. M.; Murray, D. K.; Bant, A. M.; Kazarians, G.; Moyers, M. F.; Nelson, G. A.; Tran, D. T.

    2001-01-01

    The RBE of protons has been assumed to be equivalent to that of photons. The objective of this study was to determine whether radiation-induced DNA and chromosome damage, apoptosis, cell killing and cell cycling in organized epithelial cells was influenced by radiation quality. Thyroid-stimulating hormone-dependent Fischer rat thyroid cells, established as follicles, were exposed to gamma rays or proton beams delivered acutely over a range of physical doses. Gamma-irradiated cells were able to repair DNA damage relatively rapidly so that by 1 h postirradiation they had approximately 20% fewer exposed 3' ends than their counterparts that had been irradiated with proton beams. The persistence of free ends of DNA in the samples irradiated with the proton beam implies that either more initial breaks or a quantitatively different type of damage had occurred. These results were further supported by an increased frequency of chromosomal damage as measured by the presence of micronuclei. Proton-beam irradiation induced micronuclei at a rate of 2.4% per gray, which at 12 Gy translated to 40% more micronuclei than in comparable gamma-irradiated cultures. The higher rate of micronucleus formation and the presence of larger micronuclei in proton-irradiated cells was further evidence that a qualitatively more severe class of damage had been induced than was induced by gamma rays. Differences in the type of damage produced were detected in the apoptosis assay, wherein a significant lag in the induction of apoptosis occurred after gamma irradiation that did not occur with protons. The more immediate expression of apoptotic cells in the cultures irradiated with the proton beam suggests that the damage inflicted was more severe. Alternatively, the cell cycle checkpoint mechanisms required for recovery from such damage might not have been invoked. Differences based on radiation quality were also evident in the alpha components of cell survival curves (0.05 Gy(-1) for gamma rays, 0

  9. Genotoxic Effects of Exposure to Gasoline Fumes on Petrol Pump Workers

    OpenAIRE

    Amrin Shaikh; Darshana Barot; Divya Chandel

    2018-01-01

    Background: Petrol pump workers are occupationally exposed to gasoline and its fumes consisting of several mutagenic chemicals. Objective: To evaluate the genotoxic effects of exposure to gasoline fumes on petrol pump workers. Methods: The study groups included 70 petrol pump workers (exposed group) and 70 healthy age-matched individuals with no known exposure (comparison group). Buccal micronucleus cytome assay (BMCyt) was performed to check the genotoxicity caused due to inhalation ...

  10. Comet assay and micronucleus tests on Oreochromis niloticus (Perciforme: Cichlidae) exposed to raw sugarcane vinasse and to phisicochemical treated vinasse by pH adjustment with lime (CaO).

    Science.gov (United States)

    Correia, Jorge E; Christofoletti, Cintya Ap; Ansoar-Rodríguez, Yadira; Guedes, Thays A; Fontanetti, Carmem S

    2017-04-01

    In Brazil vinasse, a main sugarcane distillery residue, stands out because every liter of alcohol generates 10-15 L of vinasse as waste. An alternative for the disposal of this waste is the fertirrigation of the sugarcane culture itself. However, the high amount released can saturate the soil and through leaching/percolation contaminate water resources. The aim of this study is verifying the toxic potential of vinasse in tilapias and effectiveness of the physicalchemical treatment of this waste with pH adjustment with lime (CaO). The comet assay and the micronucleus test were applied on animals exposed to dilutions of raw vinasse and vinasse adjusted to neutral pH. Bioassays with raw vinasse dilutions indicated a toxic and genotoxic potential; fish exposed to the highest concentration died less than 48 h after the exposure; the incidence of micronucleus was significantly higher when compared to negative control for all dilutions. For the comet assay, the scores of damage were statistically higher for all dilutions, with the exception of the 1% dillution. However, in the bioassay with the chemically treated vinasse (neutral pH), most fish in the 10% dilution survived and there was no significant difference when compared to the control. Damage scores in the comet assay were similar to the results of the untreated vinasse. The chemical treatment of vinasse with lime to neutralize the pH proved to be an effective alternative for the toxicity reduction of this residue, since it reduced the mortality of fish at higher concentrations and the incidence of damage to DNA. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Lack of micronucleus induction activity of ethyl tertiary-butyl ether in the bone marrow of F344 rats by sub-chronic drinking-water treatment, inhalation exposure, or acute intraperitoneal injection.

    Science.gov (United States)

    Noguchi, Tadashi; Kamigaito, Tomoyuki; Katagiri, Taku; Kondou, Hitomi; Yamazaki, Kazunori; Aiso, Shigetoshi; Nishizawa, Tomoshi; Nagano, Kasuke; Fukushima, Shoji

    2013-01-01

    Ethyl tertiary-butyl ether (ETBE) is an oxygenated gasoline additive synthesized from ethanol and isobutene that is used to reduce CO2 emissions. To support the Kyoto Protocol, the production of ETBE has undergone a marked increase. Previous reports have indicated that exposure to ETBE or methyl tertiary-butyl ether resulted in liver and kidney tumors in rats and/or mice. These reports raise concern about the effects of human exposure being brought about by the increased use of ETBE. The present study was conducted to evaluate the genotoxicity of ETBE using micronucleus induction of polychromatic erythrocytes in the bone marrow of male and female rats treated with ETBE in the drinking-water at concentrations of 0, 1,600, 4,000 or 10,000 ppm or exposed to ETBE vapor at 0, 500, 1,500 or 5,000 ppm for 13 weeks. There were no significant increases in micronucleus induction in either the drinking water-administered or inhalation-administered groups at any concentration of ETBE; although, in both groups red blood cells and hemoglobin concentration were slightly reduced in the peripheral blood in rats administered the highest concentration of ETBE. In addition, two consecutive daily intraperitoneal injections of ETBE at doses of 0, 250, 500 or 1,000 mg/kg did not increase the frequency of micronucleated bone marrow cells in either sex; all rats receiving intraperitoneal injections of ETBE at a dose of 2,000 mg/kg died after treatment day 1. These data suggest that ETBE is not genotoxic in vivo.

  12. Environmental and human risk assessment of landfill leachate: An integrated approach with the use of cytotoxic and genotoxic stress indices in mussel and human cells

    Energy Technology Data Exchange (ETDEWEB)

    Toufexi, Eirini; Tsarpali, Vasiliki [Section of Animal Biology, Department of Biology, School of Natural Sciences, University of Patras, GR 26500 Patras (Greece); Efthimiou, Ioanna; Vidali, Maria-Sophia; Vlastos, Dimitris [Department of Environmental and Natural Resources Management, University of Patras, 2 Seferi Str., GR 30100 Agrinio (Greece); Dailianis, Stefanos, E-mail: sdailianis@upatras.gr [Section of Animal Biology, Department of Biology, School of Natural Sciences, University of Patras, GR 26500 Patras (Greece)

    2013-09-15

    Highlights: • Landfill leachate poses a threat for aquatic biota and humans. • Leachate induces cytotoxic and oxidative effects on mussel hemocytes. • Increased levels of DNA damage were observed both in vivo and in vitro in hemocytes. • Leachate low doses enhance MN formation in human lymphocyte cultures. • Potential leachate aneugenic activity was detected in human lymphocytes. -- Abstract: The present study investigates leachate hazardous effects on marine biota and human cells, with the use of a battery of assays, both under in vivo and in vitro conditions. According to the results, mussels exposed for 4 days to 0.01 and 0.1% (v/v) of leachate showed increased levels of DNA damage and micronuclei (MN) frequencies in their hemocytes. Similarly, enhanced levels of DNA damage were also observed in hemocytes treated in vitro with relevant concentrations of leachate, followed by a significant enhancement of both superoxide anions (·O{sub 2}{sup −}) and lipid peroxidation products (malondialdehyde/MDA). On the other hand, human lymphocyte cultures treated with such a low concentrations of leachate (0.1, 0.2 and 1%, v/v), showed increased frequencies of MN formation and large MN size ratio, as well as decreased cell proliferation, as indicated by the use of the cytokinesis block micronucleus (CBMN) assay and Cytokinesis Block Proliferation Index (CBPI) respectively. These findings showed the clear-cut genotoxic and cytotoxic effects of leachate on both cellular types, as well as its potential aneugenic activity in human lymphocytes.

  13. Influence of processing and storage of integral grape juice (Vitis labrusca L.) on its physical and chemical characteristics, cytotoxicity, and mutagenicity in vitro.

    Science.gov (United States)

    Düsman, E; Almeida, I V; Pinto, E P; Lucchetta, L; Vicentini, V E P

    2017-05-31

    Integral grape juice is extracted from the grape through processes that allow the retention of their natural composition. However, due to the severity of some processes, fruit juices can undergo changes in their quality. The present study evaluated the cytotoxic and mutagenic effects of integral grape juice by a cytokinesis-blocked micronucleus assay in Rattus norvegicus hepatoma cells (HTC) in vitro. Vitis labrusca L. (variety Concord) were produced organically and by a conventional system, and their juice was extracted by a hot extraction process. The organic grapes were subjected to ultraviolet-type C radiation (UV-C). Experiments were performed after production and after 6 months in storage. Physicochemical analyses revealed that UV-C irradiation of organic grapes, the juice production process, and storage resulted in nutraceutical alterations. However, none of the juice concentrations were cytotoxic to HTC cells by the cytokinesis-blocked proliferation index results or were mutagenic, because the formation of micronucleated cells was not induced. In general, juice induced cell proliferation, possibly due to the presence of vitamins and sugar content (total soluble solid). The data increased the understanding of food technology and confirmed the quality and safety consumption of these juices.

  14. Environmental and human risk assessment of landfill leachate: An integrated approach with the use of cytotoxic and genotoxic stress indices in mussel and human cells

    International Nuclear Information System (INIS)

    Toufexi, Eirini; Tsarpali, Vasiliki; Efthimiou, Ioanna; Vidali, Maria-Sophia; Vlastos, Dimitris; Dailianis, Stefanos

    2013-01-01

    Highlights: • Landfill leachate poses a threat for aquatic biota and humans. • Leachate induces cytotoxic and oxidative effects on mussel hemocytes. • Increased levels of DNA damage were observed both in vivo and in vitro in hemocytes. • Leachate low doses enhance MN formation in human lymphocyte cultures. • Potential leachate aneugenic activity was detected in human lymphocytes. -- Abstract: The present study investigates leachate hazardous effects on marine biota and human cells, with the use of a battery of assays, both under in vivo and in vitro conditions. According to the results, mussels exposed for 4 days to 0.01 and 0.1% (v/v) of leachate showed increased levels of DNA damage and micronuclei (MN) frequencies in their hemocytes. Similarly, enhanced levels of DNA damage were also observed in hemocytes treated in vitro with relevant concentrations of leachate, followed by a significant enhancement of both superoxide anions (·O 2 − ) and lipid peroxidation products (malondialdehyde/MDA). On the other hand, human lymphocyte cultures treated with such a low concentrations of leachate (0.1, 0.2 and 1%, v/v), showed increased frequencies of MN formation and large MN size ratio, as well as decreased cell proliferation, as indicated by the use of the cytokinesis block micronucleus (CBMN) assay and Cytokinesis Block Proliferation Index (CBPI) respectively. These findings showed the clear-cut genotoxic and cytotoxic effects of leachate on both cellular types, as well as its potential aneugenic activity in human lymphocytes

  15. Evaluation of the cytotoxic and genotoxic potential of lecithin/chitosan nanoparticles

    Science.gov (United States)

    Taner, Gökçe; Yeşilöz, Recep; Özkan Vardar, Deniz; Şenyiğit, Taner; Özer, Özgen; Degen, Gisela H.; Başaran, Nurşen

    2014-02-01

    Nanoparticles-based drug targeting delivery systems have been introduced in the treatment for various diseases because of their effective properties, although there have been conflicting results on the toxicity of nanoparticles. In the present study, the aim was to evaluate the cytotoxicity and the genotoxicity of different concentrations of lecithin/chitosan nanoparticles with and without clobetasol-17-propionate (CP) by neutral red uptake (NRU) cytotoxicity assay and single cell gel electrophoresis (Comet) and cytokinesis-blocked micronucleus assays. The IC50 values of lecithin/chitosan nanoparticles with/without CP were found as 1.9 and 1.8 %, respectively, in the NRU cytotoxicity test. High concentrations of lecithin/chitosan nanoparticles induced DNA damage in human lymphocytes as evaluated by comet assay. The micronucleus frequency was increased by the lecithin/chitosan treatment in a dose-dependent manner. Also at the two highest concentrations, a significant increase in micronucleus formation was observed. Lecithin/chitosan nanoparticles with CP did not increase the frequency of micronucleus and also did not induce additional DNA damage when compared with lecithin/chitosan nanoparticles without CP; therefore, CP itself has not found to be genotoxic at the studied concentration.

  16. Study on cell survival, induction of apoptosis and micronucleus formation in SCL-II and RTiV3 cells after exposure to the Auger electron emitter Tc-99m

    International Nuclear Information System (INIS)

    Kadenbach, K.; Kriehuber, R.; Weiss, D.G.

    2003-01-01

    Full text: Cell survival, induction of apoptosis and micronucleus (MN) formation have been investigated in the human squamous cell carcinoma cell line SCL-II and in the rat tracheal cell line RTiV3 after exposure to the Auger electron emitter Tc-99m. Cells were either acutely gamma(Co-60)-irradiated (0.78 Gy/min) or exposed to Tc-99m-Pertechnetate (25-300 MBq/20ml) for 24 h under cell culture conditions and assayed for cell survival (Colony-forming assay), micronucleus formation (Cytochalasin B assay) and the frequency of apoptotic cells (Fluorescence microscopy). Analytical dosimetrical models have been applied to derive the absorbed dose corresponding to the accumulated decays of Tc-99m. Absorbed doses up to 1.3 Gy could be achieved after Tc-99m exposure leading to no significant cell killing in this dose range except at one dose point (0.25 Gy) in SCL-II cells. MN formation was consistently lower when compared to Co-60 irradiated cells and showed a linear dose-response. The apoptotic response in SCL-II cells after Tc-99m exposure was described best by a 3rd order polynomial and increased apoptosis induction could be observed at much lower doses (0.25 Gy) in comparison to the reference radiation (0.8 Gy). The relative biological effectiveness (RBE) has been determined for MN formation and apoptosis induction and was found to be in the range of 0.1- 1.3 for both investigated biological endpoints, depending on which mathematical model for describing the dose-effect curve was used. Up-take experiments revealed an activity concentration ratio cells vs. medium of 1.2 after 16 h up to 24 h of exposure. No increased biological effectiveness of Tc-99m applied as Sodium-Pertechnetate could be observed in the investigated cell lines in comparison to gamma-irradiation. Induction of apoptosis is slightly increased after Tc-99m exposure in SCL-II cells and it has to be further evaluated, if this is due to the emitted Auger-component. A passive up-take mechanism of Tc-99m is

  17. Glutathione S-Transferase Gene Polymorphisms: Modulator of Genetic Damage in Gasoline Pump Workers.

    Science.gov (United States)

    Priya, Kanu; Yadav, Anita; Kumar, Neeraj; Gulati, Sachin; Aggarwal, Neeraj; Gupta, Ranjan

    2015-01-01

    This study investigated genetic damage in gasoline pump workers using the cytokinesis blocked micronucleus (CBMN) assay. Blood and urine samples were collected from 50 gasoline pump workers and 50 control participants matched with respect to age and other confounding factors except for exposure to benzene through gasoline vapors. To determine the benzene exposure, phenol was analyzed in urinary samples of exposed and control participants. Urinary mean phenol level was found to be significantly high (P gasoline pump workers (6.70 ± 1.78) when compared to control individuals (2.20 ± 0.63; P gasoline vapors can increase genotoxic risk in gasoline pump workers. © The Author(s) 2015.

  18. Studies on bystander effects of 14MeV neutrons in human blood lymphocytes using CBMN assay

    International Nuclear Information System (INIS)

    Bakkiam, D.; Arul Anantha Kumar, A.; Sonwani, Swetha; Alaguraja, E.; Mathiyarasu, R.; Baskaran, R.; Venkatraman, B.

    2018-01-01

    Radiation induced Bystander Effects (RIBE) in cells generally describes the phenomenon that non-irradiated cells respond as if they have themselves been irradiated upon receiving signals from directly irradiated cells, either through partnering or medium transfer. While it has been well established that bystander effects could be induced by gamma radiation and alpha-particle radiation it is still a question whether neutrons induce bystander effects or not. In view of this, experiments were carried out to quantify cytogenetic damage in human blood lymphocytes induced by neutron directly and indirectly i.e. RIBE through medium transfer method. Cytokinesis Blocked MicroNucleus (CBMN) assay was used to study DNA damage events wherein micronuclei (MN) were scored in binucleated cells. Results of MN frequency in neutron direct and indirect irradiated blood lymphocytes (bystander samples) are compared

  19. Assessment of in vitro genotoxic and cytotoxic effects of flurbiprofen on human cultured lymphocytes.

    Science.gov (United States)

    Timocin, Taygun; Ila, Hasan Basri; Dordu, Tuba; Husunet, Mehmet Tahir; Tazehkand, Mostafa Norizadeh; Valipour, Ebrahim; Topaktas, Mehmet

    2016-01-01

    Flurbiprofen is non-steroidal anti-inflammatory drug which is commonly used for its analgesic, antipyretic, and anti-inflammatory effects. The purpose of the study was to explore the genotoxic and cytotoxic effects of flurbiprofen in human cultured lymphocytes by sister chromatid exchange, chromosome aberration, and cytokinesis-blocked micronucleus tests. 10, 20, 30, and 40 μg/mL concentrations of flurbiprofen (solvent is DMSO) were used to treatment of human cultured lymphocytes at two different treatment periods (24 and 48 h). Flurbiprofen had no significant genotoxic effect in any of these tests. But exposing to flurbiprofen for 24 and 48 h led to significant decrease on proliferation index, mitotic index, and nuclear division index (NDI). Also, all decreases were concentration-dependent (except NDI at 24 h treatment period). Consequently, the findings of this research showed that flurbiprofen had cytotoxic effects in human blood lymphocytes.

  20. Assessment of individual radiosensitivity in human lymphocytes of cancer patients and its correlation with adverse side effects to radiation therapy

    CERN Document Server

    Di Giorgio, M; Busto, E; Mairal, L; Menendez, P; Roth, B; Sardi, M; Taja, M R; Vallerga, M B

    2003-01-01

    Background and purpose: Individual radiosensitivity is an inherent characteristic, associated with an increased reaction to ionizing radiation on the human body. Biological endpoints such as clonogenic survival, chromosome aberration formation and repair capacity of radiation-induced damage have been applied to evaluate individual radiosensitivity in vitro. 5%-7% of cancer patients develop adverse side effects to radiation therapy in normal tissues within the treatment field, which are referred as 'clinical radiation reactions' and include acute effects, late effects and cancer induction. It has been hypothesized that the occurrence and severity of these reactions are mainly influenced by genetic susceptibility to radiation. Additionally, the nature of the genetic disorders associated with hypersensitivity to radiotherapy suggests that DNA repair mechanisms are involved. Consequently, the characterization of DNA repair in lymphocytes through cytokinesis blocked micronucleus (MN) and alkaline single-cell micro...

  1. Chemical composition and in vitro cytotoxic, genotoxic effects of essential oil from Urtica dioica L.

    Science.gov (United States)

    Gül, Süleyman; Demirci, Betül; Başer, Kemal Hüsnü Can; Akpulat, H Aşkin; Aksu, Pinar

    2012-05-01

    The aim of this study was to determine the chemical composition of Urtica dioica essential oil, and to evaluate its cytotoxic and genotoxic effects, using cytogenetic tests such as the cytokinesis-block micronucleus assay and chromosomal aberration analysis in human lymphocyte cultures in vitro. GC-MS analysis of U. dioica essential oil identified 43 compounds, representing 95.8% of the oil. GC and GC-MS analysis of the essential oil of U. dioica revealed that carvacrol (38.2%), carvone (9.0%), naphthalene (8.9%), (E)-anethol (4.7%), hexahydrofarnesyl acetone (3.0%), (E)-geranyl acetone (2.9%), (E)-β-ionone (2.8%) and phytol (2.7%) are the main components, comprising 72.2% of the oil. A significant correlation was found between the concentration of essential oil and the following: chromosomal aberrations, micronuclei frequency, apoptotic cells, necrotic cells, and binucleated cells.

  2. Cytogenetics data in adult men involved in the recycling of electronic wastes

    Directory of Open Access Journals (Sweden)

    Yanan Du

    2018-04-01

    Full Text Available In this data article, 146 villagers (exposed group were randomly selected from the workers who involved in the e-wastes recycling directly as a daily job in Tianjin. Control group, including 121 villagers, came from another town without e-waste disposal sites. Chromosomal aberrations (CA and cytokinesis blocking micronucleus (CBMN were performed to detect the cytogenetic effect for each subject. DNA damage was detected using comet assay; the DNA percentage in the comet tail (TDNA%, tail moment (TM, and Olive tail moment (OTM were recorded to describe DNA damage to lymphocytes and spermatozoa. Routine semen analysis, spermatozoa motility and morphology were analyzed. The RT2Profiler PCR array was used to measure levels of expression of 84 genes related to quality of DNA. It showed significant relationships between CA, CBMN, DNA damage and exposure time in exposure subjects. The alteration of sperm motility rate, abnormality rate and total sperm counts had association with exposure time and age.

  3. In situ biomonitoring of the genotoxic effects of mixed industrial emissions using the Tradescantia micronucleus and pollen abortion tests with wild life plants: Demonstration of the efficacy of emission controls in an eastern European city

    Energy Technology Data Exchange (ETDEWEB)

    Misik, Miroslav [Department of Botany, Comenius University in Bratislava, Faculty of Natural Sciences, Revova 39, SK 811 02 Bratislava 1 (Slovakia); Micieta, Karol [Department of Botany, Comenius University in Bratislava, Faculty of Natural Sciences, Revova 39, SK 811 02 Bratislava 1 (Slovakia); Solenska, Martina [Department of Botany, Comenius University in Bratislava, Faculty of Natural Sciences, Revova 39, SK 811 02 Bratislava 1 (Slovakia); Misikova, Katarina [Department of Botany, Comenius University in Bratislava, Faculty of Natural Sciences, Revova 39, SK 811 02 Bratislava 1 (Slovakia); Pisarcikova, Helena [Department of Botany, Comenius University in Bratislava, Faculty of Natural Sciences, Revova 39, SK 811 02 Bratislava 1 (Slovakia); Knasmueller, Siegfried [Institute of Cancer Research, Department of Inner Medicine I, Medical University of Vienna, Borschkegasse 8a, A-1090 Vienna (Austria)]. E-mail: siegfried.knasmueller@meduniwien.ac.at

    2007-01-15

    Aim of the study was to monitor changes of genotoxic activity of urban air caused by an incinerator and a petrochemical plant in Tradescantia micronucleus (Trad-MCN) and pollen fertility assays with wild plants (Chelidonium majus, Clematis vitalba, Cichorium intybus, Linaria vulgaris, Robinia pseudoacacia). While in the first sampling period (1997-2000) significantly (on average 80%) more MN were found at the polluted site in comparison to controls from a rural area, no significant effects were observed during a later period (between 2003 and 2005). A similar pattern was observed in the pollen abortion assays in which the most pronounced effects were found in chicory and false acacia. The differences of the results obtained in the two periods can be explained by a substantial reduction of air pollution by use of new technologies. In particular the decrease of SO{sub 2} emissions may account for the effects seen in the present study. - Air pollution caused by industrial emissions induced micronuclei in Tradescantia and increased pollen abortion in wild plant species.

  4. Selective accumulation of 147Pm in organism on induction of PCE's micronucleus and SCE of bone marrow cells as well as the chromosome aberrations on fetal liver and spleen

    International Nuclear Information System (INIS)

    Zhu Shoupeng; Zheng Siying; Wang Liuyi; Lu Zhongyan; Yang Shuqin

    1989-01-01

    Study of accumulation peculiarity of 147 Pm showed that I.V. different doses of 147 Pm were the same selectively localized in skeleton and liver. Retention of 147 Pm in skeleton and liver was elevated when the radioactive doses of 147 Pm were increased. At the same time absorption does of 147 Pm radiation was heightened. The ability of 147 Pm to induce sister chromatid exchanges (SCEs) has been investigated by IdU labelling methods. A statistically significant elevation of SCEs was observed after 147 Pm intake.In mice the number of SCEs per cell in bone marrow cells was always higher when the animals were maintained on the doses of 37 Bq/g. The injurious effects of 147 Pm, using PCE's micronucleus rates in bone marrow cells were observed. 147 Pm was dominantly deposited on maternal liver. Deposition of 147 Pm in maternal spleen was about quandrantal of the maternal liver. Studies indicated that maternal contamination of 147 Pm could induced chromosome aberrations in fetal liver and spleen cells. Among the type of aberrations induced by 147 Pm, chromatid breakage were predominant. The incidence of chromosome aberrations on fetal liver cells induced by 147 Pm was higher on fetal spleen cells

  5. Predictive cytogenetic biomarkers for colorectal neoplasia in medium risk patients.

    Science.gov (United States)

    Ionescu, E M; Nicolaie, T; Ionescu, M A; Becheanu, G; Andrei, F; Diculescu, M; Ciocirlan, M

    2015-01-01

    DNA damage and chromosomal alterations in peripheral lymphocytes parallels DNA mutations in tumor tissues. The aim of our study was to predict the presence of neoplastic colorectal lesions by specific biomarkers in "medium risk" individuals (age 50 to 75, with no personal or family of any colorectal neoplasia). We designed a prospective cohort observational study including patients undergoing diagnostic or opportunistic screening colonoscopy. Specific biomarkers were analyzed for each patient in peripheral lymphocytes - presence of micronuclei (MN), nucleoplasmic bridges (NPB) and the Nuclear Division Index (NDI) by the cytokinesis-blocked micronucleus assay (CBMN). Of 98 patients included, 57 were "medium risk" individuals. MN frequency and NPB presence were not significantly different in patients with neoplastic lesions compared to controls. In "medium risk" individuals, mean NDI was significantly lower for patients with any neoplastic lesions (adenomas and adenocarcinomas, AUROC 0.668, p 00.5), for patients with advanced neoplasia (advanced adenoma and adenocarcinoma, AUROC 0.636 p 0.029) as well as for patients with adenocarcinoma (AUROC 0.650, p 0.048), for each comparison with the rest of the population. For a cut-off of 1.8, in "medium risk" individuals, an NDI inferior to that value may predict any neoplastic lesion with a sensitivity of 97.7%, an advanced neoplastic lesion with a sensitivity of 97% and adenocarcinoma with a sensitivity of 94.4%. NDI score may have a role as a colorectal cancer-screening test in "medium risk" individuals. DNA = deoxyribonucleic acid; CRC = colorectal cancer; EU = European Union; WHO = World Health Organization; FOBT = fecal occult blood test; CBMN = cytokinesis-blocked micronucleus assay; MN = micronuclei; NPB = nucleoplasmic bridges; NDI = Nuclear Division Index; FAP = familial adenomatous polyposis; HNPCC = hereditary non-polypoid colorectal cancer; IBD = inflammatory bowel diseases; ROC = receiver operating

  6. Cytogenetic damage after 131-iodine treatment for hyperthyroidism and thyroid cancer

    International Nuclear Information System (INIS)

    Gutierrez, S.; Carbonell, E.; Creus, A.; Marcos, R.

    1999-01-01

    To detect the incidence and persistence of potential chromosome damage induced by iodine-131 therapy, we applied the cytokinesis-block micronucleus assay to peripheral blood lymphocytes from hyperthyroidism and thyroid cancer patients treated with 131 I. Two groups of patients were evaluated in a longitudinal study; one group was composed of 47 hyperthyroid patients and the other of 39 thyroid cancer patients. In the hyperthyroidism group, the micronuclei frequency was determined before 131 I therapy and 1 week, 1 month and 3 months after it. Furthermore, an additional sample was taken from a subgroup of 17 hyperthyroidism patients 6 months after treatment. In the thyroid cancer group, the analysis was also conducted over time, and four samples were studied: before treatment and 1 week, 6 months and 1 year later. Simultaneously, a cross-sectional study was performed with 70 control subjects and 54 thyroid cancer patients who had received the last therapeutic dose 1-6 years before the present study. In the hyperthyroidism group a significant increase in the micronuclei average was found over time. In the sample obtained 6 months after therapy, the micronuclei mean frequency was practically the same as in the sample taken 3 months before. In the thyroid cancer group a twofold increase in the frequency of micronuclei was seen 1 week after therapy. Although this value decreased across time, the micronuclei frequency obtained 1 year after 131 I therapy remained higher than the value found before it. Concerning the data from the cross-sectional study, a significant increase in the frequency of micronuclei was detected in the subgroup of thyroid cancer patients treated between 1 and 3 years before the current study. These results indicate that exposure to 131 I therapy induces chromosome damage in peripheral lymphocytes and that the cytokinesis-block micronucleus assay is sensitive enough to detect the genetic damage by exposure to sufficiently high levels of radiation

  7. Cytogenetic damage after 131-iodine treatment for hyperthyroidism and thyroid cancer

    Energy Technology Data Exchange (ETDEWEB)

    Gutierrez, S.; Carbonell, E.; Creus, A.; Marcos, R. [Universitat Autonoma de Barcelona (Spain). Dept. de Genetica i de Microbiologia; Galofre, P. [Servei de Medicina Nuclear, Ciutat Sanitaria i Universitaria Vall d' Hebron, Barcelona (Spain)

    1999-12-01

    To detect the incidence and persistence of potential chromosome damage induced by iodine-131 therapy, we applied the cytokinesis-block micronucleus assay to peripheral blood lymphocytes from hyperthyroidism and thyroid cancer patients treated with {sup 131}I. Two groups of patients were evaluated in a longitudinal study; one group was composed of 47 hyperthyroid patients and the other of 39 thyroid cancer patients. In the hyperthyroidism group, the micronuclei frequency was determined before {sup 131}I therapy and 1 week, 1 month and 3 months after it. Furthermore, an additional sample was taken from a subgroup of 17 hyperthyroidism patients 6 months after treatment. In the thyroid cancer group, the analysis was also conducted over time, and four samples were studied: before treatment and 1 week, 6 months and 1 year later. Simultaneously, a cross-sectional study was performed with 70 control subjects and 54 thyroid cancer patients who had received the last therapeutic dose 1-6 years before the present study. In the hyperthyroidism group a significant increase in the micronuclei average was found over time. In the sample obtained 6 months after therapy, the micronuclei mean frequency was practically the same as in the sample taken 3 months before. In the thyroid cancer group a twofold increase in the frequency of micronuclei was seen 1 week after therapy. Although this value decreased across time, the micronuclei frequency obtained 1 year after {sup 131}I therapy remained higher than the value found before it. Concerning the data from the cross-sectional study, a significant increase in the frequency of micronuclei was detected in the subgroup of thyroid cancer patients treated between 1 and 3 years before the current study. These results indicate that exposure to {sup 131}I therapy induces chromosome damage in peripheral lymphocytes and that the cytokinesis-block micronucleus assay is sensitive enough to detect the genetic damage by exposure to sufficiently high

  8. Graphistrength© C100 MultiWalled Carbon Nanotubes (MWCNT): thirteen-week inhalation toxicity study in rats with 13- and 52-week recovery periods combined with comet and micronucleus assays

    International Nuclear Information System (INIS)

    Régnier, Jean-François; Pothmann-Krings, Daniela; Simar, Sophie; Dony, Eva; Net, Jean-Loïc Le; Beausoleil, Julien

    2017-01-01

    Graphistrength© C100 provides superior electrical and mechanical properties for various applications and is one of the industrial MWCNT referenced in the OECD sponsorship program for the safety testing of nanomaterials. Graphistrength© C100 is formed of MWCNT (ca. 12 walls, outer mean diameter ca. 12 nm, length ca. 1 µm) agglomerated in particles with a granulometry centered on 400 µm. A general feature of MWCNT after inhalation or intratracheal exposures is the induction of an inflammatory reaction in the lungs sometimes associated with local genotoxic effects. Most of the in vitro and in vivo genotoxicity data available on Graphistrength© C100 are negative. However, a weak DNA damage activity in the in vitro and in vivo FPG-modified Comet assays and a weak clastogenic effect in the in vitr o micronucleus test were reported. After investigating different parameters for the aerosol generation, male and female Wistar rats were exposed by nose-only inhalation (6h/day, 5d/week) to target concentrations of 0.05, 0.25 and 5.0 mg/m 3 air of a respirable aerosol (MMAD < 3 µm) and sacrificed immediately after 4 and 13 weeks of exposure and 13 and 52 weeks of recovery after the 13-week exposure. Clinical, biological and histological evaluations were performed according to the OECD TG 413. Broncho-alveolar lavage fluid (BALF) was collected and analysed for cytokines and inflammatory parameters. Immediately after 13 weeks of exposure, chromosomal aberrations in the bone marrow cells of males and females were evaluated by the micronucleus test (OECD TG 474) and DNA damage in the lung, kidney and liver cells of males were assessed by both the standard and the human 8-oxoguanine DNA N-glycosylase 1 (hOGG1)-modified comet assay (OECD TG 489). Concentration-related deposition of black particles (MWCNT) was observed in lungs. At all sacrifice periods, an inflammatory lung reaction was observed in rats exposed to 5.0 mg/m 3 associated with changes in the differential white

  9. Graphistrength© C100 MultiWalled Carbon Nanotubes (MWCNT): thirteen-week inhalation toxicity study in rats with 13- and 52-week recovery periods combined with comet and micronucleus assays

    Science.gov (United States)

    Régnier, Jean-François; Pothmann-Krings, Daniela; Simar, Sophie; Dony, Eva; Le Net, Jean-Loïc; Beausoleil, Julien

    2017-06-01

    Graphistrength© C100 provides superior electrical and mechanical properties for various applications and is one of the industrial MWCNT referenced in the OECD sponsorship program for the safety testing of nanomaterials. Graphistrength© C100 is formed of MWCNT (ca. 12 walls, outer mean diameter ca. 12 nm, length ca. 1 µm) agglomerated in particles with a granulometry centered on 400 µm. A general feature of MWCNT after inhalation or intratracheal exposures is the induction of an inflammatory reaction in the lungs sometimes associated with local genotoxic effects. Most of the in vitro and in vivo genotoxicity data available on Graphistrength© C100 are negative. However, a weak DNA damage activity in the in vitro and in vivo FPG-modified Comet assays and a weak clastogenic effect in the in vitro micronucleus test were reported. After investigating different parameters for the aerosol generation, male and female Wistar rats were exposed by nose-only inhalation (6h/day, 5d/week) to target concentrations of 0.05, 0.25 and 5.0 mg/m3 air of a respirable aerosol (MMAD males and females were evaluated by the micronucleus test (OECD TG 474) and DNA damage in the lung, kidney and liver cells of males were assessed by both the standard and the human 8-oxoguanine DNA N-glycosylase 1 (hOGG1)-modified comet assay (OECD TG 489). Concentration-related deposition of black particles (MWCNT) was observed in lungs. At all sacrifice periods, an inflammatory lung reaction was observed in rats exposed to 5.0 mg/m3 associated with changes in the differential white blood cells counts. The lung inflammation was characterized by changes in the cytological, biochemical and cytokine parameters of the BALF, an increase of the lung weight, an interstitial inflammation mainly around the alveolar ducts at the bronchiole-alveolar junction and a cell hypertrophy/hyperplasia in the terminal and respiratory bronchioles. The slight changes in BALF parameters observed at 0.25 mg/m3 recovered after

  10. The chromosome damage induced by x-ray radiation doses. Comparison between dicentric chromosomes, micronuclei and Sister Chromatid Exchanges analyses. Valoracion de dao cromosomico originado por una dosis de rayos X. Comparacion de los analisis de cromosomas dicentricos, micronucleos e intercambios entre cromatidas hermanas

    Energy Technology Data Exchange (ETDEWEB)

    Fernandez, J.L.; Losada, C.; Losada, G.; Veiras, C. (Centro Oncologico de Galicia. La Corua (Spain)); Goyanes, V.J. (Hospital ' ' Teresa Herrera' ' . La Corua (Spain))

    1993-01-01

    Exposure to ionizing radiations is a well-known source of chromosome damage. Here we present a comparison among three different methodologies employed to recognize cytogenetic damage, after an acute exposure of human lymphocytes to 3 Gy of X-rays (100kVp). Scoring of dicentric chromosomes, present in first mitosis ''in vitro'', was the method of preference as dicentrics increased 937.5 times with respect to background. Micronucleus scoring in binucleated-cytokinesis blocked cells showed an increase of 32.5 times, while it was only of 1.46 times when Sister Chromatid Exchanges (SCEs) were analyzed. The estimated probability of an acentric fragment becoming a micronucleus was around 0.25. Intercellular distribution of dicentrics agree with Poisson, while micronucleus were overdispersed. When analyzed at second cycle after damage induction, the dicentrics yield as well as the level of cells with unstable cromosome aberrations, decreased around a half. Finally, SCEs level was similar in cells with or without unstable structural chromosome aberrations. (Author)

  11. Séroprévalence du cytomégalovirus chez les donneurs de sang au ...

    African Journals Online (AJOL)

    Objective: The aim of this study was to determine the seroprevalence of CMV-IgG and CMV-IgM among voluntary blood donors in national centre of blood transfusion (CNTS) of Lomé, main town of Togo, West Africa. This is a view to making recommendations on donor screening and transfusion protocols among susceptible ...

  12. Correlation between micronuclei frequency in peripheral blood lymphocytes and retention of 131-I in thyroid cancer patients

    International Nuclear Information System (INIS)

    Vrndic, O.B.; Milosevic-Djordjevic, O.M.; Mijatovic Teodorovic, L.C.; Zivancevic Simonovic, S.T.; Jeremic, M.Z.; Stosic, I.M.; Grujicic, D.V.

    2013-01-01

    Differentiated thyroid cancers (DTCs) derive from thyroid follicular cells and include papillary and follicular cancers. In patients with DTCs, the initial treatment includes thyroidectomy and radioactive iodine (131-I) therapy. The objective of this study was to examine whether the intensity of DNA damage in peripheral blood lymphocytes (PBLs) of DTC patients depends on the amount of 131-I retained in the selected regions of interest (thyroid and abdominal region) as well as in the whole-body 72 hours after therapy. In addition, the possible influence of other factors that may affect micronuclei (MN) frequency, such as age, gender, smoking habits, and histological type of tumour was analyzed. The study population consisted of 22 DTC patients and 20 healthy donors. Data on the distribution of 131-I were obtained from the whole-body scans. MN frequency and cytokinesis-block proliferation index (CBPI) were measured using cytokinesis-block micronucleus assay. 131-I therapy significantly increased the MN frequency (19.50±6.90 vs. 27.10±19.50 MN) and significantly decreased the CBPI (1.52±0.20 vs. 1.38±0.17) in patients' lymphocytes. There was a clear correlation between the increased MN frequency and 131-I accumulation in the thyroid region in patients without metastases. The MN values did not differ in relation to the factors that could affect MN, such as age, gender, smoking habits, and histological type of tumour. In conclusion, the MN frequency in PBLs of DTC patients without metastases depends on the accumulation of 131-I in the thyroid region and does not depend on the other factors examined. (author)

  13. Compounds used to produce cloned animals are genotoxic and mutagenic in mammalian assays in vitro and in vivo

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, R.J. [Programa de Pós-Graduação em Biologia Celular e Molecular, Instituto de Biociências de Rio Claro, Universidade Estadual Paulista, Rio Claro, SP (Brazil); Centro de Estudos em Células Tronco, Terapia Celular e Genética Toxicológica, Núcleo de Hospital Universitário, Universidade Federal de Mato Grosso do Sul, Campo Grande, MS (Brazil); Programa de Pós-Graduação em Saúde em Desenvolvimento na Região Centro-Oeste, Faculdade de Medicina “Dr. Hélio Mandetta”, Universidade Federal de Mato Grosso do Sul, Campo Grande, MS (Brazil); Programa de Mestrado em Farmácia, Centro de Ciências Biológicas e da Saúde, Universidade Federal de Mato Grosso do Sul, Campo Grande, MS (Brazil); Mantovani, M.S.; Silva, A.F. da [Departamento de Biologia Geral, Universidade Estadual de Londrina, Londrina, PR (Brazil); Pesarini, J.R. [Centro de Estudos em Células Tronco, Terapia Celular e Genética Toxicológica, Núcleo de Hospital Universitário, Universidade Federal de Mato Grosso do Sul, Campo Grande, MS (Brazil); Programa de Pós-Graduação em Saúde em Desenvolvimento na Região Centro-Oeste, Faculdade de Medicina “Dr. Hélio Mandetta”, Universidade Federal de Mato Grosso do Sul, Campo Grande, MS (Brazil); Mauro, M.O. [Centro de Estudos em Células Tronco, Terapia Celular e Genética Toxicológica, Núcleo de Hospital Universitário, Universidade Federal de Mato Grosso do Sul, Campo Grande, MS (Brazil); Programa de Doutorado em Biotecnologia e Biodiversidade - Rede Pró Centro-Oeste, Universidade Federal de Mato Grosso do Sul, Campo Grande, MS (Brazil); Ribeiro, L.R. [Programa de Pós-Graduação em Biologia Celular e Molecular, Instituto de Biociências de Rio Claro, Universidade Estadual Paulista, Rio Claro, SP (Brazil); Programa de Pós-Graduação em Patologia, Faculdade de Medicina de Botucatu, Universidade Estadual Paulista, Botucatu, SP (Brazil)

    2014-03-28

    The compounds 6-dimethylaminopurine and cycloheximide promote the successful production of cloned mammals and have been used in the development of embryos produced by somatic cell nuclear transfer. This study investigated the effects of 6-dimethylaminopurine and cycloheximide in vitro, using the thiazolyl blue tetrazolium bromide colorimetric assay to assess cytotoxicity, the trypan blue exclusion assay to assess cell viability, the comet assay to assess genotoxicity, and the micronucleus test with cytokinesis block to test mutagenicity. In addition, the comet assay and the micronucleus test were also performed on peripheral blood cells of 54 male Swiss mice, 35 g each, to assess the effects of the compounds in vivo. The results indicated that both 6-dimethylaminopurine and cycloheximide, at the concentrations and doses tested, were cytotoxic in vitro and genotoxic and mutagenic in vitro and in vivo, altered the nuclear division index in vitro, but did not diminish cell viability in vitro. Considering that alterations in DNA play important roles in mutagenesis, carcinogenesis, and morphofunctional teratogenesis and reduce embryonic viability, this study indicated that 6-dimethylaminopurine and cycloheximide utilized in the process of mammalian cloning may be responsible for the low embryo viability commonly seen in nuclear transfer after implantation in utero.

  14. Dose Assessment using Chromosome Aberration Analyses in Human Peripheral Blood Lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Ryu, Tae Ho; Kim, Jin-Hong; Kim, Jin Kyu [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

    2015-10-15

    The healthy five donors were recruited to establish the dose-response calibration curve for chromosomal aberrations by ionizing radiation exposure. Our cytogenetic results revealed that the mean frequency of chromosome aberration increased with increasing radiation dose. In this study, dicentric assay and CBMN assay were compared considering the sensitivity and accuracy of dose estimation. Therefore, these chromosome aberration analyses will be the foundation for biological dosimetric analysis with additional research methods such as translocation and PCC assay. The conventional analysis of dicentric chromosomes in HPBL was suggested by Bender and Gooch in 1962. This assay has been for many years, the golden standard and the most specific method for ionizing radiation damage. The dicentric assay technique in HPBL has been shown as the most sensitive biological method and reliable bio-indicator of quantifying the radiation dose. In contrast, the micronucleus assay has advantages over the dicentric assay since it is rapid and requires less specialized expertise, and accordingly it can be applied to monitor a big population. The cytokinesis-block micronucleus (CBMN) assay is a suitable method for micronuceli measurement in cultured human as well as mammalian cells. The aim of our study was to establish the dose response curve of radiation-induced chromosome aberrations in HPBL by analyzing the frequency of dicentrics and micronuclei.

  15. Compounds used to produce cloned animals are genotoxic and mutagenic in mammalian assays in vitro and in vivo

    Directory of Open Access Journals (Sweden)

    R.J. Oliveira

    2014-04-01

    Full Text Available The compounds 6-dimethylaminopurine and cycloheximide promote the successful production of cloned mammals and have been used in the development of embryos produced by somatic cell nuclear transfer. This study investigated the effects of 6-dimethylaminopurine and cycloheximide in vitro, using the thiazolyl blue tetrazolium bromide colorimetric assay to assess cytotoxicity, the trypan blue exclusion assay to assess cell viability, the comet assay to assess genotoxicity, and the micronucleus test with cytokinesis block to test mutagenicity. In addition, the comet assay and the micronucleus test were also performed on peripheral blood cells of 54 male Swiss mice, 35 g each, to assess the effects of the compounds in vivo. The results indicated that both 6-dimethylaminopurine and cycloheximide, at the concentrations and doses tested, were cytotoxic in vitro and genotoxic and mutagenic in vitro and in vivo, altered the nuclear division index in vitro, but did not diminish cell viability in vitro. Considering that alterations in DNA play important roles in mutagenesis, carcinogenesis, and morphofunctional teratogenesis and reduce embryonic viability, this study indicated that 6-dimethylaminopurine and cycloheximide utilized in the process of mammalian cloning may be responsible for the low embryo viability commonly seen in nuclear transfer after implantation in utero.

  16. Cytogenetic effects induced by radiotherapy of cancer patients

    International Nuclear Information System (INIS)

    Ekhtiar, A.; Al-Achkar, W.

