2.Mutagenic and Estrogenic Effects of Organic Compounds in Water Treated by Different Processes: A Pilot Study.
Yin LU ; Xue Min LYU ; San Hua XIAO ; Xiao Ming YANG ; Ya Zhou WANG ; Fei TANG
Biomedical and Environmental Sciences 2015;28(8):571-581
OBJECTIVEIn this study, a pilot-scale investigation was conducted to examine and compare the biotoxicity of the organic compounds in effluents from five treatment processes (P1-P5) where each process was combination of preoxidation (O3), coagulation, sedimentation, sand filtration, ozonation, granular activated carbon, biological activated carbon and chlorination (NaClO).
METHODSOrganic compounds were extracted by XAD-2 resins and eluted with acetone and dichlormethane (DCM). The eluents were evaporated and redissolved with DMSO or DCM. The mutagenicity and estrogenicity of the extracts were assayed with the Ames test and yeast estrogen screen (YES assay), respectively. The organic compounds were detected by GC-MS.
RESULTSThe results indicated that the mutation ratio (MR) of organic compounds in source water was higher than that for treated water. GC-MS showed that more than 48 organic compounds were identified in all samples and that treated water had significantly fewer types and concentrations of organic compounds than source water.
CONCLUSIONTo different extents, all water treatment processes could reduce both the mutagenicity and estrogenicity, relative to source water. P2, P3, and P5 reduced mutagenicity more effectively, while P1 reduced estrogenicity, most effectively. Water treatment processes in this pilot plant had weak abilities to remove Di-n-butyl phthalate or 1, 2-Benzene dicarboxylic acid.
Estrogens ; toxicity ; Gas Chromatography-Mass Spectrometry ; Mutagens ; toxicity ; Organic Chemicals ; toxicity ; Pilot Projects ; Water Pollutants, Chemical ; toxicity ; Water Purification ; methods
3.Scanning aneugen and clastogen by micronuclei analysis using flow cytometry.
Ming-Jie YANG ; Jian-Chang ZHOU ; Zhi LI ; Xing-Fen YANG ; Jun-Ming HUANG ; Xiao-Hua TAN ; Jia CAO ; Rui-Ping ZENG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2006;24(11):649-652
OBJECTIVETo explore a flow cytometry (FCM)-based method for discriminating aneugen- or clastogen-induced micronuclei.
METHODSCells were stained with anti-CD71-FITC and PI, and the PI fluorescent signal intensity of micronucleated reticulocyte (MN-RET) in the peripheral blood of NIH mouse treated with COL or CP was detected by flow cytometry.
RESULTSThe ratio of the median of the intensity of MN-RET fluorescent signals to that of nucleated cell was low in the cyclophosphamide treated mouse, while the median was high in the colchicine treated mouse.
CONCLUSIONThe flow cytometry-based micronucleus assay can be used to discriminate primarily smaller MN induced by the clastogen exposure from the larger MN induced by an aneugen.
Animals ; Colchicine ; toxicity ; Cyclophosphamide ; toxicity ; Flow Cytometry ; methods ; Male ; Mice ; Micronuclei, Chromosome-Defective ; Mutagens ; toxicity ; Reticulocytes ; drug effects ; ultrastructure
4.Genotoxicity study of bone cement.
Yanping HU ; Jie SONG ; Rui MA ; Xin WANG ; Jun GUO ; Xue WANG
Chinese Journal of Medical Instrumentation 2011;35(6):425-445
OBJECTIVETo investigate the genotoxicity of bone cement.
MATERIAL AND METHODSTest article was mixed of liquid and the powder extract of bone cement. Using bacterial reverse mutation test (Ames test) and mouse lymphoma Assay (MLA) with and without metabolic activation S9. Ames test was performed by the plate incorporation method for its ability to induce reverse mutations in three treatment dosage groups: 25 microl/plate, 50 microl/plate, 100 microl/plate using S. typhimurium TA98, TA100, TA1535, TA1537 and E. coli WP2 uvrA; In MLA, the mutation frequency (MF) and the percentage of small colony mutants (SC%) of L5178 tk(+/-) cells induced by bone cement at final concentration of 0.125%, 0.25% and 0.5% (V/V) were calculated and assayed with +S9/3 h, - S9/3 h, -S9/24 h.
RESULTSSlight precipitations of two dosage group 50 microl/plate and 100 microl/plate were observed after phosphoric acid buffer solution or S9mix added. Growth inhibition effect exists in TA100, TA1535 and WP2 uvrA to varying degrees. Other groups did not cause obvious increases in the mean number of revertant per plate compared with negative control (DMSO). The result of MLA indicated no significant MF or SC% increases (P>0.05) observed comparing with negative control under all test conditions.