    2008-03-01

    Ionizing radiation plays a key role in the treatment of many neoplasias. But it is well known that ionizing radiation induce wide specter of DNA damages, including SSBs, DSBs, base damage, and DNA-protein cross links. As a consequence, a second tumor may be developed after the primary tumor therapy. Attempts have been made to evaluate the genotoxicity of ionizing radiation in patients undergoing radiotherapy. In the present work, the cytogenetic damage present in peripheral blood lymphocytes of patients (29 donors) treated with fractionated partial-body radiation therapy for Head-and-neck cancer patient was followed before, during and at the end of treatment by means of the cytokinesis-block micronucleus assay. These patients had no previous chemotherapy or radiotherapy. Our results indicate that the level of spontaneous cytogenetic damage in cancer patients and smokers control (3 donors) were higher than in healthy non smoking controls (3 donor). During and after treatment, increased of micronucleus cells frequencies were observed with increasing treatment doses.(author)

  17. Detection of radioiodine-induced cytogenetic alterations in circulating lymphocytes of thyroid patients

    Energy Technology Data Exchange (ETDEWEB)

    Kasuba, V [Inst. for Medical Recearch and Occupational Health, Zagreb (Croatia). Laboratory for Mutagenesis; Konrady, A; Koeteles, G J [Frederic Joliot-Curie National Research Institute for Radiobiology and Radiohygiene, Budapest (Hungary); Kusic, Z [Clinical Hospital Sestre Milosrdnice, Zagreb (Croatia). Dept. of Oncology and Nuclear Medicine

    1994-10-01

    Radioiodines are often used for experimental purposes and for diagnosis and therapy in clinical practice. Human population might also be exposed to radioiodines in nuclear accidents. The ionizing energy of radioiodine affects not only the thyroid where it concentrates but also other tissues, especially the lymphocytes during their circulation through and around the gland containing the radioisotopes. Therefore, it seemed to be of interest to carry out investigations concerning the cytogenetic alterations in blood lymphocytes of patients treated with iodine-131. The method of choice was the relatively easily performable micronucleus assay in cytokinesis-blocked cultures of human peripheral lymphocytes. The test was performed on blood samples of 30 patients before the radioisotope treatment and one, two and four days after, one as well as 6 and - in a few cases - 12 weeks later. The amounts of iodine-131 injected were dependent on the clinical practices to reach the therapeutic radiation doses for hyperthyroidism and adenomas and were in the range of 220 and 5180 MBq. it was observed that the micronucleus frequency increased in the treated hyperthyroid patients while in patients with toxic adenomas the radioiodine did not result in an increase or even as compared to the pretreatment values in a few cases decreased values were seen. The results suggest individual differences in radiosensitivity as well as that the frequency of cytogenetic alterations depend on the physiological or pathological conditions of the thyroid. The significance of this observation will be discussed for dose assessments by cytogenetic techniques due to internal radioiodine. (author).

  18. Induction of micronuclei by 2-hydroxypyridine in water and elimination of solution genotoxicity by UVC (254 nm) photolysis

    International Nuclear Information System (INIS)

    Skoutelis, Charalambos G.; Vlastos, Dimitris; Kortsinidou, Marianna C.; Theodoridis, Ioannis T.; Papadaki, Maria I.

    2011-01-01

    Highlights: ► 2-Hydroxypyridine (2-HPY) is the major metabolite of 2-halogenated pyridines photolysis. ► We examine the genotoxicity of 2-HPY in cultured human lymphocytes applying the micronucleus assay. ► 2-HPY was found to be genotoxic. ► Aqueous solutions of 2-HPY were irradiated by UV at 254 nm. ► Solution genotoxicity can be completely removed after prolonged phototreatment. - Abstract: 2-Hydroxypyridine (2-HPY) is a major first-stage product formed upon the photolytic destruction of 2-halogenated pyridines. Genotoxicity of 2-HPY in water was studied as a function of concentration. Aqueous solutions of 2-HPY were irradiated by ultraviolet (UV) at 254 nm. 2-HPY concentration, solution total organic carbon (TOC) concentration and solution genotoxicity were measured as a function of treatment time and their profile as a function of time is presented in this work. 2-HPY was found to be genotoxic at all concentrations in the range of 5–400 μg ml −1 . 2-HPY mineralises completely upon prolonged UV irradiation. All untreated and irradiated solution samples, taken at different photo-treatment times, were tested in cultured human lymphocytes applying the cytokinesis block micronucleus (CBMN) assay. The genotoxicity of the solution was reduced near to the control level after prolonged UV irradiation.

  19. Can Spirulina maxima reduce the mutagenic potential of sibutramine?

    Science.gov (United States)

    Araldi, R P; Santos, N P; Mendes, T B; Carvalho, L B; Ito, E T; de-Sá-Júnior, P L; Souza, E B

    2015-12-28

    The worldwide obesity pandemic requires the use of anti-obesity drugs. Sibutramine is an anti-obesity drug that has been used worldwide but is indiscriminately consumed in Brazil. Several studies have demonstrated that sibutramine promotes weight loss and weight maintenance, but several side effects have been associated with its systematic consumption. For this reason, sibutramine was withdrawn from the European and American markets, but still remains legal for use in Brazil. Studies have shown that a 5-10% reduction in body weight results in outstanding health benefits for obese patients. However, in order to promote significant weight loss, it is necessary to use sibutramine for at least 2 years. This long-term exposure has carcinogenic potential, as sibutramine causes DNA damage. Thus, this study evaluated the in vivo mutagenic potential of sibutramine alone (5, 7, 10, 15, and 20 mg/kg) and in association with Spirulina maxima (150 and 300 mg/kg), a cyanobacterium with antioxidant potential, using the polychromatic erythrocyte micronucleus test. Our results reinforced the mutagenic potential of sibutramine alone, which showed a time-dependent action. Combinatory treatments with S. maxima were not able to reduce the genotoxicity of sibutramine. These results were confirmed in vitro with the cytokinesis-blocked micronucleus test. In conclusion, our data showed that new alternative anti-obesity treatments are needed since the consumption of sibutramine can increase the risk of cancer in overweight patients.

  20. Compounds used to produce cloned animals are genotoxic and mutagenic in mammalian assays in vitro and in vivo

    International Nuclear Information System (INIS)

    Oliveira, R.J.; Mantovani, M.S.; Silva, A.F. da; Pesarini, J.R.; Mauro, M.O.; Ribeiro, L.R.

    2014-01-01

    The compounds 6-dimethylaminopurine and cycloheximide promote the successful production of cloned mammals and have been used in the development of embryos produced by somatic cell nuclear transfer. This study investigated the effects of 6-dimethylaminopurine and cycloheximide in vitro, using the thiazolyl blue tetrazolium bromide colorimetric assay to assess cytotoxicity, the trypan blue exclusion assay to assess cell viability, the comet assay to assess genotoxicity, and the micronucleus test with cytokinesis block to test mutagenicity. In addition, the comet assay and the micronucleus test were also performed on peripheral blood cells of 54 male Swiss mice, 35 g each, to assess the effects of the compounds in vivo. The results indicated that both 6-dimethylaminopurine and cycloheximide, at the concentrations and doses tested, were cytotoxic in vitro and genotoxic and mutagenic in vitro and in vivo, altered the nuclear division index in vitro, but did not diminish cell viability in vitro. Considering that alterations in DNA play important roles in mutagenesis, carcinogenesis, and morphofunctional teratogenesis and reduce embryonic viability, this study indicated that 6-dimethylaminopurine and cycloheximide utilized in the process of mammalian cloning may be responsible for the low embryo viability commonly seen in nuclear transfer after implantation in utero

  1. Physico-chemical characteristics and cyto-genotoxic potential of ZnO and TiO{sub 2} nanoparticles on human colon carcinoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Barone, F; Bizzarri, L; Andreoli, C; Zijno, A; De Angelis, I [Department of Environment and Primary Prevention, Istituto Superiore di Sanita, Viale Regina Elena 299, 00161 Rome (Italy); De Berardis, B [Department of Technology and Health, Istituto Superiore di Sanita, Viale Regina Elena 299, 00161 Rome (Italy); Degan, P, E-mail: barone@iss.it [Molecular Mutagenesis and DNA Repair, Istituto Nazionale per la Ricerca sul Cancro, L.go R. Benzi 10, 16132 Genova (Italy)

    2011-07-06

    The aim of the present study is to investigate the role of the physico-chemical properties of ZnO and TiO{sub 2} NPs in the potential cytotoxicity, genotoxicity and oxidative DNA damage induction on Caco-2 cell line. As negative control, fine TiO{sub 2} particles were used. The characterization of particles was carried out by electron microscopy (SEM, TEM) using a Soft Imaging System. To evaluate the effects of ZnO and TiO{sub 2} NPs induced on Caco-2 viability, Neutral Red assay was performed after treatment with different particle concentrations. Our results showed a significant dose and time dependent effect after treatment with ZnO NPs. On the contrary, no effect was observed on Caco-2 cells exposed to TiO{sub 2} particles either in micro-and in nano-size. The role of surface in the cytotoxicity induced on Caco-2 was also considered. The levels of DNA 8-oxodG, as the main marker of oxidative DNA damage, were measured by high-performance liquid chromatography with electrochemical detection (HPLC/EC). A significant increase in the 8-oxodG levels was observed after 6 h exposure for both NPs. The estimation of the potential genotoxicity of the two NPs is ongoing by the cytokinesis-block micronucleus assay. Our preliminary results showed that a slight micronucleus increase in binucleated cells was detected in the dose range applied only for ZnO.

  2. Genotoxicity assessment of membrane concentrates of landfill leachate treated with Fenton reagent and UV-Fenton reagent using human hepatoma cell line.

    Science.gov (United States)

    Wang, Guifang; Lu, Gang; Yin, Pinghe; Zhao, Ling; Yu, Qiming Jimmy

    2016-04-15

    Membrane concentrates of landfill leachates contain organic and inorganic contaminants that could be highly toxic and carcinogenic. In this paper, the genotoxicity of membrane concentrates before and after Fenton and UV-Fenton reagent was assessed. The cytotoxicity and genotoxicity was determined by using the methods of methyltetrazolium (MTT), cytokinesis-block micronucleus (CBMN) and comet assay in human hepatoma cells. MTT assay showed a cytotoxicity of 75% after 24h of exposure to the highest tested concentration of untreated concentrates, and no cytotoxocity for UV-Fenton and Fenton treated concentrates. Both CBMN and comet assays showed increased levels of genotoxicity in cells exposed to untreated concentrates, compared to those occurred in cells exposed to UV-Fenton and Fenton reagent treated concentrates. There was no significant difference between negative control and UV-Fenton treated concentrates for micronucleus and comet assay parameters. UV-Fenton and Fenton treatment, especially the former, were effective methods for degradation of bisphenol A and nonylphenol in concentrates. These findings showed UV-Fenton and Fenton reaction were effective methods for treatment of such complex concentrates, UV-Fenton reagent provided toxicological safety of the treated effluent, and the genotoxicity assays were found to be feasible tools for assessment of toxicity risks of complex concentrates. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Estudo fitoquímico e análise mutagênica das folhas e inflorescências de Erythrina mulungu (Mart. ex Benth. através do teste de micronúcleo em roedores Phytochemical and mutagenic analysis of leaves and inflorescences of Erythrina mulungu (Mart. Ex Benth through micronucleus test in rodents

    Directory of Open Access Journals (Sweden)

    A.P De Bona

    2012-01-01

    Full Text Available Este trabalho teve como objetivo investigar a composição química, estabelecer a dose letal média (DL50 e avaliar os potenciais efeitos mutagênicos do extrato hidroalcoólico de folhas e inflorescências de Erythrina mulungu Mart. ex Benth por meio do teste de micronúcleo em medula óssea de camundongos. Os ensaios fitoquímicos foram realizados através de reações preliminares com mudança de coloração e/ou formação de precipitado; a DL50, por meio da administração intraperitoneal de três concentrações dos extratos, avaliando-se o número de óbitos após 48 horas e o teste de micronúcleo foi feito por meio do método do esfregaço, após exposição dos animais a cinco dias de tratamento. Os resultados fitoquímicos demonstraram presença de açúcares redutores, fenóis e taninos, proteínas e aminoácidos, flavonóides, alcalóides, depsídeos e depsidonas e derivados de cumarina em ambos os órgãos; saponinas espumídicas e esteróides e triterpenóides nas folhas e glicosídeos cardiotônicos e antraquinônicos e alcalóides nas inflorescências. Para a DL50 a folha demonstrou-se atóxica e a inflorescência moderadamente tóxica. Para o teste de micronúcleo, os resultados indicaram ausência de citotoxicidade e genotoxicidade dose-dependente para as folhas e independente da dose para as inflorescências. Assim, esses resultados sugerem que a planta, nas condições analisadas, possui potencial para induzir danos ao DNA.This study aimed to investigate the chemical composition, to establish the mean lethal dose (LD50 and to assess the potential mutagenic effects of hydroalcoholic extract of leaves and inflorescences of Erythrina mulungu Mart. ex Benth by using micronucleus test in bone marrow of mice. Phytochemical assays were carried out through preliminary reactions with color change and/or precipitate formation; the LD50 was obtained by intraperitoneal administration of three concentrations of the extracts, assessing

  4. Comparación entre dos biomodelos murinos (ratones Balb/c y ratas Sprague Dawley en el ensayo de micronúcleos transplacentarios Comparison between two murine biomodles (Balb/c mice and Sprague Dawley rats in a transplacental micronucleus assay

    Directory of Open Access Journals (Sweden)

    Daniel Francisco Arencibia Arrebola

    2012-03-01

    at 14th, 15th and 16th days of gestation, and 24 h after the last inoculation in mice and 48 h in rats, it proceeded to perform euthanasia in the pregnant animals to obtain the fetal liver samples. Results: the fetuses from Sprague Dawley exhibited smaller cytotoxicity and genotoxicity indexes, and the lowest endogenous micronucleus results. The best results of the cytotoxicity and genotoxicity induction for the high micronucleus formation with cyclophosphamide were found in Sprague Dawley rat fetuses, being more susceptible to the genotoxic damage by this mutagen. The clastogenic transplacental power of cyclophosphamide was confirmed whereas this genotoxicity assay was linked to reproduction toxicology. Conclusions: these results suggest that the Sprague Dawley rats fetuses could be better used as biomodels in this assay when cyclophosphamide is employed as positive control through the way of administration and the studied dosage. It could be similarly used in the evaluation of new antigenotoxic drugs with antigenotoxic effect through transplacental administration

  5. In vitro safety and efficacy evaluations of a complex botanical mixture of Eugenia dysenterica DC. (Myrtaceae): Prospects for developing a new dermocosmetic product.

    Science.gov (United States)

    Moreira, Larissa Cleres; de Ávila, Renato Ivan; Veloso, Danillo Fabrini Maciel Costa; Pedrosa, Tatiana Nascimento; Lima, Emerson Silva; do Couto, Renê Oliveira; Lima, Eliana Martins; Batista, Aline Carvalho; de Paula, José Realino; Valadares, Marize Campos

    2017-12-01

    In the context of developing a new natural product-based cosmetic, the in vitro efficacy and safety evaluations of a complex botanical mixture based on Eugenia dysenterica leaf hydroalcoholic extract (EDE) (2.5-1000μg/mL) were carried out. Chromatographic analysis demonstrated the presence of the tannin (ellagic acid) and flavonoids (quercetin and gallic acid) which characterize the EDE as a polyphenol-rich mixture. Using HFF-1 fibroblasts, it was shown that EDE promoted cell regeneration after UVA exposure. It also led to the inhibition of the collagenase, elastase and tyrosinase enzymes, which are involved in skin-related disorders. In terms of toxicological evaluation, the EDE was classified as non-phototoxic through the 3T3 Neutral Red Uptake Phototoxicity Test (OECD N° 432, 2004) and non-eye irritant by Bovine Corneal Opacity and Permeability (OECD N° 437, 2013) assay, in conjunction with corneal histomorphometric analysis. Furthermore, the EDE has no skin sensitization potential as demonstrated by a two-out-of-three prediction model [protein-binding/haptenization (OECD N° 442C, 2015), keratinocyte and dendritic cell activations]. In addition, it was shown that the EDE seems to be non-genotoxic through the cytokinesis-block micronucleus assay (OECD N° 487, 2014) using HepG2 cells. When considered together, these findings support the use of EDE botanical mixture in cosmetic/pharmaceutical products. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. Evaluation of in vitro cytoxicity and genotoxicity of size-fractionated air particles sampled during road tunnel construction.

    Science.gov (United States)

    Dominici, Luca; Guerrera, Elena; Villarini, Milena; Fatigoni, Cristina; Moretti, Massimo; Blasi, Paolo; Monarca, Silvano

    2013-01-01

    In tunnel construction, workers exposed to dust from blasting, gases, diesel exhausts, and oil mist have shown higher risk for pulmonary diseases. A clear mechanism to explain how these pollutants determine diseases is lacking, and alveolar epithelium's capacity to ingest inhaled fine particles is not well characterized. The objective of this study was to assess the genotoxic effect exerted by fine particles collected in seven tunnels using the cytokinesis-block micronuclei test in an in vitro model on type II lung epithelium A549 cells. For each tunnel, five fractions with different aerodynamic diameters of particulate matter were collected with a multistage cascade sampler. The human epithelial cell line A549 was exposed to 0.2 m(3)/mL equivalent of particulate for 24 h before testing. The cytotoxic effects of particulate matter on A549 cells were also evaluated in two different viability tests. In order to evaluate the cells' ability to take up fine particles, imaging with transmission electron microscopy of cells after exposure to particulate matter was performed. Particle endocytosis after 24 h exposure was observed as intracellular aggregates of membrane-bound particles. This morphologic evidence did not correspond to an increase in genotoxicity detected by the micronucleus test.

  7. Moderate acute intake of de-alcoholized red wine, but not alcohol, is protective against radiation-induced DNA damage ex vivo -- results of a comparative in vivo intervention study in younger men.

    Science.gov (United States)

    Greenrod, W; Stockley, C S; Burcham, P; Abbey, M; Fenech, M

    2005-12-11

    Moderate intake of wine is associated with reduced risk of cardiovascular disease and possibly cancer however it remains unclear whether the potential health benefits of wine intake are due to alcohol or the non-alcoholic fraction of wine. We therefore tested the hypothesis that the non-alcoholic fraction of wine protects against genome damage induced by oxidative stress in a crossover intervention study involving six young adult males aged 21-26 years. The participants adhered to a low plant phenolic compound diet for 48 h prior to consuming 300 mL of complete red wine, de-alcoholized red wine or ethanol on separate occasions 1 week apart. Blood samples were collected 0.5, 1.0 and 2.0 h after beverage consumption. Baseline and radiation-induced genome damage was measured using the cytokinesis-block micronucleus assay and total plasma catechin concentration was measured. Consumption of de-alcoholized red wine significantly decreased the gamma radiation-induced DNA damage at 1 and 2 h post-consumption by 20%. In contrast alcohol tended to increase radiation-induced genome damage and complete wine protected against radiation-induced genome damage relative to alcohol. The observed effects were only weakly correlated with the concentration of total plasma catechin (R=-0.23). These preliminary data suggest that only the non-alcoholic fraction of red wine protects DNA from oxidative damage but this effect cannot be explained solely by plasma catechin.

  8. Moderate acute intake of de-alcoholised red wine, but not alcohol, is protective against radiation-induced DNA damage ex vivo-Results of a comparative in vivo intervention study in younger men

    Energy Technology Data Exchange (ETDEWEB)

    Greenrod, W. [CSIRO Health Sciences and Nutrition, Genome Health and Nutrigenomics Laboratory, PO Box 10041, Adelaide BC, SA 5000 (Australia); Department of Clinical and Experimental Pharmacology, University of Adelaide, South Australia (Australia); Stockley, C.S. [Australian Wine Research Institute, South Australia (Australia); Burcham, P. [Department of Clinical and Experimental Pharmacology, University of Adelaide, South Australia (Australia); Abbey, M. [CSIRO Health Sciences and Nutrition, Genome Health and Nutrigenomics Laboratory, PO Box 10041, Adelaide BC, SA 5000 (Australia); Fenech, M. [CSIRO Health Sciences and Nutrition, Genome Health and Nutrigenomics Laboratory, PO Box 10041, Adelaide BC, SA 5000 (Australia)]. E-mail: michael.fenech@hsn.csiro.au

    2005-12-11

    Moderate intake of wine is associated with reduced risk of cardiovascular disease and possibly cancer however it remains unclear whether the potential health benefits of wine intake are due to alcohol or the non-alcoholic fraction of wine. We therefore tested the hypothesis that the non-alcoholic fraction of wine protects against genome damage induced by oxidative stress in a crossover intervention study involving six young adult males aged 21-26 years. The participants adhered to a low plant phenolic compound diet for 48 h prior to consuming 300 mL of complete red wine, dealcoholised red wine or ethanol on separate occasions 1 week apart. Blood samples were collected 0.5, 1.0 and 2.0 h after beverage consumption. Baseline and radiation-induced genome damage was measured using the cytokinesis-block micronucleus assay and total plasma catechin concentration was measured. Consumption of dealcoholised red wine significantly decreased the gamma radiation-induced DNA damage at 1 and 2 h post-consumption by 20%. In contrast alcohol tended to increase radiation-induced genome damage and complete wine protected against radiation-induced genome damage relative to alcohol. The observed effects were only weakly correlated with the concentration of total plasma catechin (R = -0.23). These preliminary data suggest that only the non-alcoholic fraction of red wine protects DNA from oxidative damage but this effect cannot be explained solely by plasma catechin.

  9. Correlation between serum anyloid a low density lipoprotein and genotoxicity in smokers

    International Nuclear Information System (INIS)

    Jamil, A.; Rashid, A.; Majeed, A.; Naveed, A.K.

    2018-01-01

    Objective:To investigate the relation between serum amyloid A-low density lipoprotein (SAA-LDL) and genotoxicity in smokers. Study Design:An experimental study. Place and Duration of Study:Army Medical College, Rawalpindi and National Institute of Health (NIH), Islamabad, from June 2014 to February 2015. Methodology:Seventy healthy Sprague Dawley rats were purchased from NIH and exposed to cigarette smoke in smoke chamber for three months. Blood samples were drawn from each rat at the end of the study period. SAA-LDL was determined by enzyme-linked immunosorbent assay (ELISA). Genotoxicity was assessed by cytokinesis block micronucleus (CBMN) assay. Pearson correlation was used to find correlation between SAA-LDL and genotoxicity. Results:Strong positive correlation was found between SAA-LDL and micronuclei frequency in smoke-exposed rats (r=0.799, N=70, p <0.01). Conclusion:Statistically significant strong positive correlation between SAA-LDL and genotoxicity in smoke-exposed rats shows that changes in one is associated with changes in other and vice versa. (author)

  10. Radioprotective effect of sesamol on γ-radiation induced DNA damage, lipid peroxidation and antioxidants levels in cultured human lymphocytes

    International Nuclear Information System (INIS)

    Prasad, N. Rajendra; Menon, Venugopal P.; Vasudev, V.; Pugalendi, K.V.

    2005-01-01

    Sesamol pretreated (1, 5 and 10 μg/ml) lymphocytes were exposed to different doses of γ-radiation, i.e., 1, 2 and 4 Gray (Gy) and the cellular changes were estimated by using cytokinesis blocked micronucleus assay (MN), dicentric aberration (DC), thiobarbituric acid reactive substances (TBARS), reduced glutathione (GSH) and the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). Radiation significantly increased MN, DC frequencies, TBARS levels and decreased GSH and antioxidant enzyme levels in a dose dependent manner. The highest damage to lymphocytes was observed at 4 Gy irradiation. On the other hand, sesamol pretreatment significantly decreased MN, DC frequencies, TBARS levels and increased GSH levels and SOD, CAT and GPx activities in a concentration dependent manner. At 1 Gy irradiation all concentrations of sesamol (1, 5 and 10 μg/ml) significantly protects the lymphocytes from radiation damage. At 2 Gy irradiation 5 and 10 μg/ml of sesamol shows significant radioprotection. Since the highest damage was observed at 4 Gy irradiation both 1 and 5 μg/ml of sesamol pretreatment were not sufficient to protect the lymphocytes from radiation damage but 10 μg/ml of sesamol significantly (p < 0.05) protects the lymphocytes from radiation effect. Thus, sesamol pretreatment gives significant protection to cultured human lymphocytes against γ-radiation induced cellular damage. The possible mechanism involved in the radioprotective influence of sesamol is discussed

  11. Assay of micronuclei in peripheral blood lymphocytes as a biological indicator of radiation dose

    International Nuclear Information System (INIS)

    Sreedevi, B.; Rao, B.S.

    1994-01-01

    Chromosomal aberration analysis (CA) has regularly been used as a biological dosemeter to evaluate suspected overexposures to ionising radiations. Recently, the micronucleus (MN) assay has been suggested as an alternative method. An attempt has been made to explore the dose response parameters of MN assay in cytokinesis-blocked lymphocytes. Whole blood was irradiated with 60 Co gamma rays or 250 kV p X rays. A dose-dependent increase in micronuclei yield was observed. The dose response could be best described by a linear-quadratic relationship for both gamma rays and X rays. The α and β coefficients were found to be 1.9 x 10 -2 Gy -1 and 5.7 x 10 -2 Gy -2 for gamma rays and 6.3 x 10 -2 Gy -1 and 4.3 x 10 -2 Gy -2 for X rays, respectively. In the low dose region X rays were three times more efficient in inducing micronuclei. The background value derived for 25 samples from healthy individuals ranged from 6-18 micronuclei per 1000 cells, with a mean value of 12 ± 4 x 10 -3 . Biological dose estimates for individuals exposed in the range 0.1-1 Gy made by MN and CA methods yielded similar results for doses ≥ 0.5 Gy. Due to the uncertainties in the background incidence of MN, at present this technique cannot provide reliable estimates at low doses. (author)

  12. In vitro cytogenetic evaluation of the particular combination of flurbiprofen and roxithromycin.

    Science.gov (United States)

    Timocin, Taygun; Husunet, Mehmet Tahir; Valipour, Ebrahim; Norizadeh Tazehkand, Mostafa; Celik, Rima; Topaktas, Mehmet; Ila, Hasan B

    2017-07-01

    Flurbiprofen (FLB) (anti-inflammatory and analgesic drug) and roxithromycin (RXM) (antibiotic) were widely used in world wide. This study deals with investigation of genotoxicity, cytotoxicity, and oxidative stress effects of a particular combination of these drugs in human cultured lymphocytes. Also, DNA damaging-protective effects of combination of these drugs were analyzed on plasmid DNA. Human lymphocytes were treated with different concentrations (FLB + RXM; 10 μg/mL + 25 μg/mL, 15 μg/mL + 50 μg/mL, and 20 μg/mL + 100 μg/mL) of the drugs following by study of their genotoxic and cytotoxic effects by analysis of cytokinesis-block micronucleus test and nuclear division index, respectively. The effect of the combination in aspect of anti-oxidative and DNA damaging activity was evaluated on Pet-22b plasmid. According to our results, the combination of FLB and RXM did not show a notable genotoxic effect on cells. Although each of the substances had been shown as a cytotoxic agent by previous researchers, in this research, the combination of these drugs did not exhibit any adverse effect on cell division. FLB had DNA protection effect against H 2 O 2 while in combination with RXM had not the same effect on the plasmid.

  13. Clastogenic effects in human lymphocytes exposed to low and high dose rate X-ray irradiation and vitamin C

    International Nuclear Information System (INIS)

    Konopacka, M; Rogolinski, J.

    2011-01-01

    In the present work we investigated the ability of vitamin C to modulate clastogenic effects induced in cultured human lymphocytes by X-irradiation delivered at either high (1 Gy/min) or low dose rate (0.24 Gy/min). Biological effects of the irradiation were estimated by cytokinesis-block micronucleus assay including the analysis of the frequency of micronuclei (MN) and apoptotic cells as well as calculation of nuclear division index (NDI). The numbers of micronucleated binucleate lymphocytes (MN-CBL) were 24.85 ± 2.67% and 32.56 ± 3.17% in cultures exposed to X-rays (2 Gy) delivered at low and high dose rates, respectively. Addition of vitamin C (1-20 μg/ml) to the medium of cultures irradiated with the low dose rate reduced the frequency of micronucleated lymphocytes with multiple MN in a concentration-dependent manner. Lymphocytes exposed to the high dose rate radiation showed a U-shape response: low concentration of vitamin C significantly reduced the number of MN, whereas high concentration influenced the radiation-induced total number of micronucleated cells insignificantly, although it increased the number of cells with multiple MN. Addition of vitamin C significantly reduced the fraction of apoptotic cells, irrespective of the X-ray dose rate. These results indicate that radiation dose rate is an important exposure factor, not only in terms of biological cell response to irradiation, but also with respect to the modulating effects of antioxidants. (authors)

  14. Mutagenic Potential ofBos taurus Papillomavirus Type 1 E6 Recombinant Protein: First Description

    Directory of Open Access Journals (Sweden)

    Rodrigo Pinheiro Araldi

    2015-01-01

    Full Text Available Bovine papillomavirus (BPV is considered a useful model to study HPV oncogenic process. BPV interacts with the host chromatin, resulting in DNA damage, which is attributed to E5, E6, and E7 viral oncoproteins activity. However, the oncogenic mechanisms of BPV E6 oncoprotein per se remain unknown. This study aimed to evaluate the mutagenic potential of Bos taurus papillomavirus type 1 (BPV-1 E6 recombinant oncoprotein by the cytokinesis-block micronucleus assay (CBMNA and comet assay (CA. Peripheral blood samples of five calves were collected. Samples were subjected to molecular diagnosis, which did not reveal presence of BPV sequences. Samples were treated with 1 μg/mL of BPV-1 E6 oncoprotein and 50 μg/mL of cyclophosphamide (positive control. Negative controls were not submitted to any treatment. The samples were submitted to the CBMNA and CA. The results showed that BPV E6 oncoprotein induces clastogenesis per se, which is indicative of genomic instability. These results allowed better understanding the mechanism of cancer promotion associated with the BPV E6 oncoprotein and revealed that this oncoprotein can induce carcinogenesis per se. E6 recombinant oncoprotein has been suggested as a possible vaccine candidate. Results pointed out that BPV E6 recombinant oncoprotein modifications are required to use it as vaccine.

  15. Effect of Hecogenin on DNA instability

    Directory of Open Access Journals (Sweden)

    Marina Sampaio Cruz

    Full Text Available Hecogenin is a sapogenin found in Agave species in high quantities and is responsible for the many therapeutic effects of these medicinal plants. In addition, this compound is also widely used in the pharmaceutical industry as a precursor for the synthesis of steroidal hormones and anti-inflammatory drugs. Despite Hecogenin being widely used, little is known about its toxicological properties. Therefore, the present study aimed to investigate the cytotoxic, genotoxic and mutagenic effects of Hecogenin on HepG2 cells. Cytotoxicity was analyzed using the MTT test. Then, genotoxic and mutagenic potentials were assessed by comet assay and cytokinesis-block micronucleus assay, respectively. Cytotoxic effect was observed only when cells were exposed to concentrations of Hecogenin equal or higher than 100 μM. Although a lower concentration of Hecogenin caused DNA damage, a reduction on nuclear mutagenic markers in HepG2 cells was observed. The results indicated that Hecogenin treatment generated DNA damage, but in fact it would be repaired, avoiding dissemination of the damage throughout the cell division. Further studies need to be performed to confirm the observed protective effect of Hecogenin against genomic instability. Keywords: Hecogenin, CBMN, Genotoxicity, Comet assay

  16. Protective Effect of Onion Extract on Bleomycin-Induced Cytotoxicity and Genotoxicity in Human Lymphocytes

    Directory of Open Access Journals (Sweden)

    Yoon Hee Cho

    2016-02-01

    Full Text Available Following one of the world’s largest nuclear accidents, occured at Fukushima, Japan in 2011, a significant scientific effort has focused on minimizing the potential adverse health effects due to radiation exposure. The use of natural dietary antioxidants to reduce the risk of radiation-induced oxidative DNA damage is a simple strategy for minimizing radiation-related cancer rates and improving overall health. The onion is among the richest sources of dietary flavonoids and is an important food for increasing their overall intake. Therefore, we examined the effect of an onion extract on cyto- and geno-toxicity in human lymphocytes treated with bleomycin (BLM, a radiomimetic agent. In addition, we measured the frequency of micronuclei (MN and DNA damage following treatment with BLM using a cytokinesis-blocked micronucleus assay and a single cell gel electrophoresis assay. We observed a significant increase in cell viability in lymphocytes treated with onion extract then exposed to BLM compared to cells treated with BLM alone. The frequency of BLM induced MN and DNA damage increased in a dose-dependent manner; however, when lymphocytes were pretreated with onion extract (10 and 20 μL/mL, the frequency of BLM-induced MN was decreased at all doses of BLM and DNA damage was decreased at 3 μg/mL of BLM. These results suggest that onion extract may have protective effects against BLM-induced cyto- and genotoxicity in human lymphocytes.

  17. Genotoxicity evaluation of HMG CoA reductase inhibitor rosuvastatin.

    Science.gov (United States)

    Berber, Ahmet Ali; Celik, Mustafa; Aksoy, Hüseyin

    2014-07-01

    The genotoxic potential of rosuvastatin as one of the statin drugs was assessed by chromosomal aberrations (CAs), micronucleus (MN) and DNA damage by comet assay in the human peripheral blood lymphocytes. Rosuvastatin was used at concentrations of 0.0625, 0.125, 0.25, 0.5 and 1 µg/mL for these in vitro assays. In all assays, a negative and positive control were also included. CA frequencies were significantly increased in all concentrations at 24 hours and significantly increased in all concentrations except 0.0625 µg/mL at 48 hours, compared to the negative control. Rosuvastatin has a decreased mitotic index (MI) at 0.5- and 1-µg/mL concentrations at 24 hours and at 0.25, 0.5 and 1 µg/mL at 48 hours. A significant increase was observed for induction of MN in all treatments, compared to the negative control. Cytokinesis-block proliferation indices were not affected by treatments with rosuvastatin. In the comet assay, significant increases in comet tail length and tail moment were observed at 0.0625-, 0.5- and 1-µg/mL concentrations. Comet intensity was significantly increased in all concentrations except 0.0625 µg/mL. According to these results, rosuvastatin is cytotoxic and clastogenic/aneugenic in human peripheral lymphocytes. Further studies should be conducted in other test systems to evaluate the full genotoxic potential of rosuvastatin.

  18. A cytogenetic approach to the effects of low levels of ionizing radiations on occupationally exposed individuals

    Energy Technology Data Exchange (ETDEWEB)

    Zakeri, Farideh [National Radiation Protection Department, Iranian Nuclear Regulatory Authority, Nuclear Science and Research Institute-Agriculture, Medicine and Industry Research School, Tehran (Iran, Islamic Republic of); Graduate School of Science and Technology, Chiba University, Inage-ku, Chiba 263-8522 (Japan)], E-mail: fzakeri@aeoi.org.ir; Hirobe, Tomohisa [Graduate School of Science and Technology, Chiba University, Inage-ku, Chiba 263-8522 (Japan); Radiation Effect Mechanism Research Group, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan)

    2010-01-15

    The aim of the present study was to assess occupationally induced chromosomal damage in hospital workers exposed to low levels of ionizing radiation. Thirty-two interventional cardiologists, 36 nuclear medicine physicians and 33 conventional radiologists were included in this study, along with 35 healthy age- and sex-matched individuals as the control group. We used conventional metaphase chromosome aberration (CA) analysis, cytokinesis-block micronucleus (MN) assay as important biological indicators of ionizing radiation exposure. Occupational dosimetry records were collected over the last year (ranged from 0.25 to 48 mSv) and their whole life exposure (ranged from 1.5 to 147 mSv). The results showed significantly higher frequencies of dicentric and acentric CAs (p < 0.001) and MN (p < 0.01) in all exposed groups than in the controls. Taking all the confounding factors into account, no obvious trend of increased chromosomal damages as a function of either duration of employment, exposed dose, sex or age was observed. Interventional cardiologists had the highest rates of CA and MN frequencies between the worker groups, though the differences were not significant. These results indicate that long term exposure to low dose ionizing radiation could result in DNA damage. Hence, the personnel who work in the hospitals should carefully apply the radiation protection procedures.

  19. Radioprotective effect of Haberlea rhodopensis (Friv.) leaf extract on gamma-radiation-induced DNA damage, lipid peroxidation and antioxidant levels in rabbit blood.

    Science.gov (United States)

    Georgieva, Svetlana; Popov, Borislav; Bonev, Georgi

    2013-01-01

    Different concentrations of H. rhodopensis total extract (HRE; 0.03, 0.06 and 0.12 g/kg body weight) were injected im, into rabbits 2 h before collecting the blood samples. The whole blood samples were exposed in vitro to 2.0 Gy 60Co gamma-radiation. The radiation-induced changes were estimated by using the chromosome aberration test (CA) and cytokinesis blocked micronucleus assay (CBMN) in peripheral lymphocytes, and by determining the malondialdehyde levels (MDA) in blood plasma and the superoxide dismutase (SOD) and catalase (CAT) activity in erythrocytes. Radiation significantly increased the chromosome aberration and micronuclei frequencies as well as MDA levels and decreased the antioxidant enzyme activity. On the other hand, the HRE pretreatment significantly decreased the CA, MN frequencies and MDA levels and increased the SOD and CAT activity in a concentration dependent manner. The most effective was the highest concentration of HRE (0.12 g/kg body weight). The results suggest that HRE as a natural product with a nantioxidant capacity could play a modulatory role against the cellular damage induced by gamma-irradiation. The possible mechanism involved in the radioprotective potential of HRE is discussed.

  20. Radioprotective effect of Haberlea rhodopensis (Friv.) leaf extract on γ-radiation-induced DNA damage, lipid peroxidation and antioxidant levels in rabbit blood

    International Nuclear Information System (INIS)

    Georgieva, Svetlana; Bonev, Georgi; Popov, Borislav

    2013-01-01

    Different concentrations of H. rhodopensis total extract (HRE; 0.03, 0.06 and 0.12 g/kg body weight) were injected im, into rabbits 2 h before collecting the blood samples. The whole blood samples were exposed in vitro to 2.0 Gy 60 Co γ-radiation. The radiation-induced changes were estimated by using the chromosome aberration test (CA) and cytokinesis blocked micronucleus assay (CBMN) in peripheral lymphocytes, and by determining the malondialdehyde levels (MDA) in blood plasma and the superoxide dismutase (SOD) and catalase (CAT) activity in erythrocytes. Radiation significantly increased the chromosome aberration and micronuclei frequencies as well as MDA levels and decreased the antioxidant enzyme activity. On the other hand, the HRE pretreatment significantly decreased the CA, MN frequencies and MDA levels and increased the SOD and CAT activity in a concentration dependent manner. The most effective was the highest concentration of HRE (0.12 g/kg body weight). The results suggest that HRE as a natural product with an antioxidant capacity could play a modulatory role against the cellular damage induced by γ-irradiation. The possible mechanism involved in the radioprotective potential of HRE is discussed. (author)

  1. The use of genotoxicity biomarkers in molecular epidemiology: applications in environmental, occupational and dietary studies

    Directory of Open Access Journals (Sweden)

    Carina Ladeira

    2017-08-01

    Full Text Available Molecular epidemiology is an approach increasingly used in the establishment of associations between exposure to hazardous substances and development of disease, including the possible modulation by genetic susceptibility factors. Environmental chemicals and contaminants from anthropogenic pollution of air, water and soil, but also originating specifically in occupational contexts, are potential sources of risk of development of disease. Also, diet presents an important role in this process, with some well characterized associations existing between nutrition and some types of cancer. Genotoxicity biomarkers allow the detection of early effects that result from the interaction between the individual and the environment; they are therefore important tools in cancer epidemiology and are extensively used in human biomonitoring studies. This work intends to give an overview of the potential for genotoxic effects assessment, specifically with the cytokinesis blocked micronucleus assay and comet assay in environmental and occupational scenarios, including diet. The plasticity of these techniques allows their inclusion in human biomonitoring studies, adding important information with the ultimate aim of disease prevention, in particular cancer, and so it is important that they be included as genotoxicity assays in molecular epidemiology.

  2. Three new labdanes isolated from Eragrostis viscosa

    Energy Technology Data Exchange (ETDEWEB)

    Sebastiao, N' Soki N. [Chemistry Department, Agostinho Neto University, Luanda (Angola); Fernandes, Nelson; Vieira, Liliana; Mendonca, Dina I.M.D. de [Textile and Paper Materials Center and Chemistry Department, University of Beira Interior, Covilha (Portugal); Mendonca, Antonio J.G. [CICS-UBI - Health Sciences Research Centre, University of Beira Interior, Covilha, (Portugal); Gaspar, Jorge F.; Martins, Celia; Rueff, Jose, E-mail: disabel@ubi.pt [Genetics Department, Faculty of Medical Sciences, New University of Lisbon, Lisboa (Portugal); Diakanamwa, Carlos [Biology Department, Agostinho Neto University, Luanda (Angola)

    2012-10-15

    Three new labdanes with 8{alpha},15-epoxy ring [methyl 8{alpha},15-epoxylabdan-16{beta}-oate, 8{alpha},15-epoxylabdan-16{beta}-ol and 8{alpha},15-epoxy-16-norlabdan-13{beta}-ol] and five known compounds [8{alpha},15-epoxy-16-norlabdan-13-one, 8{alpha},15-epoxylabdan-16{beta}-oic acid, 3{beta}-(3''4''dihydroxy)-(E)-cinnamoyloxylup-20(29)-ene, 3-(2',3',4',6'-tetra-O-acetyl-{beta}-D-glucopyranosyloxy)-{beta}-sitosterol and 16-acetoxy-8{alpha},15-epoxylabdane] were isolated from toluene and dichloromethane extracts of aerial parts of Eragrostis viscosa. The structures of all the compounds were established based on their spectroscopic data and X-ray diffraction analysis of 8{alpha},15-epoxylabdan-16{beta}-ol. It was also studied the genotoxicity of E. viscosa, particularly compounds 16-acetoxy-8{alpha},15-epoxylabdane, 8{alpha},15-epoxy-16-norlabdan-13-one and 8{alpha},15-epoxilabdan-16{beta}-ol, using a cytokinesis-block micronucleus assay and the Ames test to assess mutagenicity. Both assays were negative. Cytotoxicity was also analyzed using an MTT assay, and 8{alpha},15-epoxy-16{beta}-ol was shown to be the most cytotoxic of the compounds tested. E. viscosa extracts were also tested to determine their antioxidant capacities, peroxide values and total phenolic contents. (author)

  3. Cytogenetic Dosimetry: Applications in Preparedness for and Response to Radiation Emergencies - Training Materials

    International Nuclear Information System (INIS)

    2013-01-01

    These materials are designed for use at a four day training course on the application of cytogenetic dosimetry in preparedness for and response to radiation emergencies. They contain information on: (1) Basics of biological effects of ionizing radiation: Parts 1+2; (2) Basics of dosimetry; (3) dicentric assay; (4) Retrospective dosimetry by translocation analysis; (5) Premature chromosome condensation analysis; (6) Cytokinesis block micronucleus assay; (7) Applied statistics for biodosimetry; (8) Automatic analysis of chromosomal assays; (9) Biodosimetry in mass casualty events; (10) Safety of laboratory staff and quality programmes; (11) Examples of accident investigations; (12) Cytogenetic dose estimation in the criticality accident in Tokaimura; (13) Radiological accidents in Latin America; (14) Radiological accidents in Georgia. Additionally, the CD contains two working sessions with the reference materials for use and a standard training programme. This training course consists of lectures and work sessions that can easily be utilized by a State to build a basic capability in biodosimetry application in a nuclear or radiological emergency

  4. Incidence of chromosomal aberrations and micronuclei in cave tour guides.

    Science.gov (United States)

    Bilban, M; Bilban-Jakopin, C; Vrhovec, S

    2001-01-01

    An analysis of structural chromosomal aberrations (SCA) and micronucleus tests (MN) were performed in 38 subjects, cave tour guides and in appropriate control group. The dominant type of chromosomal aberrations in tourist guides were chromosomal breaks (0.013 per cell) and acentric fragments (0.011 per cell). In the control group, these aberrations were present up to 0.008 on cells. Considering the analysed cells of the guides in total (33,556), the incidence of dicentric and rings range is below 0.0008 on cells, even though three dicentric and ring chromosoms were found already in the first 1000 in vitro metaphases of some guides. Only 0.0003 dicentrics and neither other translocations were found in control group (ambiental exposure). The incidence of micronuclei in cytokinesis blocked lymphocytes ranged from 12-32 per 500 CB cells in the cave tour guides and from 4-11 per 500 CB cells in control group. Measurements of radon and its daughters were performed at different locations in the cave. Annual doses from 40-60 mSv were estimated per 2000 work hours for cave guides. The changes found in the genome of somatic cells may be related to the exposure doses of radon and its daughters, although smoking should not be ignored.

  5. Lycopene: An antioxidant and radioprotector against γ-radiation-induced cellular damages in cultured human lymphocytes

    International Nuclear Information System (INIS)

    Srinivasan, M.; Devipriya, N.; Kalpana, K.B.; Menon, Venugopal P.

    2009-01-01

    The present study aimed to evaluate the radioprotective effect of lycopene, a naturally occurring dietary carotenoid on γ-radiation-induced toxicity. The cellular changes were estimated by using lipid peroxidative indices like thiobarbituric acid reactive substances (TBARS), hydroperoxides (HP), the antioxidants superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and reduced glutathione (GSH). The DNA damage was analyzed by cytokinesis blocked micronucleus assay (CBMN), dicentric aberration (DC) and translocation frequency. The γ-radiation at different doses (1, 2 and 4 Gy) resulted in a significant increase in the number of micronuclei (MN), DC, translocation frequency, TBARS and HP level, whereas the levels of GSH and antioxidant enzymes were significantly decreased when compared with normal control. The maximum damage to lymphocytes was observed at 4 Gy irradiation. Lycopene pretreatment (1, 5 and 10 μg/ml) significantly decreased the frequency of MN, DC and translocation when compared with γ-radiation control. The levels of TBARS, HP were also decreased and activities of SOD, CAT and GPx were significantly increased along with GSH levels when compared with γ-radiation control. The dose of 5 μg/ml of lycopene was found to be more effective than the other two doses. Thus, our result shows that pretreatment with lycopene offers protection to normal lymphocytes against γ-radiation-induced cellular damage.

  6. Occupational Exposure in Nuclear Medicine - a Case Study

    International Nuclear Information System (INIS)

    Rakic, B.; Pajic, J.; Prokic, V.; Mirkov, Z.; Marinkovic, O.; Jovicic, D.

    2011-01-01

    Chromosomal aberrations (CA) analysis and cytokinesis-block micronucleus (CBMN) test are standardised techniques used in dose assessment of subjects professionally, medically and accidentally exposed to ionising radiation. At this time, there is no best applicable method for dose assessment in case of internal contamination with radionuclides. Available bioassays are more or less sensitive, depending on particular irradiation. Existing physical methods for dose assessment of subjects operating open sources in most cases are not enough for accurate dose assessment, and there is a need for implementing additional methods, namely, extremity dosimetry. There is, very often, disagreement in dose assessment in physical and biodosimetry in case of exposure to low doses of IR. However, in certain situations, both bioassays and available physical dosimetry methods seem to be quite enough in detecting internal contamination. In this paper are presented values for personal equivalent doses Hp(10) and Hp(0.07) (from Personal Dosimetry Department - SIOH) and results of CA and MN analyses after first and control examinations from Cytogenetic Laboratory of same Institute. This paper also showed that CBMN test is more sensitive technique in case of internal contamination than CA analysis. (author)

  7. Moderate acute intake of de-alcoholised red wine, but not alcohol, is protective against radiation-induced DNA damage ex vivo-Results of a comparative in vivo intervention study in younger men

    International Nuclear Information System (INIS)

    Greenrod, W.; Stockley, C.S.; Burcham, P.; Abbey, M.; Fenech, M.

    2005-01-01

    Moderate intake of wine is associated with reduced risk of cardiovascular disease and possibly cancer however it remains unclear whether the potential health benefits of wine intake are due to alcohol or the non-alcoholic fraction of wine. We therefore tested the hypothesis that the non-alcoholic fraction of wine protects against genome damage induced by oxidative stress in a crossover intervention study involving six young adult males aged 21-26 years. The participants adhered to a low plant phenolic compound diet for 48 h prior to consuming 300 mL of complete red wine, dealcoholised red wine or ethanol on separate occasions 1 week apart. Blood samples were collected 0.5, 1.0 and 2.0 h after beverage consumption. Baseline and radiation-induced genome damage was measured using the cytokinesis-block micronucleus assay and total plasma catechin concentration was measured. Consumption of dealcoholised red wine significantly decreased the gamma radiation-induced DNA damage at 1 and 2 h post-consumption by 20%. In contrast alcohol tended to increase radiation-induced genome damage and complete wine protected against radiation-induced genome damage relative to alcohol. The observed effects were only weakly correlated with the concentration of total plasma catechin (R = -0.23). These preliminary data suggest that only the non-alcoholic fraction of red wine protects DNA from oxidative damage but this effect cannot be explained solely by plasma catechin

  8. Effects of genistein following fractionated lung irradiation in mice

    International Nuclear Information System (INIS)

    Para, Andrea E.; Bezjak, Andrea; Yeung, Ivan W.T.; Van Dyk, Jake; Hill, Richard P.

    2009-01-01

    Background and purpose: This study investigated protection of lung injury by genistein following fractionated doses of radiation and its effect on tumor response. Material and methods: C3H/HeJ mice were irradiated (100 kVp X-rays) with 9 fractions of 3.1 Gy over 30 days (approximately equivalent to 10 Gy single dose) and were maintained on a genistein diet (∼10 mg/kg). Damage was assessed over 28 weeks in lung cells by a cytokinesis block micronucleus (MN) assay and by changes in breathing rate and histology. Tumor protection was assessed using a colony assay to determine cell survival following in situ irradiation of small lung nodules (KHT fibrosarcoma). Results: Genistein caused about a 50% reduction in the MN damage observed during the fractionated radiation treatment and this damage continued to decrease at later times to background levels by 16 weeks. In mice not receiving Genistein MN levels remained well above background out to 28 weeks after irradiation. Genistein reduced macrophage accumulation by 22% and reduced collagen deposition by 28%. There was minimal protection against increases in breathing rate or severe morbidity during pneumonitis. No tumor protection by genistein treatment was observed. Conclusions: Genistein at the dose levels used in this study partially reduced the extent of fibrosis developing in mouse lung caused by irradiation but gave minimal protection against pneumonitis. There was no evidence that genistein caused protection of small tumors growing in the lung.

  9. Water mutagenic potential assessment on a semiarid aquatic ecosystem under influence of heavy metals and natural radioactivity using micronuclei test.

    Science.gov (United States)

    Chaves, Luiz Cláudio Cardozo; Navoni, Julio Alejandro; de Morais Ferreira, Douglisnilson; Batistuzzo de Medeiros, Silvia; Ferreira da Costa, Thomas; Petta, Reinaldo Antônio; Souza do Amaral, Viviane

    2016-04-01

    The contamination of water bodies by heavy metals and ionizing radiation is a critical environmental issue, which can affect water quality and, thus, human health. This study aimed to evaluate the water quality of the Boqueirão de Parelhas Dam in the Brazilian semiarid region. A 1-year study (2013-2014) was performed through the assessment of physicochemical parameters, heavy metal content, and radioactivity along with the mutagenicity potential of water using micronuclei test in Orechromis niloticus (in vivo) and the cytokinesis-block micronucleus (CBMN) assay in human lymphocytes (in vitro). A deterioration of water organoleptics characteristics by the presence of high levels of sulfate and total solids was observed. High concentrations of aluminum, nickel, silver, and lead along with the alpha particle content were higher than the limits suggested by the World Health Organization and Brazilian legislation for drinking water. An increase in the frequency of micronuclei and nuclear abnormalities was observed in both experimental models. The results obtained confirmed the mutagenic potential present in water samples. This study highlights that geogenic agents affect water quality becoming a human health concern to be taken into account due to the relevance that this water reservoir has in the region.

  10. Three new labdanes isolated from Eragrostis viscosa

    International Nuclear Information System (INIS)

    Sebastiao, N'Soki N.; Fernandes, Nelson; Vieira, Liliana; Mendonca, Dina I.M.D. de; Mendonca, Antonio J.G.; Gaspar, Jorge F.; Martins, Celia; Rueff, Jose; Diakanamwa, Carlos

    2012-01-01

    Three new labdanes with 8α,15-epoxy ring [methyl 8α,15-epoxylabdan-16β-oate, 8α,15-epoxylabdan-16β-ol and 8α,15-epoxy-16-norlabdan-13β-ol] and five known compounds [8α,15-epoxy-16-norlabdan-13-one, 8α,15-epoxylabdan-16β-oic acid, 3β-(3''4''dihydroxy)-(E)-cinnamoyloxylup-20(29)-ene, 3-(2',3',4',6'-tetra-O-acetyl-β-D-glucopyranosyloxy)-β-sitosterol and 16-acetoxy-8α,15-epoxylabdane] were isolated from toluene and dichloromethane extracts of aerial parts of Eragrostis viscosa. The structures of all the compounds were established based on their spectroscopic data and X-ray diffraction analysis of 8α,15-epoxylabdan-16β-ol. It was also studied the genotoxicity of E. viscosa, particularly compounds 16-acetoxy-8α,15-epoxylabdane, 8α,15-epoxy-16-norlabdan-13-one and 8α,15-epoxilabdan-16β-ol, using a cytokinesis-block micronucleus assay and the Ames test to assess mutagenicity. Both assays were negative. Cytotoxicity was also analyzed using an MTT assay, and 8α,15-epoxy-16β-ol was shown to be the most cytotoxic of the compounds tested. E. viscosa extracts were also tested to determine their antioxidant capacities, peroxide values and total phenolic contents. (author)

  11. Ficus racemosa Stem Bark Extract: A Potent Antioxidant and a Probable Natural Radioprotector

    Directory of Open Access Journals (Sweden)

    V. P. Veerapur

    2009-01-01

    Full Text Available Ethanol extract (FRE and water extract (FRW of Ficus racemosa (family: Moraceae were subjected to free radical scavenging both by steady state and time resolved methods such as nanosecond pulse radiolysis and stopped-flow spectrophotometric analyses. FRE exhibited significantly higher steady state antioxidant activity than FRW. FRE exhibited concentration dependent DPPH, ABTS•-, hydroxyl radical and superoxide radical scavenging and inhibition of lipid peroxidation with IC50 comparable with tested standard compounds. In vitro radioprotective potential of FRE was studied using micronucleus assay in irradiated Chinese hamster lung fibroblast cells (V79. Pretreatment with different doses of FRE 1h prior to 2 Gy γ-radiation resulted in a significant (P < 0.001 decrease in the percentage of micronucleated binuclear V79 cells. Maximum radioprotection was observed at 20 μg/ml of FRE. The radioprotection was found to be significant (P < 0.01 when cells were treated with optimum dose of FRE (20 μg/ml 1 h prior to 0.5, 1, 2, 3 and 4 Gy γ-irradiation compared to the respective radiation controls. The cytokinesis-block proliferative index indicated that FRE does not alter radiation induced cell cycle delay. Based on all these results we conclude that the ethanol extract of F. racemosa acts as a potent antioxidant and a probable radioprotector.

  12. Phyllanthus emblica Fruit Extract Activates Spindle Assembly Checkpoint, Prevents Mitotic Aberrations and Genomic Instability in Human Colon Epithelial NCM460 Cells

    Directory of Open Access Journals (Sweden)

    Xihan Guo

    2016-09-01

    Full Text Available The fruit of Phyllanthus emblica Linn. (PE has been widely consumed as a functional food and folk medicine in Southeast Asia due to its remarkable nutritional and pharmacological effects. Previous research showed PE delays mitotic progress and increases genomic instability (GIN in human colorectal cancer cells. This study aimed to investigate the similar effects of PE by the biomarkers related to spindle assembly checkpoint (SAC, mitotic aberrations and GIN in human NCM460 normal colon epithelial cells. Cells were treated with PE and harvested differently according to the biomarkers observed. Frequencies of micronuclei (MN, nucleoplasmic bridge (NPB and nuclear bud (NB in cytokinesis-block micronucleus assay were used as indicators of GIN. Mitotic aberrations were assessed by the biomarkers of chromosome misalignment, multipolar division, chromosome lagging and chromatin bridge. SAC activity was determined by anaphase-to- metaphase ratio (AMR and the expression of core SAC gene budding uninhibited by benzimidazoles related 1 (BubR1. Compared with the control, PE-treated cells showed (1 decreased incidences of MN, NPB and NB (p < 0.01; (2 decreased frequencies of all mitotic aberration biomarkers (p < 0.01; and (3 decreased AMR (p < 0.01 and increased BubR1 expression (p < 0.001. The results revealed PE has the potential to protect human normal colon epithelial cells from mitotic and genomic damages partially by enhancing the function of SAC.

  13. Ultrasound-mediated structural changes in cells revealed by FTIR spectroscopy: A contribution to the optimization of gene and drug delivery

    Science.gov (United States)

    Grimaldi, Paola; Di Giambattista, Lucia; Giordani, Serena; Udroiu, Ion; Pozzi, Deleana; Gaudenzi, Silvia; Bedini, Angelico; Giliberti, Claudia; Palomba, Raffaele; Congiu Castellano, Agostina

    2011-12-01

    Ultrasound effects on biological samples are gaining a growing interest concerning in particular, the intracellular delivery of drugs and genes in a safe and in a efficient way. Future progress in this field will require a better understanding of how ultrasound and acoustic cavitation affect the biological system properties. The morphological changes of cells due to ultrasound (US) exposure have been extensively studied, while little attention has been given to the cells structural changes. We have exposed two different cell lines to 1 MHz frequency ultrasound currently used in therapy, Jurkat T-lymphocytes and NIH-3T3 fibroblasts, both employed as models respectively in the apoptosis and in the gene therapy studies. The Fourier Transform Infrared (FTIR) Spectroscopy was used as probe to reveal the structural changes in particular molecular groups belonging to the main biological systems. The genotoxic damage of cells exposed to ultrasound was ascertained by the Cytokinesis-Block Micronucleus (CBMN) assay. The FTIR spectroscopy results, combined with multivariate statistical analysis, regarding all cellular components (lipids, proteins, nucleic acids) of the two cell lines, show that Jurkat cells are more sensitive to therapeutic ultrasound in the lipid and protein regions, whereas the NIH-3T3 cells are more sensitive in the nucleic acids region; a meaningful genotoxic effect is present in both cell lines only for long sonication times while in the Jurkat cells also a significant cytotoxic effect is revealed for long times of exposure to ultrasound.

  14. Discrimination of uranium chemo-toxic and radio-toxic effects: definition of biological markers for evaluating professional risks in nuclear industry

    International Nuclear Information System (INIS)

    Darolles, Carine

    2010-01-01

    Uranium (U) is a heavy metal that is also considered as an alpha emitter. Thus the origin of U toxicity is both chemical and radiological. The identification of bio-markers to discriminate chemical and radiological toxicity for a given U compound is required to assess accurately the health effects of isotopic mixtures such as depleted U in 235 U with a low specific activity. Data from the literature show that the best candidates are cytogenetic markers. In the present work, the assessment of bio-markers of U contamination was performed on three cellular models (mouse fibroblasts, rat lymphocytes and human lymphocytes) that were exposed to different isotopic mixtures of U. The cytokinesis-block micronucleus (MN) centromere assay was performed to discriminate the chemo-toxic and radio-toxic effects of U. This study showed that the evaluation of micronuclei in bi-nucleated cells could not assess U genotoxicity accurately. Instead, the assessment of centromere-negative micronuclei and nucleo-plasmic bridges correlated with the radio-toxic effects of U. The evaluation of centromere-positive micronuclei and micronuclei in mono-nucleated cells correlated with the chemo-toxic effects of U. These cytogenetic markers should be validated on different biological models and could be proposed to discriminate radiological and chemical toxicity of a given isotopic mixture of U. These four cytogenetic markers could be a useful complement of the classical dosimetric bio-markers for the assessment of internal uranium contamination. (author)

  15. Cytotoxicity and genotoxicity of Agaricus blazei methanolic extract fractions assessed using gene and chromosomal mutation assays

    Directory of Open Access Journals (Sweden)

    Marilanda Ferreira Bellini

    2008-01-01

    Full Text Available Functional food investigations have demonstrated the presence of substances that could be beneficial to human health when consumed. However, the toxic effects of some substances contained in foods have been determined. Reported medicinal and nutritive properties have led to the extensive commercialization of the basidiomycete fungi Agaricus blazei Murrill (sensu Heinemann, also known as Agaricus brasiliensis Wasser et al., Agaricus subrufescens Peck or the Brazilian medical mushroom (BMM. Different methanolic extract fractions (ME of this mushroom were submitted to the cytokinesis-block micronucleus (CBMN clastogenic assay and the hypoxanthine-guanine phosphoribosyl transferase locus (HGPRT assay for gene mutation, both using Chinese hamster ovary cells clone K1 (CHO-K1. The results suggest that all the fractions tested possess cytotoxic and mutagenic potential but no clastogenic effects. Further information is needed on the biochemical components of the A. blazei methanol fractions to identify any substances with cytotoxic and/or mutagenicity potential. These findings indicate that A. blazei methanolic extract should not be used due to their genotoxicity and care should be taken in the use of A. blazei by the general population until further biochemical characterization of this fungi is completed.

  16. Evaluation of genotoxic and anti-mutagenic properties of cleistanthin A and cleistanthoside A tetraacetate.

    Science.gov (United States)

    Himakoun, Lakana; Tuchinda, Patoomratana; Puchadapirom, Pranom; Tammasakchai, Ratigon; Leardkamolkarn, Vijittra

    2011-01-01

    Cleistanthin A (CleinA) and cleistanthoside A (CleisA) isolated from plant Phyllanthus taxodiifolius Beille have previously shown potent anticancer effects. To promote their medicinal benefits, CleisA was modified to cleistanthoside A tetraacetate (CleisTA) and evaluated for genotoxic and anti-mutagenic properties in comparison with CleinA. Both compounds showed no significant mutagenic activity to S. typhimulium bacteria and no cytotoxic effect to normal mammalian cells. The non genotoxic effect of CleinA was further confirmed by un-alteration of cytokinesis-block proliferation index (CBPI) and micronucleus (MN) frequency assays in Chinese hamster lung fibroblast (V79) cells, and of CleisTA was confirmed by un-changes of human peripheral blood lymphocytes (HPBL) chromosomal structure assay. Moreover, the metabolic form of CleinA efficiently demonstrated cytostasis effect to V79 cell and prevented mutagen induced Salmonella TA98 and TA100 reversion, whereas both metabolic and non-metabolic forms of CleisTA reduced HPBL mitotic index (%M.I) in a concentration-dependent relationship. The results support CleinA and CleisTA as the new lead compounds for anti-cancer drug development.

  17. Monitoramento da ação genotóxica em trabalhadores de sapatarias através do teste de micronúcleos, Pelotas, Rio Grande do Sul Monitoring the genotoxic action in shoe workers by micronucleus test, Pelotas, Rio Grande do Sul State

    Directory of Open Access Journals (Sweden)

    Natália Silveira Corrêa

    2009-12-01

    Full Text Available Neste trabalho, investigou-se a frequência de micronúcleos em células esfoliadas da mucosa bucal de trabalhadores de sapatarias, na cidade de Pelotas (RS. O estudo constou de 54 trabalhadores de sapatarias expostos à cola e solventes e 54 controles. Avaliou-se a incidência de células com micronúcleos(CMN, binucleadas(CBN, núcleos ligados(CNL e total de anomalias(TA, em 2.000 células por indivíduo. Elaborou-se um banco de dados no programa SPSS "for Windows" pelo teste de Mann-Whitney U, pIn this paper it was investigated the micronuclei frequency in exfoliated oral mucosa cells in shoe shop workers in the city of Pelotas, RS. The study counted on 54 shoe workers exposed to glue and solvents and 54 controls. It was evaluated the incidence of cells with micronucleus (CMN, bi-nucleus (CBN, linked nucleus (CLN and total amount of anomalies (TAA, in 2000 cells per person. A database was created in the SPSS "for Windows" software using the Mann-Whitney U, p<0.05 test. The average of anomalies among shoe workers was 8.69±6.49CMN; 8.85±4.92CBN; 5.78±4.78CNL; 23.31±10.01TA, in the controlled 4.00±5.05CMN; 4.63±4.35CBN; 4.76±5.00CNL; 13.39±9.43TA (p=0.0001; p=0.0001; p=0.144 and p=0.0001 respectively. It was also evaluated the age, gender, time of work, family income, smoke, alcohol beverages, the influence of dermatological, ophthalmological, respiratory and central nervous system (CNS diseases in the number of cell anomalies. These items did not have any influence. It was only observed that among the age group of 15 to 29 years old the number of CNL was bigger than among the age group of 45 to 72. Among those with time of work of 0.1 and 10 years presented a higher CNM than in the other group range.

  18. Genotoxicity assessment of membrane concentrates of landfill leachate treated with Fenton reagent and UV-Fenton reagent using human hepatoma cell line

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Guifang [Department of Chemistry, Jinan University, Guangzhou 510632 (China); Lu, Gang [Key Laboratory of Water/Soil Toxic Pollutants Control and Bioremediation of Guangdong Higher Education Institutes, Department of Environmental Engineering, Jinan University, Guangzhou 510632 (China); Yin, Pinghe, E-mail: tyinph@jnu.edu.cn [Research Center of Analysis and Test, Jinan University, Guangzhou 510632 (China); Zhao, Ling, E-mail: zhaoling@jnu.edu.cn [Key Laboratory of Water/Soil Toxic Pollutants Control and Bioremediation of Guangdong Higher Education Institutes, Department of Environmental Engineering, Jinan University, Guangzhou 510632 (China); Jimmy Yu, Qiming [Griffith School of Engineering, Griffith University, Nathan Campus, Brisbane, Queensland 4111 (Australia)

    2016-04-15

    Highlights: • Membrane concentrates have a threat to human health and environment. • Untreated membrane concentrates induces cytotoxic and genotoxic to HepG2 cells. • Both methods were effective method for degradation of BPA and NP in concentrates. • Both methods were efficient in reducing genotoxic effects of concentrates. • UV-Fenton reagent had higher removal efficiency and provides toxicological safety. - Abstract: Membrane concentrates of landfill leachates contain organic and inorganic contaminants that could be highly toxic and carcinogenic. In this paper, the genotoxicity of membrane concentrates before and after Fenton and UV-Fenton reagent was assessed. The cytotoxicity and genotoxicity was determined by using the methods of methyltetrazolium (MTT), cytokinesis-block micronucleus (CBMN) and comet assay in human hepatoma cells. MTT assay showed a cytotoxicity of 75% after 24 h of exposure to the highest tested concentration of untreated concentrates, and no cytotoxocity for UV-Fenton and Fenton treated concentrates. Both CBMN and comet assays showed increased levels of genotoxicity in cells exposed to untreated concentrates, compared to those occurred in cells exposed to UV-Fenton and Fenton reagent treated concentrates. There was no significant difference between negative control and UV-Fenton treated concentrates for micronucleus and comet assay parameters. UV-Fenton and Fenton treatment, especially the former, were effective methods for degradation of bisphenol A and nonylphenol in concentrates. These findings showed UV-Fenton and Fenton reaction were effective methods for treatment of such complex concentrates, UV-Fenton reagent provided toxicological safety of the treated effluent, and the genotoxicity assays were found to be feasible tools for assessment of toxicity risks of complex concentrates.

  19. Radiation induced chromosomal instability in lymphocytes of cancer patients

    International Nuclear Information System (INIS)

    Sudo, H.; Sagara, M.; Ban, S.; Noda, S.; Iwakawa, M.; Harada, Y.; Imai, T.; Cologne, J.B.

    2003-01-01

    Full text: Cytokinesis-blocked micronucleus (CBMN) assay has been extensively used to evaluate the radiation sensitivity of human individuals. Using the CBMN assay, Scott et al (1998, 1999) demonstrated that a fraction of radiosensitive individuals in breast cancer case population was larger than in normal individual population. However, Vral et al were very skeptical about the Scott et al's findings (2002). Under the approval from the ethical committee of NIRS, peripheral blood was obtained from 46 normal healthy females, 131 breast cancer patients, 32 cervical cancer patients and 7 female head and neck cancer patients. Radiosensitivity of T-lymphocytes was assessed by using a CBMN assay. The frequencies of MN per binucleated cell in healthy donors were 0.031(±0.010) and 0.151(±0.066) for cells treated before and after X-ray-irradiation (2Gy), respectively. Spontaneous MN frequencies in cancer patients were significantly higher than healthy donors (p < 0.001). Radiation sensitivities of breast- and head and neck-cancer patients were significantly higher than normal individuals (p < 0.001). Cervical cancer patients were more resistant to irradiation than healthy donors, though the number of cases for statistical analysis was small. (p < 0.001). We are considering that the HPV infection affected the radiosensitivity of cervical cancer cases. Because it is widely believed that one key mechanism which leads to spontaneous micronucleus formation involves an imbalance of chromosomal segregation and a chromosomal instability in patients' lymphocytes might be greater than that in normal individuals' lymphocytes. Recently, Kuschel et al (2002) demonstrated that ratios in two SNPs on XRCC3 were significantly different between cancer patients and healthy females. Then, we can suppose that the radiation-related genes with low penetrance may be involved in tumorigenesis of mammary- and head and neck-cells, and also, in patients' radiation susceptibility

  20. Bystander effects of exposure to low-dose-rate 125I seeds on human lung cancers cells in vitro

    International Nuclear Information System (INIS)

    Jia Rongfei; Chen Honghong; Yu Lei; Zhao Meijia; Shao Chunlin; Cheng Wenying

    2007-01-01

    The bystander effects induced by continuous low-dose-rate (LDR) 125 I seeds radiation on damage of human lung cancer cells were investigated. Human adenocarcinoma cell line A549 and human small cell lung cancer cell line NCI-H446, which have different sensitivities to high-dose rate (HDR) external irradiation, were exposed directly to 125 I seeds in vitro and co-cultured with unirradiated cells for 24 h. Using cytokinesis-blocking micronucleus method and γ H2AX fluorescence immunoassay, bystander effects induced by 2Gy and 4Gy 125 I seed irradiation on micronucleus formation and DNA double-strand breaks (DSBs) of human lung cancer cells were detected and evaluated. The results showed that irradiation with 125 I seeds can induce medium-mediated bystander effects in A549 cells and NCI-H446 cells, exhibiting that both micronuclei formation and γ H2AX focus formation in bystander cells were increased significantly compared with non-irradiated cells. The extent of DNA damage induced by bystander effects was correlated with accumulated radiation dose and radiosensitive of tumor cells. NCI-H446 cells that were sensitive to HDR γ irradiation were more sensitive to continuous LDR irradiation and bystander effects than A549. However, a comparison between the bystander effects and direct effects elicits the intensity of bystander responses of A549 cells was higher than that of NCI-H446 cells. A dose-related reduction in bystander responses was observed both in A549 cells and NCI-H446 cells, suggesting that the signaling factors involved in the bystander signaling pathways may decrease with the increase of cell damages. (authors)

  1. Occupational health risks among trichloroethylene-exposed workers in a clock manufacturing factory.

    Science.gov (United States)

    Singthong, Siriporn; Pakkong, Pannee; Choosang, Kantima; Wongsanit, Sarinya

    2014-08-22

    Trichloroethylene (TCE) is an important volatile organic compound once widely used in industry throughout the world. Occupational exposure to TCE can cause a number of health hazards such as allergic reactions and genetic damage. The purpose of this study was to evaluate occupational exposure to TCE, by analysis of the air in the breathing zone and of urine from workers employed in a clock manufacturing factory. A subjective symptom survey was conducted by using a self-administered questionnaire to evaluate the health hazards. Micronucleus (MN) frequency, based on the cytokinesis-block micronucleus assay (CBMN) in peripheral blood lymphocytes, (PBLs) was used as a biomarker for chromosome damage. A total of 244 participants, including 171 workers occupationally exposed to TCE and 73 non-exposed control employees, working mainly in office jobs in the same factory, were enrolled in this study. Analyses of airborne TCE concentrations in the workplace, and of urinary trichloroacetic acid (TCA) of the workers and controls, were performed by Gas Chromatography-Electron Capture Detector (GC-ECD) using the modified headspace technique. The average concentration of TCE in the workplace breathing zone was 27.83 ± 6.02 ppm. The average level of urinary TCA of the exposed workers and controls was 14.84 ± 1.62, 2.95 ± 0.28 mg/L. The frequency of MN/1000BN was 7.029 ± 0.39, significantly higher than for those in the control group (3.57 ± 0.31, p = 0.001). According to multiple linear regression analysis, the results indicated that urinary TCA levels correlated with the increased MN in exposed workers (r = 0.285, p trichloroethylene exposure. The use of TCE in the factory is threatening workers' health.

  2. Protein energy-malnutrition: does the in vitro zinc sulfate supplementation improve chromosomal damage repair?

    Science.gov (United States)

    Padula, Gisel; González, Horacio F; Varea, Ana; Seoane, Analía I

    2014-12-01

    Protein-energy malnutrition (PEM) is originated by a cellular imbalance between nutrient/energy supply and body's demand. Induction of genetic damage by PEM was reported. The purpose of this study was to determine the genetic effect of the in vitro zinc sulfate (ZnSO4) supplementation of cultured peripheral blood lymphocytes from children with PEM. Twenty-four samples from 12 children were analyzed. Anthropometric and biochemical diagnosis was made. For the anthropometric assessment, height-for-age index, weight-for-age index, and weight-for-height index were calculated (WHO, 2005). Micronutrient status was evaluated. A survey for assessed previous exposure to potentially genotoxic agents was applied. Results were statistically evaluated using paired sample t test and χ (2) test. Each sample was fractionated and cultured in two separate flasks to performed two treatments. One was added with 180 μg/dl of ZnSO4 (PEMs/ZnSO4) and the other remains non-supplemented (PEMs). Cytotoxic effects and chromosomal damage were assessed using the cytokinesis-block micronucleus assay (CBMN). All participants have at least one type of malnutrition and none have anemia, nor iron, folate, vitamin A, and zinc deficiency. All PEMs/ZnSO4 samples have a significant reduction in the micronucleus (MNi) frequency compared with PEMs (t = 6.25685; p < 0.001). Nuclear division index (NDI) increase in PEMs/ZnSO4 (t = -17.4226; p < 0.001). Nucleoplasmic bridge (NPBs) frequency was four times smaller in PEMs/ZnSO4 (χ (2) = 40.82; p < 0.001). No nuclear buds (NBuds) were observed. Cytotoxic effects and chromosomal damage observed in children suffering from PEM can be repaired in vitro with zinc sulfate supplementation.

  3. Comparative study of genetic activity of chlorambucil's active metabolite steroidal esters: The role of steroidal skeleton on aneugenic potential

    International Nuclear Information System (INIS)

    Efthimiou, M.; Ouranou, D.; Stephanou, G.; Demopoulos, N.A.; Nikolaropoulos, S.S.; Alevizos, Ph.

    2010-01-01

    p-N,N-bis(2-chloroethyl)aminophenylacetic acid (PHE), a nitrogen mustard analogue and chlorambucil's active metabolite used as chemotherapeutic agent, has been shown that, in addition to its clastogenic activity, induces chromosome delay. In the present study an efford has been made (a) to investigate if the steroidal analogues of PHE (EA-92, EA-97, AK-333, AK-409 and AK-433) exert the same genetic activity as the parent compound, (b) to further analyze the aneugenic activity of nitrogen mustard analogues, (c) to investigate the mechanism by which they exert aneugenic potential and (d) to correlate the genetic activity with chemical structure. For this purpose the Cytokinesis Block Micronucleus (CBMN) assay was conducted in human lymphocytes in vitro and the micronucleus (MN) frequency was determined to investigate their genetic activity. The mechanism of micronucleation was determined in combination with Fluorescence In Situ Hybridization (FISH) using pancentromeric DNA probe. Since one of the mechanisms that chemicals cause aneuploidy is through alterations in the mitotic spindle, we also investigated the effect of the above compounds on the integrity and morphology of the mitotic spindle using double immunofluorescence of β- and γ-tubulin in C 2 C 12 mouse cell line. We found that PHE and its steroidal analogues, EA-92, EA-97, AK-333, AK-409 and AK-433, affect cell proliferation in human lymphocytes and C 2 C 12 mouse cells. All studied compounds are capable of inducing chromosome breakage events, as indicated by the enhanced C - MN frequencies. The less lipophilic compounds are the most genetically active molecules. PHE and only two of the studied analogues, AK-409 and AK-433, the most hydrophilic ones, showed aneugenic potential, by increasing the frequencies of MN containing a whole chromosome. The aneugenic potential of the above referred analogues is associated with amplification of centrosome number, since they caused high multipolar metaphase

  4. Increased levels of etheno-DNA adducts and genotoxicity biomarkers of long-term exposure to pure diesel engine exhaust.

    Science.gov (United States)

    Shen, Meili; Bin, Ping; Li, Haibin; Zhang, Xiao; Sun, Xin; Duan, Huawei; Niu, Yong; Meng, Tao; Dai, Yufei; Gao, Weimin; Yu, Shanfa; Gu, Guizhen; Zheng, Yuxin

    2016-02-01

    Etheno-DNA adducts are biomarkers for assessing oxidative stress. In this study, the aim was to detect the level of etheno-DNA adducts and explore the relationship between the etheno-DNA adducts and genotoxicity biomarkers of the diesel engine exhaust (DEE)-exposed workers. We recruited 86 diesel engine testing workers with long-term exposure to DEE and 99 non-DEE-exposed workers. The urinary mono-hydroxylated polycyclic aromatic hydrocarbons (OH-PAHs) and etheno-DNA adducts (εdA and εdC) were detected by HPLC-MS/MS and UPLC-MS/MS, respectively. Genotoxicity biomarkers were also evaluated by comet assay and cytokinesis-block micronucleus assay. The results showed that urinary εdA was significantly higher in the DEE-exposed workers (p<0.001), exhibited 2.1-fold increase compared with the non-DEE-exposed workers. The levels of urinary OH-PAHs were positively correlated with the level of εdA among all the study subjects (p<0.001). Moreover, we found that the increasing level of εdA was significantly associated with the increased olive tail moment, percentage of tail DNA, or frequency of micronucleus in the study subjects (p<0.01). No significant association was observed between the εdC level and any measured genotoxicity biomarkers. In summary, εdA could serve as an indicator for DEE exposure in the human population. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Genotoxicity assessment of membrane concentrates of landfill leachate treated with Fenton reagent and UV-Fenton reagent using human hepatoma cell line

    International Nuclear Information System (INIS)

    Wang, Guifang; Lu, Gang; Yin, Pinghe; Zhao, Ling; Jimmy Yu, Qiming

    2016-01-01

    Highlights: • Membrane concentrates have a threat to human health and environment. • Untreated membrane concentrates induces cytotoxic and genotoxic to HepG2 cells. • Both methods were effective method for degradation of BPA and NP in concentrates. • Both methods were efficient in reducing genotoxic effects of concentrates. • UV-Fenton reagent had higher removal efficiency and provides toxicological safety. - Abstract: Membrane concentrates of landfill leachates contain organic and inorganic contaminants that could be highly toxic and carcinogenic. In this paper, the genotoxicity of membrane concentrates before and after Fenton and UV-Fenton reagent was assessed. The cytotoxicity and genotoxicity was determined by using the methods of methyltetrazolium (MTT), cytokinesis-block micronucleus (CBMN) and comet assay in human hepatoma cells. MTT assay showed a cytotoxicity of 75% after 24 h of exposure to the highest tested concentration of untreated concentrates, and no cytotoxocity for UV-Fenton and Fenton treated concentrates. Both CBMN and comet assays showed increased levels of genotoxicity in cells exposed to untreated concentrates, compared to those occurred in cells exposed to UV-Fenton and Fenton reagent treated concentrates. There was no significant difference between negative control and UV-Fenton treated concentrates for micronucleus and comet assay parameters. UV-Fenton and Fenton treatment, especially the former, were effective methods for degradation of bisphenol A and nonylphenol in concentrates. These findings showed UV-Fenton and Fenton reaction were effective methods for treatment of such complex concentrates, UV-Fenton reagent provided toxicological safety of the treated effluent, and the genotoxicity assays were found to be feasible tools for assessment of toxicity risks of complex concentrates.

  6. GENETIC POLYMORPHISM AND CYTOGENETIC CHANGES IN THE PERIPHERAL BLOOD T-LYMPHOCYTES OF PATIENTS WITH ARTHRITIS ASSOCIATED WITH IXODES TICK-BORNE BORRELIOSIS IN THE NORTHERN REGIONS OF SIBERIA

    Directory of Open Access Journals (Sweden)

    N. N. Ilyinskikh

    2017-01-01

    Full Text Available The authors have previously conducted studies that demonstrate the increased level of cytogenetic disturbances in patients with Ixodes tick-borne borreliosis (ITB. The severity of arthritis associated with ITB (AITB is also ascertained to depend on whether the patient has certain HLA-DRB1 alleles.Objective: to assess the association between HLA-DRB1 gene polymorphism and cytogenetic changes in the peripheral blood T lymphocytes of patients with AITB.Subjects and methods. 146 patients with AITB, 100 clinically healthy convalescents with ITB (CITB, and a control group of 98 healthy blood donors (HBDs without a history of tick-borne infections were examined using cytogenetic (micronucleus analysis of cytokinesis-blocked peripheral blood T lymphocytes and molecular genetic (PCR analysis of HLA-DRB1 gene polymorphism methods.Results and discussion. The frequency of cytokinesis-blocked lymphocytes with micronuclei in the AITB group was significantly higher than that in the CITB and HBD groups (p<0.01 with the exception of the results obtained in the subgroup of patients with AITB who had the DRB1*10 allele (p>0.05. The highest levels of lymphocytes with micronuclei were observed in AITB patients with the DRB1*17(03, *01, and *04 alleles as compared to those in the CITB and HBD groups (p<0.001. The CITB group showed the most significant increase in the detection rate of lymphocytes with micronuclei in people with the DRB1*01, DRB1*04, or *17(03 alleles. At the same time, there were no significant differences in the number of lymphocytes with micronuclei in the HBD group, depending on the HLA-DRB1 gene alleles (p>0.05. Thus, the patients with AITB had the highest frequency of cytogenetic disorders with the exception of individuals with the DRB1*10 allele.

  7. Micronuclei, nucleoplasmic bridges, and nuclear buds induced in human lymphocytes by the fungicide signum and its active ingredients (boscalid and pyraclostrobin).

    Science.gov (United States)

    Çayır, Akin; Coskun, Munevver; Coskun, Mahmut

    2014-05-01

    The aim of this study was to investigate the genotoxic and cytotoxic potential of the Signum fungicide and its active ingredients (boscalid and pyraclostrobin) on human peripheral blood lymphocytes using the cytokinesis-block micronucleus (CBMN) assay. Micronuclei (MNi), nucleoplasmic bridges (NPBs), nuclear bud (NBUDs) formations, and the cytokinesis-block proliferation index (CBPI) were evaluated in treated lymphocytes in Go (cells were treated and then kept in culture without stimulation for 24 h) and proliferation phases (cells were treated after 44 h culture in medium containing phytohemagglutinin). MN formation in lymphocytes treated in G0 statistically increased at doses of 2, 6, and 25 μg/mL signum; 0.5 and 2 μg/mL boscalid; and 0.5, 1.5, and 2 μg/mL pyraclostrobin; while NPB formation increased at a dose of 0.25 μg/mL pyraclostrobin. All concentrations of each fungicide did not statistically increase NBUD formation, while the cytotoxicity increased the dependent on concentration in lymphocytes treated in G0 . Doses of 0.5, 1, 1.5, and 3 μg/mL signum; 0.5, 1, and 1.5 μg/mL boscalid; and 0.75 μg/mL pyraclostrobin statistically increased the MN formation in proliferating lymphocytes. NPB formation increased in proliferating lymphocytes at doses of 1, 1.5, 2, and 3 μg/mL signum and at a dose of 0.75 μg/mL pyraclostrobin. In addition, a dose of 0.75 μg/mL pyraclostrobin increased NBUD frequencies. Cytotoxicity increased with increasing concentrations of each fungicide. It is concluded that signum, boscalid, and pyraclostrobin may be genotoxic and cytotoxic in vitro human peripheral blood lymphocytes in consideration of each of the two protocols. © 2012 Wiley Periodicals, Inc. Environ Toxicol 29: 723-732, 2014. Copyright © 2012 Wiley Periodicals, Inc., a Wiley company.

  8. Significance of micronucleus in cervical intraepithelial lesions and carcinoma

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    B N Gayathri

    2012-01-01

    Conclusions: MN scoring on the epithelial cells of cervix could be used as a biomarker in cancer screening. This is an easy, simple, reliable, reproducible and objective test which can be performed on routinely stained pap smears.

  9. Micronucleus Assay and Heavy Metals Characterization of E-waste ...

    African Journals Online (AJOL)

    ADOWIE PERE

    2018-03-28

    Mar 28, 2018 ... Ba) in the sediments, water, leachate and aquatic fauna (Tilapia guineensis, Callinectes amnicola and Cardiosoma ..... (2003) limit standard (0.01mg/L) for treated waste water .... the erythrocyte of grey mullet (Mugil cephalus).

  10. Cytogenetic biomonitoring in petrol station attendants: A micronucleus study

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    Medhini Singaraju

    2012-01-01

    Conclusion: The MN test in exfoliated epithelial cells seems to be a useful biomarker of occupational exposure to genotoxic chemicals. Phenol is the principal metabolite of benzene. Therefore, phenol concentration in the urine of exposed workers can be used as a biomarker of external exposure.

  11. 40 CFR 799.9539 - TSCA mammalian erythrocyte micronucleus test.

    Science.gov (United States)

    2010-07-01

    ... sampling time, unless acceptable inter-animal variability and frequencies of cells with micronuclei are...., Wehr, C.M., and Langlois, R.G. A Simple Fluorescent Staining Procedure for Micronuclei and RNA in...

  12. Micronucleus test in fish genome: A sensitive monitor for aquatic ...

    African Journals Online (AJOL)

    The aquatic environment makes up the major part of our environment and resources, therefore its safety is directly related to the safety our health. In this study, three tilapia species (Oreochromis niloticus, Oreochromis aureus and Tilapia zilli) and Clarias gariepinus were employed to estimate water pollution using ...

  13. Micronucleus test in mice fed on irradiated whole diet

    International Nuclear Information System (INIS)

    Reddy, P.P.; Reddi, O.S.; Pentiah, P.R.; Rani, M.V.U.; Devi, K.R.; Goud, S.N.

    1981-01-01

    Eight week old Swiss albino male mice were fed on freshly irradiated or unirradiated whole diet for one week. (Exposure was to 75 or 200 kR γ rays from a 1000 Ci 60 Co γ source at a dose rate of 584 R/min.) On the seventh day, six hours after feeding, the mice were killed and bone marrow preparations were made by the Schmid technique. From each group three animals were taken and from each animal 2000 polychromatic and normochromatic erythrocytes were scored. It was evident from the data obtained that the irradiated whole diet failed to induce any significant increase in the incidence of micronuclei in polychromatic erythrocytes. Similarly, there was no significant increase in the frequency of micronuclei in normochromatic erythrocytes when compared with control data. The polychromatic to normochromatic ratio was also unaffected. The diet consisted of wheat flour (60%). groundnut cake (20%), fish meal (8%), Bengal gram flour (8%), dried yeast (3%), salt/mineral mixture (1%) and traces of vitamins. (U.K.)

  14. Genotoxicity of nanomaterials: DNA damage and micronuclei induced by carbon nanotubes and graphite nanofibres in human bronchial epithelial cells in vitro.

    Science.gov (United States)

    Lindberg, Hanna K; Falck, Ghita C-M; Suhonen, Satu; Vippola, Minnamari; Vanhala, Esa; Catalán, Julia; Savolainen, Kai; Norppa, Hannu

    2009-05-08

    Despite the increasing industrial use of different nanomaterials, data on their genotoxicity are scant. In the present study, we examined the potential genotoxic effects of carbon nanotubes (CNTs; >50% single-walled, approximately 40% other CNTs; 1.1 nm x 0.5-100 microm; Sigma-Aldrich) and graphite nanofibres (GNFs; 95%; outer diameter 80-200 nm, inner diameter 30-50 nm, length 5-20 microm; Sigma-Aldrich) in vitro. Genotoxicity was assessed by the single cell gel electrophoresis (comet) assay and the micronucleus assay (cytokinesis-block method) in human bronchial epithelial BEAS 2B cells cultured for 24h, 48h, or 72h with various doses (1-100 microg/cm(2), corresponding to 3.8-380 microg/ml) of the carbon nanomaterials. In the comet assay, CNTs induced a dose-dependent increase in DNA damage at all treatment times, with a statistically significant effect starting at the lowest dose tested. GNFs increased DNA damage at all doses in the 24-h treatment, at two doses (40 and 100 microg/cm(2)) in the 48-h treatment (dose-dependent effect) and at four doses (lowest 10 microg/cm(2)) in the 72-h treatment. In the micronucleus assay, no increase in micronucleated cells was observed with either of the nanomaterials after the 24-h treatment or with CNTs after the 72-h treatment. The 48-h treatment caused a significant increase in micronucleated cells at three doses (lowest 10 microg/cm(2)) of CNTs and at two doses (5 and 10 microg/cm(2)) of GNFs. The 72-h treatment with GNFs increased micronucleated cells at four doses (lowest 10 microg/cm(2)). No dose-dependent effects were seen in the micronucleus assay. The presence of carbon nanomaterial on the microscopic slides disturbed the micronucleus analysis and made it impossible at levels higher than 20 microg/cm(2) of GNFs in the 24-h and 48-h treatments. In conclusion, our results suggest that both CNTs and GNFs are genotoxic in human bronchial epithelial BEAS 2B cells in vitro. This activity may be due to the fibrous nature

  15. Genotoxic Effects of Exposure to Gasoline Fumes on Petrol Pump Workers.

    Science.gov (United States)

    Shaikh, Amrin; Barot, Darshana; Chandel, Divya

    2018-04-01

    Petrol pump workers are occupationally exposed to gasoline and its fumes consisting of several mutagenic chemicals. To evaluate the genotoxic effects of exposure to gasoline fumes on petrol pump workers. The study groups included 70 petrol pump workers (exposed group) and 70 healthy age-matched individuals with no known exposure (comparison group). Buccal micronucleus cytome assay (BMCyt) was performed to check the genotoxicity caused due to inhalation of gasoline fumes. The frequencies of micronucleated cells, nuclear bud, condensed chromatin cells, karyorrhectic cells, pyknotic cells, and karyolytic cells were significantly higher in the exposed workers compared to the comparison group. Exposure to gasoline fumes is associated with increased frequency of cell abnormalities. This may lead to various health consequences including cancer in those occupationally exposed to gasoline fumes.

  16. Genotoxic Effects of Exposure to Gasoline Fumes on Petrol Pump Workers

    Directory of Open Access Journals (Sweden)

    Amrin Shaikh

    2018-04-01

    Full Text Available Background: Petrol pump workers are occupationally exposed to gasoline and its fumes consisting of several mutagenic chemicals. Objective: To evaluate the genotoxic effects of exposure to gasoline fumes on petrol pump workers. Methods: The study groups included 70 petrol pump workers (exposed group and 70 healthy age-matched individuals with no known exposure (comparison group. Buccal micronucleus cytome assay (BMCyt was performed to check the genotoxicity caused due to inhalation of gasoline fumes. Results: The frequencies of micronucleated cells, nuclear bud, condensed chromatin cells, karyorrhectic cells, pyknotic cells, and karyolytic cells were significantly higher in the exposed workers compared to the comparison group. Conclusion: Exposure to gasoline fumes is associated with increased frequency of cell abnormalities. This may lead to various health consequences including cancer in those occupationally exposed to gasoline fumes.

  17. Advantages of micronuclei analysis through images autocapturing and screen scoring

    International Nuclear Information System (INIS)

    González, J.E.; Martínez-López, W.

    2015-01-01

    The cytokinesis-block micronucleus (CBMN) test is a quantitative assay for genetic toxicity assessment. One of the advantages of the MN assay is its amenability for automation. Different type of cells has been used to evaluate genetic damage through MN assay, such as, human lymphocytes and rodent cell lines (i.e. CHO, V79, CHL and L5178Y). The MN quantification is a time consuming process and several efforts has been conducted for its automation. Some of them include an operator checking step, like PathFinder CellScan System, or are fully automated such as MNScore from MetaSytems. Usually, fully automated systems detect two or three times less MN than visual scoring. In some cases, the impact of false positive detection is reduced with a visual detection step. In the present work we have tested a combination of image autocapturing of CHOK1 cells previously treated with bleomycin (0, 2.5, 5.0 and 10.0 μg/ml) or UVC (0, 4, 8 and 16 J/m”2 ) with a screen scoring. Capturing images using the AutoCapture option from Metafer 4 from MetaSystems (GmbH, Germany) plus screen scoring render similar results in terms of MN cells frequency than microscopic live scoring. The resultant bias from the Bland–Altman analysis was -1.1% with confidence intervals between -2.2% and -0.1%, indicating an acceptable agreement between both MN scoring method. However, the mean time devoted to live microscope scoring per sample was 159 minutes compared to 39 minutes for microscope images autocapturing and screen scoring. Therefore, it become advantageous to combine autocapturing of microscope images plus screen scoring when many samples have to be analyzed for radiological biodosimetry purposes. (authors)

  18. Influence of age, sex and life style factors (smoking habits) on the spontaneous and radiation induced micronuclei frequencies

    International Nuclear Information System (INIS)

    Di Giorgio, M.; Nasazzi, N.; Heredia, M.L.