CONCLUSIONSThe bone cement did not induce reverse mutation at his(-)/trp(-) and there was no obvious damage effect to gene tk(+/-) or chromosome under this study condition.
Animals ; Bone Cements ; toxicity ; Cell Line, Tumor ; DNA Damage ; Mice ; Micronucleus Tests ; Mutagens ; toxicity ; Mutation ; Mutation Rate ; Salmonella
5.Protective effect of epigallocatechin gallate against sperm abnormality in mice.
Liu-Cai SUI ; Yi-Feng GE ; Juan-Juan XU ; Rong-Hua WU ; Hai-Yan FU ; Bing YAO
National Journal of Andrology 2014;20(12):1068-1072
OBJECTIVETo investigate the protective effect of epigallocatechin gallate (EGCG) on mouse sperm in vivo.
METHODSA total of 64 six-week-old male Kuming mice were randomly divided into eight groups of equal number to be treated with normal saline (negative control), Cyclophosphamide (CP) at 30 mg/kg (positive control), and CP followed by EGCG (experimental) at 20, 40, and 80 mg/kg, respectively, given every other day for 10 days. At 4 and 5 weeks after treatment, the bilateral testes of the mice were harvested for examination of sperm abnormality.
RESULTSEGCG did not increase the rate of CP-induced sperm abnormality in the mice, but reduced it instead with the prolonged time of treatment.
CONCLUSIONEGCG protects mouse sperm in vivo.
Animals ; Catechin ; analogs & derivatives ; pharmacology ; Cyclophosphamide ; toxicity ; Male ; Mice ; Mutagens ; toxicity ; Random Allocation ; Spermatozoa ; drug effects ; Time Factors
6.Mutagen sensitivity in peripheral blood lymphocytes among coke-oven workers.
Juan CHENG ; Shu-guang LENG ; Hai-shan LI ; Yong NIU ; Yao-feng SUN ; Hua-wei DUAN ; Ping BIN ; Lin-yuan ZHANG ; Xue-miao LIANG ; Han LIN ; Zhong-xu WANG ; Yu-fei DAI ; Bin LI ; Yu-xin ZHENG
Chinese Journal of Preventive Medicine 2008;42(5):307-311
OBJECTIVETo investigate the sensitivity to bleomycin (BLM) in peripheral blood lymphocytes (PBL) among coke-oven workers.
METHODSNinty-four coke-oven workers with exposure to a high level of polycyclic aromatic hydrocarbons and 64 non-coke-oven workers (control) were recruited into this study. PBL was challenged by 8 microg/ml BLM, a known carcinogen, to induce certain amount of DNA damage, the difference of olive tail moment (TM) measured by comet assay before and after BLM treatment reflected the sensitivity towards mutagens.
RESULTSThe distribution of age, sex, and prevalence of smoking and drinking were not significantly different between these two groups. The geometric mean of urinary 1-hydroxypyrene (1-OHP) was significantly higher in coke-oven workers than in controls (9.0 versus 1.5 microg/L, t = -9.317, P < 0.01). The coke-oven workers showed significantly higher sensitivity to BLM than controls (17.7 versus 14.9, t = -2.583, P = 0.01). A large inter-group difference in sensitivity to BLM was observed in both controls and coke-oven workers. Stratification analysis revealed the significant association between high 1-OHP level (> 9.0 microg/L) and increased sensitivity to BLM (F = 4.001, P = 0.05) among coke-oven workers. Smoking subjects showed a significant higher value of sensitivity than nonsmokers in controls but not in coke-oven workers. No significant difference was observed between age, drinking status, coking history or external exposure class and BLM sensitivity.
CONCLUSIONExposure to coke oven emission could increase the sensitivity to mutagens, which might be a reason of high incidence of lung cancer among coke-oven workers.
Adult ; Benzo(a)pyrene ; toxicity ; Bleomycin ; toxicity ; Coke ; Comet Assay ; DNA Damage ; DNA Repair ; Female ; Humans ; Lymphocytes ; drug effects ; Male ; Middle Aged ; Mutagens ; toxicity ; Occupational Exposure
7.Relationship between urinary 1-hydroxypyrene level and peripheral blood lymphocyte chromosomal damage among coke oven workers.
Wei-jun GUAN ; Nan LIU ; Shu-lian PANG ; Xiao QI ; Guo-hui XU ; Ying-li LIU ; Qian WANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2010;28(8):561-564
OBJECTIVETo investigate the relationship between the urinary 1-hydroxypyrene level and cytokinesis-block micronucleus in peripheral blood lymphocyte in coke oven workers.
METHODSOne hundred and fifty-eight workers from a coke plant and 158 referents without occupational PAHs exposure were recruited in this study. Urnary level of 1-hydroxypyrene was measured by alkaline hydrolysis combined with high performance liquid chromatography as an internal exposure dose, and the chromosomal damage of peripheral blood lymphocyte were evaluated with cytokinesis-block micronucleus (CBMN) method. Personal information including occupational history, age, sex, smoking and alcohol drinking, was collected by questionnaire.