    1995-01-01

    Several endpoints have been used for monitoring human populations for environmental or occupational exposure to genotoxic agents, particularly ionizing radiation. The cytokinesis-block micronucleus (MN) assay in peripheral lymphocytes is a reliable method for assessing radiation induced chromosomal damage (DNA breaks and mitotic spindle disturbances) and thus, a suitable dosimeter for estimating in vivo whole body exposures. To further define the use of this assay in Biological Dosimetry, a study to determine the influence of age, sex and life style factors (smoking habit) on the spontaneous and radiation induced MN frequencies was performed. The estimation of MN frequencies was analyzed in lymphocytes cultures from 50 healthy donors aged between 4 and 62 years. On the basis of their smoking habit they were divided into 2 groups. A fraction of the sample was irradiated in vitro with γ rays in the range of 0.35 Gy to 4 Gy. A statistically significant influence on the spontaneous MN frequency was observed (R 2 = 0.59) when the variables age and smoking habit were analyzed and also a statistically significant influence on the radiation induced MN frequency was obtained (R 2 = 0.86) when dose, age and smoking habit were studied. Sex did not influence MN variability significantly but there was a greater dispersion in the results for females when compared to males, possibly due to the loss of X chromosomes. The comparison of the data from smoking donors to non smoking donors supports the convenience of taking into account the smoking habit for estimating in vivo whole body exposure to γ rays for doses below 2 Gy. (author). 8 refs., 3 figs., 1 tab

  19. Folic acid deficiency increases chromosomal instability, chromosome 21 aneuploidy and sensitivity to radiation-induced micronuclei

    International Nuclear Information System (INIS)

    Beetstra, Sasja; Thomas, Philip; Salisbury, Carolyn; Turner, Julie; Fenech, Michael

    2005-01-01

    Folic acid deficiency can lead to uracil incorporation into DNA, hypomethylation of DNA, inefficient DNA repair and increase chromosome malsegregation and breakage. Because ionising radiation increases demand for efficient DNA repair and also causes chromosome breaks we hypothesised that folic acid deficiency may increase sensitivity to radiation-induced chromosome breakage. We tested this hypothesis by using the cytokinesis-block micronucleus assay in 10 day WIL2-NS cell cultures at four different folic acid concentrations (0.2, 2, 20, and 200 nM) that span the 'normal' physiological range in humans. The study showed a significant dose-dependent increase in frequency of binucleated cells with micronuclei and/or nucleoplasmic bridges with decreasing folic acid concentration (P < 0.0001, P = 0.028, respectively). These biomarkers of chromosomal instability were also increased in cells irradiated (1.5 Gy γ-rays) on day 9 relative to un-irradiated controls (P < 0.05). Folic acid deficiency and γ-irradiation were shown to have a significant interactive effect on frequency of cells containing micronuclei (two-way ANOVA, interaction P 0.0039) such that the frequency of radiation-induced micronucleated cells (i.e. after subtracting base-line frequency of un-irradiated controls) increased with decreasing folic acid concentration (P-trend < 0.0001). Aneuploidy of chromosome 21, apoptosis and necrosis were increased by folic acid deficiency but not by ionising radiation. The results of this study show that folate status has an important impact on chromosomal stability and is an important modifying factor of cellular sensitivity to radiation-induced genome damage

  20. Increased frequency of micronuclei in adults with a history of childhood sexual abuse: a discordant monozygotic twin study.

    Directory of Open Access Journals (Sweden)

    Timothy P York

    Full Text Available Childhood sexual abuse (CSA is a traumatic life event associated with an increased lifetime risk for psychopathology/morbidity. The long-term biological consequences of CSA-elicited stress on chromosomal stability in adults are unknown. The primary aim of this study was to determine if the rate of acquired chromosomal changes, measured using the cytokinesis-block micronucleus assay on stimulated peripheral blood lymphocytes, differs in adult female monozygotic twins discordant for CSA.Monozygotic twin pairs discordant for CSA were identified from a larger population-based sample of female adult twins for whom the experience of CSA was assessed by self-report (51 individuals including a reference sample. Micronuclei (MN contain chromatin from structurally normal or abnormal chromosomes that are excluded from the daughter nuclei during cell division and serve as a biomarker to assess acquired chromosomal instability.Female twins exposed to CSA exhibited a 1.63-fold average increase in their frequency of MN compared to their nonexposed genetically identical cotwins (Paired t-test, t₁₆ = 2.65, P = 0.017. No additional effects of familial factors were detected after controlling for the effect of CSA exposure. A significant interaction between CSA history and age was observed, suggesting that the biological effects of CSA on MN formation may be cumulative.These data support a direct link between CSA exposure and MN formation measured in adults that is not attributable to genetic or environmental factors shared by siblings. Further research is warranted to understand the biological basis for the observed increase in acquired chromosomal findings in people exposed to CSA and to determine if acquired somatic chromosomal abnormalities/somatic clonal mosaicism might mediate the adult pathology associated with CSA.

  1. Increased levels of lead in the blood and frequencies of lymphocytic micronucleated binucleated cells among workers from an electronic-waste recycling site.

    Science.gov (United States)

    Wang, Qian; He, An M; Gao, Bo; Chen, Lan; Yu, Qiang Z; Guo, Huan; Shi, Bin J; Jiang, Pu; Zhang, Zeng Y; Li, Ping L; Sheng, Ying G; Fu, Mo J; Wu, Chun T; Chen, Min X; Yuan, Jing

    2011-01-01

    In recent years, adverse health effects of chemicals from electronic waste (e-waste) have been reported. However, little is known about the genotoxic effects of chemicals in e-waste. In the present study, air concentrations of the toxic metals at e-waste and control sites were analyzed using inductively-coupled plasma mass spectrometry. Levels of toxic metals (lead, copper and cadmium) in blood and urine were detected using atomic absorption spectrophotometry in 48 exposed individuals and 56 age- and sex-matched controls. The frequencies of lymphocytic micronucleated binucleated cells (MNBNCs) were determined using a cytokinesis-block micronucleus assay. Results indicated that blood lead levels were significantly higher in the exposed group (median: 11.449 μg/dL, 1st/3rd quartiles: 9.351-14.410 μg/dL) than in the control group (median: 9.104 μg/dL, 1st/3rd quartiles: 7.275-11.389 μg/dL). The exposed group had higher MNBNCs frequencies (median: 4.0 per thousand, 1st/3rd quartiles: 2.0-7.0 per thousand) compared with the controls (median: 1.0 per thousand, 1st/3rd quartiles: 0.0-2.0 per thousand). Additionally, MNBNCs frequencies and blood lead levels were positively correlated (r = 0.254, phistory of working with e-waste was a predictor for increased blood lead levels and MNBNCs frequencies in the subjects. The results suggest that both the living and occupational environments at the e-waste site may be risk factors for increased MNBNCs frequencies among those who are exposed.

  2. A mode-of-action approach for the identification of genotoxic carcinogens.

    Directory of Open Access Journals (Sweden)

    Lya G Hernández

    Full Text Available Distinguishing between clastogens and aneugens is vital in cancer risk assessment because the default assumption is that clastogens and aneugens have linear and non-linear dose-response curves, respectively. Any observed non-linearity must be supported by mode of action (MOA analyses where biological mechanisms are linked with dose-response evaluations. For aneugens, the MOA has been well characterised as disruptors of mitotic machinery where chromosome loss via micronuclei (MN formation is an accepted endpoint used in risk assessment. In this study we performed the cytokinesis-block micronucleus assay and immunofluorescence mitotic machinery visualisation in human lymphoblastoid (AHH-1 and Chinese Hamster fibroblast (V79 cell lines after treatment with the aneugen 17-β-oestradiol (E₂. Results were compared to previously published data on bisphenol-A (BPA and Rotenone data. Two concentration-response approaches (the threshold-[Td] and benchmark-dose [BMD] approaches were applied to derive a point of departure (POD for in vitro MN induction. BMDs were also derived from the most sensitive carcinogenic endpoint. Ranking comparisons of the PODs from the in vitro MN and the carcinogenicity studies demonstrated a link between these two endpoints for BPA, E₂ and Rotenone. This analysis was extended to include 5 additional aneugens, 5 clastogens and 3 mutagens and further concentration and dose-response correlations were observed between PODs from the in vitro MN and carcinogenicity. This approach is promising and may be further extended to other genotoxic carcinogens, where MOA and quantitative information from the in vitro MN studies could be used in a quantitative manner to further inform cancer risk assessment.

  3. Micronuclei frequency in albino rats exposed to high natural radiation

    International Nuclear Information System (INIS)

    Aneesh, D.; Godwin Wesley, S.

    2013-01-01

    Genotoxicity and DNA damage endpoints are used to evaluate results in the context of cell survival. Genotoxicity in mammalian cells is monitored mostly by using cytokinesis-block micronucleus (CBMN) assay. The score of micronuclei (MN) in peripheral blood lymphocytes can be used as a biomarker and also as a bio-dosimeter of radiation exposure. In the present study the effect of natural radiation on albino rats has been investigated, to find out if there is any increase in MN frequency in peripheral blood lymphocytes. Animals at the age of 2-3 weeks were exposed to natural radiation, at the dose of 10.38 μGyh -1 for a period of 6 months. A parallel control set was also maintained (0.12 μGy h -1 '). Blood samples were collected from both test (exposed to natural radiation) and control rats. Lymphocyte culture was done following 'microculture techniques' for 72 h. Cytochalasin B, at a concentration of 6.0 μg/ml, was added to the lymphocyte cultures at 44 h to block cytokinesis. The frequency of MN was evaluated by scoring a total of 1000 binucleated (BN) cells from one slide. The frequency of MN among the rats exposed to natural radiation was found to be 1.83±0.05 per 1000 BN cells and in the control it was 1.82±0.07 per 1000 BN cells. No statistically significant difference in the MN frequencies of exposed and control groups (p>0.05) was seen. The lower MN frequency in natural radiation exposed rats could be an indication of adaptive response. (author)

  4. Pulsed radiobiology with laser-driven plasma accelerators

    Science.gov (United States)

    Giulietti, Antonio; Grazia Andreassi, Maria; Greco, Carlo

    2011-05-01

    Recently, a high efficiency regime of acceleration in laser plasmas has been discovered, allowing table top equipment to deliver doses of interest for radiotherapy with electron bunches of suitable kinetic energy. In view of an R&D program aimed to the realization of an innovative class of accelerators for medical uses, a radiobiological validation is needed. At the present time, the biological effects of electron bunches from the laser-driven electron accelerator are largely unknown. In radiobiology and radiotherapy, it is known that the early spatial distribution of energy deposition following ionizing radiation interactions with DNA molecule is crucial for the prediction of damages at cellular or tissue levels and during the clinical responses to this irradiation. The purpose of the present study is to evaluate the radio-biological effects obtained with electron bunches from a laser-driven electron accelerator compared with bunches coming from a IORT-dedicated medical Radio-frequency based linac's on human cells by the cytokinesis block micronucleus assay (CBMN). To this purpose a multidisciplinary team including radiotherapists, biologists, medical physicists, laser and plasma physicists is working at CNR Campus and University of Pisa. Dose on samples is delivered alternatively by the "laser-linac" operating at ILIL lab of Istituto Nazionale di Ottica and an RF-linac operating for IORT at Pisa S. Chiara Hospital. Experimental data are analyzed on the basis of suitable radiobiological models as well as with numerical simulation based on Monte Carlo codes. Possible collective effects are also considered in the case of ultrashort, ultradense bunches of ionizing radiation.

  5. Whole thorax irradiation of non-human primates induces persistent nuclear damage and gene expression changes in peripheral blood cells.

    Directory of Open Access Journals (Sweden)

    Shanaz A Ghandhi

    Full Text Available We investigated the cytogenetic and gene expression responses of peripheral blood cells of non-human primates (NHP, Macaca mulatta that were whole-thorax irradiated with a single dose of 10 Gy. In this model, partial irradiation of NHPs in the thoracic region (Whole Thorax Lung Irradiation, WTLI allows the study of late radiation-induced lung injury, while avoiding acute radiation syndromes related to hematopoietic and gastrointestinal injury. A transient drop in circulating lymphocytes and platelets was seen by 9 days, followed by elevations in respiratory rate, circulating neutrophils, lymphocytes, and monocytes at 60-100 days, corresponding to computed tomography (CT and histologic evidence of pneumonitis, and elective euthanasia of four animals. To evaluate long-term DNA damage in NHP peripheral blood lymphocytes after 10 Gy WTLI, we used the cytokinesis-block micronucleus (CBMN assay to measure chromosomal aberrations as post-mitotic micronuclei in blood samples collected up to 8 months after irradiation. Regression analysis showed significant induction of micronuclei in NHP blood cells that persisted with a gradual decline over the 8-month study period, suggesting long-term DNA damage in blood lymphocytes after WTLI. We also report transcriptomic changes in blood up to 30 days after WTLI. We isolated total RNA from peripheral blood at 3 days before and then at 2, 5 and 30 days after irradiation. We identified 1187 transcripts that were significantly changed across the 30-day time course. From changes in gene expression, we identified biological processes related to immune responses, which persisted across the 30-day study. Response to oxygen-containing compounds and bacteria were implicated by gene-expression changes at the earliest day 2 and latest, day 30 time-points. Gene expression changes suggest a persistent altered state of the immune system, specifically response to infection, for at least a month after WTLI.

  6. Comparison of Radiation-Induced Bystander Effect in QU-DB Cells after Acute and Fractionated Irradiation: An In Vitro Study.

    Science.gov (United States)

    Soleymanifard, Shokouhozaman; Bahreyni Toossi, Mohammad Taghi; Kamran Samani, Roghayeh; Mohebbi, Shokoufeh

    2016-01-01

    Radiation effects induced in non-irradiated cells are termed radiation-induced bystander effects (RIBE). The present study intends to examine the RIBE response of QU-DB bystander cells to first, second and third radiation fractions and compare their cumulative outcome with an equal, single acute dose. This experimental study irradiated three groups of target cells for one, two and three times with(60)Co gamma rays. One hour after irradiation, we transferred their culture media to non-irradiated (bystander) cells. We used the cytokinesis block micronucleus assay to evaluate RIBE response in the bystander cells. The numbers of micronuclei generated in bystander cells were determined. RIBE response to single acute doses increased up to 4 Gy, then decreased, and finally at the 8 Gy dose disappeared. The second and third fractions induced RIBE in bystander cells, except when RIBE reached to the maximum level at the first fraction. We split the 4 Gy acute dose into two fractions, which decreased the RIBE response. However, fractionation of 6 Gy (into two fractions of 3 Gy or three fractions of 2 Gy) had no effect on RIBE response. When we split the 8 Gy acute dose into two fractions we observed RIBE, which had disappeared following the single 8 Gy dose. The impact of dose fractionation on RIBE induced in QU-DB cells de- pended on the RIBE dose-response relationship. Where RIBE increased proportion- ally with the dose, fractionation reduced the RIBE response. In contrast, at high dos- es where RIBE decreased proportionally with the dose, fractionation either did not change RIBE (at 6 Gy) or increased it (at 8 Gy).

  7. Neutron Exposures in Human Cells: Bystander Effect and Relative Biological Effectiveness

    Science.gov (United States)

    Seth, Isheeta; Schwartz, Jeffrey L.; Stewart, Robert D.; Emery, Robert; Joiner, Michael C.; Tucker, James D.

    2014-01-01

    Bystander effects have been observed repeatedly in mammalian cells following photon and alpha particle irradiation. However, few studies have been performed to investigate bystander effects arising from neutron irradiation. Here we asked whether neutrons also induce a bystander effect in two normal human lymphoblastoid cell lines. These cells were exposed to fast neutrons produced by targeting a near-monoenergetic 50.5 MeV proton beam at a Be target (17 MeV average neutron energy), and irradiated-cell conditioned media (ICCM) was transferred to unirradiated cells. The cytokinesis-block micronucleus assay was used to quantify genetic damage in radiation-naïve cells exposed to ICCM from cultures that received 0 (control), 0.5, 1, 1.5, 2, 3 or 4 Gy neutrons. Cells grown in ICCM from irradiated cells showed no significant increase in the frequencies of micronuclei or nucleoplasmic bridges compared to cells grown in ICCM from sham irradiated cells for either cell line. However, the neutron beam has a photon dose-contamination of 5%, which may modulate a neutron-induced bystander effect. To determine whether these low doses of contaminating photons can induce a bystander effect, cells were irradiated with cobalt-60 at doses equivalent to the percent contamination for each neutron dose. No significant increase in the frequencies of micronuclei or bridges was observed at these doses of photons for either cell line when cultured in ICCM. As expected, high doses of photons induced a clear bystander effect in both cell lines for micronuclei and bridges (pbystander effect in these cells. Finally, neutrons had a relative biological effectiveness of 2.0±0.13 for micronuclei and 5.8±2.9 for bridges compared to cobalt-60. These results may be relevant to radiation therapy with fast neutrons and for regulatory agencies setting standards for neutron radiation protection and safety. PMID:24896095

  8. Genotoxic Effects of Low- and High-LET Radiation on Human Epithelial Cells Grown in 2-D Versus 3-D Culture

    Science.gov (United States)

    Patel, Z. S.; Cucinotta, F. A.; Huff, J. L.

    2011-01-01

    Risk estimation for radiation-induced cancer relies heavily on human epidemiology data obtained from terrestrial irradiation incidents from sources such as medical and occupational exposures as well as from the atomic bomb survivors. No such data exists for exposures to the types and doses of high-LET radiation that will be encountered during space travel; therefore, risk assessment for space radiation requires the use of data derived from cell culture and animal models. The use of experimental models that most accurately replicate the response of human tissues is critical for precision in risk projections. This work compares the genotoxic effects of radiation on normal human epithelial cells grown in standard 2-D monolayer culture compared to 3-D organotypic co-culture conditions. These 3-D organotypic models mimic the morphological features, differentiation markers, and growth characteristics of fully-differentiated normal human tissue and are reproducible using defined components. Cultures were irradiated with 2 Gy low-LET gamma rays or varying doses of high-LET particle radiation and genotoxic damage was measured using a modified cytokinesis block micronucleus assay. Our results revealed a 2-fold increase in residual damage in 2 Gy gamma irradiated cells grown under organotypic culture conditions compared to monolayer culture. Irradiation with high-LET particle radiation gave similar results, while background levels of damage were comparable under both scenarios. These observations may be related to the phenomenon of "multicellular resistance" where cancer cells grown as 3-D spheroids or in vivo exhibit an increased resistance to killing by chemotherapeutic agents compared to the same cells grown in 2-D culture. A variety of factors are likely involved in mediating this process, including increased cell-cell communication, microenvironment influences, and changes in cell cycle kinetics that may promote survival of damaged cells in 3-D culture that would

  9. Assessment of in vivo and in vitro genotoxicity of glibenclamide in eukaryotic cells.

    Directory of Open Access Journals (Sweden)

    Juliane Rocha de Sant'Anna

    Full Text Available Glibenclamide is an oral hypoglycemic drug commonly prescribed for the treatment of type 2 diabetes mellitus, whose anti-tumor activity has been recently described in several human cancer cells. The mutagenic potential of such an antidiabetic drug and its recombinogenic activity in eukaryotic cells were evaluated, the latter for the first time. The mutagenic potential of glibenclamide in therapeutically plasma (0.6 μM and higher concentrations (10 μM, 100 μM, 240 μM and 480 μM was assessed by the in vitro mammalian cell micronucleus test in human lymphocytes. Since the loss of heterozygosity arising from allelic recombination is an important biologically significant consequence of oxidative damage, the glibenclamide recombinogenic activity at 1 μM, 10 μM and 100 μM concentrations was evaluated by the in vivo homozygotization assay. Glibenclamide failed to alter the frequency of micronuclei between 0.6 μM and 480 μM concentrations and the cytokinesis block proliferation index between 0.6 μM and 240 μM concentrations. On the other hand, glibenclamide changed the cell-proliferation kinetics when used at 480 μM. In the homozygotization assay, the homozygotization indices for the analyzed markers were lower than 2.0 and demonstrated the lack of recombinogenic activity of glibenclamide. Data in the current study demonstrate that glibenclamide, in current experimental conditions, is devoid of significant genotoxic effects. This fact encourages further investigations on the use of this antidiabetic agent as a chemotherapeutic drug.

  10. Nuclear Division Index may Predict Neoplastic Colorectal Lesions.

    Science.gov (United States)

    Ionescu, Mirela E; Ciocirlan, Mihai; Becheanu, Gabriel; Nicolaie, Tudor; Ditescu, Cristina; Teiusanu, Adriana G; Gologan, Serban I; Arbanas, Tudor; Diculescu, Mircea M

    2011-07-01

    Colorectal cancer (CRC) develops by accumulation of multiple genetic damages leading to genetic instability that can be evaluated by cytogenetic methods. In the current study we used Cytokinesis-Blocked Micronucleus Assay (CBMN) technique to assess the behavior of Nuclear Division Index(NDI) in peripheral lymphocytes of patients with CRC and polyps versus patients with normal colonoscopy. Blood samples were collected from patients after informed consent. By CBMN technique we assessed the proportion of mono-nucleated, bi-nucleated, tri-nucleated and tetra-nucleated cells/500 cells, to calculate NDI. Data were statistically analyzed using the SPSS 11.0 package. 45 patients were available for analysis, 23 men and 22 women, with a mean age of 58.7±13.5. 17 had normal colonoscopy, 17 colonic polyps and 11 CRC. The mean NDI values were significantly smaller for patients with CRC or polyps than in patients with normal colonoscopy (1.57 vs 1.73, p=0.013). The difference persisted for patients with neoplastic lesions (adenomas and carcinomas) when compared with patients with normal colonoscopy or non neoplastic (hyperplastic) polyps (1.56 vs.1.71, p=0.018). The NDI cut-off value to predict the presence of adenomas or carcinomas was equal to 1.55 with a 54.2% sensitivity and 81% specificity of lower values (p=0.019). The NDI cut off value to predict the presence of advanced adenomas or cancer was 1.525 for a sensitivity of 56.3% and a specificity of 82.8% (p=0.048). NDI may be useful in screening strategies for colorectal cancer as simple, noninvasive, inexpensive cytogenetic biomarker.

  11. Telomere dysfunction and chromosome structure modulate the contribution of individual chromosomes in abnormal nuclear morphologies

    Energy Technology Data Exchange (ETDEWEB)

    Pampalona, J.; Soler, D.; Genesca, A. [Department of Cell Biology, Physiology and Immunology, Universitat Autonoma de Barcelona, Bellaterra E-08193 (Spain); Tusell, L., E-mail: laura.tusell@uab.es [Department of Cell Biology, Physiology and Immunology, Universitat Autonoma de Barcelona, Bellaterra E-08193 (Spain)

    2010-01-05

    The cytokinesis-block micronucleus assay has emerged as a biomarker of chromosome damage relevant to cancer. Although it was initially developed to measure micronuclei, it is also useful for measuring nucleoplasmic bridges and nuclear buds. Abnormal nuclear morphologies are frequently observed in malignant tissues and short-term tumour cell cultures. Changes in chromosome structure and number resulting from chromosome instability are important factors in oncogenesis. Telomeres have become key players in the initiation of chromosome instability related to carcinogenesis by means of breakage-fusion-bridge cycles. To better understand the connection between telomere dysfunction and the appearance of abnormal nuclear morphologies, we have characterised the presence of micronuclei, nucleoplasmic bridges and nuclear buds in human mammary primary epithelial cells. These cells can proliferate beyond the Hayflick limit by spontaneously losing expression of the p16{sup INK4a} protein. Progressive telomere shortening leads to the loss of the capping function, and the appearance of end-to-end chromosome fusions that can enter into breakage-fusion-bridge cycles generating massive chromosomal instability. In human mammary epithelial cells, different types of abnormal nuclear morphologies were observed, however only nucleoplasmatic bridges and buds increased significantly with population doublings. Fluorescent in situ hybridisation using centromeric and painting specific probes for chromosomes with eroded telomeres has revealed that these chromosomes are preferentially included in the different types of abnormal nuclear morphologies observed, thus reflecting their common origin. Accordingly, real-time imaging of cell divisions enabled us to determine that anaphase bridge resolution was mainly through chromatin breakage and the formation of symmetric buds in daughter nuclei. Few micronuclei emerged in this cell system thus validating the scoring of nucleoplasmic bridges and

  12. Telomere dysfunction and chromosome structure modulate the contribution of individual chromosomes in abnormal nuclear morphologies

    International Nuclear Information System (INIS)

    Pampalona, J.; Soler, D.; Genesca, A.; Tusell, L.

    2010-01-01

    The cytokinesis-block micronucleus assay has emerged as a biomarker of chromosome damage relevant to cancer. Although it was initially developed to measure micronuclei, it is also useful for measuring nucleoplasmic bridges and nuclear buds. Abnormal nuclear morphologies are frequently observed in malignant tissues and short-term tumour cell cultures. Changes in chromosome structure and number resulting from chromosome instability are important factors in oncogenesis. Telomeres have become key players in the initiation of chromosome instability related to carcinogenesis by means of breakage-fusion-bridge cycles. To better understand the connection between telomere dysfunction and the appearance of abnormal nuclear morphologies, we have characterised the presence of micronuclei, nucleoplasmic bridges and nuclear buds in human mammary primary epithelial cells. These cells can proliferate beyond the Hayflick limit by spontaneously losing expression of the p16 INK4a protein. Progressive telomere shortening leads to the loss of the capping function, and the appearance of end-to-end chromosome fusions that can enter into breakage-fusion-bridge cycles generating massive chromosomal instability. In human mammary epithelial cells, different types of abnormal nuclear morphologies were observed, however only nucleoplasmatic bridges and buds increased significantly with population doublings. Fluorescent in situ hybridisation using centromeric and painting specific probes for chromosomes with eroded telomeres has revealed that these chromosomes are preferentially included in the different types of abnormal nuclear morphologies observed, thus reflecting their common origin. Accordingly, real-time imaging of cell divisions enabled us to determine that anaphase bridge resolution was mainly through chromatin breakage and the formation of symmetric buds in daughter nuclei. Few micronuclei emerged in this cell system thus validating the scoring of nucleoplasmic bridges and nuclear

  13. Telomere dysfunction and chromosome structure modulate the contribution of individual chromosomes in abnormal nuclear morphologies.

    Science.gov (United States)

    Pampalona, J; Soler, D; Genescà, A; Tusell, L

    2010-01-05

    The cytokinesis-block micronucleus assay has emerged as a biomarker of chromosome damage relevant to cancer. Although it was initially developed to measure micronuclei, it is also useful for measuring nucleoplasmic bridges and nuclear buds. Abnormal nuclear morphologies are frequently observed in malignant tissues and short-term tumour cell cultures. Changes in chromosome structure and number resulting from chromosome instability are important factors in oncogenesis. Telomeres have become key players in the initiation of chromosome instability related to carcinogenesis by means of breakage-fusion-bridge cycles. To better understand the connection between telomere dysfunction and the appearance of abnormal nuclear morphologies, we have characterised the presence of micronuclei, nucleoplasmic bridges and nuclear buds in human mammary primary epithelial cells. These cells can proliferate beyond the Hayflick limit by spontaneously losing expression of the p16(INK4a) protein. Progressive telomere shortening leads to the loss of the capping function, and the appearance of end-to-end chromosome fusions that can enter into breakage-fusion-bridge cycles generating massive chromosomal instability. In human mammary epithelial cells, different types of abnormal nuclear morphologies were observed, however only nucleoplasmatic bridges and buds increased significantly with population doublings. Fluorescent in situ hybridisation using centromeric and painting specific probes for chromosomes with eroded telomeres has revealed that these chromosomes are preferentially included in the different types of abnormal nuclear morphologies observed, thus reflecting their common origin. Accordingly, real-time imaging of cell divisions enabled us to determine that anaphase bridge resolution was mainly through chromatin breakage and the formation of symmetric buds in daughter nuclei. Few micronuclei emerged in this cell system thus validating the scoring of nucleoplasmic bridges and nuclear

  14. A cytogenetic bio-monitoring of industrial radiographers occupationally exposed to low levels of ionizing radiation by using CBMN assay

    International Nuclear Information System (INIS)

    Shakeri, Mahsa; Changizi, Vahid; Zakeri, Farideh; Rajabpour, Mohammad Reza; Farshidpour, Mohammad Reza

    2017-01-01

    Industrial radiography is the process of using either gamma-emitting radionuclide sources or X-ray machines to examine the safety of industrial materials. The average annual effective dose in industrial radiography is one of the highest among radiation workers. The aim of this study was to investigate the cytogenetic effects of ionizing radiation in the peripheral blood lymphocytes of 60 industrial radiographers and 40 non-exposed individuals as the control group by using cytokinesis-block micronucleus (CBMN) assay. Totally, the frequencies of micronuclei (MN), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) were significantly higher in the industrial radiographers than in the controls (p = 0.000). The mean MN frequency per 1000 binucleated cells in the industrial radiographers with last 5-y radiation dose of >100 mSv was significantly higher than those with ≤100 mSv (34.81 ± 12.70 vs. 26.33 ± 7.940, p = 0.024). The effect of age was observed in the control group and subjects with the age of >30 y showed significantly higher MN frequency compared with the subjects with the age of ≤30 y (9.45 ± 3.710 vs. 6.81 ± 3.050, p = 0.02). No obvious trend of increased MN as a function of either duration of employment or age or smoking status was observed in the industrial radiographers. The results show the increased levels of cytogenetic damages in the industrial radiographers. Even the workers exposed to the permissible doses are subjected to elevated frequencies of DNA damages. These findings confirm the importance of cytogenetic bio-monitoring program beside physical dosimetry, surveying radiation safety of equipment and periodic training of workers for improvement of safety and radiation protection. (authors)

  15. Chronic obstructive pulmonary disease among lung cancer-free smokers: The importance of healthy controls.

    Science.gov (United States)

    Karpman, Michelle D; Eldridge, Ronald; Follis, Jack L; Etzel, Carol J; Shete, Sanjay; El-Zein, Randa A

    2018-01-01

    The prevalence of chronic obstructive pulmonary disease (COPD) in smokers enrolled as "healthy" controls in studies is 10-50%. The COPD status of ideal smoker populations for lung cancer case-control studies should be checked via spirometry; however, this is often not feasible, because no medical indications exist for asymptomatic smokers to undergo spirometry prior to study enrollment. Therefore, there is an unmet need for robust, cost effective assays for identifying undiagnosed lung disease among asymptomatic smokers. Such assays would help excluding unhealthy smokers from lung cancer case-control studies. We used the cytokinesis-blocked micronucleus (CBMN) assay (a measure of genetic instability) to identify undiagnosed lung disease among asymptomatic smokers. We used a convenience population from an on-going lung cancer case-control study including smokers with lung cancer (n = 454), smoker controls (n = 797), and a self-reported COPD (n = 200) contingent within the smoker controls. Significant differences for all CBMN endpoints were observed when comparing lung cancer to All controls (which included COPD) and Healthy controls (with no COPD). The risk ratio (RR) was increased in the COPD group vs. Healthy controls for nuclear buds (RR 1.28, 95% confidence interval 1.01-1.62), and marginally increased for micronuclei (RR 1.06, 0.98-1.89) and nucleoplasmic bridges (RR 1.07, 0.97-1.15). These findings highlight the importance of using truly healthy controls in studies geared toward assessment of lung cancer risk. Using genetic instability biomarkers would facilitate the identification of smokers susceptible to tobacco smoke carcinogens and therefore predisposed to either disease. Copyright © 2017 The Japanese Respiratory Society. All rights reserved.

  16. In-Vitro Radio protective Role of Ferulic Acid in Cultured Lymphocytes

    International Nuclear Information System (INIS)

    Ahmed, M.M.; Al Fateh, N.M.; Tawfik, S.S.

    2010-01-01

    Ferulic acid (FA), C 10 H 10 O 4 is the most abundant, ubiquitous hydroxycinnamic acid derived from photochemical phenolic compounds. It is a major constituent of fruits and vegetables such as orange, tomato, carrot, sweet corn and rice bran. Gamma rays generate hydroxyl radicals in cells and cellular DNA damage which leads to genotoxicity and chromosome aberrations. To establish most effective protective support, we used two different concentrations of FA (5 and 10 μg/ ml) and 2 Gy dose of gamma-radiation. Cytogenetic analysis was evaluated using the analysis of structural chromosome aberration (CA) and cytokinesis block micronucleus assay (CBMN). The level of lipid peroxidation analyzed as thiobarbituric acid reactive substances (TBARS), total glutathione (GSH), the enzyme activities of lymphocytes defence mechanism: Superoxide dismutase (SOD), Catalase (CAT) and Glutathione peroxidase (GPx) were determined. The result obtained by all endpoints indicates acceptable toxicity profiles of FA in-vitro when compared with normal lymphocytes; irradiation at 2 Gy increased the MN and CA frequencies. Treatment with FA for 30 min before radiation exposure resulted in a significant decline both of MN and CA yields as FA concentration increased. The levels of TBARS and GSH were altered significantly whereas the levels of the enzymatic antioxidants were decreased in gamma-irradiated lymphocytes. Pretreatment with 10 μg/ ml of FA has attenuated the toxic effects of radiation more than FA (5 μg/ ml) by reduction in the TBARS level, restoration GSH contents and prevented the decreases in the radiation-induced SOD, CAT and GPx activities. These results lead us to the conclusion that FA has antimutagenic effect and benefit as a radio protector against oxidative stress involved by gamma-rays exposure

  17. Induction of adaptive response in human blood lymphocytes exposed to radiofrequency radiation.

    Science.gov (United States)

    Sannino, Anna; Sarti, Maurizio; Reddy, Siddharth B; Prihoda, Thomas J; Vijayalaxmi; Scarfì, Maria Rosaria

    2009-06-01

    The incidence of micronuclei was evaluated to assess the induction of an adaptive response to non-ionizing radiofrequency (RF) radiation in peripheral blood lymphocytes collected from five different human volunteers. After stimulation with phytohemagglutinin for 24 h, the cells were exposed to an adaptive dose of 900 MHz RF radiation used for mobile communications (at a peak specific absorption rate of 10 W/kg) for 20 h and then challenged with a single genotoxic dose of mitomycin C (100 ng/ml) at 48 h. Lymphocytes were collected at 72 h to examine the frequency of micronuclei in cytokinesis-blocked binucleated cells. Cells collected from four donors exhibited the induction of adaptive response (i.e., responders). Lymphocytes that were pre-exposed to 900 MHz RF radiation had a significantly decreased incidence of micronuclei induced by the challenge dose of mitomycin C compared to those that were not pre-exposed to 900 MHz RF radiation. These preliminary results suggested that the adaptive response can be induced in cells exposed to non-ionizing radiation. A similar phenomenon has been reported in cells as well as in animals exposed to ionizing radiation in several earlier studies. However, induction of adaptive response was not observed in the remaining donor (i.e., non-responder). The incidence of micronuclei induced by the challenge dose of mitomycin C was not significantly different between the cells that were pre-exposed and unexposed to 900 MHz RF radiation. Thus the overall data indicated the existence of heterogeneity in the induction of an adaptive response between individuals exposed to RF radiation and showed that the less time-consuming micronucleus assay can be used to determine whether an individual is a responder or non-responder.

  18. Cytogenetic characterization of low-dose hyper-radiosensitivity in Cobalt-60 irradiated human lymphoblastoid cells

    Energy Technology Data Exchange (ETDEWEB)

    Joshi, Gnanada S. [Department of Biological Sciences, Wayne State University, Detroit, MI 48202 (United States); Joiner, Michael C. [Department of Radiation Oncology, Wayne State University, Detroit, MI 48201 (United States); Tucker, James D., E-mail: jtucker@biology.biosci.wayne.edu [Department of Biological Sciences, Wayne State University, Detroit, MI 48202 (United States)

    2014-12-15

    Highlights: • Human cells were irradiated in G1 or G2 and evaluated for micronuclei and bridges. • Cells irradiated in G2 but not in G1 exhibit low dose hyper-radiosensitivity. • Response curves of cells irradiated in G2 do not fit a linear-no-threshold model. • Response curves of cells irradiated in G1 fit a linear-no-threshold model. - Abstract: The dose-effect relationships of cells exposed to ionizing radiation are frequently described by linear quadratic (LQ) models over an extended dose range. However, many mammalian cell lines, when acutely irradiated in G2 at doses ≤0.3 Gy, show hyper-radiosensitivity (HRS) as measured by reduced clonogenic cell survival, thereby indicating greater cell lethality than is predicted by extrapolation from high-dose responses. We therefore hypothesized that the cytogenetic response in G2 cells to low doses would also be steeper than predicted by LQ extrapolation from high doses. We tested our hypothesis by exposing four normal human lymphoblastoid cell lines to 0–400 cGy of Cobalt-60 gamma radiation. The cytokinesis block micronucleus assay was used to determine the frequencies of micronuclei and nucleoplasmic bridges. To characterize the dependence of the cytogenetic damage on dose, univariate and multivariate regression analyses were used to compare the responses in the low- (HRS) and high-dose response regions. Our data indicate that the slope of the response for all four cell lines at ≤20 cGy during G2 is greater than predicted by an LQ extrapolation from the high-dose responses for both micronuclei and bridges. These results suggest that the biological consequences of low-dose exposures could be underestimated and may not provide accurate risk assessments following such exposures.

  19. Cytogenetic Analysis In Blood Lymphocyte From Workers Occupationally Exposed To Low Levels Of Radiation

    International Nuclear Information System (INIS)

    Rahimah Abdul Rahim; Mohd Rodzi Ali; Noraisyah Mohd Yusof; Juliana Mahamad Napiah; Yahaya Talib; Shafii Khamis

    2016-01-01

    Whether it comes from the ground, the sky, or medical treatment, humans are constantly exposed to ionizing radiation from the world around them. This is a normal occurrence, and has always been the case. According to the IAEA International Basic Safety Standard, the radiation dose for public is not more than 1 mSv per year. That is just an average though, and the actual figure may fluctuate widely per person depending on where they live and the medical procedures they had that year. The international standard is to allow people who work with and around radioactive material (researchers, nuclear power plant workers, X-ray technicians and others) to have exposures of not more than 20 mSv total per year. The 20 mSv annual dose is considered to be safe and not significantly increase the risk for radiation-related health effects. Biological dosimetry based on the analysis of micronuclei in the cytokinesis-block micronucleus (CBMN) assay can be used as an alternative method for scoring dicentric chromosomes in the field of radiation protection. Bio dosimetry is mainly performed, in addition to the physical dosimetry, with the aim of individual dose assessment. The aim of the present study was to perform a cytogenetic analysis in peripheral blood lymphocyte of 30 individuals occupationally exposed to low level of ionizing radiation and compare the result with 30 controls using CBMN assay. Number of bi-nucleated cell and micronuclei were scored and statistical analysis was done to see the effect of micronuclei with gender, age and occupation. In conclusion, scoring of micronuclei is a useful cytogenetic monitoring for radiation workers and assessment of genetic damage. (author)

  20. Chromosomal radiosensitivity of prostate cancer patients

    International Nuclear Information System (INIS)

    McRobbie, M.L.; Riches, A.; Baxby, K.

    2003-01-01

    Full text: Radiosensitivity of peripheral blood lymphocytes from prostate cancer patients is being investigated using the G2 assay and the Cytokinesis Block Micronucleus(CBMN)assay. The G2 assay evaluates chromosomal damage caused by irradiating cells in the G2 phase of the cell cycle. The CBMN assay quantifies the post mitotic micronuclei, which are the expression of damage incurred during G0. An association between hypersensitivity to the chromosome damaging effects of ionising radiation and cancer predispostion has been demonstrated in a number of heritable conditions by using the aforementioned techniques. Recently, increased chromosomal radiosensitivity has been demonstrated in a significant proportion of patients with no obvious family history of malignancy. The aim of this study is to establish whether a group of prostatic carcinoma patients exists and if so whether there are any correlations between their G2 and G0 sensitivities. The study has shown there is no correlation between G2 and G0 sensitivity, confirming the general trend that individuals exhibiting chromosomal radiosensitivity are defective in only one mechanism and G2 and G0 sensitivity are largely independent. Current data indicates that there is an identifiable group of men within the prostate cancer population with increased chromosomal radiosensitivity. Using the G2 assay and the 90th percentile of the controls as a cut off point for sensitivity, no significant difference between the controls and the patient population has been found. However, using the CBMN assay and again the 90th percentile, approximately 11% of the control group are sensitive compared with approximately 40% of the carcinoma cases. The implications of this increased radiosensitivity are as yet unclear, but it is indicative of increased chromosomal fragility and therefore, possibly associated with malignant transformation. Hence, it may prove a useful tool in identifying individuals at increased risk of developing

  1. Investigations on potential co-mutagenic effects of formaldehyde

    Energy Technology Data Exchange (ETDEWEB)

    Speit, Günter, E-mail: guenter.speit@uni-ulm.de; Linsenmeyer, Regina; Duong, Giang; Bausinger, Julia

    2014-02-15

    Highlights: • A549 cells were exposed to formaldehyde in combination with various mutagens. • Formaldehyde did not affect the induction and removal of DNA damage (comet assay). • Formaldehyde did not affect the induction of micronuclei by the mutagens tested. • The expression of the O{sup 6}-methylguanine-DNA methyltransferase was not affected. - Abstract: The genotoxicity and mutagenicity of formaldehyde (FA) has been well-characterized during the last years. Besides its known direct DNA-damaging and mutagenic activity in sufficiently exposed cells, FA at low concentrations might also enhance the mutagenic and carcinogenic effects of other environmental mutagens by interfering with the repair of DNA lesions induced by these mutagens. To further assess potential co-mutagenic effects of FA, we exposed A549 human lung cells to FA in combination with various mutagens and measured the induction and removal of DNA damage by the comet assay and the production of chromosomal mutations by the cytokinesis-block micronucleus assay (CBMN assay). The mutagens tested were ionizing radiation (IR), (±)-anti-B[a]P-7,8-dihydrodiol-9,10-epoxide (BPDE), N-nitroso-N-methylurea (methyl nitrosourea; MNU) and methyl methanesulfonate (MMS). FA (10–75 μM) did not enhance the genotoxic and mutagenic activity of these mutagens under the test conditions applied. FA alone and in combination with MNU or MMS did not affect the expression (mRNA level) of the gene of the O{sup 6}-methylguanine-DNA methyltransferase (MGMT) in A549 cells. The results of these experiments do not support the assumption that low FA concentrations might interfere with the repair of DNA damage induced by other mutagens.

  2. Investigations of antioxidant-mediated protection and mitigation of radiation-induced DNA damage and lipid peroxidation in murine skin.

    Science.gov (United States)

    Jelveh, Salomeh; Kaspler, Pavel; Bhogal, Nirmal; Mahmood, Javed; Lindsay, Patricia E; Okunieff, Paul; Doctrow, Susan R; Bristow, Robert G; Hill, Richard P

    2013-08-01

    Radioprotection and mitigation effects of the antioxidants, Eukarion (EUK)-207, curcumin, and the curcumin analogs D12 and D68, on radiation-induced DNA damage or lipid peroxidation in murine skin were investigated. These antioxidants were studied because they have been previously reported to protect or mitigate against radiation-induced skin reactions. DNA damage was assessed using two different assays. A cytokinesis-blocked micronucleus (MN) assay was performed on primary skin fibroblasts harvested from the skin of C3H/HeJ male mice 1 day, 1 week and 4 weeks after 5 Gy or 10 Gy irradiation. Local skin or whole body irradiation (100 kVp X-rays or caesium (Cs)-137 γ-rays respectively) was performed. DNA damage was further quantified in keratinocytes by immunofluorescence staining of γ-histone 2AX (γ-H2AX) foci in formalin-fixed skin harvested 1 hour or 1 day post-whole body irradiation. Radiation-induced lipid peroxidation in the skin was investigated at the same time points as the MN assay by measuring malondialdehyde (MDA) with a Thiobarbituric acid reactive substances (TBARS) assay. None of the studied antioxidants showed significant mitigation of skin DNA damage induced by local irradiation. However, when EUK-207 or curcumin were delivered before irradiation they provided some protection against DNA damage. In contrast, all the studied antioxidants demonstrated significant mitigating and protecting effects on radiation-induced lipid peroxidation at one or more of the three time points after local skin irradiation. Our results show no evidence for mitigation of DNA damage by the antioxidants studied in contrast to mitigation of lipid peroxidation. Since these agents have been reported to mitigate skin reactions following irradiation, the data suggest that changes in lipid peroxidation levels in skin may reflect developing skin reactions better than residual post-irradiation DNA damage in skin cells. Further direct comparison studies are required to confirm

  3. Whole thorax irradiation of non-human primates induces persistent nuclear damage and gene expression changes in peripheral blood cells.

    Science.gov (United States)

    Ghandhi, Shanaz A; Turner, Helen C; Shuryak, Igor; Dugan, Gregory O; Bourland, J Daniel; Olson, John D; Tooze, Janet A; Morton, Shad R; Batinic-Haberle, Ines; Cline, J Mark; Amundson, Sally A

    2018-01-01

    We investigated the cytogenetic and gene expression responses of peripheral blood cells of non-human primates (NHP, Macaca mulatta) that were whole-thorax irradiated with a single dose of 10 Gy. In this model, partial irradiation of NHPs in the thoracic region (Whole Thorax Lung Irradiation, WTLI) allows the study of late radiation-induced lung injury, while avoiding acute radiation syndromes related to hematopoietic and gastrointestinal injury. A transient drop in circulating lymphocytes and platelets was seen by 9 days, followed by elevations in respiratory rate, circulating neutrophils, lymphocytes, and monocytes at 60-100 days, corresponding to computed tomography (CT) and histologic evidence of pneumonitis, and elective euthanasia of four animals. To evaluate long-term DNA damage in NHP peripheral blood lymphocytes after 10 Gy WTLI, we used the cytokinesis-block micronucleus (CBMN) assay to measure chromosomal aberrations as post-mitotic micronuclei in blood samples collected up to 8 months after irradiation. Regression analysis showed significant induction of micronuclei in NHP blood cells that persisted with a gradual decline over the 8-month study period, suggesting long-term DNA damage in blood lymphocytes after WTLI. We also report transcriptomic changes in blood up to 30 days after WTLI. We isolated total RNA from peripheral blood at 3 days before and then at 2, 5 and 30 days after irradiation. We identified 1187 transcripts that were significantly changed across the 30-day time course. From changes in gene expression, we identified biological processes related to immune responses, which persisted across the 30-day study. Response to oxygen-containing compounds and bacteria were implicated by gene-expression changes at the earliest day 2 and latest, day 30 time-points. Gene expression changes suggest a persistent altered state of the immune system, specifically response to infection, for at least a month after WTLI.

  4. Harmonization of radiobiological assays: why and how?

    International Nuclear Information System (INIS)

    Prasanna, Pataje G.

    2014-01-01

    The International Atomic Energy Agency has made available a technical manual for cytogenetic biodosimetry assays (dicentric chromosome aberration (DCA) and cytokinesis-block micronucleus (CBMN) assays) used for radiation dose assessment in radiation accidents. The International Standardization Organization, which develops standards and guidelines, also provides an avenue for laboratory accreditation, has developed guidelines and recommendations for performing cytogenetic biodosimetry assays. Harmonization of DCA and CBMN assays, has improved their accuracy. Double-blinded inter-laboratory comparison studies involving several networks have further validated DCA and CBMN assays and improved the confidence in their potential use for radiation dose assessment in mass casualties. This kind of international harmonization is lacking for pre-clinical radiobiology assays. The widely used pre-clinical assays that are relatively important to set stage for clinical trials include clonogenic assays, flow-cytometry assays, apoptotic assays, and tumor regression and growth delay assays. However, significant inter-laboratory variations occur with respect to data among laboratories. This raises concerns on the reliability and reproducibility of preclinical data that drives further development and translation. Lack of reproducibility may stem from a variety of factors such as poor scientist training, less than optimal experimental design, inadequate description of methodology, and impulse to publish only the positive data etc. Availability of technical manuals, standard operating procedures, accreditation avenues for laboratories performing such assays, inter-laboratory comparisons, and use of standardized protocols are necessary to enhance reliability and reproducibility. Thus, it is important that radiobiological assays are harmonized for laboratory protocols to ensure successful translation of pre-clinical research on radiation effect modulators to help design clinic trials with

  5. DNA damage and cytotoxicity in pathology laboratory technicians exposed to organic solvents

    Directory of Open Access Journals (Sweden)

    TATIANE DE AQUINO

    2016-03-01

    Full Text Available The aim of this study was to evaluate potential DNA damage and cytotoxicity in pathology laboratory technicians exposed to organic solvents, mainly xylene. Peripheral blood and buccal cells samples were collected from 18 technicians occupationally exposed to organic solvents and 11 non-exposed individuals. The technicians were sampled at two moments: Monday and Friday. DNA damage and cytotoxicity were evaluated using the Comet Assay and the Buccal Micronucleus Cytome assay. Fifteen subjects (83.5% of the exposed group to solvents complained about some symptom probably related to contact with vapours of organic solvents. DNA damage in the exposed group to solvents was nearly 2-fold higher on Friday than on Monday, and in both moments the individuals of this group showed higher levels of DNA damage in relation to controls. No statistical difference was detected in buccal cell micronucleus frequency between the laboratory technicians and the control group. However, in the analysis performed on Friday, technicians presented higher frequency (about 3-fold of karyolytic and apoptotic-like cells (karyorrhectic and pyknotic in relation to control group. Considering the damage frequency and the working time, a positive correlation was found in the exposed group to solvents (r=0.468; p=0.05. The results suggest that pathology laboratory workers inappropriately exposed to organic solvents have increased levels of DNA damage.

  6. A three-dimensional in vitro HepG2 cells liver spheroid model for genotoxicity studies.

    Science.gov (United States)

    Shah, Ume-Kulsoom; Mallia, Jefferson de Oliveira; Singh, Neenu; Chapman, Katherine E; Doak, Shareen H; Jenkins, Gareth J S

    2018-01-01

    The liver's role in metabolism of chemicals makes it an appropriate tissue for toxicity testing. Current testing protocols, such as animal testing and two-dimensional liver cell systems, offer limited resemblance to in vivo liver cell behaviour, in terms of gene expression profiles and metabolic competence; thus, they do not always accurately predict human toxicology. In vitro three-dimensional liver cell models offer an attractive alternative. This study reports on the development of a 3D liver model, using HepG2 cells, by a hanging-drop technique, with a focus on evaluating spheroid growth characteristics and suitability for genotoxicity testing. The cytokinesis-blocked micronucleus assay protocol was adapted to enable micronucleus (MN) detection in the 3D spheroid models. This involved evaluating the difference between hanging vs non-hanging drop positions for dosing of the test agents and comparison of automated Metafer scoring with manual scoring for MN detection in HepG2 spheroids. The initial seeding density, used for all experiments, was 5000 cells/20 μl drop hanging spheroids, harvested on day 4, with >75% cell viability. Albumin secretion (7.8 g/l) and both CYP1A1 and CYP1A2 gene expression were highest in the 3D environment at day 4. Exposure to metabolically activated genotoxicants for 24 h resulted in a 6-fold increase in CYP1A1 enzyme activity (3 μM B[a]P) and a 30-fold increase in CYP1A2 enzyme activity (5 μM PhIP) in 3D hanging spheroids. MN inductions in response to B[a]P or PhIP were 2-fold and 3-fold, respectively, and were greater in 3D hanging spheroids than in 2D format, showing that hanging spheroids are more sensitive to genotoxic agents. HepG2 hanging-drop spheroids are an exciting new alternative system for genotoxicity studies, due to their improved structural and physiological properties, relative to 2D cultures. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Is mobile phone radiation genotoxic? An analysis of micronucleus frequency in exfoliated buccal cells.

    Science.gov (United States)

    de Oliveira, F M; Carmona, A M; Ladeira, C

    2017-10-01

    Electromagnetic fields (EMF) are classified as "possibly carcinogenic" by the International Agency for Research on Cancer (IARC). Some publications have reported associations between EMF exposure and DNA damage, but many other studies contradict such findings. Cytomorphological changes, such as micronuclei (MN), indicative of genomic damage, are biomarkers of genotoxicity. To test whether mobile phone-associated EMF exposure affects the MN frequency in exfoliated buccal cells, we obtained cells smears from the left and right inner cheeks of healthy mobile phone users, aged 18-30 (n=86), who also completed a characterization survey. MN frequencies were tested for potential confounding factors and for duration of phone use and preferential side of mobile phone use. No relationship was observed between MN frequency and duration of mobile phone use in daily calls. Cells ipsilateral to mobile phone use did not present a statistically significantly higher MN frequency, compared to cells contralateral to exposure. A highly statistically significant (pphone-associated EMF do not to induce MN formation in buccal cells at the observed exposure levels. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Ultrastructural changes of the macronucleus of Paramecium bursaria after UV irradiation of the micronucleus

    Energy Technology Data Exchange (ETDEWEB)

    Fokin, S.I.; Borchsenius, O.N. (Leningradskij Gosudarstvennyj Univ. (USSR). Biologicheskij Nauchno-Issledovatel' skij Inst.)

    1982-01-01

    Dynamics of appearance of macronucleus (MA) fragments in vegetative cells of Paramecium bursaria after irradiation of their micronuclei (MI) with ultraviolet rays (exposure dose of 306 J/m/sup 2/) has been studied. Formation of invaginations of MA shell began in cells fixed already in 0.5-7 h after irradiation. In 2 and 6 days in the shell of cell MA- descendants of irradiated P. bursaria - deep invaginations appeared which resulted in the formation of marked MA blades. In the structure of MA chromatin elements visible changes have not been found during mentioned fixation periods. In 30 days after the irradiation of irradiated cell posterity MA often fragmentize, packing density and sizes of chromatin bands increasing. Near-nucleus areas of cytoplasma in 2.6 and 30 days are saturated with cisterns and channels of endoplasmic reticulum, obviously representing a certain kind of material ''depot'' necessary for increasing MA shell area in connection with its fragmentation. The MA fragmentation in P. bursaria descendants with irradiated MI points to the existence of interaction of these heteromorphous nuclei.

  9. Ultrastructural changes of the macronucleus of Paramecium bursaria after UV irradiation of the micronucleus

    International Nuclear Information System (INIS)

    Fokin, S.I.; Borchsenius, O.N.

    1982-01-01

    Dynamics of appearance of macronucleus (MA) fragments in vegetative cells of Paramecium bursaria after irradiation of their micronuclei (MI) with ultraviolet rays (exposure dose of 306 J/m 2 ) has been studied. Formation of invaginations of MA shell began in cells fixed already in 0.5-7 h after irradiation. In 2 and 6 days in the shell of cell MA- descendants of irradiated P. bursaria - deep invaginations appeared which resulted in the formation of marked MA blades. In the structure of MA chromatin elements visible changes have not been found during mentioned fixation periods. In 30 days after the irradiation of irradiated cell posterity MA often fragmentize, packing density and sizes of chromatin bands increasing. Near-nucleus areas of cytoplasma in 2.6 and 30 days are saturated with cisterns and channels of endoplasmic reticulum, obviously representing a certain kind of material ''depot'' necessary for increasing MA shell area in connection with its fragmentation. The MA fragmentation in P. bursaria descendants with irradiated MI points to the existence of interaction of these heteromorphous nuclei

  10. The use of computer in the biological dosimetry of radiation injury with lymphocyte micronucleus

    International Nuclear Information System (INIS)

    Jiang Benrong; Yao Buo

    1994-01-01

    A new approach for determining by computer the radiation dosage in early diagnosis of radiation injury was presented, with a purpose to widen its practical application. A set of programs designed in Clanguage for computing and drawing was compiled. The technical details discussed here can be used for compiling other kinds of practical programs. It is a valuable attempt in improving the efficiency of the computer-aid clinical diagnosis

  11. Association between cervical lesion grade and micronucleus frequency in the Papanicolaou test

    Directory of Open Access Journals (Sweden)

    Caroline Tanski Bueno

    2014-09-01

    Full Text Available The aim of this study was to evaluate the association between the frequency of micronuclei (MN and the cellular changes detected in the conventional Papanicolaou test. One hundred and seventy-four Papanicolaou test smears with cellular changes were examined. MN screening was done in cytopathological smears by counting 1,000 cervical cells in a light microscope. MN frequencies were significantly higher in the group with cellular changes compared to the control group (p < 0.001. The mean MN frequencies were 0.95 ± 1.12 (mean ± SD in the control group (n = 223, 2.98 ± 1.20 in individuals with atypical squamous cells of undetermined significance (ASC-US (n = 50, 4.04 ± 1.45 in cervical intraepithelial neoplasia (CIN I (n = 52, 5.97 ± 1.83 in CIN II (n = 30, 7.29 ± 1.55 in CIN III (n = 17 and 8.64 ± 1.55 in invasive cancer (n = 25. These findings suggest that MN monitoring should be included as an additional criterion for the early detection of cytogenetic damage in routine examinations. This monitoring should be done in the same smear as used for cytopathological examination. More specific and systematic studies are necessary to confirm this proposal.

  12. The micronucleus assay in mammalian cells in vitro to assess health benefits of various phytochemicals.

    Science.gov (United States)

    Meschini, Roberta; Berni, Andrea; Filippi, Silvia; Pepe, Gaetano; Grossi, Maria Rosaria; Natarajan, Adayapalam T; Palitti, Fabrizio

    2015-11-01

    We evaluated the protective effects of Gentiana lutea extracts (GLEx) and 6-Gingerol (6-G) on clastogenicity of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 7,12-dimethylbenz(α) anthracene (DMBA) in vitro on HepG2 cells using the frequencies of induced micronuclei (MN) as the end point. Pre-, post- and simultaneous treatments with GLEx or 6-G and the carcinogens were carried out. Both GLEx post- and simultaneous treatments reduced the frequencies of MN induced by MNNG and DMBA. Probably this effect is due to an increase of cytostasis and a physico-chemical interaction between GLEx and DMBA under simultaneous treatment. Pre- and simultaneous treatments with 6-G significantly reduced the yield of MNNG-induced micronuclei without affecting % of cytostasis. Simultaneous treatment with 6-G plus DMBA resulted in reduction in the frequency of MN and an increase in cytotoxicity compared to sample treated alone with DMBA, whereas a post-treatment, caused a significant decrease in the yield of MN compared with DMBA alone without any cytotoxic effect. These results are compared with our earlier data obtained in the same system with other phytochemicals. It is concluded that for a critical evaluation of the protective effects of phytochemicals, both the influence on the induced MN and induced cytostasis have to be considered. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Differences in micronucleus frequency and acrylamide adduct levels with hemoglobin between vegetarians and non-vegetarians.

    Science.gov (United States)

    Kotova, Natalia; Frostne, Cecilia; Abramsson-Zetterberg, Lilianne; Tareke, Eden; Bergman, Rolf; Haghdoost, Siamak; Paulsson, Birgit; Törnqvist, Margareta; Segerbäck, Dan; Jenssen, Dag; Grawé, Jan

    2015-10-01

    Nutrients and food constituents can prevent or contribute to genotoxicity. In this study, the possible influence of a vegetarian/non-vegetarian diet on genotoxic effects was investigated in 58 non-smoking healthy vegetarians (V) and non-vegetarians (NV), age 21-37 years from the Stockholm area in Sweden. Physical activity and dietary habits were similar in both groups, with the exception of the intake of meat and fish. Using flow cytometry, we determined the formation of micronuclei (MN) in transferrin-positive immature peripheral blood reticulocytes (Trf-Ret) (Total: n = 53; V: n = 27; NV: n = 26). Dietary exposure to acrylamide was measured through hemoglobin (Hb) adducts in peripheral erythrocytes (Total: n = 53; V: n = 29; NV: n = 24). Hb adducts of both acrylamide and its genotoxic metabolite glycidamide were monitored as a measure of the corresponding in vivo doses. Our data demonstrated that compared with the non-vegetarians, the vegetarians exhibited lower frequencies of MN (fMN) in the Trf-Ret (p vegetarians and non-vegetarians. Furthermore, there were no significant relationships between the adduct levels and fMN in the individuals. The ratio of the Hb adduct levels from glycidamide and acrylamide, however, showed a significant difference (p vegetarian diet might be beneficial in lowering genomic instability in healthy individuals. The measured Hb adduct levels indicate that the total intake of acrylamide does not differ between the two studied groups and does not contribute to the observed difference in fMN, although an influence of the diet on the metabolic rates of acrylamide was indicated. In addition, the observed significant difference in the background fMN in the two groups demonstrated that the MN analysis method has a sensitivity applicable to the biomonitoring of human lifestyle factors.

  14. Use of micronucleus test in evaluating radioprotective action of mixed tremella preparation

    International Nuclear Information System (INIS)

    Lu Jiaben; Huang Yinmei; Zhu Bingchai

    1993-01-01

    The radioprotective action of mixed tremella preparation (Yinshen Yin) on whole body gamma irradiated dogs has been studied. 25 male dogs were divided into five groups: tremella, mercaminum, Yinshen Yin at high dose, Yinshen Yin at low dose and control. All the dogs were given above drugs in 10 successive days before acute exposure of 1 Gy radiation. The results showed that the prophylactic use of Yinshen Yin had good anti-irradiation effect, and the radioprotective effect was similar to the mercaminum, but significantly higher than tremella. Thereby it was considered as a good radioprotective agent. The dose reduction factor in different groups were: Yinshen Yin high dose 2.01, Yinshen Yin low dose group 1.72, mercaminum group 1.55 and tremella group 1.11

  15. Assessment of the Genotoxicity of olive mill waste water (OMWW) with the Vicia faba Micronucleus test

    International Nuclear Information System (INIS)

    El Hajjouji, H.; Pinelli, E.; Revel, J. C.; Hafidi, M.

    2009-01-01

    Olive mill waste water (OMW) can cause serious environmental hazards in olive producing countries, especially around the Mediterranean basin. In Morocco, olive mills are noe of the foremost polluters: the volume of OMW produced annually is estimated at 250 000 m 3 during the season of production. the present study concerns the genotoxicity of OMW generated in mills producing olive oil in Morocco. (Author)

  16. Tradescantia micronucleus test indicates genotoxic potential of traffic emissions in European cities

    DEFF Research Database (Denmark)

    Klumpp, A.; Ansel, W.; Klumpp, G.

    2006-01-01

    Urban atmospheres contain complex mixtures of air pollutants including mutagenic and carcinogenic substances such as benzene, diesel soot, heavy metals and polycyclic aromatic hydrocarbons. In the frame of a European network for the assessment of air quality by the use of bioindicator plants...

  17. CYTOGENETIC STUDIES OF THREE TRIAZINE HERBICIDES II. IN VIVO MICRONUCLEUS STUDIES IN MOUSE BONE MARROW

    Science.gov (United States)

    Atrazine, simazine, and cyanazine are widely used preemergence and postemergence triazine herbicides that have made their way into the potable water supply of many agricultural communities. There are several contradictory studies in the literature. Our previous in vitro studies...

  18. A preliminary study on micronucleus analysis and nuclear anomalies in Pelophylax ridibundus (Pallas,

    Directory of Open Access Journals (Sweden)

    Mert Gürkan

    2015-12-01

    Full Text Available Bu çalışmada Vize (Kırklareli ve Kazdağı (Yenice, Çanakkale civarında toplanan Pelophylax ridibundus örneklerine ait eritrositlerde mikronukleus analizi yapıldı ve nuklear anomaliler tespit edildi. Bu amaçla Nisan 2011’de Vize’den 9 (5♂♂, 4♀♀, Mayıs 2011’de Kazdağı’ndan 5 (3♂♂, 2♀♀ adet P. ridibundus örneği toplandı. Vize’den toplanan örneklerde ortalama mikronukleus sayısı 6±4,17, frekans % 0,3; Kazdağı’ndan toplanan örneklerde ise 0,6±0,43, frekans % 0,03 olarak hesaplandı. İstatistiksel analizler neticesinde iki lokalite arasında toplam mikronukleus sayısı bakımından önemli fark bulunduğu belirlendi (p≤0,05. Çalışmada binuklear, çentikli, tomurcuklu ve küçük loplu olmak üzere 4 tip nuklear anomali saptandı. Vize ve Kazdağı örnekleri arasında toplam nuklear anomali sayısı bakımından fark tespit edilmedi (p=0,31. Araştırmamız sonunda elde edilen bulgulara göre, Vize örneklerinde mikronukleus frekansının daha yüksek bulunması, bölgedeki yoğun tarımsal faaliyetlerde kullanılan genotoksik pestisitlere maruz kalınmayla ilişkilendirilebilir

  19. Studies of the in vivo radiosensitivity of human skin fibroblasts

    International Nuclear Information System (INIS)

    Hill, Richard P.; Kaspler, Pavel; Griffin, Anthony M.; O'Sullivan, Brian; Catton, Charles; Alasti, Hamideh; Abbas, Ahmar; Heydarian, Moustafa; Ferguson, Peter; Wunder, Jay S.; Bell, Robert S.

    2007-01-01

    Background and purpose: To examine the radiosensitivity of skin cells obtained directly from the irradiated skin of patients undergoing fractionated radiation treatment prior to surgery for treatment of soft tissue sarcoma (STS) and to determine if there was a relationship with the development of wound healing complications associated with the surgery post-radiotherapy. Methods: Micronucleus (MN) formation was measured in cells (primarily dermal fibroblasts) obtained from human skin at their first division after being removed from STS patients during post-radiotherapy surgery (2-9 weeks after the end of the radiotherapy). At the time of radiotherapy (planned tumor dose - 50 Gy in 25 daily fractions) measurements were made of surface skin dose at predetermined marked sites. Skin from these sites was obtained at surgery and cell suspensions were prepared directly for the cytokinesis-blocked MN assay. Cultured strains of the fibroblasts were also established from skin nominally outside the edge of the radiation beam and DNA damage (MN formation) was examined following irradiation in vitro for comparison with the results from the in situ irradiations. Results: Extensive DNA damage (MN) was detectable in fibroblasts from human skin at extended periods after irradiation (2-9 weeks after the end of the 5-week fractionated radiotherapy). Analysis of skin receiving a range of doses demonstrated that the level of damage observed was dose dependent. There was no clear correlation between the level of damage observed after irradiation in situ and irradiation of cell strains in culture. Similarly, there was no correlation between the extent of MN formation following in situ irradiation and the propensity for the patient to develop wound healing complications post-surgery. Conclusions: Despite the presence of DNA damage in dermal fibroblasts weeks after the end of the radiation treatment, there was no relationship between this damage and wound healing complications following

  20. First evaluation of the biologic effectiveness factors of boron neutron capture therapy (BNCT) in a human colon carcinoma cell line.

    Science.gov (United States)

    Dagrosa, Maria Alejandra; Crivello, Martín; Perona, Marina; Thorp, Silvia; Santa Cruz, Gustavo Alberto; Pozzi, Emiliano; Casal, Mariana; Thomasz, Lisa; Cabrini, Romulo; Kahl, Steven; Juvenal, Guillermo Juan; Pisarev, Mario Alberto

    2011-01-01

    DNA lesions produced by boron neutron capture therapy (BNCT) and those produced by gamma radiation in a colon carcinoma cell line were analyzed. We have also derived the relative biologic effectiveness factor (RBE) of the neutron beam of the RA-3- Argentine nuclear reactor, and the compound biologic effectiveness (CBE) values for p-boronophenylalanine ((10)BPA) and for 2,4-bis (α,β-dihydroxyethyl)-deutero-porphyrin IX ((10)BOPP). Exponentially growing human colon carcinoma cells (ARO81-1) were distributed into the following groups: (1) BPA (10 ppm (10)B) + neutrons, (2) BOPP (10 ppm (10)B) + neutrons, (3) neutrons alone, and (4) gamma rays ((60)Co source at 1 Gy/min dose-rate). Different irradiation times were used to obtain total absorbed doses between 0.3 and 5 Gy (±10%) (thermal neutrons flux = 7.5 10(9) n/cm(2) sec). The frequency of micronucleated binucleated cells and the number of micronuclei per micronucleated binucleated cells showed a dose-dependent increase until approximately 2 Gy. The response to gamma rays was significantly lower than the response to the other treatments (p irradiations with neutrons alone and neutrons + BOPP showed curves that did not differ significantly from, and showed less DNA damage than, irradiation with neutrons + BPA. A decrease in the surviving fraction measured by 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolium bromide (MTT) assay as a function of the absorbed dose was observed for all the treatments. The RBE and CBE factors calculated from cytokinesis block micronucleus (CBMN) and MTT assays were, respectively, the following: beam RBE: 4.4 ± 1.1 and 2.4 ± 0.6; CBE for BOPP: 8.0 ± 2.2 and 2.0 ± 1; CBE for BPA: 19.6 ± 3.7 and 3.5 ± 1.3. BNCT and gamma irradiations showed different genotoxic patterns. To our knowledge, these values represent the first experimental ones obtained for the RA-3 in a biologic model and could be useful for future experimental studies for the application of BNCT to colon carcinoma

  1. Evaluation of genotoxic effects of surface waters using a battery of bioassays indicating different mode of action.

    Science.gov (United States)

    Han, Yingnan; Li, Na; Oda, Yoshimitsu; Ma, Mei; Rao, Kaifeng; Wang, Zijian; Jin, Wei; Hong, Gang; Li, Zhiguo; Luo, Yi

    2016-11-01

    With the burgeoning contamination of surface waters threatening human health, the genotoxic effects of surface waters have received much attention. Because mutagenic and carcinogenic compounds in water cause tumors by different mechanisms, a battery of bioassays that each indicate a different mode of action (MOA) is required to evaluate the genotoxic effects of contaminants in water samples. In this study, 15 water samples from two source water reservoirs and surrounding rivers in Shijiazhuang city of China were evaluated for genotoxic effects. Target chemical analyses of 14 genotoxic pollutants were performed according to the Environmental quality standards for surface water of China. Then, the in vitro cytokinesis-block micronucleus (CBMN) assay, based on a high-content screening technique, was used to detect the effect of chromosome damage. The SOS/umu test using strain TA1535/pSK1002 was used to detect effects on SOS repair of gene expression. Additionally, two other strains, NM2009 and NM3009, which are highly sensitive to aromatic amines and nitroarenes, respectively, were used in the SOS/umu test to avoid false negative results. In the water samples, only two of the genotoxic chemicals listed in the water standards were detected in a few samples, with concentrations that were below water quality standards. However, positive results for the CBMN assay were observed in two river samples, and positive results for the induction of umuC gene expression in TA1535/pSK1002 were observed in seven river samples. Moreover, positive results were observed for NM2009 with S9 and NM3009 without S9 in some samples that had negative results using the strain TA1535/pSK1002. Based on the results with NM2009 and NM3009, some unknown or undetected aromatic amines and nitroarenes were likely in the source water reservoirs and the surrounding rivers. Furthermore, these compounds were most likely the causative pollutants for the genotoxic effect of these water samples. Therefore

  2. Investigation of olive mill wastewater (OMW) ozonation efficiency with the use of a battery of selected ecotoxicity and human toxicity assays

    Energy Technology Data Exchange (ETDEWEB)

    Siorou, Sofia [Section of Animal Biology, Department of Biology, Faculty of Sciences, University of Patras, GR-26500 Patras (Greece); Vgenis, Theodoros T.; Dareioti, Margarita A. [Laboratory of Biochemical Engineering and Environmental Technology, Department of Chemical Engineering, University of Patras, 1 Karatheodori Str., University Campus, GR-26500 Patras (Greece); Vidali, Maria-Sophia; Efthimiou, Ioanna [Department of Environmental and Natural Resources Management, University of Patras, 2 Seferi Str., GR-30100 Agrinio (Greece); Kornaros, Michael [Laboratory of Biochemical Engineering and Environmental Technology, Department of Chemical Engineering, University of Patras, 1 Karatheodori Str., University Campus, GR-26500 Patras (Greece); Vlastos, Dimitris [Department of Environmental and Natural Resources Management, University of Patras, 2 Seferi Str., GR-30100 Agrinio (Greece); Dailianis, Stefanos, E-mail: sdailianis@upatras.gr [Section of Animal Biology, Department of Biology, Faculty of Sciences, University of Patras, GR-26500 Patras (Greece)

    2015-07-15

    Highlights: • Raw- and ozonated-olive mill wastewater (OMW) toxic effects were investigated. • A battery of biological assays and toxic endpoints were used. • Ozonation for up to 300 min attenuates OMW toxicity, following phenols’ reduction. • Further OMW ozonation (>300 min) could enhance OMW toxicity. • OMW ozonation efficacy depends on OMW-derived intermediates and high NO{sub 3}{sup −}–N levels. - Abstract: The effects of olive mill wastewater (OMW) on a battery of biological assays, before and during the ozonation process, were investigated in order to assess ozone’s efficiency in removing phenolic compounds from OMW and decreasing the concomitant OMW toxicity. Specifically, ozonated-OMW held for 0, 60, 120, 300, 420, 540 min in a glass bubble reactor, showed a drastic reduction of OMW total phenols (almost 50%) after 300 min of ozonation with a concomitant decrease of OMW toxicity. In particular, the acute toxicity test primarily performed in the fairy shrimp Thamnocephalus platyurus (Thamnotoxkit F™ screening toxicity test) showed a significant attenuation of OMW-induced toxic effects, after ozonation for a period of 120 and in a lesser extent 300 min, while further treatment resulted in a significant enhancement of ozonated-OMW toxic effects. Furthermore, ozonated-OMW-treated mussel hemocytes showed a significant attenuation of the ability of OMW to cause cytotoxic (obtained by the use of NRRT assay) effects already after an ozonation period of 120 and to a lesser extent 300 min. In accordance with the latter, OMW-mediated oxidative (enhanced levels of superoxide anions and lipid peroxidation by-products) and genotoxic (induction of DNA damage) effects were diminished after OMW ozonation for the aforementioned periods of time. The latter was also revealed by the use of cytokinesis block micronucleus (CBMN) assay in human lymphocytes exposed to different concentrations of both raw- and ozonated-OMW for 60, 120 and 300 min. Those findings

  3. Application of translocation, γ-H2AX, and Sam68 as a biological indicators for the assessment of radiation exposure in nuclear power plant workers

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Kwang Hee; Park, Hyung Sun; Nam, Seon Young [Korea Hydro Nuclear Power Co., Seoul (Korea, Republic of)

    2014-05-15

    protection programs of many Member States. However, a recognized drawback of the dicentric and cytokinesis-block micronucleus (CBMN) assays is that the damage is unstable. For retrospective biological dosimetry, FISH (fluorescence in situ hybridization) method is possible because stable aberrations such as a reciprocal translocation will pass successfully through mitosis and into the daughter cells.

  4. Vincristine-induced bystander effect in human lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Testi, Serena; Azzarà, Alessia; Giovannini, Caterina; Lombardi, Sara [Unità di Genetica, Dipartimento di Biologia, Pisa University, Via Derna 1, 56126 Pisa (Italy); Piaggi, Simona [Dipartimento di Ricerca Traslazionale e delle Nuove Tecnologie in Medicina e Chirurgia, Pisa University, Via Savi 10, 56126 Pisa (Italy); Facioni, Maria Sole [Unità di Genetica, Dipartimento di Biologia, Pisa University, Via Derna 1, 56126 Pisa (Italy); Scarpato, Roberto, E-mail: roberto.scarpato@unipi.it [Unità di Genetica, Dipartimento di Biologia, Pisa University, Via Derna 1, 56126 Pisa (Italy); Research Center of Nutraceuticals and Food for Health, University of Pisa, Pisa (Italy)

    2016-07-15

    Highlights: • We studied whether or not vincristine induced a bystander response in human lymphocytes. • Vincristine significantly increased MN frequencies in mononucleated recipient cells. • ROS or soluble proteins (IL-32 and TGF-β) may account for the observed response. - Abstract: Bystander effect is a known radiobiological effect, widely described using ionizing radiations and which, more recently, has also been related to chemical mutagens. In this study, we aimed to assess whether or not a bystander response can be induced in cultured human peripheral lymphocytes by vincristine, a chemotherapeutic mutagen acting as spindle poison, and by mitomycin-C, an alkylating agent already known to induce this response in human lymphoblastoid cells. Designing a modified ad hoc protocol for the cytokinesis blocked micronucleus (MN) assay, we detected the presence of a dose-dependent bystander response in untreated cultures receiving the conditioned medium (CM) from mitomycin-C (MMC) or vincristine (VCR) treated cultures. In the case of MMC, MN frequencies, expressed as micronucleated binucleates, were: 13.5 ± 1.41 at 6 μM, 22 ± 2.12 at 12 μM or 28.25 ± 5.13 at 15 μM vs. a control value of 4.75 ± 1.59. MN levels for VCR, expressed as micronucleated mononucleates were: 2.75 ± 0.88 at 0.0 μM, 27.25 ± 2.30 at 0.4 μM, 46.25 ± 1.94 at 0.8 μM, 98.25 ± 7.25 at 1.6 μM. To verify that no mutagen residual was transferred to recipient cultures together with the CM, we evaluated MN levels in cultures receiving the medium immediately after three washings following the chemical treatment (unconditioned medium). We further confirmed these results using a cell-mixing approach where untreated lymphocytes were co-cultured with donor cells treated with an effect-inducing dose of MMC or VCR. A distinct production pattern of both reactive oxygen species and soluble mediator proteins by treated cells may account for the differences observed in the manifestation of the

  5. Characterization of coal fly ash nanoparticles and induced oxidative DNA damage in human peripheral blood mononuclear cells

    International Nuclear Information System (INIS)

    Dwivedi, Sourabh; Saquib, Quaiser; Al-Khedhairy, Abdulaziz A.; Ali, Al-Yousef Sulaiman; Musarrat, Javed

    2012-01-01

    The nano-sized particles present in coal fly ash (CFA) were characterized through the X-ray diffraction (XRD), transmission and scanning electron microscopy (TEM, SEM), atomic force microscopy (AFM) and Fourier transform infrared spectroscopy (FTIR) analyses. The XRD data revealed the average crystallite size of the CFA nanoparticles (CFA-NPs) as 14 nm. TEM and SEM imaging demonstrated predominantly spherical and some polymorphic structures in the size range of 11 to 25 nm. The amount of heavy metal associated with CFA particles (μg/g) were determined as Fe (34160.0 ± 1.38), Ni (150.8 ± 0.78), Cu (99.3 ± 0.56) and Cr (64.0 ± 0.86). However, the bioavailability of heavy metals in terms of percent release was in the order as Cr > Ni > Cu > Fe in CFA-dimethyl sulfoxide (DMSO) extract. The comet and cytokinesis blocked micronucleus (CBMN) assays revealed substantial genomic DNA damage in peripheral blood mononuclear (PBMN) cells treated with CFA-NPs in Aq and DMSO extracts. About 1.8 and 3.6 strand breaks per unit of DNA were estimated through alkaline unwinding assay at 1:100 DNA nucleotide/CFA ppm ratios with the Aq and DMSO extracts, respectively. The DNA and mitochondrial damage was invariably greater with CFA-DMSO extract vis-à-vis -Aq extract. Generation of superoxide anions (O 2 • − ) and intracellular reactive oxygen species (ROS) through metal redox-cycling, alteration in mitochondrial potential and 8-oxodG production elucidated CFA-NPs induced oxidative stress as a plausible mechanism for CFA-induced genotoxicity. -- Highlights: ► CFA consists of spherical crystalline nanoparticles in size range of 11–25 nm. ► Alkaline unwinding assay revealed single-strandedness in CFA treated ctDNA. ► CFA nanoparticles exhibited the ability to induce ROS and oxidative DNA damage. ► Comet and CBMN assays revealed DNA and chromosomal breakage in PBMN cells. ► CFA-NPs resulted in mitochondrial membrane damage in PBMN cells.

  6. Characterization of coal fly ash nanoparticles and induced oxidative DNA damage in human peripheral blood mononuclear cells

    Energy Technology Data Exchange (ETDEWEB)

    Dwivedi, Sourabh; Saquib, Quaiser; Al-Khedhairy, Abdulaziz A. [Department of Zoology, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451 (Saudi Arabia); Ali, Al-Yousef Sulaiman [Department of Medical Laboratory Sciences, College of Applied Medical Science, University of Dammam, P.O. Box 1683, Hafr Al Batin-31991 (Saudi Arabia); Musarrat, Javed, E-mail: musarratj1@yahoo.com [Department of Zoology, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451 (Saudi Arabia); Department of Agricultural Microbiology, Faculty of Agricultural Sciences, AMU, Aligarh202002 (India)

    2012-10-15

    The nano-sized particles present in coal fly ash (CFA) were characterized through the X-ray diffraction (XRD), transmission and scanning electron microscopy (TEM, SEM), atomic force microscopy (AFM) and Fourier transform infrared spectroscopy (FTIR) analyses. The XRD data revealed the average crystallite size of the CFA nanoparticles (CFA-NPs) as 14 nm. TEM and SEM imaging demonstrated predominantly spherical and some polymorphic structures in the size range of 11 to 25 nm. The amount of heavy metal associated with CFA particles ({mu}g/g) were determined as Fe (34160.0 {+-} 1.38), Ni (150.8 {+-} 0.78), Cu (99.3 {+-} 0.56) and Cr (64.0 {+-} 0.86). However, the bioavailability of heavy metals in terms of percent release was in the order as Cr > Ni > Cu > Fe in CFA-dimethyl sulfoxide (DMSO) extract. The comet and cytokinesis blocked micronucleus (CBMN) assays revealed substantial genomic DNA damage in peripheral blood mononuclear (PBMN) cells treated with CFA-NPs in Aq and DMSO extracts. About 1.8 and 3.6 strand breaks per unit of DNA were estimated through alkaline unwinding assay at 1:100 DNA nucleotide/CFA ppm ratios with the Aq and DMSO extracts, respectively. The DNA and mitochondrial damage was invariably greater with CFA-DMSO extract vis-a-vis -Aq extract. Generation of superoxide anions (O{sub 2} Bullet {sup -}) and intracellular reactive oxygen species (ROS) through metal redox-cycling, alteration in mitochondrial potential and 8-oxodG production elucidated CFA-NPs induced oxidative stress as a plausible mechanism for CFA-induced genotoxicity. -- Highlights: Black-Right-Pointing-Pointer CFA consists of spherical crystalline nanoparticles in size range of 11-25 nm. Black-Right-Pointing-Pointer Alkaline unwinding assay revealed single-strandedness in CFA treated ctDNA. Black-Right-Pointing-Pointer CFA nanoparticles exhibited the ability to induce ROS and oxidative DNA damage. Black-Right-Pointing-Pointer Comet and CBMN assays revealed DNA and chromosomal

  7. Vincristine-induced bystander effect in human lymphocytes

    International Nuclear Information System (INIS)

    Testi, Serena; Azzarà, Alessia; Giovannini, Caterina; Lombardi, Sara; Piaggi, Simona; Facioni, Maria Sole; Scarpato, Roberto

    2016-01-01

    Highlights: • We studied whether or not vincristine induced a bystander response in human lymphocytes. • Vincristine significantly increased MN frequencies in mononucleated recipient cells. • ROS or soluble proteins (IL-32 and TGF-β) may account for the observed response. - Abstract: Bystander effect is a known radiobiological effect, widely described using ionizing radiations and which, more recently, has also been related to chemical mutagens. In this study, we aimed to assess whether or not a bystander response can be induced in cultured human peripheral lymphocytes by vincristine, a chemotherapeutic mutagen acting as spindle poison, and by mitomycin-C, an alkylating agent already known to induce this response in human lymphoblastoid cells. Designing a modified ad hoc protocol for the cytokinesis blocked micronucleus (MN) assay, we detected the presence of a dose-dependent bystander response in untreated cultures receiving the conditioned medium (CM) from mitomycin-C (MMC) or vincristine (VCR) treated cultures. In the case of MMC, MN frequencies, expressed as micronucleated binucleates, were: 13.5 ± 1.41 at 6 μM, 22 ± 2.12 at 12 μM or 28.25 ± 5.13 at 15 μM vs. a control value of 4.75 ± 1.59. MN levels for VCR, expressed as micronucleated mononucleates were: 2.75 ± 0.88 at 0.0 μM, 27.25 ± 2.30 at 0.4 μM, 46.25 ± 1.94 at 0.8 μM, 98.25 ± 7.25 at 1.6 μM. To verify that no mutagen residual was transferred to recipient cultures together with the CM, we evaluated MN levels in cultures receiving the medium immediately after three washings following the chemical treatment (unconditioned medium). We further confirmed these results using a cell-mixing approach where untreated lymphocytes were co-cultured with donor cells treated with an effect-inducing dose of MMC or VCR. A distinct production pattern of both reactive oxygen species and soluble mediator proteins by treated cells may account for the differences observed in the manifestation of the

  8. A cytogenetic biomonitoring of industrial radiographers occupationally exposed to low levels of ionizing radiation by using cbmn assay.

    Science.gov (United States)

    Shakeri, Mahsa; Zakeri, Farideh; Changizi, Vahid; Rajabpour, Mohammad Reza; Farshidpour, Mohammad Reza

    2017-06-15

    Industrial radiography is the process of using either gamma-emitting radionuclide sources or X-ray machines to examine the safety of industrial materials. The average annual effective dose in industrial radiography is one of the highest among radiation workers. The aim of this study was to investigate the cytogenetic effects of ionizing radiation in the peripheral blood lymphocytes of 60 industrial radiographers and 40 non-exposed individuals as the control group by using cytokinesis-block micronucleus (CBMN) assay. Totally, the frequencies of micronuclei (MN), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) were significantly higher in the industrial radiographers than in the controls (p = 0.000). The mean MN frequency per 1000 binucleated cells in the industrial radiographers with last 5-y radiation dose of >100 mSv was significantly higher than those with ≤100 mSv (34.81 ± 12.7‰ vs. 26.33 ± 7.94‰, p = 0.024). The effect of age was observed in the control group and subjects with the age of >30 y showed significantly higher MN frequency compared with the subjects with the age of ≤30 y (9.45 ± 3.71‰ vs. 6.81 ± 3.05‰, p = 0.02). No obvious trend of increased MN as a function of either duration of employment or age or smoking status was observed in the industrial radiographers. The results show the increased levels of cytogenetic damages in the industrial radiographers. Even the workers exposed to the permissible doses are subjected to elevated frequencies of DNA damages. These findings confirm the importance of cytogenetic biomonitoring program beside physical dosimetry, surveying radiation safety of equipment and periodic training of workers for improvement of safety and radiation protection. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  9. The principal phenolic and alcoholic components of wine protect human lymphocytes against hydrogen peroxide- and ionising radiation-induced DNA damage in vitro

    International Nuclear Information System (INIS)

    Fenech, M.; Greenrod, W.

    2003-01-01

    We have tested the hypothesis that the alcoholic and phenolic components of wine are protective against the DNA damaging and cytotoxic effects of hydrogen peroxide and gamma radiation in vitro. The components of wine tested were ethanol, glycerol, a mixture of the phenolic compounds catechin and caffeic acid, and tartaric acid, all at concentrations that were 2.5% or 10.0% of the concentration in a typical Australian white wine Riesling. These components were tested individually or combined as a mixture and compared to a white wine stripped of polyphenols as well as a Hanks balanced salt solution control which was the diluent for the wine components. The effect of the components was tested in lymphocytes, using the cytokinesis-block micronucleus assay, after 30 minutes incubation in plasma or whole blood for the hydrogen peroxide or gamma-radiation challenge respectively. The results obtained showed that ethanol, glycerol, the catechin-caffeic acid mixture, the mixture of all components, and the stripped white wine significantly reduced the DNA damaging effects of hydrogen peroxide and gamma radiation (ANOVA P = 0.043 - 0.001). The strongest protective effect against DNA damage by gamma irradiation was observed for the catechin-caffeic acid mixture and mixture of all components (30% and 32% reduction respectively). These two treatments as well as ethanol produced the strongest protective effects against DNA damage by hydrogen peroxide (24%, 25% and 18% respectively) . The protection provided by the mixture did not account for the expected additive protective effects of the individual components suggesting that the components may be exerting their effects through similar mechanisms which are saturated at the concentrations tested. Ethanol was the only component that significantly increased base-line DNA damage rate, however, this effect was negated in the mixture. In conclusion our results suggest that the main phenolic and alcoholic components of wine can reduce

  10. The effect of dietary estimates calculated using food frequency questionnaires on micronuclei formation in European pregnant women

    DEFF Research Database (Denmark)

    Vande Loock, Kim; Botsivali, Maria; Zangogianni, Marina

    2014-01-01

    ) and the cytokinesis blocked proliferation index using a semi-automated image analysis system. Food frequency questionnaires (FFQs) were used to estimate intake of fatty acids and a broad range of immunotoxic and genotoxic/carcinogenic compounds through the diet. Pooled difference based on delivery type revealed...

  11. Predictive radiosensitivity tests in human lymphocytes

    International Nuclear Information System (INIS)

    Di Giorgio, Marina; Vallerga, Maria B.; Taja, Maria R.; Sardi, M.; Busto, E.; Mairal, L.; Roth, B.; Menendez, P.; Bonomi, M.

    2004-01-01

    Individual radiosensitivity is an inherent characteristic, associated with an abnormally increased reaction to ionising radiation of both the whole body and cells derived from body tissues. Human population is not uniform in its radiation sensitivity. Radiosensitive sub-groups exist, which would suffer an increased incidence of both deterministic and stochastic effects. Clinical studies have suggested that a large part of the spectrum of normal tissue reaction may be due to differences in individual radiosensitivity. The identification of such sub-groups should be relevant for radiation therapy and radiation protection purposes. It is suggested that DNA repair mechanisms are involved. Consequently, the characterization of DNA repair in lymphocytes through cytokinesis blocked micronucleus (MN) and alkaline single-cell microgel electrophoresis (comet) assays could be a suitable approache to evaluate individual radiosensitivity in vitro. The aims of this study were: 1) To assess the in vitro radiosensitivity of peripheral blood lymphocytes from two groups of cancer patients (prospectively and retrospectively studied), using MN and comet assays, in comparison with the clinical radiation reaction and 2) To test the predictive potential of both techniques for the identification of radiosensitivity sub-groups. 38 cancer patients receiving radiation therapy were enrolled in this study. 19 patients were evaluated prior, mid-way and on completion of treatment (prospective group) and 19 patients were evaluated about 6-18 month after radiotherapy (retrospective group). Cytogenetic data from the prospective group were analysed using a mathematical model to evaluate the attenuation of the cytogenetic effect as a function of the time between a single exposure and blood sampling, estimating a cytogenetic recovery factor k. In the retrospective group, blood samples were irradiated in vitro with 0 (control) or 2 Gy and evaluated using MN test. Cytogenetic data were analysed

  12. Biological dosimetry of low doses (0.05 - 0.4 Gy) with micronucleus dicentrics and chromosome fragments

    International Nuclear Information System (INIS)

    Rodriguez, L.; Sanchez, E.; Linares, C.; Navlet, J.