RESULTSThe lymphocyte chromosomal damage level expressed as frequency of CBMN in coke oven workers was significantly higher than that of controls (3.32 ± 2.90 vs 0.57 ± 0.88, P < 0.01) after adjusting for sex, age, smoking and alcohol drinking, and correlation between urinary 1-hydroxypyrene concentrations and frequency of CBMN was found (Spearman Partial correlation coefficient = 0.28, P < 0.05) in coke oven workers. Three hundreds and sixteen subjects were divided into three groups by their urine 1-hydroxypyrene level (expressed as 0.11 ∼ 0.70, 0.71 ∼ 4.09 and 4.10 ∼ 24.74 µmol/mol Cr). After adjusting for age, sex, smoking and alcohol drinking by multiple nonparametric analysis of covariance, the frequency of CBMN in the groups of 0.71 ∼ 4.09 and 4.10 ∼ 24.74 µmol/mol C were 1.89 ± 2.37 and 3.29 ± 2.36, significantly higher than that in the group of 0 ∼ 0.70 µmol/mol Cr (0.56 ± 0.89).
CONCLUSIONSUnder present PAHs exposure levels, the Cytokinesis-block micronucleus test could detect PAHs-induced genotoxicity in coke oven workers.
Adult ; Case-Control Studies ; Coke ; toxicity ; Cytokinesis ; DNA Damage ; Female ; Humans ; Lymphocytes ; cytology ; Male ; Micronucleus Tests ; Middle Aged ; Mutagens ; toxicity ; Occupational Exposure ; Pyrenes ; toxicity ; Urinalysis ; Urine ; chemistry
10.Endonuclease modified comet assay for oxidative DNA damage induced by detection of genetic toxicants.
Jian ZHAO ; Hongli LI ; Qingfeng ZHAI ; Yugang QIU ; Yong NIU ; Yufei DAI ; Yuxin ZHENG ; Huawei DUAN
Chinese Journal of Preventive Medicine 2014;48(3):208-212
OBJECTIVEThe aim of this study was to investigate the use of the lesion-specific endonucleases-modified comet assay for analysis of DNA oxidation in cell lines.
METHODSDNA breaks and oxidative damage were evaluated by normal alkaline and formamidopyrimidine-DNA-glycosylase (FPG) modified comet assays. Cytotoxicity were assessed by MTT method. The human bronchial epithelial cell (16HBE) were treated with benzo (a) pyrene (B(a)P), methyl methanesulfonate (MMS), colchicine (COL) and vincristine (VCR) respectively, and the dose is 20 µmol/L, 25 mg/ml, 5 mg/L and 0.5 mg/L for 24 h, respectively. Oxidative damage was also detected by levels of reactive oxygen species in treated cells.
RESULTSFour genotoxicants give higher cytotoxicity and no significant changes on parameters of comet assay treated by enzyme buffer. Cell survival rate were (59.69 ± 2.60) %, (54.33 ± 2.81) %, (53.11 ± 4.00) %, (51.43 ± 3.92) % in four groups, respectively. There was the direct DNA damage induced by test genotoxicants presented by tail length, Olive tail moment (TM) and tail DNA (%) in the comet assay. The presence of FPG in the assays increased DNA migration in treated groups when compared to those without it, and the difference was statistically significant which indicated that the clastogen and aneugen could induce oxidative damage in DNA strand. In the three parameters, the Olive TM was changed most obviously after genotoxicants treatment. In the contrast group, the Olive TM of B(a) P,MMS, COL,VCR in the contrast groups were 22.99 ± 17.33, 31.65 ± 18.86, 19.86 ± 9.56 and 17.02 ± 9.39, respectively, after dealing with the FPG, the Olive TM were 34.50 ± 17.29, 43.80 ± 10.06, 33.10 ± 12.38, 28.60 ± 10.53, increased by 58.94%, 38.48%, 66.86% and 68.21%, respectively (t value was 3.91, 3.89, 6.66 and 3.87, respectively, and all P < 0.05), and the correlation between Olive TM and reactive oxygen species was better than other parameters (r = 0.77, P < 0.05).
CONCLUSIONThis study indicates that FPG-comet assay appears more specific for detecting oxidative DNA damage induced by genotoxicants exposure, and the application of comet assay will be expanded. The endonuclease modified comet assay will be used widely in the toxicology and molecular epidemiology study.
Cell Line ; Comet Assay ; methods ; DNA Damage ; Endonucleases ; Humans ; Mutagens ; toxicity ; Oxidation-Reduction ; Oxidative Stress ; Reactive Oxygen Species ; metabolism