    1997-01-01

    This study was carried out within the framework of the agreement between the National Nuclear Safety Council and the University of Alcala of Hernares and in co-operation with the Radiological Service and the Radiation Protection Unit of the Gregorio Maranon General Hospital of Madrid, where irradiations were performed. Blood samples were taken of 4 individuals irradiated with 6 doses of gamma rays between 0.05 Gy and 0.40 Gy, leaving an aliquot dose of 0 Gy. Cultures of lymphocytes for the study of dicentric chromosomes (DC) and chromosomal fragments (fr), stopping the first mitosis after postirradiation with Colcemid. A study of micronuclei (MN) in binuclear cells was also performed, interrupting the first cytokinesis after postirradiation with citocalasina B. After the corresponding studies with optical microscope, statistical analysis was made on the observed data on DC, fr and MN. We made a multiple linear regression analysis of the data of the 4 individuals. We obtained the average of the 4 individuals for each variable and dose and performed the variance analysis. According to our study, neither the DC nor the MN are valid dosemeters for lower doses up to 0.4 Gy. Nevertheless there are indications that the fragments are correlated with the dose to these levels. By increasing both the points of dose and the number of metaphases studied, we believe that a dose curve can be done that would allow to estimate with a reasonable degree of confidence the dose received by a sample irradiated

  13. Evaluation of DNA by the micronucleus test of the samples Allium cepa germinated in the presence of uranium

    Energy Technology Data Exchange (ETDEWEB)

    Souza, Ayrton Nascimento de; Oliveira, Andressa L.; Gennari, Roseli F.; Maffei, Eliane M.D.; Campos, Simara S., E-mail: simaracampos@gmail.com [Universidade Estadual do Sudoeste da Bahia (UESB), Vitoria da Conquista, BA, (Brazil). Departamento de Ciencias Exatas e Tecnologicas; Universidade de Sao Paulo (USP), Sao Paulo, SP (Brazil). Instituto de Fisica. Departamento de Fisica Nuclear

    2017-11-01

    Although Brazil composes the seventh position in the world ranking of uranium reserves, the ore is still little explored in Brazil, considering its vast existing arsenal. On the one hand, despite nuclear energy has brought great benefits, technological and socio-economic development, it generates controversy about environmental contamination and risks to public health. Studies on this subject indicate that areas where uranium ore concentration is high, natural environmental radiation exposure levels are already higher than in other regions. The aim of this study is to observe the simple germination of the bio-indicator (Allium cepa), typically used to assess potential chromosomal aberrations, suffer any adverse effect caused by natural radiation uranium. The choice of this bio-indicator is based on its potential for evaluating the mutation caused by innumerous chemical compounds. Four treatments with three replicates were designed. In each treatment, 10 seeds of onion A. cepa without any pesticides were packed in the Petri dish lined with germination paper and room temperature (25°C) was kept until the root reaches approximately 1cm long and 0, 5 g of the U. As a result 3000 cells were counted during the interphase, and structural changes were observed in the chromosomes, elongated nuclei and loose chromosomes of the spindle fiber, concluding that U possibly affected the cell cycle da Allium cepa. (author)

  14. Assessment of the mutagenic and antimutagenic activity of Synadenium umbellatum Pax latex by micronucleus test in mice

    Directory of Open Access Journals (Sweden)

    PR. Melo-Reis

    Full Text Available Synadenium umbellatum Pax, popularly known as "cola-nota", is a medicinal plant that grows in tropical regions. The latex of this plant is used against various diseases, such as diabetes mellitus, leprosy, tripanosomiasis, leukemia, and several malignant tumors. The mutagenic, antimutagenic, and cytotoxic effects of the latex of this plant were investigated by measuring the frequency of micronuclei in mice bone marrow cells. To evaluate mutagenicity, the animals were treated with four doses of latex (10, 30, 50, and 100 mg/kg body weight. To study the antimutagenic activity, the animals were simultaneously treated with latex and mitomycin C (4 mg/kg. The cytotoxicity was evaluated by polychromatic and normochromatic erythrocytes ratio. Our results showed a significant increase of frequency of micronucleated polychromatic erythrocytes (MNPCE compared to the negative control group (p 0.05 was detected at the doses of 50 and 100 mg/kg. Under our experimental conditions, the results obtained indicate strong mutagenic and cytotoxic activity of S. umbellatum latex except the dose of 10 mg/kg and moderate antimutagenic effect at lower doses.

  15. Evaluation of chromosome aberration and micronucleus frequencies in blood lymphocytes of workers exposed to low concentrations of benzene.

    Science.gov (United States)

    Lovreglio, Piero; Maffei, Francesca; Carrieri, Mariella; D'Errico, Maria N; Drago, Ignazio; Hrelia, Patrizia; Bartolucci, Giovanni B; Soleo, Leonardo

    2014-08-01

    The frequency of chromosome aberrations (CA) and micronuclei (MN) was investigated in the peripheral lymphocytes of workers occupationally exposed to low or very low concentrations of benzene. The study included 43 exposed workers (all males), namely 19 fuel-tanker drivers and 24 filling-station attendants, and 31 male subjects with no occupational exposure to the toxicant (controls). Benzene exposure was verified by means of environmental monitoring with passive personal samplers (Radiello(®)), and through biological monitoring, i.e. by measurement of urinary trans,trans-muconic acid, S-phenylmercapturic acid and benzene. The frequency of CA and MN in peripheral lymphocytes was determined according to standard procedures. Exposure to benzene was found to be significantly higher for fuel-tanker drivers (median 246.6 μg/m(3)) than for filling-station attendants (median 19.9 μg/m(3)). Both groups had significantly higher exposure than controls (median 4.3 μg/m(3)). No increased frequency of CA and MN was observed in either fuel-tanker drivers or filling-station attendants compared with controls. In all subjects examined as a single group, the frequency of MN was significantly dependent on age. Only in the fuel-tanker drivers was the frequency of MN found to depend not only on age, but also on exposure to benzene. In conclusion, the frequency of MN, but not of CA, could be influenced by exposure to benzene concentrations of up to one order of magnitude lower than the threshold limit value (time-weighted average). Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Comparison of radiation sensitivity for three cell lines as measured by the cloning assay and the micro-nucleus test

    NARCIS (Netherlands)

    Stap, J.; Aten, J. A.

    1990-01-01

    The correlation between cell killing and the induction of micro-nuclei was studied for three cell lines after treatment with gamma radiation to investigate whether the frequency of micro-nucleated cells can be used to determine the radiation sensitivity of a cell type. R1 rat rhabdomyosarcoma cells

  17. DNA Damage Potential of Engine Emissions Measured In Vitro by Micronucleus Test in Human Bronchial Epithelial Cells

    Czech Academy of Sciences Publication Activity Database

    Červená, Tereza; Rössnerová, Andrea; Sikorová, Jitka; Beránek, V.; Vojtíšek-Lom, M.; Cigánek, M.; Topinka, Jan; Rössner ml., Pavel

    2017-01-01

    Roč. 121, SI (2017), s. 102-108 ISSN 1742-7835 R&D Projects: GA MŠk(CZ) LO1508; GA MŠk(CZ) LM2015073; GA ČR(CZ) GA13-01438S Institutional support: RVO:68378041 Keywords : saliva DNA * methylation patterns * cancer-patients Subject RIV: DN - Health Impact of the Environment Quality OBOR OECD: Public and environmental health Impact factor: 3.176, year: 2016

  18. Multi-micronucleus cells related with viral diseases, detected in the study of children affected by the Chernobyl accident

    International Nuclear Information System (INIS)

    Garcia L, O.; Lamadrid, A.I.; Manzano, J.

    1996-01-01

    Cells with multiple chromosome aberrations have been observed in human peripheral blood lymphocytes. Different explanation have proposed, included hot particle induction in persons related to the Chernobyl accident. The frequency of chromosome aberration and micronuclei were established in 14 Ukrainian children with different hematological disorders. They arrived in Cuba thanks to the program by means of which medical attention is offered to children from areas affected by the Chernobyl accident. At least 500 metaphases and bi-nucleate cells were analyzed in each case. The detection of 4 cells with 7-11 micronuclei in a 14 year old boy with cat scratch disease was the most significant cytogenetical finding. The viral origin of the cat scratch disease has been reported, this suggested a viral etiology of the cells with multiple micronuclei. No rogue cells were detected. Cells with multiple micronuclei or rogue cells were not found in other patients from this group. (authors). 7 refs., 3 tabs

  19. Region-specific chromatin decondensation and micronucleus formation induced by 5-azacytidine in human TIG-7 cells.

    Science.gov (United States)

    Satoh, T; Yamamoto, K; Miura, K F; Sofuni, T

    2004-01-01

    A human diploid lung fibroblast cell strain, TIG-7, has a heteromorphic chromosome 15 with an extra short arm carrying a homogeneously staining region (15p+hsr). We demonstrated previously that the 15p+hsr consists of an inactive and G+C-rich rDNA cluster characterized by fluorescence in situ hybridization (FISH) and various chromosome banding techniques. Thus, it was suggested that the region could contain highly methylated DNA. To observe methylation status on the target region directly under the microscope, we used a demethylating agent, 5-azacytidine (5-azaC), to induce decondensation of the chromatin containing methylated DNA. At 24 h after treatment with 0.5 microM 5-azaC, marked decondensation of the 15p+hsr was observed in almost all of the metaphases. Furthermore, we observed micronuclei, which were equivalent to the rDNA of the 15p+hsr demonstrated by FISH in the same preparation. In contrast, the DNA cross-linking agent mitomycin C (MMC) preferentially induced 15p+hsr-negative micronuclei. These findings indicated that chromatin decondensation and subsequent DNA strand breakage induced by the demethylating effect of 5-azaC led specifically to 15p+hsr-positive micronuclei. Copyright 2003 S. Karger AG, Basel

  20. Whole-genome gene expression modifications associated with nitrosamine exposure and micronucleus frequency in human blood cells

    DEFF Research Database (Denmark)

    Hebels, Dennie G A J; Jennen, Danyel G J; van Herwijnen, Marcel H M

    2011-01-01

    association between MN frequency and urinary NOCs (r = 0.41, P = 0.025) and identified modifications in among others cytoskeleton remodeling, cell cycle, apoptosis and survival, signal transduction, immune response, G-protein signaling and development pathways, which indicate a response to NOC......-induced genotoxicity. Moreover, we established a network of genes, the most important ones of which include FBXW7, BUB3, Caspase 2, Caspase 8, SMAD3, Huntingtin and MGMT, which are involved in processes relevant in carcinogenesis. The modified genetic processes and genes found in this study may be of interest...

  1. Evaluation of DNA by the micronucleus test of the samples Allium cepa germinated in the presence of uranium

    International Nuclear Information System (INIS)

    Souza, Ayrton Nascimento de; Oliveira, Andressa L.; Gennari, Roseli F.; Maffei, Eliane M.D.; Campos, Simara S.; Universidade de Sao Paulo

    2017-01-01

    Although Brazil composes the seventh position in the world ranking of uranium reserves, the ore is still little explored in Brazil, considering its vast existing arsenal. On the one hand, despite nuclear energy has brought great benefits, technological and socio-economic development, it generates controversy about environmental contamination and risks to public health. Studies on this subject indicate that areas where uranium ore concentration is high, natural environmental radiation exposure levels are already higher than in other regions. The aim of this study is to observe the simple germination of the bio-indicator (Allium cepa), typically used to assess potential chromosomal aberrations, suffer any adverse effect caused by natural radiation uranium. The choice of this bio-indicator is based on its potential for evaluating the mutation caused by innumerous chemical compounds. Four treatments with three replicates were designed. In each treatment, 10 seeds of onion A. cepa without any pesticides were packed in the Petri dish lined with germination paper and room temperature (25°C) was kept until the root reaches approximately 1cm long and 0, 5 g of the U. As a result 3000 cells were counted during the interphase, and structural changes were observed in the chromosomes, elongated nuclei and loose chromosomes of the spindle fiber, concluding that U possibly affected the cell cycle da Allium cepa. (author)

  2. Mutagenic potential assessment associated with human exposure to natural radioactivity.

    Science.gov (United States)

    Marcon, Alexandre Endres; Navoni, Julio Alejandro; de Oliveira Galvão, Marcos Felipe; Garcia, Anuska Conde Fagundes Soares; do Amaral, Viviane Souza; Petta, Reinaldo Antônio; Campos, Thomas Ferreira da Costa; Panosso, Renata; Quinelato, Antônio Luiz; de Medeiros, Sílvia Regina Batistuzzo

    2017-01-01

    Lucrécia city, known to harbor a high cancer rate, is located in a semiarid region characterized by the presence of mineral reservoirs, facing a high exposure to metal and natural radioactivity. The present study aimed to assess the environmental scenario at a semiarid region located in Northeastern Brazil. Metal concentration, alpha and beta radiation, and cyanobacteria content in tap water along with indoor radon and gamma emitters (U, K and Th) concentrations were measured. In addition, mutagenic and nuclear instability effects were assessed using buccal micronucleus cytome assay. The study included five samplings corresponding to a period between 2007 and 2009. Drinking water from Lucrécia city presented levels of Mn, Ni and Cr along with cyanobacteria in concentrations one to four times higher than regulatory guidelines considered. Furthermore, high levels of all the tested radionuclides were found. A high percentage of the houses included in this study presented indoor radon concentrations over 100 Bq m -3 . The mean annual effective dose from Lucrécia houses was six times higher than observed in a control region. The levels of exposure in most of the Lucrécia houses were classified as middle to high. A significant mutagenic effect, represented as an increase of micronuclei (MN) frequency and nuclear abnormalities as nuclear buds (NB), binucleated cells (BN), and pyknotic cells (PYC) were found. The results obtained highlight the role of high background radioactivity on the observed mutagenic effect and could help to explain the exacerbated cancer rate reported in this locality. Copyright © 2016. Published by Elsevier Ltd.

  3. Increased Micronuclei Frequency in Oral and Lingual Epithelium of Treated Diabetes Mellitus Patients

    Directory of Open Access Journals (Sweden)

    Jesús Emilo Quintero Ojeda

    2018-01-01

    Full Text Available Diabetes mellitus (DM is a metabolic disease characterized by persistent high levels of glucose in plasma. Chronic hyperglycemia is thought to increase oxidative stress and the formation of free radicals that in turn damage cells. Thus, we decided to determine the frequency of nuclear abnormalities in epithelial cells from cheek and tongue mucosa of DM patients with type 1 (DM1, treated only with insulin and type 2 (DM2, treated with metformin using the buccal micronucleus cytome (BMCyt assay. Micronuclei frequency in cheek epithelial cells was higher in both DM1 (0.75 ± 0.31, P<0.001 and DM2 (0.52 ± 0.27, P<0.001 patients, as compared to healthy controls (0.07  ±  0.06. Similarly, micronuclei frequency in tongue epithelium was increased in DM1 (0.81  ±  0.22, P<0.001 and DM2 (0.41  ±  0.21, P<0.001 groups, in comparison to controls (0.06  ±  0.05. Besides, we found a positive correlation between micronuclei frequency and the onset time of DM2 in both cheek (ρ = 0.69, P<0.001 and tongue epithelial cells (ρ = 0.71, P<0.001, but not with onset time of DM1 or age of the patients. Considering all this, we pose that BMCyt could serve as a fast and easily accessible test to assess genotoxic damage during dental visits of DM patients, helping to monitor their disease.

  4. Occupational exposure to anesthetics leads to genomic instability, cytotoxicity and proliferative changes

    International Nuclear Information System (INIS)

    Souza, Kátina M.; Braz, Leandro G.; Nogueira, Flávia R.; Souza, Marajane B.; Bincoleto, Lahis F.; Aun, Aline G.; Corrente, José E.; Carvalho, Lídia R.; Braz, José Reinaldo C.; Braz, Mariana G.

    2016-01-01

    Highlights: • Anesthesiologists exposed to the most commonly used anesthetic gases were evaluated. • No alterations were detected for lymphocyte DNA damage detected by the comet assay. • Decreased frequencies of basal cells were detected in exfoliated buccal cells (BMCyt). • Increased frequencies of micronucleus and cytotoxicity were observed in BMCyt assay. • Anesthesiologists have genomic instability due to occupational exposure. - Abstract: Data on the genotoxic and mutagenic effects of occupational exposure to the most frequently used volatile anesthetics are limited and controversial. The current study is the first to evaluate genomic instability, cell death and proliferative index in exfoliated buccal cells (EBC) from anesthesiologists. We also evaluated DNA damage and determined the concentrations of the anesthetic gases most commonly used in operating rooms. This study was conducted on physicians who were allocated into two groups: the exposed group, which consisted of anesthesiologists who had been exposed to waste anesthetic gases (isoflurane, sevoflurane, desflurane and nitrous oxide − N 2 O) for at least two years; and the control group, which consisted of non-exposed physicians matched for age, sex and lifestyle with the exposed group. Venous blood and EBC samples were collected from all participants. Basal DNA damage was evaluated in lymphocytes by the comet assay, whereas the buccal micronucleus (MN) cytome (BMCyt) assay was applied to evaluate genotoxic and cytotoxic effects. The concentrations of N 2 O and anesthetics were measured via a portable infrared spectrophotometer. The average concentration of waste gases was greater than 5 parts per million (ppm) for all of the halogenated anesthetics and was more than 170 ppm for N 2 O, expressed as a time-weighted average. There was no significant difference between the groups in relation to lymphocyte DNA damage. The exposed group had higher frequencies of MN, karyorrhexis and pyknosis, and

  5. Occupational exposure to anesthetics leads to genomic instability, cytotoxicity and proliferative changes

    Energy Technology Data Exchange (ETDEWEB)

    Souza, Kátina M.; Braz, Leandro G.; Nogueira, Flávia R.; Souza, Marajane B.; Bincoleto, Lahis F.; Aun, Aline G. [Faculdade de Medicina de Botucatu, UNESP − Univ Estadual Paulista, Departamento de Anestesiologia, Botucatu (Brazil); Corrente, José E.; Carvalho, Lídia R. [Instituto de Biociências de Botucatu, UNESP − Univ Estadual Paulista, Departamento de Bioestatística, Botucatu (Brazil); Braz, José Reinaldo C. [Faculdade de Medicina de Botucatu, UNESP − Univ Estadual Paulista, Departamento de Anestesiologia, Botucatu (Brazil); Braz, Mariana G., E-mail: mgbraz@hotmail.com [Faculdade de Medicina de Botucatu, UNESP − Univ Estadual Paulista, Departamento de Anestesiologia, Botucatu (Brazil)

    2016-09-15

    Highlights: • Anesthesiologists exposed to the most commonly used anesthetic gases were evaluated. • No alterations were detected for lymphocyte DNA damage detected by the comet assay. • Decreased frequencies of basal cells were detected in exfoliated buccal cells (BMCyt). • Increased frequencies of micronucleus and cytotoxicity were observed in BMCyt assay. • Anesthesiologists have genomic instability due to occupational exposure. - Abstract: Data on the genotoxic and mutagenic effects of occupational exposure to the most frequently used volatile anesthetics are limited and controversial. The current study is the first to evaluate genomic instability, cell death and proliferative index in exfoliated buccal cells (EBC) from anesthesiologists. We also evaluated DNA damage and determined the concentrations of the anesthetic gases most commonly used in operating rooms. This study was conducted on physicians who were allocated into two groups: the exposed group, which consisted of anesthesiologists who had been exposed to waste anesthetic gases (isoflurane, sevoflurane, desflurane and nitrous oxide − N{sub 2}O) for at least two years; and the control group, which consisted of non-exposed physicians matched for age, sex and lifestyle with the exposed group. Venous blood and EBC samples were collected from all participants. Basal DNA damage was evaluated in lymphocytes by the comet assay, whereas the buccal micronucleus (MN) cytome (BMCyt) assay was applied to evaluate genotoxic and cytotoxic effects. The concentrations of N{sub 2}O and anesthetics were measured via a portable infrared spectrophotometer. The average concentration of waste gases was greater than 5 parts per million (ppm) for all of the halogenated anesthetics and was more than 170 ppm for N{sub 2}O, expressed as a time-weighted average. There was no significant difference between the groups in relation to lymphocyte DNA damage. The exposed group had higher frequencies of MN, karyorrhexis and

  6. Biochemical and genetic studies on cardiometabolic syndrome

    OpenAIRE

    Supriya Simon, A.; Dinesh Roy, D.; Jayapal, V.; Vijayakumar, T.

    2010-01-01

    Cardiometabolic syndrome is one of the major public health issues of this century which describes a cluster of clinical characteristics. Seventy two patients with coronary artery disease (CAD) and cardiometabolic syndrome and forty healthy age and sex matched normal controls were selected for this study. Detailed clinical epidemiological and anthropometric characteristics were noted. Lipid profile and Cytokinesis-block micronuclei (CBMN) assay using cytochalasin B were carried out in all the ...

  7. Normal tissue adverse side effects in radiotherapy cancer patients and applicability of predictive radiosensitivity tests for new radiation treatment decision

    International Nuclear Information System (INIS)

    Di Giorgio, Marina; Vallerga, Maria B.; Radl, Analia; Sardi, Mabel

    2008-01-01

    Full text: Around 5 % -7 % of cancer patients develop adverse side effects, which include acute effects, late effects and cancer induction to radiation therapy in normal tissues in the treatment field. Such effects are of particular interest as the cancer patient population that reaches prolonged survival has increased with the improvements in cancer therapy and health care. These adverse reactions are mainly influenced by deficiencies in DNA repair pathways. However, tissue response to IR could be modified by several treatment- and patient- related factors. Numerous studies have been carried out to evaluate the correlation between clinical and cellular radiosensitivity, by in vitro tests. Previous own studies, characterizing DNA repair capacity in peripheral lymphocytes of cancer patients through cytokinesis blocked micronucleus test and alkaline single-cell microgel electrophoresis (comet), indicated that such assays correlated with the clinical radiation signs of radiosensitivity and showed the predictive potential of both techniques in the identification of radiosensitivity subgroups. In this paper, retrospective studies are conducted in 10 representative cases, which had developed acute or late toxicity in previous treatments and at present require new radiation treatments due to secondary malignancies or recurrence. Samples were in vitro irradiated with 2 Gy. MN data were analyzed comparing expected MN frequencies with values observed after in vitro irradiation. DNA repair capacity was evaluated through comet assay for initial damage and after specific times of repair (0-120 minutes). Captured images were analyzed by CASP image analysis software. Repair capacity was quantified by the Olive tail moment. Weibull alpha parameter was applied to describe DNA damage at the different evaluated repair times after in vitro irradiation and fitted by a mono-exponential model to describe the kinetic profile. In every evaluated patient a correlation between mean half

  8. Micronuclei: sensitivity for the detection of radiation induced damage

    International Nuclear Information System (INIS)

    Di Giorgio, M.; Nasazzi, N.B.; Taja, M.R.

    1998-01-01

    The in vitro cytokinesis-block (CB) micronucleus (MN) assay for human peripheral blood has been used extensively for the assessment of chromosomal damage induced by ionizing radiation and chemicals and considered a suitable biological dosimeter for estimating in vivo whole body exposures, particularly in the case of large scale radiation accidents. One of the major drawbacks of the MN assay is its reduced sensitivity for the detection of damage induced by low doses of low LET radiation, due to the high variability among the spontaneous MN frequencies. It is suggested that age, smoking habit and sex are the main confounding factors that contribute to the observed variability. Previous work in our laboratory, shows a significant positive correlation of the spontaneous and radiation induced MN frequencies with age and smoking habit, the latter being the strongest confounder. These findings led to in vitro studies of the dose-response relationships for smoking and non smoking donors evaluated separately, using 60 Co γ rays. The objectives of the present work are: 1-To increase the amount of data of the dose-response relationships, using γ rays from a 60 Co source, for smoking and non smoking donors, in order to find, if applicable, a correction factor for the calibration curve that takes into account the smoking habit of the individual in the case of accidental overexposure dose assessment, particularly in the low dose range. 2-To establish general conclusions on the current state of the technique. The sample for smoking and non smoking calibration curves was enlarged in the range of 0Gy to 2Gy. The fitting of both curves, performed up to the 2Gy dose, resulted in a linear quadratic model. MN distribution among bi nucleated cells was found to be over dispersed with respect to Poisson distribution, the average ratio of variance to mean being 1.13 for non smokers and 1.17 for smokers. Each fitted calibration curve, for smoking and non smoking donors, fell within the 95

  9. Assessment of individual radiosensitivity in human lymphocytes of cancer patients and its correlation with adverse side effects to radiation therapy

    International Nuclear Information System (INIS)

    Di Giorgio, Marina; Vallerga, Maria B.; Taja, Maria R.; Sardi, M.; Busto, E.; Roth, B.; Menendez, P.; Bonomi, M.; Mairal, L.

    2003-01-01

    Background and purpose: Individual radiosensitivity is an inherent characteristic, associated with an increased reaction to ionizing radiation on the human body. Biological endpoints such as clonogenic survival, chromosome aberration formation and repair capacity of radiation-induced damage have been applied to evaluate individual radiosensitivity in vitro. 5%-7% of cancer patients develop adverse side effects to radiation therapy in normal tissues within the treatment field, which are referred as 'clinical radiation reactions' and include acute effects, late effects and cancer induction. It has been hypothesized that the occurrence and severity of these reactions are mainly influenced by genetic susceptibility to radiation. Additionally, the nature of the genetic disorders associated with hypersensitivity to radiotherapy suggests that DNA repair mechanisms are involved. Consequently, the characterization of DNA repair in lymphocytes through cytokinesis blocked micronucleus (MN) and alkaline single-cell micro gel electrophoresis (comet) assays could be suitable approaches to evaluate individual radiosensitivity in vitro. The MN assay is an established cytogenetic technique to evaluate intrinsic cell radiosensitivity in tumor cells and lymphocytes; comet assay is a sensitive and rapid method for measuring DNA damage and repair in individual cells. The aims of this study were: 1) To assess the in vitro radiosensitivity of peripheral blood lymphocytes from two groups of cancer patients (retrospectively and prospectively studied), using MN and comet assays, in comparison with the observed clinical response; and 2) To test the predictive potential of both techniques. Materials and methods: 38 cancer patients receiving radiation therapy were enrolled in this study. The tumor sites were: head and neck (n 25) and cervix (n = 13). Nineteen patients were evaluated about 6-18 month after radiotherapy (retrospective group) and 19 patients were evaluated prior, mid-way and on

  10. Study protocol for an approach based on diagnosis and therapy of cutaneous radiation induced lesions

    International Nuclear Information System (INIS)

    Di Giorgio, Marina; Vallerga, Maria B.; Radl, Analia; Portas, Mercedes

    2008-01-01

    Full text: In the frame of an agreement between the 'Hospital de Quemados del Gobierno de la Ciudad de Buenos Aires'-Burn Center- (a reference hospital of the Medical Radiological Emergency Response Network of Argentina) and the Nuclear Regulatory Authority, a research project for diagnostic and therapeutic approach of cutaneous radiation syndrome (CRS) is in progress. Sixty seven persons, which developed acute and/or late CRS, were included in this protocol from 1997 to 2007, treated with an equivalent therapeutic scheme and evaluated through clinical follow-up, serial photographic record and complementary tests (tele-thermography and high frequency ultrasonography). There exist individual variations that could condition the response to ionizing radiation (IR) in not only accidental but also planned exposures (such as radiotherapy and interventional radiology). Deficiencies in DNA repair mechanisms would be involved on hypersensitivity to deterministic effects of IR. Consequently, the characterization of DNA repair capacity in lymphocytes through cytokinesis blocked micronucleus (MN) and alkaline single-cell microgel electrophoresis (comet) assays could be suitable approaches to evaluate in vitro individual radiosensitivity. Under this context, individual radiosensitivity assessment was conducted in patients included in this research protocol that showed acute and/or late cutaneous reactions with grades 3 and 4 of the Toxicity Criteria of the Radiation Therapy Oncology Group and the European Organization for Research and Treatment of Cancer. DNA repair capacity was evaluated through MN and comet assay for initial damage and after specific times of repair (0-120 minutes). DNA damage and repair capacity were quantified by the Olive tail moment. Previous own studies have identified three subpopulations, characterized by the mean values of their repair mean half-time: healthy controls (2.6 ± 0.3 minutes), average-reactor cancer patients (4.7 ± 2.9 minutes) and over

  11. Study Protocol for an Approach Based on Diagnosis and Therapy of Cutaneous Radiation Induced Lesions

    International Nuclear Information System (INIS)

    Di Giorgio, M.; Vallerga, M.B.; Radl, A.; Portas, M.

    2011-01-01

    In the frame of an agreement between the 'Hospital de Quemados del Gobierno de la Ciudad de Buenos Aires' - Burn Center - (a reference hospital of the Medical Radiological Emergency Response Network of Argentina) and the Nuclear Regulatory Authority, a research project for diagnostic and therapeutic approach of cutaneous radiation syndrome (CRS) is in progress. Sixty seven persons, which developed acute and/or late CRS, were included in this protocol from 1997 to 2007, treated with an equivalent therapeutic scheme and evaluated through clinical follow-up, serial photographic record and complementary tests (telethermography and high frequency ultrasonography). There exist individual variations that could condition the response to ionizing radiation (IR) in not only accidental but also planned exposures (such as radiotherapy and interventional radiology). Deficiencies in DNA repair mechanisms would be involved on hypersensitivity to deterministic effects of IR. Consequently, the characterization of DNA repair capacity in lymphocytes through cytokinesis blocked micronucleus (MN) and alkaline single-cell microgel electrophoresis (comet) assays could be suitable approaches to evaluate in vitro individual radiosensitivity. Under this context, individual radiosensitivity assessment was conducted in patients included in this research protocol that showed acute and/or late cutaneous reactions with grades 3 and 4 of the Toxicity Criteria of the Radiation Therapy Oncology Group and the European Organization for Research and Treatment of Cancer. DNA repair capacity was evaluated through MN and comet assay for initial damage and after specific times of repair (0-120 minutes). DNA damage and repair capacity were quantified by the Olive tail moment. Previous own studies have identified three subpopulations, characterized by the mean values of their repair mean half-time: healthy controls (2.6 ± 0.3 minutes), average-reactor cancer patients (4.7 ± 2.9 minutes) and over

  12. Normal tissue adverse side effects in radiotherapy cancer patients and applicability of predictive radiosensitivity tests for new radiation treatment decision

    International Nuclear Information System (INIS)

    Di Giorgio, M.; Vallerga, M.B.; Radl, A.; Sardi, M.

    2011-01-01

    Around 5%-7% of cancer patients develop adverse side effects, which include acute effects, late effects and cancer induction to radiation therapy in normal tissues in the treatment field. Such effects are of particular interest as the cancer patient population that reaches prolonged survival has increased with the improvements in cancer therapy and health care. These adverse reactions are mainly influenced by deficiencies in DNA repair pathways. However, tissue response to IR could be modified by several treatment- and patient- related factors. Numerous studies have been carried out to evaluate the correlation between clinical and cellular radiosensitivity, by in vitro tests. Previous own studies, characterizing DNA repair capacity in peripheral lymphocytes of cancer patients through cytokinesis blocked micronucleus test and alkaline single-cell microgel electrophoresis (comet), indicated that such assays correlated with the clinical radiation signs of radiosensitivity and showed the predictive potential of both techniques in the identification of radiosensitivity subgroups. In this paper, retrospective studies are conducted in 10 representative cases, which had developed acute or late toxicity in previous treatments and at present require new radiation treatments due to secondary malignancies or recurrence. Samples were in vitro irradiated with 2 Gy. MN data were analyzed comparing expected MN frequencies with values observed after in vitro irradiation. DNA repair capacity was evaluated through comet assay for initial damage and after specific times of repair (0-120 minutes). Captured images were analyzed by CASP image analysis software. Repair capacity was quantified by the Olive tail moment. Weibull alpha parameter was applied to describe DNA damage at the different evaluated repair times after in vitro irradiation and fitted by a mono-exponential model to describe the kinetic profile. In every evaluated patient a correlation between mean half-time (T1/2) and

  13. Chromosomal aberrations and DNA damage in human populations exposed to the processing of electronics waste.

    Science.gov (United States)

    Liu, Qiang; Cao, Jia; Li, Ke Qiu; Miao, Xu Hong; Li, Guang; Fan, Fei Yue; Zhao, Yong Cheng

    2009-05-01

    It has been known that the pollutants of electronic wastes (E-wastes) can lead to severe pollution to the environment. It has been reported that about 50% to 80% of E-wastes from developed countries are exported to Asia and Africa. It has become a major global environmental problem to deal with 'E-wastes'. E-waste recycling has remained primitive in Jinghai, China. This not only produces enormous environmental pollution but also can bring about toxic or genotoxic effects on the human body, threatening the health of both current residents and future generations living in the local environment. The concentration of lead in the blood of children in the E-waste polluted area in China is higher than that of the control area. But little is known about the cytogenetic effect to human beings caused by the pollution of E-wastes. In the present study, experiments have been performed to investigate the genetics of permanent residents of three villages with numerous E-waste disposal sites and to analyze the harmful effects of exposure to E-wastes. In total, 171 villagers (exposed group) were randomly selected from permanent residents of three villages located in Jinghai County of Tianjin, China, where there has been massive disposal of E-wastes. Thirty villagers were selected from the neighboring towns without E-waste disposal sites to serve as controls. Chromosomal aberrations and cytokinesis blocking micronucleus were performed to detect the cytogenetic effect, dic + r (dicentric and ring chromosome), monomer, fragments (acentric fragments, minute chromosomes, and acentric rings), translocation, satellite, quadriradial, total aberrations, and micronuclear rate were scored for each subject. DNA damage was detected using comet assay; the DNA percentage in the comet tail (TDNA%), tail moment (TM), and Olive tail moment (OTM) were recorded to describe DNA damage to lymphocytes. The total chromosome aberration rates (5.50%) and micronuclear rates (16.99%) of the exposure group

  14. Integrative content-driven concepts for bioinformatics “beyond the cell”

    Indian Academy of Sciences (India)

    Prakash

    2006-10-10

    Oct 10, 2006 ... systems, we have to acquire a comprehensive knowledge about their ..... type) is documented by making use of the Cytomer ontology. (see below) for .... and GPCR PD were added for covering the whole space of human ...

  15. Radiation-induced micronucleus frequencies in female peripheral blood lymphocytes collected during the first and second half of the menstrual cycle

    International Nuclear Information System (INIS)

    Krol, M.; Lankoff, A.; Buraczewska, I.; Derezinska, E.; Wojcik, A.

    2007-01-01

    Biological dosimetry relies on the assessment of dose in peripheral blood lymphocytes (PBL) of a victim. Variability in the individual radiosensitivity of PBL has an impact on the precision of dose estimate and radiation-induced micronuclei show a strong individual variability. A factor which can influence the radiosensitivity of PBL is the hormonal status of female donors, which shows a regular pattern during the menstrual cycle. The aim of the present investigation was to verify whether the position within the menstrual cycle has an impact on the level of micronuclei in PBL. Blood was collected from 19 donors during the first and second half of the menstrual cycle and exposed to 2 Gy. Although statistically significant differences between the MN frequencies in PBL collected during the different time points were observed in the case of some donors, no reproducible trend that could find application in biological dosimetry could be detected. (authors)

  16. Radioprotective effects of selenium and vitamin-E against 6MV X-rays in human blood lymphocytes by micronucleus assay

    OpenAIRE

    Rostami, Aram; Moosavi, Seyed Akbar; Changizi, Vahid; Abbasian Ardakani, Ali

    2016-01-01

    Background: Critical macromolecules of cells such as DNA are in exposure to damage of free radicals that induced from the interaction of ionizing radiation with biological systems. Selenium and vitamin-E are natural compounds that have been shown to be a direct free radical scavenger. The aim of this study was to investigate the radioprotective effect of selenium and vitamin-E separately and synergistically against genotoxicity induced by 6MV x-rays irradiation in blood lymphocytes. Methods: ...

  17. Xanthium strumarium L. extracts produce DNA damage mediated by cytotoxicity in in vitro assays but does not induce micronucleus in mice.

    Science.gov (United States)

    Piloto Ferrer, Janet; Cozzi, Renata; Cornetta, Tommaso; Stano, Pasquale; Fiore, Mario; Degrassi, Francesca; De Salvia, Rosella; Remigio, Antonia; Francisco, Marbelis; Quiñones, Olga; Valdivia, Dayana; González, Maria L; Pérez, Carlos; Sánchez-Lamar, Angel

    2014-01-01

    Xanthium strumarium L. is a member of the Asteraceae commonly used in Cuba, mainly as diuretic. Some toxic properties of this plant have also been reported and, to date, very little is known about its genotoxic properties. The present work aims was to evaluate the potential cytotoxic and genotoxic risk of whole extract from Xanthium strumarium L. whole extract of aerial parts. No positive response was observed in a battery of four Salmonella typhimurium strains, when exposed to concentrations up to 5 mg/plate, with and without mammalian metabolic activation (liver microsomal S9 fraction from Wistar rats). In CHO cells, high concentrations (25-100 μg/mL) revealed significant reduction in cell viability. Results from sister chromatid exchanges, chromosome aberrations, and comet assay showed that X. strumarium extract is genotoxic at the highest concentration used, when clear cytotoxic effects were also observed. On the contrary, no increase in micronuclei frequency in bone marrow cells was observed when the extract was orally administered to mice (100, 500, and 2000 mg/Kg doses). The data presented here constitute the most complete study on the genotoxic potential of X. strumarium L. and show that the extract can induce in vitro DNA damage at cytotoxic concentrations.

  18. Xanthium strumarium L. Extracts Produce DNA Damage Mediated by Cytotoxicity in In Vitro Assays but Does Not Induce Micronucleus in Mice

    Directory of Open Access Journals (Sweden)

    Janet Piloto Ferrer

    2014-01-01

    Full Text Available Xanthium strumarium L. is a member of the Asteraceae commonly used in Cuba, mainly as diuretic. Some toxic properties of this plant have also been reported and, to date, very little is known about its genotoxic properties. The present work aims was to evaluate the potential cytotoxic and genotoxic risk of whole extract from Xanthium strumarium L. whole extract of aerial parts. No positive response was observed in a battery of four Salmonella typhimurium strains, when exposed to concentrations up to 5 mg/plate, with and without mammalian metabolic activation (liver microsomal S9 fraction from Wistar rats. In CHO cells, high concentrations (25–100 μg/mL revealed significant reduction in cell viability. Results from sister chromatid exchanges, chromosome aberrations, and comet assay showed that X. strumarium extract is genotoxic at the highest concentration used, when clear cytotoxic effects were also observed. On the contrary, no increase in micronuclei frequency in bone marrow cells was observed when the extract was orally administered to mice (100, 500, and 2000 mg/Kg doses. The data presented here constitute the most complete study on the genotoxic potential of X. strumarium L. and show that the extract can induce in vitro DNA damage at cytotoxic concentrations.

  19. MICRONUCLEUS STUDIES IN THE PERIPHERAL BLOOD AND BONE MARROW OF MICE TREATED WITH JET FUELS, JP-8 AND JET-A

    Science.gov (United States)

    The potential adverse effects of dermal and inhalation exposure of jet fuels are important for health hazard evaluation in humans. In an animal model, the genotoxic potential of jet fuels, JP-8 and Jet-A, was investigated. Mice were treated dermally with either a single or multip...

  20. MEETING IN VANCOUVER, B.C.: MICRONUCLEUS STUDIES IN THE PERIPHERAL BLOOD AND BONE MARROW OF MICE TREATED WITH JET FUELS, JP-8 AND JET-A

    Science.gov (United States)

    The potential adverse effects of dermal and inhalation exposure of jet fuels are important for health hazard evaluation in humans. In an animal model, the genotoxic potential of jet fuels, JP-8 and Jet-A, was investigated. Mice were treated dermally with either a single or multip...

  1. Effect of sodium benzoate on DNA breakage, micronucleus formation and mitotic index in peripheral blood of pregnant rats and their newborns

    Directory of Open Access Journals (Sweden)

    Cetin Saatci

    2016-11-01

    Full Text Available Sodium benzoate (SB is one of the most widely used additives in food products in the world. The aim of this study was to assess the effect of three different concentrations of SB on the DNA breakage in liver cells and on the micronuclei formation and the mitotic index in lymphocytes of pregnant rats and their fetuses, as well as to evaluate the effects of SB on the fetus development. The results showed that general genomic injuries were present in almost all the liver cell samples obtained from the SB group compared with the control (non-treated group. This indicates that SB usage may cause DNA damage and increase micronuclei formation. We recommend that pregnant women should avoid consuming foodstuffs containing SB as an additive.

  2. In vivo micronucleus studies with 6 titanium dioxide materials (3 pigment-grade & 3 nanoscale) in orally-exposed rats.

    Science.gov (United States)

    Donner, E M; Myhre, A; Brown, S C; Boatman, R; Warheit, D B

    2016-02-01

    Six pigment-grade (pg) or ultrafine (uf)/nanoscale (anatase and/or rutile) titanium dioxide (TiO2) particulates were evaluated for in vivo genotoxicity (OECD 474 Guidelines) in male and female rats by two different laboratories. All test materials were robustly characterized. The BET surface areas of the pg and uf samples ranged from 7 to 17 m(2)/g and 50 to 82 m(2)/g respectively. The materials were assessed for induction of micronuclei and toxicity in bone marrow by analyzing peripheral blood reticulocytes (RETs) by flow cytometry. Single oral gavage doses of 500, 1000 or 2000 mg/kg body weight (bw) of each material were implemented with concurrent negative (water) and positive controls (cyclophosphamide). Approximately 48 and 72 h after exposure, blood samples were collected and 20,000 RETs per animal were analyzed. For each of the six tests, there were no biologically or toxicologically relevant increases in the micronucleated RET frequency in any TiO2 exposed group at either time point at any dose level. In addition, there were a lack of biologically relevant decreases in %RETs among total erythrocytes. All six TiO2 test substances were negative for in vivo genotoxicity effects; however, it is noted that the exposure to target tissues was likely negligible. One pigment grade and one ultrafine material each were evaluated for potential systemic exposure/uptake from the gastrointestinal tract by analysis of TiO2 into blood and liver. No significant increases in TiO2 over controls were measured in blood (48 or 72 h) or liver (72 h) following exposures to 2000 mg/kg bw TiO2. These data indicate that there was no absorption of the test material from the gastrointestinal tract into the blood circulation and the lack of genotoxic effects is therefore attributed to a lack of exposure due to the inability of the test material to migrate from the gastrointestinal tract into the blood and then into target tissues. Copyright © 2015 Elsevier Inc. All rights reserved.

  3. Effect of radiosensitizer BSO on the incidence of micronuclei in cultured cells

    International Nuclear Information System (INIS)

    Jin Yizun; Cai Rongmei; Ding Li; Shen Zhifen; Xu Liming; Yang Jiakuan

    1992-01-01

    The effects of BSO, a potent radiosensitizing and chemical sensitizing chemical, on the incidence of micronuclei in four different cell lines have been studied using the cytokinesis-block (CB) method. The number of micronuclei in cultured human peripheral lymphocytes, Chinese hamster cells and human breast cancer cells were not affected by 0.1-2 mmol/L BSO treatment alone. However, significant increase in the incidence of micronuclei in these cells could be detected when BSO was used in combination with γ-irradiation. Linear relationship between the incidence of micronuclei and the radiation dose was observed

  4. 86 original article association of cytomegalo virus with type

    African Journals Online (AJOL)

    boaz

    But : L'association entre le cytomégalovirus et DT1 sucre a été étudiée avec une comparaison aux sujets sains et pour corréler son ... the association between cytomegalovirus and type 1 diabetes mellitus ..... American Diabetes Association.

  5. Immunohistochemical detection of Her-2/neu overexpression in ...

    African Journals Online (AJOL)

    Materials and Methods: Immunohistochemical staining for Her-2/neu was performed on 10% formalin-fixed, paraffinembedded primary carcinoma of the breast from 83 patients, between 2003 and 2007 using anti-Her-2/neu rabbit polyclonal antibody (DakoCytomation, CA, USA) and reactivity detected by an avidin-biotin ...

  6. High-resolution cytometry represents the main technology used in the laboratory of molecular cytology and cytometry

    Czech Academy of Sciences Publication Activity Database

    Kozubek, Stanislav; Bártová, Eva; Lukášová, Emilie; Falk, Martin; Ondřej, Vladan; Kozubek, Michal; Kroupová, Jana; Matula, Pe.; Matula, Pa.

    2006-01-01

    Roč. 39, č. 5 (2006), s. 341-341 ISSN 0960-7722. [Cytomics Emerging from Cytometry 16th Annual Meeting of the german Society for cytometry. 18.10.2006-21.10.2006, Leipzig] Institutional research plan: CEZ:AV0Z50040507 Keywords : high-resolution cytometry * cytogenetics * epigenetics Subject RIV: BO - Biophysics

  7. Combined assessment of cyto-, geno-, and embryotoxicity of silica nanoparticles in experiments in vitro

    International Nuclear Information System (INIS)

    Protasova, G A; Popov, V B; Shabasheva, L V; Strekalovskii, I V; Panferova, Y A; Khlebnikova, N S; Radilov, A A

    2017-01-01

    A complex of express methods for cyto-, geno-, and embryotoxicity assessment of nanoparticles (NPs), based on the simultaneous use of three models: the cultures of pre- and post-implantation embryos of mice (egg cleavage, blastocyst formation, 1−5 day of development) and rats (head fold−neurulation stages, 30 somite pairs, 9.5−11.5 day of development), as well as human peripheral blood lymphocytes, is developed. Pathogenic (dysmorphogenic) effects of SiO 2 NPs in concentrations of 100 and 200 µg/mL on early embryo development processes were established. The cytogenetic methods used for cyto-, geno-, and mutagenicity assessment (micronuclear test under cytokinesis-block conditions, chromosomal aberrations test, and DNA comet assay) revealed no effects of SiO 2 NPs in concentrations 200, 100, and 20 µg/mL. (paper)

  8. Low dose response analysis through a cytogenetic end-point

    International Nuclear Information System (INIS)

    Bojtor, I.; Koeteles, G.J.

    1998-01-01

    The effects of low doses were studied on human lymphocytes of various individuals. The frequency of micronuclei in cytokinesis-blocked cultured lymphocytes was taken as end-point. The probability distribution of radiation-induced increment was statistically proved and identified as to be asymmetric when the blood samples had been irradiated with doses of 0.01-0.05 Gy of X-rays, similarly to that in unirradiated control population. On the contrary, at or above 1 Gy the corresponding normal curve could be accepted only reflecting an approximately symmetrical scatter of the increments about their mean value. It was found that the slope as well as the closeness of correlation of the variables considerably changed when lower and lower dose ranges had been selected. Below approximately 0.2 Gy even an unrelatedness was found betwen the absorbed dose and the increment

  9. Environmental lead exposure increases micronuclei in children

    DEFF Research Database (Denmark)

    Kapka, Lucyna; Baumgartner, Adolf; Siwińska, Ewa

    2007-01-01

    The objective of this pilot study was to investigate the contribution of environmental exposures to lead in the development of cytogenetic damage detected as the frequency of micronuclei (MN) in children. The other aim was to apply the MN assay in combination with fluorescence in situ hybridization...... age from an unexposed recreational area. Exposure to lead was assessed by determination of lead concentrations in blood (PbB) by atomic absorption spectrophotometry, whereas the level of selenium (Se) in serum was detected by using graphite furnace atomic-absorption spectrometry. The frequency of MN...... was determined by the cytokinesis-block MN assay and fluorescence in situ hybridization performed using a specific pan-centromeric probe. Environmental exposure to lead resulted in significantly increased levels of PbB (5.29 +/- 2.09 versus 3.45 +/- 1.20 microg/dl in controls), although the average level...

  10. The study on the effect of artesunate on the radio-sensitivity of human cervical cancer

    International Nuclear Information System (INIS)

    Geng Chong; Cao Jianping; Ni Qianying

    2011-01-01

    To investigate the effect of artesunate on radio-sensitivity of human cervical cancer cells in vitro. The human cervical cancer cells HeLa and Siha were used as the experimental cells. MTT assay was used to determine the most appropriate drug concentration in the subsequent experiment, and the effect of human cervical cancer cells treated with artesunate and irradiation of 60 Co γ-rays was studied by using conventional chromosomal aberration analysis and cytokinesis block method (CB method). The results show that when the concentration of artesunate in this experiment was 2.0 μmol/L for HeLa cell and 4.0 μmol/L for Siha cell respectively, the chromosome aberration, micronuclei cell and micronuclei rates of HeLa cells treated with artesunate were more serious than that of the only irradiation, but there is almost no change with Siha cells. (authors)

  11. Effect of oxygen on formation of micronuclei and binucleated cells and cell survival in γ-irradiated 3T3 cells

    International Nuclear Information System (INIS)

    Zhang Peng; Zheng Xiulong

    1991-01-01

    Formation of micronuclei and binucleate cells and their relationships with cell survival were studied in the aerobically- and anaerobically-irradiated 3T3 cells. The results showed taht frequency of micronuclei, percentage of micronucleus cells and percentage of binucleate cells increased linearly with the radiation dose in certain range. Oxygen enhancement ratios (OER) of micronucleus frequency, percentage of micronucleus cells, percentage of binucleate cells and cell survival were 2.02, 1.96, 1.87 and 1.83 respectively. The percentage of micronucleus cells or the percentage of micronucleus cells plus binucleate cells correlated negatively well with cell survival. The mechanism of oxygen effect in the radiation response of 3T3 cells and the significance of formation of micronuclei and binucleate cells were discussed

  12. In vivo evaluation of anticlastogenicity of extracts from medicinal plants of Miconia genus using the micronucleus test Avaliação in vivo da anticlastogenicidade de extratos de plantas medicinais do gênero Miconia através do teste do micronúcleo

    Directory of Open Access Journals (Sweden)

    Ilce Mara S. Cólus

    2008-10-01

    Full Text Available The genus Miconia is comprised of approximately 1000 species. For some of them, biological activities have already been described such as the analgesic and the anti-microbial ones. The purpose of this work was to evaluate the possible protective and cytotoxic effects of the methanolic extract from M. albicans, M. cabucu, M. rubiginosa and M. stenostachya and the chloroformic extract from M. albicans in mice bone marrow cells in 540 mg/kg p.c. dose. The extracts were administered by means of forced feeding and the cyclophosphamide (CPA was applied intraperitonially one hour after supplementation with extracts. All animals were submitted to euthanasia 30 hours after the treatment. The analyzed cells were extracted from mice bone marrow according to protocol described by Schmid (1975. The cytotoxicity of the extracts was evaluated through the percentage of polychromatic erythrocytes (PCE in 200 erythrocytes (PCE + NCE. Two thousand PCEs of each animal were analyzed and the micronucleated polychromatic erythrocytes (MNPCEs frequencies were scored. The results obtained indicated that none of the extracts associated with the CPA showed cytotoxic effect and only the extracts of M. rubiginosa and M. stenostachya showed protective effect to DNA. Chemical analyses of the extracts showed that the four species studied contain mainly flavonoids, phenolic compounds and tannins. The phytochemical characterization of these extracts could contribute to elucidate the protective effect presented only by the species M. rubiginosa and M. stenostachya, besides making it possible the study of other therapeutic activities. O gênero Miconia possui aproximadamente 1000 espécies, e para algumas, já foram descritas atividades biológicas como a analgésica e antimicrobiana. Esse trabalho teve como objetivo avaliar os possíveis efeitos protetores e citotóxicos dos extratos metanólicos de M. albicans, M. cabucu, M. rubiginosa e M. stenostachya e do extrato clorofórmico de M. albicans em células da medula óssea de camundongos na dose de 540 mg/kg p.c. Os extratos foram administrados via gavage e a ciclofosfamida (CPA foi aplicada intraperitonealmente 1h, após a suplementação com os extratos. Todos os animais foram submetidos à eutanásia 30h após o tratamento. As células analisadas foram retiradas da medula óssea de acordo com protocolo descrito por Schmid (1975. A citotoxicidade dos extratos foi avaliada pela percentagem de eritrócitos policromáticos (PCE em 200 eritrócitos (PCE + NCE. Foram analisados 2000 PCEs por animal e anotadas as freqüências de MNPCEs. Os resultados obtidos mostraram que nenhum dos extratos associados à CPA apresentou efeito citotóxico e somente os extratos de M. rubiginosa, M. stenostachya mostraram efeito protetor ao DNA. A análise química dos extratos mostrou que as quatro espécies estudadas contêm, principalmente, flavonóides, compostos fenólicos e taninos. A caracterização fitoquímica desses extratos poderia contribuir para elucidação do efeito protetor apresentado somente pelas espécies M. rubiginosa e M. stenostachya, além de possibilitar o estudo de outras possíveis atividades terapêuticas.

  13. Dose rate effect on micronuclei induction in human blood lymphocytes exposed to single pulse and multiple pulses of electrons.

    Science.gov (United States)

    Acharya, Santhosh; Bhat, N N; Joseph, Praveen; Sanjeev, Ganesh; Sreedevi, B; Narayana, Y

    2011-05-01

    The effects of single pulses and multiple pulses of 7 MV electrons on micronuclei (MN) induction in cytokinesis-blocked human peripheral blood lymphocytes (PBLs) were investigated over a wide range of dose rates per pulse (instantaneous dose rate). PBLs were exposed to graded doses of 2, 3, 4, 6, and 8 Gy of single electron pulses of varying pulse widths at different dose rates per pulse, ranging from 1 × 10(6) Gy s(-1) to 3.2 × 10(8) Gy s(-1). Different dose rates per pulse were achieved by changing the dose per electron pulse by adjusting the beam current and pulse width. MN yields per unit absorbed dose after irradiation with single electron pulses were compared with those of multiple pulses of electrons. A significant decrease in the MN yield with increasing dose rates per pulse was observed, when dose was delivered by a single electron pulse. However, no reduction in the MN yield was observed when dose was delivered by multiple pulses of electrons. The decrease in the yield at high dose rates per pulse suggests possible radical recombination, which leads to decreased biological damage. Cellular response to the presence of very large numbers of chromosomal breaks may also alter the damage.

  14. Radioprotective Effect of Achillea millefolium L Against Genotoxicity Induced by Ionizing Radiation in Human Normal Lymphocytes

    Directory of Open Access Journals (Sweden)

    Somayeh Shahani

    2015-04-01

    Full Text Available The radioprotective effect of Achillea millefolium L (ACM extract was investigated against genotoxicity induced by ionizing radiation (IR in human lymphocytes. Peripheral blood samples were collected from human volunteers and incubated with the methanolic extract of ACM at different concentrations (10, 50, 100, and 200 μg/mL for 2 hours. At each dose point, the whole blood was exposed in vitro to 2.5 Gy of X-ray and then the lymphocytes were cultured with mitogenic stimulation to determine the micronuclei in cytokinesis-blocked binucleated cell. Antioxidant capacity of the extract was determined using free radical-scavenging method. The treatment of lymphocytes with the extract showed a significant decrease in the incidence of micronuclei binucleated cells, as compared with similarly irradiated lymphocytes without any extract treatment. The maximum protection and decrease in frequency of micronuclei were observed at 200 μg/mL of ACM extract which completely protected genotoxicity induced by IR in human lymphocytes. Achillea millefolium extract exhibited concentration-dependent radical-scavenging activity on 1,1-diphenyl-2-picryl hydrazyl free radicals. These data suggest that the methanolic extract of ACM may play an important role in the protection of normal tissues against genetic damage induced by IR.

  15. Differential Micronuclei Induction in Human Lymphocyte Cultures by Imidacloprid in the Presence of Potassium Nitrate

    Directory of Open Access Journals (Sweden)

    Polychronis Stivaktakis

    2010-01-01

    Full Text Available Humans are exposed to pesticides as a consequence of their application in farming or their persistence in a variety of media, including food, water, air, soil, plants, animals, and smoke. The interaction of pesticides with environmental factors may result in the alteration of their physicochemical properties. Square wave cathodic stripping voltammetry (SW-CSV, a technique that simulates electrodynamically the cellular membrane, is used to investigate whether the presence of potassium nitrate (KNO3 in the culture medium interferes with the genotoxic behavior of imidacloprid. The cytokinesis block micronuclei (CBMN method is used to evaluate imidacloprid's genotoxicity in the absence or presence of KNO3 in the culture medium and, as a consequence, its adsorption by lymphocytes. Comparing micronuclei (MN frequencies in control and imidacloprid-treated blood cell cultures, statistically significant differences were not detected. KNO3 did not induce MN frequencies compared to control. Statistically significant differences in MN frequencies were observed when blood cell cultures were treated with imidacloprid in the presence of increasing concentrations of KNO3. SW-CSV revealed that by increasing KNO3 molarity, imidacloprid's concentration in the culture medium decreased in parallel. This finding indicates that imidacloprid is adsorbed by cellular membranes. The present study suggests a novel role of a harmless environmental factor, such as KNO3, on the genotoxic behavior of a pesticide, such as imidacloprid. KNO3 rendered imidacloprid permeable to lymphocytes, resulting in elevated MN frequencies.

  16. Genotoxic evaluation of terbinafine in human lymphocytes in vitro.

    Science.gov (United States)

    Tolomeotti, Danielle; de Castro-Prado, Marialba Avezum Alves; de Sant'Anna, Juliane Rocha; Martins, Ana Beatriz Tozzo; Della-Rosa, Valter Augusto

    2015-01-01

    Terbinafine is an antimycotic drug usually used against several superficial fungal infections and with a potential application in the treatment of human cancers. Since to date there are few data on the genotoxic effects of terbinafine in mammalian cells, current study evaluated the potential genotoxic of such antifungal agent in cultured human peripheral blood lymphocytes. Terbinafine was used at the peak plasma concentration (1.0 μg/ml) and in four additional concentrations higher than the human plasmatic peak (5.0 μg/ml, 25.0 μg/ml, 50.0 μg/ml and 100.0 μg/ml). Chromosomal aberrations (CA), sister chromatid exchanges (SCE), micronuclei (MN), nucleoplasmic bridges (NP) and nuclear buds (NB) were scored as genetic endpoints. In all analysis no significant differences (α = 0.05, Kruskal-Wallis test) were observed. Complementary criterion adopted to obtain the final response in cytogenetic agreed with statistical results. Therefore, results of this study showed that terbinafine neither induced CA, SCE, MN, NP and NB nor affected significantly mitotic, replication and cytokinesis-block proliferation indices in any of the tested concentrations. It may be assumed that terbinafine was not genotoxic or cytotoxic to cultured human peripheral blood lymphocytes in our experimental conditions.

  17. Cytogenetic damage in circulating lymphocytes and buccal mucosa cells of head-and-neck cancer patients undergoing radiotherapy

    International Nuclear Information System (INIS)

    Minicucci, E.M.; Ribeiro, L.R.; Camargo, J.L.V. de; Salvadori, D.M.F.

    2005-01-01

    This study evaluated cytogenetic damage by measuring the frequency of micronucleated cells (MNC) in peripheral blood and buccal mucosa of head-and-neck cancer patients undergoing radiotherapy. MNC frequencies were assessed in 31 patients before, during, and after radiotherapy, and in 17 healthy controls matched for gender, age, and smoking habits. Results showed no statistically significant difference between patients and controls prior to radiotherapy in cytokinesis-blocked lymphocytes or buccal mucosa cells. During treatment, increased MNC frequencies were observed in both cell types. Micronucleated lymphocyte levels remained high in samples collected 30 to 140 days after the end of treatment, while MNC frequency in buccal mucosa decreased to values statistically similar to baseline values. There is controversy over the effects of age, smoking habit, tumor stage, and/or metastasis on MNC frequency. However, increased frequency of micronucleated buccal mucosa cells was seen in patients under 60 years old and in those with tumors >4 cm. In conclusion, the data show that radiotherapy has a potent clastogenic effect in circulating lymphocytes and buccal mucosa cells of head-and-neck cancer patients, and that the baseline MNC frequency in these two tissues is not a sensitive marker for head-and neck neoplasm. (author)

  18. 179 ORIGINAL ARTICLE

    African Journals Online (AJOL)

    boaz

    été conçue pour déterminer les sous – ensembles de lymphocyte T (CD4 + cellules T, CD8 + cellules T et le rapport CD4/CD8) dans les sujets atteints de cancer de la prostate et sujets témoins. CD4 + cellules T ( l/count) et CD8 + cellules T ont été estimées en utilisant la méthode de cytométrique en flux par Partec tandis ...

  19. Effects of zerumbone on cisplatin-induced clastogenesis in Sprague ...

    African Journals Online (AJOL)

    Administrator

    2011-06-20

    Jun 20, 2011 ... bol ester-induced papilloma formation in mouse skin which is another indication of its ... zerumbet plant. The rhizomes were obtained from the wet market in ... erythrocyte micronucleus test), guideline No. 474 (OECD 1997).

  20. African Journal of Biotechnology - Vol 7, No 5 (2008)

    African Journals Online (AJOL)

    ... practices on yield and profitability in kenaf (Hibiscus cannabinus L.) fibre cultivation ... Micronucleus test in fish genome: A sensitive monitor for aquatic pollution ... Response of microalgae from mud-flats to petroleum hydrocarbons in the ...

  1. (UVB)-induced DNA damage

    African Journals Online (AJOL)

    Jane

    2011-08-17

    dependent cytogenetic lesions were assessed by the micronucleus test (MNT). It was found that POE effectively reduced the extent of DNA breakages and cytogenetic lesions upon exposure to UVB (erythemal ultraviolet (EUV);.

  2. Combined effects of γ-ray radiation and high atmospheric pressure on peripheral blood lymphocytes

    International Nuclear Information System (INIS)

    Zhu Bingchai; Lu Jiaben; Wang Zongwu; Chen Tiehe

    1989-01-01

    The combined effects of γ-ray radiation and high atmospheric pressure on chromosome aberration, micronucleus and transformation frequency in peripheral blood lymphocytes have been studied. The results indicated that there were no significant influence for effects of high atmospheric pressure on chromosome aberrations, transformation frequency in peripheral blood lymphocytes induced γ-ray radiation, and that high atmospheric pressure increased effect of micronucleus in human peripheral blood lymphocytes in vitro induced γ-ray radiation

  3. Detection of Chlamydophila psittaci from feral pigeons in environmental samples: problems with currently available techniques.

    Science.gov (United States)

    Geigenfeind, Ila; Haag-Wackernagel, Daniel

    2010-03-01

    Chlamydophila psittaci (Lillie, 1930) Everett et al., 1999, the pathogenic agent of human ornithosis, is widespread in feral pigeon populations and many cases of transmission from feral pigeons to humans have been reported. The aim of the present study was to detect C. psittaci in environmental samples to find out more about possible transmission routes and, therefore, to assess the zoonotic risk for humans. Fecal samples were collected from nest boxes in a feral pigeon loft. Additionally, samples were taken from the feather dust film covering the water surface of public fountains where pigeons regularly bathe. The samples were tested for the presence of chlamydial antigen using an antigen enzyme-linked immunosorbent assay to prove shedding of C. psittaci by feral pigeons. This test detects a genus specific lipopolysaccharide in the outer membrane of the chlamydial bacteria. Samples were tested using the IDEIA PCE Chlamydia Test kit (DakoCytomation) and positive results were verified with IDEIA Chlamydia Blocking Reagents (DakoCytomation). The IDEIA PCE Chlamydia Test yields a high proportion of positive results. However, when IDEIA Chlamydia Blocking was performed, most of the positive results turned out to be negative or could not be interpreted. We conclude that antigen-enzyme-linked immunosorbent assay tests are not suitable for detecting C. psittaci in environmental samples. Previous publications where no blocking test was used should be reconsidered critically. © 2010 ISZS, Blackwell Publishing and IOZ/CAS.

  4. Evaluation of genotoxicity of nitrile fragrance ingredients using in vitro and in vivo assays.

    Science.gov (United States)

    Bhatia, S P; Politano, V T; Api, A M

    2013-09-01

    Genotoxicity studies were conducted on a group of 8 fragrance ingredients that belong to the nitrile family. These nitriles are widely used in consumer products however there is very limited data in the literature regarding the genotoxicity of these nitriles. The 8 nitriles were assessed for genotoxicity using an Ames test, in vitro chromosome aberration test or in vitro micronucleus test. The positive results observed in the in vitro tests were further investigated using an in vivo micronucleus test. The results from these different tests were compared and these 8 nitriles are not considered to be genotoxic. Dodecanitrile and 2,2,3-trimethylcyclopent-3-enylacetonitrile were negative in the in vitro chromosome aberration test and in vitro micronucleus test, respectively. While citronellyl nitrile, 3-methyl-5-phenylpentanenitrile, cinnamyl nitrile, and 3-methyl-5-phenylpent-2-enenitrile revealed positive results in the in vitro tests, but confirmatory in vivo tests determined these nitriles to be negative in the in vivo micronucleus assay. The remaining two nitriles (benzonitrile and α-cyclohexylidene benzeneacetonitrile) were negative in the in vivo micronucleus test. This study aims to evaluate the genotoxicity potential of these nitriles as well as enrich the literature with genotoxicity data on fragrance ingredients. Copyright © 2013 Elsevier Ltd. All rights reserved.

  5. Induction of micronuclei in human and mouse lymphocytes irradiated with gamma radiation and effect of panax ginseng C. A. Meyer

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Sung Ho; Oh, Heon; Lee, Song Eun [Chonnam National Univ., Kwangju (Korea, Republic of); Lee, Yun Sil; Kim, Tae Hwan [Korea Cancer Center Hospital, Seoul (Korea, Republic of); Jeong, Kyu Sik [Korea Research Institute of Bioscience and Biotechnology, Taejon (Korea, Republic of); Ryu, Si Yun [Chungnam National Univ., Taejon (Korea, Republic of)

    1997-09-01

    The frequencies of {gamma}-ray-induced micronuclei (MN) in Cytokinesis-Blocked (CB) lymphocytes at several doses were measured in three donors of human and C57BL/6 mice. Measurements performed after irradiation showed a dose-related increases in MN frequency in each of the donors studied. The relative sensitivity of mouse in Spleen Lymphocytes (SLs) compared with human Peripheral Blood Lymphocytes (PBLs) was estimated by best fitting linear-quadratic model based on the radiation-induced MN data over the range from 0 cGy to 400 cGy. In the case of MN frequency with 0.2 per CB cell, the relative sensitivity of mouse SLs was 1.67. Compared with the radiation-induced MN formation in the PBLs of human, the SLs of mouse were more radiosensitive. Using this MN assay with human PBLs and mouse SLs, studies were performed to determine whether the water fraction of ginseng (Panax ginseng C.A.Meyer)against radiation-induced MN in human PBLs after in vitro irradiation (3Gy) and in SLs of C57BL/6 mice after in vivo irradiation (3Gy). The frequency of MN in human PBLs was reduced by water fraction of ginseng (0.5mg/ml of medium) both pre-and post treatment (p<0.01) in vitro. In addition, the frequency of MN in mouse SLs was also reduced by pretreatment of ginseng (2mg/ml of drinking water for 7 days) in vivo.

  6. Maternal diet and dioxin-like activity, bulky DNA adducts and micronuclei in mother–newborns

    International Nuclear Information System (INIS)

    Pedersen, Marie; Halldorsson, Thorhallur I.; Autrup, Herman; Brouwer, Abraham; Besselink, Harrie; Loft, Steffen; Knudsen, Lisbeth E.

    2012-01-01

    Maternal diet can contribute to carcinogenic exposures and also modify effects of environmental exposures on maternal and fetal genetic stability. In this study, associations between maternal diet and the levels of dioxin-like plasma activity, bulky DNA adducts in white blood cells and micronuclei (MN) in lymphocytes from mother to newborns were examined. From 98 pregnant women living in the greater area of Copenhagen, Denmark in 2006–2007, maternal peripheral blood and umbilical cord blood were collected, together with information on health, environmental exposure and lifestyle. Maternal diet was estimated on the basis of maternal food frequency questionnaire (FFQ) completed by the end of pregnancy. Biomarkers were detected in paired blood samples through the dioxin-responsive chemical-activated luciferase expression (CALUX) ® bioassay, 32 P-postlabelling technique and cytokinesis-block MN assay. Maternal preference for meats with dark surface were significantly associated with higher bulky DNA adducts in both maternal (β 95%CI; 0.46 (0.08, 0.84)) and cord blood (β 95%CI; 0.46 (0.05, 0.86)) before and after adjustment for potential confounders. No other significant associations between the 18 dietary variables and the biomarkers measured in maternal and fetal samples were identified. The present study suggests that maternal intake of meats with dark surface contributes to the bulky DNA adduct levels in maternal and umbilical cord blood. Relationship between food preparation and bulky DNA adducts appear to be captured by a FFQ while potential associations for other biomarkers might be more complex or need larger sample size.

  7. Toxicity of a dental adhesive compared with ionizing radiation and zoledronic acid.

    Science.gov (United States)

    Alcaraz, Miguel; Olivares, Amparo; Achel, Daniel-Giyngiri; García-Cruz, Emilio; Fondevilla-Soler, Adriana; Canteras-Jordana, Manuel

    2015-07-01

    To determine the toxicity of aqueous dilutions of a universal self-priming dental adhesive (DA) and comparing these with those elicited by exposure to ionizing radiation (IR), Zoledronic acid (Z) treatment and the synergic effects of the combined treatment with IR+Z. The genotoxic effect of DA was determined by the increase in the frequency of micronuclei in cytokinesis-blocked in cultured human lymphocytes before and after exposure to 2Gy of X-rays. The cytotoxic effect was studied by using the MTT cell viability test in normal prostate cell lines (PNT2) after exposure to different X-ray doses (0Gy-20Gy). The cell lines divided into different groups and treated with different test substances: DA in presence of O2, DA in absence of O2, Z-treated and control. An in vitro dose-dependent and time-dependent cytotoxic effect of DA, Z and IR on PNT2 cells (p>0.001) was demonstrated. DA without-O2, following the recommendations of manufacturers, had a more pronounced effect of increasing cell death than DA with-O2 (p<0.001). In the genotoxicity assay, DA at 25% of its original concentration significantly increased chromosome damage (p<0.001). The samples studied were found to be toxic, and the samples photo-polymerized in absence of O2 showed a bigger cytotoxic effect comparable to the additive toxic effect showed by the combined treatment of IR+Z. Additional effort should be carried out to develop adhesives, which would reduce the release of hazardous substances; since toxic effects are similar to that reported by other agents whose clinical use is controlled by the health authorities.

  8. Protective effect of curcumin and its analog on γ-radiation induced DNA damage and lipid peroxidation in cultured human lymphocytes and isolated rat hepatocytes in vitro

    International Nuclear Information System (INIS)

    Menon, Venugopal P.

    2007-01-01

    Ionizing radiation is known to induce oxidative stress through generation of reactive oxygen species (ROS) resulting in an imbalance of the pro-oxidant and antioxidant status in the cells, which is suggested to culminate in cell death. The present work was aimed to evaluate the radioprotective effect of curcumin and its analog on γ-radiation induced toxicity in cultured human lymphocytes and rat hepatocytes. Hepatocytes were isolated from the liver of rats by collagenase perfusion. The cellular changes were estimated using lipid peroxidative indices like thiobarbituric acid reactive substances (TBARS), the antioxidants superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and reduced glutathione (GSH). The DNA damage was analyzed by comet assay, cytokinesis blocked micro nucleus assay, dicentric aberrations and translocation frequency. Cell cycle distribution and measurement of the percentage of apoptotic cells were performed by flow cytometry analysis. To investigate whether the dietary agents like curcumin and its analog have a role on cell cycle regulation, we analyzed the changes in cell cycle profiles by using fluorescence activated cell sorter. The increase in the severity of DNA damage was observed with the increase dose (1, 2 and 4 Gy) of γ-radiation in cultured lymphocytes and hepatocytes. TBARS were increased significantly, whereas the levels of GSH and antioxidant enzymes were significantly decreased in γ-irradiated hepatocytes and lymphocytes. On pretreatment with curcumin and its analog (1, 5 and 10 μg/ml) showed a significant decrease in the levels of TBARS and DNA damage. The antioxidant enzymes were increased significantly along with the levels of GSH. The maximum protection of hepatocytes and lymphocytes was observed at 10 μg/ml curcumin and 5 μg/ml curcumin analog pretreatment. Thus, pretreatment with curcumin and its analog helps in protecting the normal hepatocytes and lymphocytes against γ-radiation induced cellular

  9. Cytogenetic evaluation of hospital workers occupationally exposed to low levels of ionizing radiation. Assessment of two cytogenetic procedures: accumulated dosimetry versus radiosensitivity

    International Nuclear Information System (INIS)

    Garcia-Sagredo, J. M.; Villalon, C.; Lopez-Abente, G.; Arranz, L.; Ferro, M. T.; Ferrando, P.; Pollan, M.; Aragones, N.; Ferrer, N.; Sastre, J. M.

    2004-01-01

    We try to establish a cumulative ionising radiation (IR) biologicla dosimetry in occupationally exposed workers as a routine health tes, analysing chromosome translocations. 100 hospital workers occupationally exposed to low levels of X-ray, g-ray and radioactive isotopes are included in this study. Blood samples were cultured for cytogenetic analysis. Chromosome translocatiosn were scored using whole chromosome paint probes cocktail (Vysis) for chromosomes 1, 2, 3, 4, 5 and 6. Furthermore, a personal detailed interview about confounding factors, as tobacco smoking X-ray examination, occupational exposure to chemotherapeutics agents and solvents, electromagnetic fields exposure, and others was done. Our results showed that there is no statistical association between cumulative doses of IRE, type of LET and chromosome translocation rate. For the contraty, we have found a translocation risk increase related with IR equivalent dose rate, independently of the time of exposure and age. Those workers receiving 1mSv/year or more vs<1mSv/year show a relative risk of 2.56 (95% confidence interval 1.10-5.95). A comparison of translocationrate and different confounding factors suggest a relative risk increase in intensive users of mobile phones. Other exposures as tobacco smoking solvents, UV radiation anaesthetic gases and any other confounding factors have not shown assocaition with translocation rate. At the same time, we try to analyse radiosensitivity through the CBMN-test (cytokinesis block micronucelus test). Results on CBMN are in progress. Finally we try to asses the two cytogentic procedures as the most suitable routine health test in radiation occupational protection. (Author)

  10. Maternal diet and dioxin-like activity, bulky DNA adducts and micronuclei in mother-newborns

    Energy Technology Data Exchange (ETDEWEB)

    Pedersen, Marie, E-mail: mpedersen@creal.cat [Section of Environmental Health, Department of Public Health, University of Copenhagen, CSS, Oester Farimagsgade, Copenhagen K (Denmark); Halldorsson, Thorhallur I., E-mail: lur@ssi.dk [Faculty of Food Science and Nutrition, School of Health Sciences, University of Iceland Reykjavik (Iceland); Center for Fetal Programming, Department of Epidemiology, Statens Serum Institute, Copenhagen (Denmark); Autrup, Herman, E-mail: ha@mil.au.dk [School of Public Health, Department of Environmental and Occupational Medicine, Aarhus University, Aarhus (Denmark); Brouwer, Abraham, E-mail: Bram.Brouwer@bds.nl [BioDetection Systems B.V., Amsterdam (Netherlands); Besselink, Harrie, E-mail: Harrie.Besselink@bds.nl [BioDetection Systems B.V., Amsterdam (Netherlands); Loft, Steffen, E-mail: stl@sund.ku.dk [Section of Environmental Health, Department of Public Health, University of Copenhagen, CSS, Oester Farimagsgade, Copenhagen K (Denmark); Knudsen, Lisbeth E., E-mail: liek@sund.ku.dk [Section of Environmental Health, Department of Public Health, University of Copenhagen, CSS, Oester Farimagsgade, Copenhagen K (Denmark)

    2012-06-01

    Maternal diet can contribute to carcinogenic exposures and also modify effects of environmental exposures on maternal and fetal genetic stability. In this study, associations between maternal diet and the levels of dioxin-like plasma activity, bulky DNA adducts in white blood cells and micronuclei (MN) in lymphocytes from mother to newborns were examined. From 98 pregnant women living in the greater area of Copenhagen, Denmark in 2006-2007, maternal peripheral blood and umbilical cord blood were collected, together with information on health, environmental exposure and lifestyle. Maternal diet was estimated on the basis of maternal food frequency questionnaire (FFQ) completed by the end of pregnancy. Biomarkers were detected in paired blood samples through the dioxin-responsive chemical-activated luciferase expression (CALUX){sup Registered-Sign} bioassay, {sup 32}P-postlabelling technique and cytokinesis-block MN assay. Maternal preference for meats with dark surface were significantly associated with higher bulky DNA adducts in both maternal ({beta} 95%CI; 0.46 (0.08, 0.84)) and cord blood ({beta} 95%CI; 0.46 (0.05, 0.86)) before and after adjustment for potential confounders. No other significant associations between the 18 dietary variables and the biomarkers measured in maternal and fetal samples were identified. The present study suggests that maternal intake of meats with dark surface contributes to the bulky DNA adduct levels in maternal and umbilical cord blood. Relationship between food preparation and bulky DNA adducts appear to be captured by a FFQ while potential associations for other biomarkers might be more complex or need larger sample size.

  11. Protection of ionizing radiation-induced cytogenetic damage by hydroalcoholic extract of Cynodon dactylon in Chinese hamster lung fibroblast cells and human peripheral blood lymphocytes.

    Science.gov (United States)

    Rao, Bola Sadashiva Satish; Upadhya, Dinesh; Adiga, Satish Kumar

    2008-01-01

    The radiomodulatory potential of hydroalcoholic extract of a medicinal plant Cynodon dactylon (family: Poaceae) against radiation-induced cytogenetic damage was analyzed using Chinese hamster lung fibroblast (V79) cells and human peripheral blood lymphocytes (HPBLs) growing in vitro. Induction of micronuclei was used as an index of cytogenetic damage, evaluated in cytokinesis blocked binucleate cells. The hydroalcoholic Cynodon dactylon extract (CDE) rendered protection against the radiation-induced DNA damage, as evidenced by the significant (p<0.001) reduction in micronucleated binucleate cells (MNBNC%) after various doses of CDE treatment in V79 cells and HPBLs. The optimum dose of CDE (40 and 50 microg/ml in HPBLs and V79 cells, respectively) with the greatest reduction in micronuclei was further used in combination with various doses of gamma radiation (0.5, 1, 2, 3, and 4 Gy) exposed 1 h after CDE treatment. A linear dose-dependent MNBNC% increase in radiation alone group was observed, while 40/50 microg/ml CDE significantly resulted in the reduction of MNBNC%, compared to the respective radiation alone groups. CDE resulted in a dose-dependent increase in free radical scavenging ability against various free radicals, viz., 2, 2-diphenyl-2-picryl-hydrazyl (DPPH); 2, 2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS); superoxide anion (O2*-); hydroxyl radical (OH*) and nitric oxide radical (NO*) generated in vitro. Also, an excellent (70%) inhibition of lipid peroxidation in vitro was observed at a dose of 300 microg/ml CDE, attaining the saturation point at higher doses. The present findings demonstrated the radioprotective effect of CDE, also rendering protection against radiation-induced genomic instability and DNA damage. The observed radioprotective effect may be partly attributed to the free radical scavenging and antilipid peroxidative potential of CDE.

  12. Analysis of chromosomal aberrations, sister-chromatid exchanges and micronuclei in peripheral lymphocytes of pharmacists before and after working with cytostatic drugs.

    Science.gov (United States)

    Roth, S; Norppa, H; Järventaus, H; Kyyrönen, P; Ahonen, M; Lehtomäki, J; Sainio, H; Sorsa, M

    1994-12-01

    The frequencies of chromosome aberrations, SCEs and micronuclei (cytokinesis-block method) in blood lymphocytes were compared among six nonsmoking female pharmacists before and after 1 year of working with cytostatic drugs. All possible precautions were taken to avoid exposure to cytostatics, including proper protective clothing and a monitored, negative-pressured working environment with vertical laminar flow cabinet. As referents, an age-matched group of six nonsmoking female hospital workers not dealing with cytostatics was simultaneously sampled twice with the same time interval. The pharmacists showed a marginally higher mean frequency of SCEs/cell (6.3; P = 0.049) after the working period than 1 year earlier (5.8). On the other hand, the referents, with no obvious exposure, had a higher mean number of cells with chromatid-type aberrations, gaps excluded, in the second sampling (2.0%; P = 0.048) than in the first one (0.5%). In addition, a slight (P = 0.055) trend towards a higher frequency of micronucleated binucleate cells was observed in the second sampling for both the exposed and control subjects. As such findings suggest technical variation in the cytogenetic parameters, the small difference observed in SCEs for the pharmacists between the two samplings was probably not related to the cytostatics exposure. No statistically significant differences were observed for any of the cytogenetic parameters in comparisons between the pharmacists and the referents. The findings suggest that caution should be exercised in comparing results obtained from two different samplings in prospective cytogenetic studies.

  13. Development of radiation biological dosimetry and treatment of radiation-induced damaged tissue

    International Nuclear Information System (INIS)

    Cho, Chul Koo; Kim, Tae Hwan; Lee, Yun Sil

    2000-04-01

    Util now, only a few methods have been developed for radiation biological dosimetry such as conventional chromosome aberration and micronucleus in peripheral blood cell. However, because these methods not only can be estimated by the expert, but also have a little limitation due to need high technique and many times in the case of radiation accident, it is very difficult to evaluate the absorbed dose of victims. Therefore, we should develop effective, easy, simple and rapid biodosimetry and its guideline(triage) to be able to be treated the victims as fast as possible. We established the apoptotic fragment assay, PCC, comet assay, and micronucleus assay which was the significant relationship between dose and cell damages to evaluate the irradiated dose as correct and rapid as possible using lymphocytes and crypt cells, and compared with chromosome dosimetry and micronucleus assay

  14. Development of radiation biological dosimetry and treatment of radiation-induced damaged tissue

    Energy Technology Data Exchange (ETDEWEB)

    Cho, Chul Koo; Kim, Tae Hwan; Lee, Yun Sil [and others

    2000-04-01

    Util now, only a few methods have been developed for radiation biological dosimetry such as conventional chromosome aberration and micronucleus in peripheral blood cell. However, because these methods not only can be estimated by the expert, but also have a little limitation due to need high technique and many times in the case of radiation accident, it is very difficult to evaluate the absorbed dose of victims. Therefore, we should develop effective, easy, simple and rapid biodosimetry and its guideline(triage) to be able to be treated the victims as fast as possible. We established the apoptotic fragment assay, PCC, comet assay, and micronucleus assay which was the significant relationship between dose and cell damages to evaluate the irradiated dose as correct and rapid as possible using lymphocytes and crypt cells, and compared with chromosome dosimetry and micronucleus assay.

  15. Chemopreventive activity of compounds extracted from Casearia sylvestris (Salicaceae) Sw against DNA damage induced by particulate matter emitted by sugarcane burning near Araraquara, Brazil

    Energy Technology Data Exchange (ETDEWEB)

    Prieto, A.M. [UNESP — Univ. Estadual Paulista, College of Pharmaceutical Sciences, Department of Clinical Analysis, Rua Expedicionários do Brasil, 1621, Araraquara (Brazil); Santos, A.G. [UNESP — Univ. Estadual Paulista, College of Pharmaceutical Sciences, Department of Natural Principles and Toxicology, Rodovia Araraquara-Jau, km 01, Araraquara (Brazil); Csipak, A.R.; Caliri, C.M.; Silva, I.C. [UNESP — Univ. Estadual Paulista, College of Pharmaceutical Sciences, Department of Clinical Analysis, Rua Expedicionários do Brasil, 1621, Araraquara (Brazil); Arbex, M.A. [UNIFESP — Federal University of São Paulo, Paulista College of Medicine, Department of Internal Medicine, Rua Pedro de Toledo, 720, São Paulo (Brazil); Silva, F.S.; Marchi, M.R.R. [UNESP — Univ. Estadual Paulista, Chemistry Institute, Department of Analytical Chemistry, Rua Francisco Degni, S/N, Araraquara (Brazil); Cavalheiro, A.J.; Silva, D.H.S.; Bolzani, V.S. [UNESP — Univ. Estadual Paulista, Chemistry Institute, Department of Organic Chemistry, Rua Francisco Degni, S/N, Araraquara (Brazil); Soares, C.P., E-mail: soarescp@hotmail.com [UNESP — Univ. Estadual Paulista, College of Pharmaceutical Sciences, Department of Clinical Analysis, Rua Expedicionários do Brasil, 1621, Araraquara (Brazil)

    2012-12-15

    Ethanolic extract of Casearia sylvestris is thought to be antimutagenic. In this study, we attempted to determine whether this extract and casearin X (a clerodane diterpene from C. sylvestris) are protective against the harmful effects of airborne pollutants from sugarcane burning. To that end, we used the Tradescantia micronucleus test in meiotic pollen cells of Tradescantia pallida, the micronucleus test in mouse bone marrow cells, and the comet assay in mouse blood cells. The mutagenic compound was total suspended particulate (TSP) from air. For the Tradescantia micronucleus test, T. pallida cuttings were treated with the extract at 0.13, 0.25, or 0.50 mg/ml. Subsequently, TSP was added at 0.3 mg/ml, and tetrads from the inflorescences were examined for micronuclei. For the micronucleus test in mouse bone marrow cells and the comet assay in mouse blood cells, Balb/c mice were treated for 15 days with the extract—3.9, 7.5, or 15.0 mg/kg body weight (BW)—or with casearin X—0.3, 0.25, or 1.2 mg/kg BW—after which they received TSP (3.75 mg/kg BW). In T. pallida and mouse bone marrow cells, the extract was antimutagenic at all concentrations tested. In mouse blood cells, the extract was antigenotoxic at all concentrations, whereas casearin X was not antimutagenic but was antigenotoxic at all concentrations. We conclude that C. sylvestris ethanolic extract and casearin X protect DNA from damage induced by airborne pollutants from sugarcane burning. -- Highlights: ► We assessed DNA protection of C. sylvestris ethanolic extract. ► We assessed DNA protection of casearin X. ► We used Tradescantia pallida micronucleus test as screening. ► We used comet assay and micronucleus test in mice. ► The compounds protected DNA against sugar cane burning pollutants.

  16. Chromosomal radiosensitivity in breast cancer patients with a known or putative genetic predisposition.

    LENUS (Irish Health Repository)

    Baeyens, A

    2002-12-02

    The chromosomal radiosensitivity of breast cancer patients with a known or putative genetic predisposition was investigated and compared to a group of healthy women. The chromosomal radiosensitivity was assessed with the G2 and the G0-micronucleus assay. For the G2 assay lymphocytes were irradiated in vitro with a dose of 0.4 Gy (60)Co gamma-rays after 71 h incubation, and chromatid breaks were scored in 50 metaphases. For the micronucleus assay lymphocytes were exposed in vitro to 3.5 Gy (60)Co gamma-rays at a high dose rate or low dose rate. 70 h post-irradiation cultures were arrested and micronuclei were scored in 1000 binucleate cells. The results demonstrated that the group of breast cancer patients with a known or putative genetic predisposition was on the average more radiosensitive than a population of healthy women, and this with the G2 as well as with the high dose rate and low dose rate micronucleus assay. With the G2 assay 43% of the patients were found to be radiosensitive. A higher proportion of the patients were radiosensitive with the micronucleus assay (45% with high dose rate and 61% with low dose rate). No correlation was found between the G2 and the G0-micronucleus chromosomal radiosensitivity. Out of the different subgroups considered, the group of the young breast cancer patients without family history showed the highest percentage of radiosensitive cases in the G2 (50%) as well as in the micronucleus assay (75-78%).

  17. Seven benzimidazole pesticides combined at sub-threshold levels induce micronuclei in vitro

    Science.gov (United States)

    Ermler, Sibylle; Scholze, Martin; Kortenkamp, Andreas

    2013-01-01

    Benzimidazoles act by disrupting microtubule polymerisation and are capable of inducing the formation of micronuclei. Considering the similarities in their mechanisms of action (inhibition of microtubule assembly by binding to the colchicine-binding site on tubulin monomers), combination effects according to the principles of concentration addition might occur. If so, it is to be expected that several benzimidazoles contribute to micronucleus formation even when each single one is present at or below threshold levels. This would have profound implications for risk assessment, but the idea has never been tested rigorously. To fill this gap, we analysed micronucleus frequencies for seven benzimidazoles, including the fungicide benomyl, its metabolite carbendazim, the anthelmintics albendazole, albendazole oxide, flubendazole, mebendazole and oxibendazole. Thiabendazole was also tested but was inactive. We used the cytochalasin-blocked micronucleus assay with CHO-K1 cells according to OECD guidelines, and employed an automated micronucleus scoring system based on image analysis to establish quantitative concentration–response relationships for the seven active benzimidazoles. Based on this information, we predicted additive combination effects for a mixture of the seven benzimidazoles by using the concepts of concentration addition and independent action. The observed effects of the mixture agreed very well with those predicted by concentration addition. Independent action underestimated the observed combined effects by a large margin. With a mixture that combined all benzimidazoles at their estimated threshold concentrations for micronucleus induction, micronucleus frequencies of ~15.5% were observed, correctly anticipated by concentration addition. On the basis of independent action, this mixture was expected to produce no effects. Our data provide convincing evidence that concentration addition is applicable to combinations of benzimidazoles that form micronuclei

  18. Acute Toxicity and Genotoxic Activity of Avocado Seed Extract (Persea americana Mill., c.v. Hass

    Directory of Open Access Journals (Sweden)

    Eduardo Padilla-Camberos

    2013-01-01

    Full Text Available The use of vegetal extracts requires toxicological and genotoxic evaluations to establish and verify safety before being added to human cosmetic, pharmaceutical medicine, or alimentary products. Persea americana seeds have been used in traditional medicine as treatment for several diseases. In this work, the ethanolic seed extract of Persea americana was evaluated with respect to its genotoxic potential through micronucleus assay in rodents. The frequency of micronuclei in groups of animals treated with avocado seed extract showed no differences compared to the negative control (vehicle; therefore, it is considered that the avocado seed extract showed no genotoxic activity in the micronucleus test.

  19. Effects of irradiation with low-energy nitrogen ion injection on root tip cells of broad bean

    International Nuclear Information System (INIS)

    Huang Yaqin; Li Jinzhe; Huang Qunce

    2012-01-01

    In order to study the cytogenetic effects of low-energy nitrogen ion irradiation, broad bean seed embryo was irradiated by different doses of nitrogen ions. Micronucleus rate, mitotic index and chromosome aberration in root-tip cells were analyzed. The results showed that the injection of ions inhibited mitosis of root tip cells, interfered the normal process of mitosis, caused aberrations of chromosome structure, behavior and number. The frequency of micronucleus and chromosomal aberrations increased with the increasing radiation dosage, while mitotic index decreased. (authors)

  20. Replication of somatic micronuclei in bovine enucleated oocytes

    Directory of Open Access Journals (Sweden)

    Canel Natalia

    2012-11-01

    Full Text Available Abstract Background Microcell-mediated chromosome transfer (MMCT was developed to introduce a low number of chromosomes into a host cell. We have designed a novel technique combining part of MMCT with somatic cell nuclear transfer, which consists of injecting a somatic micronucleus into an enucleated oocyte, and inducing its cellular machinery to replicate such micronucleus. It would allow the isolation and manipulation of a single or a low number of somatic chromosomes. Methods Micronuclei from adult bovine fibroblasts were produced by incubation in 0.05 μg/ml demecolcine for 46 h followed by 2 mg/ml mitomycin for 2 h. Cells were finally treated with 10 μg/ml cytochalasin B for 1 h. In vitro matured bovine oocytes were mechanically enucleated and intracytoplasmatically injected with one somatic micronucleus, which had been previously exposed [Micronucleus- injected (+] or not [Micronucleus- injected (−] to a transgene (50 ng/μl pCX-EGFP during 5 min. Enucleated oocytes [Enucleated (+] and parthenogenetic [Parthenogenetic (+] controls were injected into the cytoplasm with less than 10 pl of PVP containing 50 ng/μl pCX-EGFP. A non-injected parthenogenetic control [Parthenogenetic (−] was also included. Two hours after injection, oocytes and reconstituted embryos were activated by incubation in 5 μM ionomycin for 4 min + 1.9 mM 6-DMAP for 3 h. Cleavage stage and egfp expression were evaluated. DNA replication was confirmed by DAPI staining. On day 2, Micronucleus- injected (−, Parthenogenetic (− and in vitro fertilized (IVF embryos were karyotyped. Differences among treatments were determined by Fisher′s exact test (p≤0.05. Results All the experimental groups underwent the first cell divisions. Interestingly, a low number of Micronucleus-injected embryos showed egfp expression. DAPI staining confirmed replication of micronuclei in most of the evaluated embryos. Karyotype analysis revealed that all Micronucleus-injected embryos had

  1. Acute toxicity and genotoxic activity of avocado seed extract (Persea americana Mill., c.v. Hass).

    Science.gov (United States)

    Padilla-Camberos, Eduardo; Martínez-Velázquez, Moisés; Flores-Fernández, José Miguel; Villanueva-Rodríguez, Socorro

    2013-01-01

    The use of vegetal extracts requires toxicological and genotoxic evaluations to establish and verify safety before being added to human cosmetic, pharmaceutical medicine, or alimentary products. Persea americana seeds have been used in traditional medicine as treatment for several diseases. In this work, the ethanolic seed extract of Persea americana was evaluated with respect to its genotoxic potential through micronucleus assay in rodents. The frequency of micronuclei in groups of animals treated with avocado seed extract showed no differences compared to the negative control (vehicle); therefore, it is considered that the avocado seed extract showed no genotoxic activity in the micronucleus test.

  2. The influence of DNA repair inhibitors on the mutation rate

    International Nuclear Information System (INIS)

    Auzinger, Th.; Hruby, R.

    1980-12-01

    The simultaneous influence of gamma-radiation and DNA-repair inhibiting substances on the mutation frequency of mice was investigated in vivo with the micronucleus test. The detergens Tween 80, vitamin A, and the antiphlogisticum phenylbutazone were used as DNA-repair inhibiting substances. Using the same irradiation doses, a statistic significant increase of mutagenicity respectively micronucleus frequency was found in high concentrations of Tween 80 and in all used dosages of vitamin A, but not in phenylbutazone and in low concentrations of tween. (auth.)

  3. Big data analytics methods and applications

    CERN Document Server

    Rao, BLS; Rao, SB

    2016-01-01

    This book has a collection of articles written by Big Data experts to describe some of the cutting-edge methods and applications from their respective areas of interest, and provides the reader with a detailed overview of the field of Big Data Analytics as it is practiced today. The chapters cover technical aspects of key areas that generate and use Big Data such as management and finance; medicine and healthcare; genome, cytome and microbiome; graphs and networks; Internet of Things; Big Data standards; bench-marking of systems; and others. In addition to different applications, key algorithmic approaches such as graph partitioning, clustering and finite mixture modelling of high-dimensional data are also covered. The varied collection of themes in this volume introduces the reader to the richness of the emerging field of Big Data Analytics.

  4. Cytogenetic analysis of oral mucosa cells, induced by chlorhexidine, essential oils in ethanolic solution and triclosan mouthwashes

    Energy Technology Data Exchange (ETDEWEB)

    Ros-Llor, Irene; Lopez-Jornet, Pia, E-mail: majornet@um.es

    2014-07-15

    Objectives: The aim of this study was to evaluate DNA damage and cytokinetic defects, proliferative potential and cell death caused by the frequent use of mouthrinses containing chlorhexidine, triclosan and essential oils in ethanolic solution, compared to a placebo mouthwash. Study design: This double-blind, prospective, randomized clinical trial included 80 Caucasian patients. Subjects were divided into four groups: Group I used a mouthrinse, Triclosan; Group II used physiological saline; Group III used chlorhexidine; Group IV a mouthrinse with essential oils in ethanolic solution. All subjects used the mouthrinses for two weeks (15 ml, twice a day, rinsing for 30 s). Two cell samples per subject were collected, before and after mouthrinse use (on day 0 and day 15). Samples were processed as follows: cell collection from cheeks with a cytobrush; cell centrifuge; slide preparation, fixation and staining; and fluorescent microscope analysis. 2000 exfoliated cells were screened for nuclear abnormalities, particularly the presence of micronuclei by means of cytome assay. Results: No significant differences between study times (before and after use of mouthwash) were identified for any of the variables studied (p>0.05). Differences between mouthrinse groups were also compared but no significant differences were found (p>0.05). Conclusions: This study did not observe any genotoxic effect resulting from mouthrinse use. - Highlights: • Mouthrinses are used widely, mainly for their capacity to control dental plaque. • No genotoxic effects from the mouthrinses triclosan, chlorhexidine essential oils solution. • The buccal cytome assay is a sensitive, non-invasive, and low cost technique.

  5. n-TiO{sub 2} and CdCl{sub 2} co-exposure to titanium dioxide nanoparticles and cadmium: Genomic, DNA and chromosomal damage evaluation in the marine fish European sea bass (Dicentrarchus labrax)

    Energy Technology Data Exchange (ETDEWEB)

    Nigro, M.; Bernardeschi, M. [Department of Clinical and Experimental Medicine, Pisa University, Pisa (Italy); Costagliola, D. [Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, Second University of Naples, Caserta (Italy); Della Torre, C. [Department of Physical, Earth and Environmental Sciences, University of Siena, Siena (Italy); Frenzilli, G., E-mail: giada@biomed.unipi.it [Department of Clinical and Experimental Medicine, Pisa University, Pisa (Italy); Guidi, P.; Lucchesi, P. [Department of Clinical and Experimental Medicine, Pisa University, Pisa (Italy); Mottola, F.; Santonastaso, M. [Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, Second University of Naples, Caserta (Italy); Scarcelli, V. [Department of Clinical and Experimental Medicine, Pisa University, Pisa (Italy); Monaci, F.; Corsi, I. [Department of Physical, Earth and Environmental Sciences, University of Siena, Siena (Italy); Stingo, V.; Rocco, L. [Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, Second University of Naples, Caserta (Italy)

    2015-11-15

    Highlights: • European sea bass was exposed to CdCl{sub 2} and n-TiO{sub 2} alone and in combination. • Genotoxicity was evaluated by RAPD-assay, comet assay and cytome assay. • CdCl{sub 2} induced DNA primary damage but not chromosomal damage. • n-TiO{sub 2} induced chromosomal damage but not DNA primary damage. • Co-exposure effects depend on the biomarker used. - Abstract: Due to the large production and growing use of titanium dioxide nanoparticles (n-TiO{sub 2}), their release in the marine environment and their potential interaction with existing toxic contaminants represent a growing concern for biota. Different end-points of genotoxicity were investigated in the European sea bass Dicentrarchus labrax exposed to n-TiO{sub 2} (1 mg L{sup −1}) either alone and combined with CdCl{sub 2} (0.1 mg L{sup −1}) for 7 days. DNA primary damage (comet assay), apoptotic cells (diffusion assay), occurrence of micronuclei and nuclear abnormalities (cytome assay) were assessed in peripheral erythrocytes and genomic stability (random amplified polymorphism DNA-PCR, RAPD assay) in muscle tissue. Results showed that genome template stability was reduced after CdCl{sub 2} and n-TiO{sub 2} exposure. Exposure to n-TiO{sub 2} alone was responsible for chromosomal alteration but ineffective in terms of DNA damage; while the opposite was observed in CdCl{sub 2} exposed specimens. Co-exposure apparently prevents the chromosomal damage and leads to a partial recovery of the genome template stability.

  6. Cytogenetic analysis of oral mucosa cells, induced by chlorhexidine, essential oils in ethanolic solution and triclosan mouthwashes

    International Nuclear Information System (INIS)

    Ros-Llor, Irene; Lopez-Jornet, Pia

    2014-01-01

    Objectives: The aim of this study was to evaluate DNA damage and cytokinetic defects, proliferative potential and cell death caused by the frequent use of mouthrinses containing chlorhexidine, triclosan and essential oils in ethanolic solution, compared to a placebo mouthwash. Study design: This double-blind, prospective, randomized clinical trial included 80 Caucasian patients. Subjects were divided into four groups: Group I used a mouthrinse, Triclosan; Group II used physiological saline; Group III used chlorhexidine; Group IV a mouthrinse with essential oils in ethanolic solution. All subjects used the mouthrinses for two weeks (15 ml, twice a day, rinsing for 30 s). Two cell samples per subject were collected, before and after mouthrinse use (on day 0 and day 15). Samples were processed as follows: cell collection from cheeks with a cytobrush; cell centrifuge; slide preparation, fixation and staining; and fluorescent microscope analysis. 2000 exfoliated cells were screened for nuclear abnormalities, particularly the presence of micronuclei by means of cytome assay. Results: No significant differences between study times (before and after use of mouthwash) were identified for any of the variables studied (p>0.05). Differences between mouthrinse groups were also compared but no significant differences were found (p>0.05). Conclusions: This study did not observe any genotoxic effect resulting from mouthrinse use. - Highlights: • Mouthrinses are used widely, mainly for their capacity to control dental plaque. • No genotoxic effects from the mouthrinses triclosan, chlorhexidine essential oils solution. • The buccal cytome assay is a sensitive, non-invasive, and low cost technique

  7. Assessment of mutagenic, antimutagenic and genotoxicity effects of Mimosa tenuiflora

    Directory of Open Access Journals (Sweden)

    Viviane A. Silva

    2013-02-01

    Full Text Available Genotoxic effects of Mimosa tenuiflora (Willd. Poir, Fabaceae, were investigated by using both micronucleus test and bacterial reverse mutation assay in Salmonella typhimurium TA97, TA98, TA100, TA102 respectively. In respect of Ames test results show that the extract does not induce mutations in any strains of Salmonella typhimurium tested since the mutagenicity index is less than 2. In the antimutagenic effect was observed that the extract at the concentrations tested significantly decreased the mutagenicity index of all strains tested which characterized the extract as antimutagenic in these conditions. In the micronucleus test in vivo, we observed that the concentrations used did not induce an increase in the frequency of micronucleus in normochromatic erythrocytes of mice. Therefore, we concluded that the extract of M. tenuiflora is not mutagenic in the absence of exogenous metabolizing system and does not induce an increase in the frequency of the micronucleus characterized as an agent not mutagenic in these conditions. Further studies of toxicity need to be made to the use of this plant in the treatment of diseases to be stimulated.

  8. Assessment of mutagenic, antimutagenic and genotoxicity effects of Mimosa tenuiflora

    Directory of Open Access Journals (Sweden)

    Viviane A. Silva

    2013-04-01

    Full Text Available Genotoxic effects of Mimosa tenuiflora (Willd. Poir, Fabaceae, were investigated by using both micronucleus test and bacterial reverse mutation assay in Salmonella typhimurium TA97, TA98, TA100, TA102 respectively. In respect of Ames test results show that the extract does not induce mutations in any strains of Salmonella typhimurium tested since the mutagenicity index is less than 2. In the antimutagenic effect was observed that the extract at the concentrations tested significantly decreased the mutagenicity index of all strains tested which characterized the extract as antimutagenic in these conditions. In the micronucleus test in vivo, we observed that the concentrations used did not induce an increase in the frequency of micronucleus in normochromatic erythrocytes of mice. Therefore, we concluded that the extract of M. tenuiflora is not mutagenic in the absence of exogenous metabolizing system and does not induce an increase in the frequency of the micronucleus characterized as an agent not mutagenic in these conditions. Further studies of toxicity need to be made to the use of this plant in the treatment of diseases to be stimulated.

  9. Genotoxicology: Single and Joint Action of Copper and Zinc to ...

    African Journals Online (AJOL)

    Michael Horsfall

    Synodontis clarias and Tilapia nilotica using the sensitive micronucleus assay in fish genome. ... concentrations are important for the physiological ... Fish were divided into exposure groups for every concentration studied. Fish in twenties were exposed to copper and zinc, forty fish to the mixture of metals, and twenty fish ...

  10. Analysis of Genetic Damage in Lymphocytes of Former Uranium Processing Workers

    Czech Academy of Sciences Publication Activity Database

    Zölzer, F.; Havránková, R.; Freitinger Skalická, Z.; Rössnerová, Andrea; Šrám, Radim

    2015-01-01

    Roč. 147, č. 1 (2015), s. 17-23 ISSN 1424-8581 Institutional support: RVO:68378041 Keywords : alpha-radiation * genomic instability * micronucleus-centromere test * radon * uranium miners Subject RIV: DN - Health Impact of the Environment Quality Impact factor: 1.638, year: 2015

  11. CYTOGENETIC EFFECTS INDUCED BY 4-CHLOROHYDRATE-BROMO-6-METHYL-3-DIMETHYLAMINO-3-CHROMANONE IN OCIMUM BASILICUM L. SPECIES

    Directory of Open Access Journals (Sweden)

    Mihaela Florina Axente

    2006-08-01

    dimethylamino-3-chromanone. The 1/10000 dilution induces the increase of frequency of mitotic dividing cells. The cells with chromosome aberrations are in greater number in treated variants, comparatively with control. The aberration spectrum is enough large and comprises: ana-telophases with bridges, lagging chromosomes, expelled chromosomes, multipolar ana-telophases, as well as binucleate cells and interphases with micronucleuses.

  12. Cytogenetics observation and radiation influence evaluation of exposed persons in a discontinuous radiation exposure event

    International Nuclear Information System (INIS)

    Chen Ying; Liu Xiulin; Yang Guoshan; Ge Shili; Jin Cuizhen; Yao Bo

    2003-01-01

    The cytogenetics results and dose estimation of exposed and related persons in an discontinuous radiation exposure event were reported in this paper. According to dicentrics + ring and micronucleus results combined with clinical data, slight (middle) degree of subacute radiation symptom of the victim was diagnosed. A part of 52 examined persons were exposed to radiation in a certain degree

  13. Adaptive response to ionizing radiation in normal human skin fibroblasts. Enhancement of DNA repair rate and modulation of gene expression. Reponse adaptative au rayonnement ionisant des fibroblastes de peau humaine. Augmentation de la vitesse de reparation de l'ADN et variation de l'expression des genes

    Energy Technology Data Exchange (ETDEWEB)

    Toledo, S.M. de; Mitchel, R.E.J. (Atomic Energy of Canada Ltd., Chalk River, ON (Canada). Chalk River Nuclear Labs.); Azzam, E. (Atomic Energy of Canada Ltd., Chalk River, ON (Canada). Chalk River Nuclear Labs. Ottawa Univ., ON (Canada). Dept. of Biology); Raaphorst, G.P. (Ottawa Univ., ON (Canada). Dept. of Biology)

    Low doses and dose rates of ionizing radiation enhance the rate of DNA repair in human fibroblasts and protect the cells against radiation-induced micronucleus formation. Chronic exposures reduce the mRNA levels of the genes topoisomerase II and FACC-1 (Fanconi's anemia, group C). (authors). 11 refs., 1 tab., 2 figs.

  14. Adaptive response to ionizing radiation in normal human skin fibroblasts. Enhancement of DNA repair rate and modulation of gene expression

    International Nuclear Information System (INIS)

    Toledo, S.M. de; Mitchel, R.E.J.; Azzam, E.; Ottawa Univ., ON; Raaphorst, G.P.

    1994-01-01

    Low doses and dose rates of ionizing radiation enhance the rate of DNA repair in human fibroblasts and protect the cells against radiation-induced micronucleus formation. Chronic exposures reduce the mRNA levels of the genes topoisomerase II and FACC-1 (Fanconi's anemia, group C). (authors). 11 refs., 1 tab., 2 figs

  15. Micronuclei frequencies in lymphocytes and cervical cells of women with polycystic ovarian syndrome

    Directory of Open Access Journals (Sweden)

    Rengin Karataylı

    2017-09-01

    Full Text Available Objective: The aim of this study was to determine micronucleus (MN frequencies in exfoliated cervical cells and peripheral blood lymphocytes of women with polycystic ovarian syndrome (PCOS. Materials and Methods: Fifteen patients with PCOS and 11 healthy control patients were included in the study. Cervical smears and peripheral blood were collected from all patients. Specimens were analyzed for MN frequencies and compared between the groups. In addition to MN, other nuclear anomalies connected with both genotoxicity and cytotoxicity were evaluated. Results: The MN frequencies in cervical smear and peripheral blood lymphocytes were compared in patients with PCOS and normal controls. There was no statistically significant difference between the groups regarding micronucleus frequency in peripheral blood lymphocytes (p=0.239. The mean MN scores in exfoliated cervical cells of patients with PCOS and normal controls were 1.19±0.57 and 0.74±0.34, respectively. The difference regarding micronucleus frequencies in cervical cells was statistically significant between the groups (p=0.032. Conclusion: Although study group is small, our study results support that there is an increased micronucleus frequency in cervical exfoliated cells of PCOS patients; this is a determinant of genetic hazard in the disease.

  16. High throughput toxicity screening and intracellular detection of nanomaterials.

    Czech Academy of Sciences Publication Activity Database

    Collins, A. R.; Annangi, B.; Rubio, L.; Marcos, R.; Dorn, M.; Merker, C.; Estrela-Lopis, I.; Cimpan, M.R.; Ibrahim, M.; Cimpan, E.; Ostermann, M.; Sauter, A.; Yamani, N.E.; Shaposhnikov, S.; Chevillard, S.; Paget, V.; Grall, R.; Delic, J.; Goni-de-Cerio, F.; Suarez-Merino, B.; Fessard, V.; Hogeveen, K.N.; Fjellsbo, L.M.; Pran, E.R.; Brzicová, Táňa; Topinka, Jan; Silva, M.J.; Leite, P.E.; Ribeiro, A.R.; Granjeiro, J.M.; Grafström, R.; Prina-Mello, A.; Dusinská, M.

    2017-01-01

    Roč. 9, č. 1 (2017), e1413 ISSN 1939-5116 R&D Projects: GA MŠk(CZ) LD14002 Institutional support: RVO:68378041 Keywords : in-vitro micronucleus * bioelectrical-impedance analysis * bronchial epithelial-cells Subject RIV: DN - Health Impact of the Environment Quality OBOR OECD: Public and environmental health Impact factor: 4.761, year: 2016

  17. Aspects of nitrogen dioxide toxicity in environmental urban concentrations in human nasal epithelium

    International Nuclear Information System (INIS)

    Koehler, C.; Ginzkey, C.; Friehs, G.; Hackenberg, S.; Froelich, K.; Scherzed, A.; Burghartz, M.; Kessler, M.; Kleinsasser, N.

    2010-01-01

    Cytotoxicity and genotoxicity of nitrogen dioxide (NO 2 ) as part of urban exhaust pollution are widely discussed as potential hazards to human health. This study focuses on toxic effects of NO 2 in realistic environmental concentrations with respect to the current limit values in a human target tissue of volatile xenobiotics, the epithelium of the upper aerodigestive tract. Nasal epithelial cells of 10 patients were cultured as an air-liquid interface and exposed to 0.01 ppm NO 2 , 0.1 ppm NO 2 , 1 ppm NO 2 , 10 ppm NO 2 and synthetic air for half an hour. After exposure, genotoxicity was evaluated by the alkaline single-cell microgel electophoresis (Comet) assay and by induction of micronuclei in the micronucleus test. Depression of proliferation and cytotoxic effects were determined using the micronucleus assay and trypan blue exclusion assay, respectively. The experiments revealed genotoxic effects by DNA fragmentation starting at 0.01 ppm NO 2 in the Comet assay, but no micronucleus inductions, no changes in proliferation, no signs of necrosis or apoptosis in the micronucleus assay, nor did the trypan blue exclusion assay show any changes in viability. The present data reveal a possible genotoxicity of NO 2 in urban concentrations in a screening test. However, permanent DNA damage as indicated by the induction of micronuclei was not observed. Further research should elucidate the effects of prolonged exposure.

  18. Radiogenotoxicological effect of signal nuclide 134Cs on somatic and germ cells

    Institute of Scientific and Technical Information of China (English)

    ZhuShou-Peng; XiaFen; 等

    1997-01-01

    Chromosome at aberation rates in bone marrow cells and micronucleus formation in bone marrow polychromatic erythrocytes both rise with increase in radioactivities of 134Cs,and can be fitted to power functions of radioactivities of 134Cs.In spermatogonia 134Cs mainly induced chromatid breakage,and abnormalities in sperm can also be experessed as power functions of radioactivities of 134 Cs.

  19. 78 FR 66700 - Toxic Substances Control Act Chemical Testing; Receipt of Test Data

    Science.gov (United States)

    2013-11-06

    ... additive for food Rat--Up-and-Down processing, and as Procedure. ingredient in aluminum Micronucleus Test... Toxicity to Fish; Acute Toxicity to Daphnia; Toxicity to Algae; Acute Toxicity to Mammals; Bacterial..., cold Study in Zebra Fish set, and sheet-fed (Brachydanio rerio). applications. Acute Toxicity Study in...

  20. Further approaches to biological indicators of radiation injury

    International Nuclear Information System (INIS)

    Koeteles, G.J.; Kormos, C.; Kerekes, J.; Sztanyik, L.B.

    1988-01-01

    Despite of the decades-long investigations, the search for proper biological indicator of radiation injuries did not result in techniques fulfilling all the requirements. So far, the most reliable assay is the dicentric chromosome aberration analysis. New developments have been made recently on a cytogenetic technique, the micronucleus assay, and for local injuries on the application of thermography

  1. National food irradiation programme of Japan

    International Nuclear Information System (INIS)

    Fujimaki, M.

    1982-01-01

    The present state of studies on feasibility and wholesomeness of irradiated food is presented. Irradiation projects were realized of potatoes, onions, wheat, Vienna sausages, fish-paste products, and mandarine oranges. Mutagenecity tests with Salmonella or E. coli, chromosome aberration tests, dominant lethal tests, fibroblasts and micronucleus tests, and toxicity tests performed in amimals fed with irradiated food showed no positive results

  2. Biomarkers of exposition to ionizing radiation and hematology parameters in fitness for work. Case Report

    International Nuclear Information System (INIS)

    Djokovic, J.; Milacic, S.; Rakic, B.; Pajic, J.; Petrovic, D.; Vuckovic, J.

    2009-01-01

    Ionizing radiation is frequently used in medicine, especially during diagnostic procedures. The workers who are exposed to radiation have obligation for periodic check up. Presented case shows changes in hematological parameters and biomarkers of exposition to ionizing radiation (chromosome aberrations, structural changes and micronucleus test). The aim of this case report is to indicate metodology of diagnostic procedures for chronicle radiation syndrome. (author) [sr

  3. The radioprotective potential of 3,5,4'-trihydroxystilbene

    International Nuclear Information System (INIS)

    Clemente, Mary Judith Q.; Gomez, Marlyn O.

    1999-03-01

    The radioprotective potential of 3,5,4'trihydroxystilbene or resveratrol, a compound abundant in grapes, was investigated using the micronucleus test. Gamma radiation (6 Gy) was used to induce micronucleus formation in 12-week old Swiss-Webster mice. Five groups with five mice each were used. Three groups were given corresponding treatments (low, normal, high doses of reservatrol) via oral gavage for one week. The negative control group was not given any radiation nor any compound while the positive control group was exposed to radiation but was not given any compound. The mean micronucleus frequencies arranged from highest to lowest are as follows: low dose, positive control, normal dose, high dose and negative control. Using the analysis of variance-complete random design followed by the Duncan multiple range test, it was proven that resveratrol was able to inhibit micronucleus formation in polychromatic erythrocytes of 12-week old Swiss-Webster mice at the normal (60 micrograms) and high (120 micrograms) concentrations assigned. This suggests that its radioprotective potential may follow a dose-dependent pattern. (Author)

  4. Cytogenetic effects of radioiodine therapy: a 20-year follow-up study

    Energy Technology Data Exchange (ETDEWEB)

    Livingston, Gordon K. [Oak Ridge Institute for Science and Education, Radiation Emergency Assistance Center/Training Site, Oak Ridge, TN (United States); Khvostunov, Igor K. [Medical Radiological Research Center, Obninsk, Kaluga Region (Russian Federation); Gregoire, Eric [Institut de Radioprotection et de Surete Nucleaire, PRP-HOM/SRBE/LDB, BP 17, Fontenay aux roses Cedex (France); Barquinero, Joan-Francesc [Universtitat Autonoma de Barcelona, Facultat de Biociencies, Cerdanyola del Valles (Spain); Shi, Lin; Tashiro, Satoshi [Hiroshima University, Department of Cellular Biology, Research Institute for Radiation Biology and Medicine, Hiroshima (Japan)

    2016-05-15

    The purpose of this study was to compare cytogenetic data in a patient before and after treatment with radioiodine to evaluate the assays in the context of biological dosimetry. We studied a 34-year-old male patient who underwent a total thyroidectomy followed by ablation therapy with {sup 131}I (19.28 GBq) for a papillary thyroid carcinoma. The patient provided blood samples before treatment and then serial samples at monthly intervals during the first year period and quarterly intervals for 5 years and finally 20 years after treatment. A micronucleus assay, dicentric assay, FISH method and G-banding were used to detect and measure DNA damage in circulating peripheral blood lymphocytes of the patient. The results showed that radiation-induced cytogenetic effects persisted for many years after treatment as shown by elevated micronuclei and chromosome aberrations as a result of exposure to {sup 131}I. At 5 years after treatment, the micronucleus count was tenfold higher than the pre-exposure frequency. Shortly after the treatment, micronucleus counts produced a dose estimate of 0.47 ± 0.09 Gy. The dose to the patient evaluated retrospectively using FISH-measured translocations was 0.70 ± 0.16 Gy. Overall, our results show that the micronucleus assay is a retrospective biomarker of low-dose radiation exposure. However, this method is not able to determine local dose to the target tissue which in this case was any residual thyroid cells plus metastases of thyroidal origin. (orig.)

  5. PROFILE OF Nauclea diderrichii LEAF EXTRACTS

    African Journals Online (AJOL)

    pc

    Méo, M.P., (2011). Genotoxic and clastogenic activity of saponins extracted from Nauclea bark as assessed by the micronucleus and the comet assays in Chinese hamster ovary cells. Journal of Ethnopharmacology. 137: 176–183. Mustofa, Valentin, A., Benoit-Vical, F., Pélissier, Y., Koné-. Bamba, D. &Mallié, M., (2000).

  6. 75 FR 5546 - Proposed Significant New Use Rule for Multi-walled Carbon Nanotubes

    Science.gov (United States)

    2010-02-03

    ... effects of the PMN substance: A bacterial reverse mutation test (OPPTS Harmonized Test Guideline 870.5100) in vitro; a mammalian erythrocyte micronucleus test (OPPTS Harmonized Test Guideline 870.5395 or Organisation for Economic Co-operation and Development (OECD) 474 test guideline) in vivo, in bone marrow, by...

  7. Effect of Treatment Media on the Agglomeration of Titanium Dioxide Nanoparticles: Impact on Genotoxicity, Cellular Interaction, and Cell Cycle

    Science.gov (United States)

    ABSTRACT The widespread use of titanium dioxide (TiO2) nanoparticles in consumer products increases the probability of exposure to humans and the environment. Although TiO2 nanoparticles have been shown to induce DNA damage (comet assay) and chromosome damage (micronucleus ass...

  8. Micronuclei, DNA single-strand breaks and DNA-repair activity in mice exposed to 1,3-butadiene by inhalation

    Czech Academy of Sciences Publication Activity Database

    Vodička, Pavel; Štětina, R.; Šmerák, P.; Vodičková, Ludmila; Naccarati, Alessio; Bárta, I.; Hemminki, K.

    2006-01-01

    Roč. 608, - (2006), s. 49-57 ISSN 1383-5718 R&D Projects: GA ČR(CZ) GA310/01/0802 Institutional research plan: CEZ:AV0Z50390512 Keywords : Single-strand DNA breaks * Micronucleus formation * DNA-repair activity Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.122, year: 2006

  9. The relative biological effectiveness of a high energy neutron beam for micronuclei induction in T-lymphocytes of different individuals

    Energy Technology Data Exchange (ETDEWEB)

    Slabbert, J.P., E-mail: jps@tlabs.ac.z [NRF iThemba LABS (Laboratory for Accelerated Based Sciences), Somerset West (South Africa); Dept. of Medical Imaging and Clinical Oncology, University of Stellenbosch (South Africa); August, L. [NRF iThemba LABS (Laboratory for Accelerated Based Sciences), Somerset West (South Africa); Vral, A. [Dept. of Basic Medical Sciences, Ghent University (Belgium); Symons, J. [NRF iThemba LABS (Laboratory for Accelerated Based Sciences), Somerset West (South Africa)

    2010-12-15

    In assessing the radiation risk of personnel exposed to cosmic radiation fields as it pertains to radiological damage during travel in civilian aircrafts, it is particularly important to know the relative biological effectiveness (RBE) for high energy neutrons. It has been the subject of numerous investigations in recent years using different neutron energies and cytogenetic examinations. Variations in the radiosensitivity of white blood cells for different individuals are likely to influence the estimate of the relative biological effectiveness for high energy neutrons. This as such observations have been noted in the response of different cancer cell lines with varying inherent sensitivities. In this work the radiosensitivities of T-lymphocytes of different individuals to the p(66)/Be neutron beam at iThemba LABS were measured using micronuclei formations and compared to that noted following exposure to {sup 60}Co {gamma}-rays. The principle objective of this investigation was to establish if a relationship between neutron RBE and variation in biological response to {sup 60}Co {gamma}-rays for lymphocytes from different individuals could be determined. Peripheral blood samples were collected from four healthy donors and isolated lymphocytes were exposed to different doses of {sup 60}Co {gamma}-rays (1-5 Gy) and p(66)/Be neutrons (0.5-2.5 Gy). One sample per donor was not exposed to radiation and served as a control. Lymphocytes were stimulated using PHA and cultured to induce micronuclei in cytokinesis-blocked cells. Micronuclei yields were numerated using fluorescent microscopy. Radiosensitivities and RBE values were calculated from the fitted parameters describing the micronuclei frequency dose response data. Dissimilar dose response curves for different donors were observed reflecting varying inherent sensitivities to both neutron and gamma radiation. A clear reduction in the dose limiting RBE{sub M} is noted for donors with lymphocytes more sensitive to

  10. Wound healing morbidity in STS patients treated with preoperative radiotherapy in relation to in vitro skin fibroblast radiosensitivity, proliferative capacity and TGF-β activity

    International Nuclear Information System (INIS)

    Akudugu, John M.; Bell, Robert S.; Catton, Charles; Davis, Aileen M.; Griffin, Anthony M.; O'Sullivan, Brian; Waldron, John N.; Ferguson, Peter C.; Wunder, Jay S.; Hill, Richard P.

    2006-01-01

    Background and purpose: In a recent study, we demonstrated that the ability of dermal fibroblasts, obtained from soft tissue sarcoma (STS) patients, to undergo initial division in vitro following radiation exposure correlated with the development of wound healing morbidity in the patients following their treatment with preoperative radiotherapy. Transforming growth factor beta (TGF-β) is thought to play an important role in fibroblast proliferation and radiosensitivity both of which may impact on wound healing. Thus, in this study we examined the interrelationship between TGF-β activity, radiosensitivity and proliferation of cultured fibroblasts and the wound healing response of STS patients after preoperative radiotherapy to provide a validation cohort for our previous study and to investigate mechanisms. Patients and methods: Skin fibroblasts were established from skin biopsies of 46 STS patients. The treatment group consisted of 28 patients who received preoperative radiotherapy. Eighteen patients constituted a control group who were either irradiated postoperatively or did not receive radiation treatment. Fibroblast cultures were subjected to the colony forming and cytokinesis-blocked binucleation assays (low dose rate: ∼0.02 Gy/min) and TGF-β assays (high dose-rate: ∼1.06 Gy/min) following γ-irradiation. Fibroblast radiosensitivity and initial proliferative ability were represented by the surviving fraction at 2.4 Gy (SF 2.4 ) and binucleation index (BNI), respectively. Active and total TGF-β levels in fibroblast cultures were determined using a biological assay. Wound healing complication (WHC), defined as the requirement for further surgery or prolonged deep wound packing, was the clinical endpoint examined. Results: Of the 28 patients treated with preoperative radiotherapy, 8 (29%) had wound healing difficulties. Fibroblasts from patients who developed WHC showed a trend to retain a significantly higher initial proliferative ability after

  11. HL60 human premyelocitic cell line as a model system for bystander response

    International Nuclear Information System (INIS)

    Dini, V.; Tabocchini, M.A.; Belli, M.; Simone, G.; Di Carlo, B.; Sapora, O.; Superiore di Sanita, Rome

    2007-01-01

    Complete text of publication follows. Objective: to evaluate HL60 human premyelocitic cell line as a model system to study bystander response. Methods: HL60 cell line, isolated from the blood of a patient affected by premyelocitic leukemia, has 45-46 chromosomes with abnormalities mainly on chromosomes 5, 8 and X and can undergo chemical-induced in vitro differentiation. Differentiation gives rise to granulocytes, monocytes or macrophages depending on the drug used. We define as proliferative (AP) cells those in log phase of growth with less than 10 passages from thawing and as differentiated (D) cells those treated with 10 nM TPA (phorbol ester) for 72 hours. Phorbol ester treatment induces differentiation to monocytes and macrophages. Differentiation has been evaluated through the expression of differentiation cluster membrane antigens (CD95, CD9 and CD14). Results: AP cells resulted positive for CD95 and negative for CD9 and CD14, while D cells resulted positive for CD9 and negative for CD95 and CD14. Our data on AP and D cells showed that: (i) the level of intracellular reactive oxygen and nitrogen species (ROS and RNS) is lower in D cells compared to AP cells; (ii) radiation induced DNA damage (single and double strand breaks, SSB and DSB, as measured with the comet assay technique) is lower in D cells than in AP cells. This different radiosensitivity can be related to the higher degree of compactness of nuclear structure in D cells. Radiation induced bystander effect (BE) was analyzed with the medium transfer technique. The medium from irradiated, with 0.5 Gy of γ-rays, AP cells was collected after 0, 2, 4 and 24 hours from irradiation and added to non irradiated log phase cells. The frequency of micronuclei formation in bystander cells was measured by using the cytokinesis block technique by adding cytochalasin B to the non irradiated culture together with the irradiated medium. Preliminary data indicate about 1.4-fold increase in micronuclei formation in

  12. Effect of n-Butanol on Chromosomal Damage in Mice Bone Marrow Cells

    Directory of Open Access Journals (Sweden)

    Nahid Mansouri

    2016-07-01

    Full Text Available Background: n-Butanol is a four-carbon alcohol used widely in foods, cosmetics industries, biology and chemistry research laboratories, and other fields. Long time-effects of inhalation or consumption of small amounts of Butanol on human health are still unknown. On the other hand, numerous reports about the development of n-Butanol toxicity are available. The main objective of the study was to investigate the effects of inhaled and oral administration of n-Butanol as a long-term in vivo investigation.Materials and Methods: Small white laboratory, male mice (20-30 g were used in 11 groups (n=4 including experimental 1 to 6, 1 to 4 control "A” and positive control groups. Experimental groups 1-3, for 10, 20, and 40 days; 5 hours a day were inside a box with ventilation facilities exposed to air saturated with n-Butanol vapor. Experimental groups 4 to 6, received water containing n-Butanol 0.2%, 1% and 5% for 10 days. Control groups B, 1 to 3 was placed for 10, 20, and 40 days inside a similar box exposed to normal air, respectively. Control group B 4 received water without any particular substance for 10 days. The positive control group received 30µl subcutaneous vinblastine. Bone marrow cells were extracted 24 hours after treatments and stained by May-Grünwald-Giemsa staining and the number of micronucleus was counted. Vinblastine, as a positive control, increased an average of micronucleus numbers significantly compared to control group (P<0.001.Results: n-Butanol inhalation caused no significant difference in 1-3 experimental groups in the average numbers of micronucleus compared to control group, even in the 40 days treatment group, average numbers of micronucleus was decreased comparing to control group (P<0.05. Also, oral administration of 0.2% and 1% n-Butanol had no effect on the average micronucleus numbers compared to the control group, while oral administration of 5% n-Butanol caused even decrease in average numbers of micronucleus

  13. Correlation between the genotoxicity endpoints measured by two different genotoxicity assays: comet assay and CBMN assay

    Directory of Open Access Journals (Sweden)

    Carina Ladeira

    2015-06-01

    The results concerning of positive findings by micronuclei and non significant ones by comet assay, are corroborated by Deng et al. (2005 study performed in workers occupationally exposed to methotrexate, also a cytostatic drug. According to Cavallo et al. (2009, the comet assay seems to be more suitable for the prompt evaluation of the genotoxic effects, for instance, of polycyclic aromatic hydrocarbons mixtures containing volatile substances, whereas the micronucleus test seems more appropriate to evaluate the effects of exposure to antineoplastic agents. However, there are studies that observed an increase in both the comet assay and the micronucleus test in nurses handling antineoplastic drugs, although statistical significance was only seen in the comet assay, quite the opposite of our results (Maluf & Erdtmann, 2000; Laffon et al. 2005.

  14. Frequency of micronuclei in hepatocytes following X and fast-neutron irradiations--an analysis by a linear-quadratic model

    International Nuclear Information System (INIS)

    Ono, K.; Nagata, Y.; Akuta, K.; Abe, M.; Ando, K.; Koike, S.

    1990-01-01

    The usefulness of the micronucleus assay for investigating the radiation response of hepatocytes was examined. The frequency was defined as the ratio of the total number of micronuclei to the number of hepatocytes examined. The dose-response curves were curvilinear after X rays and linear after neutrons. These dose-response curves were analyzed by a linear-quadratic model, frequency = aD + bD2 + c. The a/b ratio was 3.03 +/- 1.26 Gy following X irradiation. This value is within the range of the alpha/beta ratios reported by others using the clonogenic assay of hepatocytes. While the a/b value for neutrons was 24.3 +/- 11.7 Gy, the maximum relative biological effectiveness of neutrons was 6.30 +/- 2.53. Since the micronucleus assay is simple and rapid, it may be a good tool for evaluating the radiation response of hepatocytes in vivo

  15. DNA damage and cellular death in oral mucosa cells of children who have undergone panoramic dental radiography

    International Nuclear Information System (INIS)

    Angelieri, Fernanda; Oliveira, Gabriela R. de; Sannomiya, Eduardo K.; Ribeiro, Daniel A.

    2007-01-01

    Despite wide use as a diagnostic tool in medical and dental practice, radiography can induce cytotoxic effects and genetic damage. To evaluate DNA damage (micronucleus) and cellular death (pyknosis, karyolysis and karyorrhexis) in exfoliated buccal mucosa cells taken from healthy children following exposure to radiation during dental radiography. A total of 17 children who had undergone panoramic dental radiography were included. We found no statistically significant differences (P > 0.05) between micronucleated oral mucosa cells in children before and after exposure to radiation. On the other hand, radiation did cause other nuclear alterations closely related to cytotoxicity including karyorrhexis, pyknosis and karyolysis. Taken together, these results indicate that panoramic dental radiography might not induce chromosomal damage, but may be cytotoxic. Overall, the results reinforce the importance of evaluating the health side effects of radiography and contribute to the micronucleus database, which will improve our understanding and practice of this methodology in children. (orig.)

  16. Aspects of the usefulness of five marine pollution biomarkers, with emphasis on MN and lipid content

    International Nuclear Information System (INIS)

    Koukouzika, N.; Dimitriadis, V.K.

    2008-01-01

    By using cellular biomarkers, such as the formation of the micronucleus, the morphometric alterations of lipids and the morphometric changes in the lysosomal system, we investigated effects of experimental exposure to phenanthrene, Cu, Cd, and Hg for 15 days in mussels. Concerning micronuclei, the evaluation of the total nuclear abnormalities, instead of the micronucleus only, as a biomarker of marine pollution, indicated more statistically significant differences between the control and the pollutant treated groups. Contrary to the existing knowledge showing that there is an increase in lipid content after pollutant exposure, our results showed that there was a decrease in the amount of lipid, as well as an increase in the number of neutral lipids. Furthermore, although prior studies found that fewer lysosomes formed after pollutant exposure, this was not confirmed by our work, as pollutant treated animals exhibited a decrease in the volume and an increase in the numerical density of lysosomes, compared to control groups

  17. The effect of γ-radiation on smoked fish using short-term mutagenicity assays

    International Nuclear Information System (INIS)

    Dela Rosa, A.M.; Banzon, R.B.

    1989-01-01

    The effect of γ-radiation on the mutagenicity potential of wood-smoked fish was investigated. Smoked fish were irradiated with radiation doses of 2.0, 4.0, 6.0 and 8.0 kGy. The DMSO extracts of non-radiated and irradiated smoked fish were tested for mutagenicity using the Ames plate incorporation assay, host-mediated assay, and the micronucleus test. It was observed that γ-irradiation did not induce any significant increase in the number of revertants of TA98, TA100 and TA104 as compared with the non-radiated smoked fish. Results of the host-mediated assay and the micronucleus test showed no difference in the mutagenic response of non-radiated in irradiated smoked fish. The results indicate thet γ-radiation does not introduce mutagens in smoked fish. (author). 17 refs.; 6 tabs

  18. DNA damage and cellular death in oral mucosa cells of children who have undergone panoramic dental radiography

    Energy Technology Data Exchange (ETDEWEB)

    Angelieri, Fernanda; Oliveira, Gabriela R. de [Sao Paulo Metodista University (UMESP), Department of Orthodontics, Sao Bernardo do Campo, Sao Paulo (Brazil); Sannomiya, Eduardo K. [Sao Paulo Metodista University (UMESP), Department of Dento-Maxillofacial Radiology, Sao Bernardo do Campo, Sao Paulo (Brazil); Ribeiro, Daniel A. [Federal University of Sao Paulo (UNIFESP), Department of Health Sciences, Santos, Sao Paulo (Brazil); Universidade Federal de Sao Paulo (UNIFESP), Departamento de Ciencias da Saude, Santos, Sao Paulo (Brazil)

    2007-06-15

    Despite wide use as a diagnostic tool in medical and dental practice, radiography can induce cytotoxic effects and genetic damage. To evaluate DNA damage (micronucleus) and cellular death (pyknosis, karyolysis and karyorrhexis) in exfoliated buccal mucosa cells taken from healthy children following exposure to radiation during dental radiography. A total of 17 children who had undergone panoramic dental radiography were included. We found no statistically significant differences (P > 0.05) between micronucleated oral mucosa cells in children before and after exposure to radiation. On the other hand, radiation did cause other nuclear alterations closely related to cytotoxicity including karyorrhexis, pyknosis and karyolysis. Taken together, these results indicate that panoramic dental radiography might not induce chromosomal damage, but may be cytotoxic. Overall, the results reinforce the importance of evaluating the health side effects of radiography and contribute to the micronucleus database, which will improve our understanding and practice of this methodology in children. (orig.)

  19. Micronuclei rate in workers of Novi Han radioactive waste repository

    International Nuclear Information System (INIS)

    Atamasova, P.; Hadjidekova, V.

    2003-01-01

    A cytogenetic study has been carried out by the micronucleus test of lymphocytes from the peripheral blood of 30 workers from the Novi Han radioactive waste repository. The results are compared to the results of a control group of 6 persons from the administrative staff, and to outside group of 39 healthy persons. All persons are questioned through a special questionnaire about their occupational, medical, and social status. The rate of the cells with micronuclei and the total number of the micronuclei are analysed in the peripheral blood lymphocytes using the cytogenesis-block micronucleus test. The comparison of the results does not show an increase of the lymphocytes with micronuclei in the workers

  20. Bystander effects on mammalian cells induced by carbon ions

    International Nuclear Information System (INIS)

    Wang Jufang; Zhao Jing; Ma Qiufeng; Chinese Academy of Sciences, Beijing; Li Weijian; Zhou Guangming; Dang Bingrong; Mao Limin; Feng Yan

    2004-01-01

    Bystander effects on unirradiated V79 cells were observed by irradiated conditioned medium (ICM) method and co-cultured with carbon-ion-irradiated V79 cells. The results showed that the colony formation efficiency of unirradiated cells is obviously decreased by ICM. After co-culture with carbon-ion-irradiated cells for some time, the colony formation efficiency of co-cultured cells was lower than expected results assuming no bystander effects. The micronucleus frequency and hprt gene mutation rate was almost the same as expected results. Cytotoxic factor(s), which was effective for cell growth but not for micronucleus and mutation on unirradiated cells, might be released by irradiated cells. (authors)

  1. The cytogenetic effects of black tea and green tea on cultured human lymphocytes

    Directory of Open Access Journals (Sweden)

    Halil Erhan Eroğlu

    2011-12-01

    Full Text Available In this study, the cytogenetic effects of black tea and green tea were determined in cultured peripheral blood lymphocytes. Results showed that black tea and green tea induced the mitotic and replication indexes and decreased micronuclei. But these data were not statistically significant for green tea. The effects of black tea on the micronucleus formation and mitotic index were statistically significant. The decrease in micronucleus counts indicated that black tea and green tea had considerable anticlastogenic and antigenotoxic effects as observed in vitro in human lymphocytes. Thus, it could be concluded that tea polyphenols protected the normal cells from genotoxic or carcinogenic agents, which indicated the therapeutic and antioxidative role of catechins, flavonoids or other tea compounds.

  2. Safety and toxicology assessment of chicken breast for high-dose irradiation

    International Nuclear Information System (INIS)

    Zhu Jiating; Feng Min; Yan Jianmin; Yang Ping; Wang Dening; Gao Meixu; Ha Yiming; Liu Chunquan

    2011-01-01

    Feeding wholesomeness tests of irradiated chicken breast were studied by using acute oral toxicology, Ames, micronucleus of born marrow cell, sperm shape abnormality in mice and 30 d feeding test. The LD 50 of all the rats and mice were more than 10 g/kg · BW, which means that the pet foods belonged to actually non-toxic grade; ames test, and the tests of micronucleus of born marrow cell, sampan shape abnormality in mice were all negative results; 30 d feeding test in rats demonstrated that it had no distinctive effects on routine blood, body weight and biochemical index. It is concluded that pet foods irradiated up to 25 kGy high dose were no safety and toxicology problems. (authors)

  3. EFSA CEF Panel (EFSA Panel on Food Contact Materials, Enzymes, Flavourings and Processing Aids), 2015. Scientific Opinion on Flavouring Group Evaluation 208 Revision 1 (FGE.208Rev1): Consideration of genotoxicity data on representatives for 10 alicyclic aldehydes with the a,b-unsaturation in ring / side-chain and precursors from chemical subgroup 2.2 of FGE.19

    DEFF Research Database (Denmark)

    Beltoft, Vibe Meister; Nørby, Karin Kristiane

    genotoxicity studies on p-mentha-1,8-dien-7-al [FL-no: 05.117], the representative substance for FGE.19 subgroup 2.2. This substance was tested in vivo in a combined micronucleus assay in bone marrow and Comet assay in liver and duodenum. It did not induce any increase in micronucleated polychromatic...... erythrocytes of the bone marrow of male rats in the micronucleus test and it did not induce DNA damage in duodenum of the same animals as analysed by the Comet assay. The Comet assay performed in liver shows a positive result and therefore the Panel concluded that p-mentha-1,8-dien-7-al [FL-no: 05...

  4. Micronuclei in lymphocytes from currently active uranium miners

    International Nuclear Information System (INIS)

    Zoelzer, Friedo; Freitinger Skalicka, Zuzana; Havrankova, Renata; Hon, Zdenek; Rosina, Jozef; Navratil, Leos; Skopek, Jiri

    2012-01-01

    Micronuclei can be used as markers of past radiation exposure, but only few studies have dealt with uranium miners. In this paper, we report on micronuclei in lymphocytes from individuals currently working at Rozna, Czech Republic, the last functioning uranium mine in the European Union. A modified micronucleus-centromere test was applied to assess the occurrence of micronuclei in stimulated lymphocytes, as well as their content in terms of whole chromosomes or fragments. Compared with unexposed individuals, the miners had higher frequencies of micronucleus-containing lymphocytes and higher percentages of micronuclei without centromeres, and the differences were significant for both parameters (0.74 ± 0.60 vs. 0.50 ± 0.42, p = 0.017 and 49 ± 44 vs. 12 ± 21, p = 0.0002; means ± standard deviations). There were also significant correlations between one or other of these parameters on the one hand and various dose values on the other, in particular with a 'retrievable' dose, that is, a dose whose effect should still be recognisable in lymphocytes assuming a half-life of 3 years. The 'retrievable' dose at which a doubling of the micronucleus frequency was observed was around 35 mSv, corresponding to a total dose of 90 mSv received while working in the mines. Altogether, our data show that the micronucleus-centromere test is a valuable tool for the assessment of past radiation exposure in uranium miners. The scatter in the data is of course far too great to allow individual dosimetry, but for groups of a few dozen exposed individuals, the method can be used to monitor doses clearly below 100 mSv. (orig.)