Detection of Cytogenetic Effects in Peripheral Lymphocytes of Students Exposed to Formaldehyde with Cytokinesis-Blocked Micronucleus Assay

Cytokinesis-blocked micronucleus assay was applied as a biological dosimeter to detect abnormalities in human peripheral lymphocytes of thirteen students exposed to formaldehyde (FA) during a 12-week (10 h per week) anatomy class. Breathing-zone air samples colleeted during dissection procedures showed a mean concentration of 2. 37 ppm (3. 17mg/m3 ). Ten students from the same school but without FA exposure served as controls. Chromosome aberrations (CA) and sister chromatid exchanges (SCE) were detected in both groups. The micronuclei (MN) rate (6. 38 ± 2. 50‰ ) and CA rate (5. 92 ±2. 40‰ ) in the FA-exposed group showed a significant increase (P< 0. 01 ) when compared with those of the controls (3. 15 ±1. 46‰and 3. 40 ± 1. 57 % respectively). A correlation between MN and CA in individuals was observed. SCE in the exmpd group were also increased (P< 0. 05), but not so greatly as MN or CA. The results indicated that FA might damage the chromosomes of human lymphocytes.

Effect of various drinking water on human micronucleus frequency in high-risk population of PHC

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TESTING THE MUTAGENICITY OF FIVE PESTICIDES WITH MICRONUCLEUS TECHNIQUE OF TRADESCANTIA PALUDOSA

Applying the micronucleus technique T.adescanlia paludosa, the mutagenicity of five pesticides, Decis, Sumicid, Cypermethrin, Trichlorphon and Diazinon were tested. The testing results showed that 0.64%, 0.32%, 0.16%, 0.08% of Decis, 0.64%, 0.32%, 0. 16%, 0.04% of Trichlorphon and 0.32%, 0.16%, 0.08%, 0.04% of Diazinon all had mutagenicity on Tradescantia paludosa, but Sumicid and Cypermethrin had no such characteristic.

The Cytokinesis-Block Micronucleus Assay on Peripheral Blood Lymphocytes as a Prospective Biological Test-System to Estimate the Influence of Radon on the Human Organism: Recent Progress and Future Prospects

This paper discusses the problem of assessing the negative after-effects of low doses of ionising radiation exposure in humans. Radon and its decay daughter products are the most widespread source of such irradiation. Miners (in both uranium and non-uranium mines) as well as laypeople in domestic life may be exposed to radon, making the problem of assessing the cytogenetic effects of exposure extremely crucial. One of the more promising test systems to assess the effect of radon is the cytokinesis-block micronucleus assay (CBMN) on peripheral blood lymphocytes, which has a number of advantages over other cytogenetic techniques. Recent progress and future prospects of this cytogenetic method are discussed here.

Application of Micronucleus Technique in Testing River Pollution in the Qinhuang River, Binzhou City

To figure out water pollution of the Qinhuang River in Binzhou City,broad bean root-tip micronucleus technique was applied to test water quality,water sample from different sampling points was taken to detect the contamination of broad bean root tips,so as to reflect water pollution using the pol ution indexes.The results showed that water in the Qinhuang River was polluted in different degrees,and water pollution grew more serious from the upper to the lower reaches.Water pollution sources include domestic sewage and solid waste,thus pollution discharge into the river must be strictly controlled to restore the polluted water and the ecological functions of waterscapes.

The Chromosomal Effect of Birchen Dust as Determined by the Micronucleus Test

In a wood processing factory, the measured air concentration of birchen dust was 1. 26 ±0. 41 mg/m3, and the micronucleus frequency of peripheral blood lymphocytes in 83 workersexposed to wood dust was 1. 13 ± 2. 83%, which was significantly higher (P < 0.01 ) thanthat of control group (0. 51 ± 1. 41% ). The number of exposed workers with positive mi-cronucleus test was 9. 6 %, which was higher than that of control group (4. 5 % ), but thedifference was not significant (P >0. 05 ). The micronucleus test in mice treated with waterextracts of unsteamed and unbaked birchen dust showed that the micronucleus frequencies inall treated groups were significantly higher than that of contro group (P < 0. 01 ) and therewas also a doseresponse correlation (r = 0. 96, P < 0. 0005 ). The results of steamed andbaked birchen dust extracts were significantly lower than those of the unsteamed and unbakedones at the same doses (P< 0. 001 ). This suggests that when the birchen dust is steamed atthe temperature of 100℃ for 24h or baked at the temperature of 80℃, its inducing effect inmicronucleus test could be lowered

Evaluation of Protective Effects of Bioactive Phytochemicals Against Methotrexate in Salmonella typhimurium TA1535/pSK1002 Coupled with Micronucleus Assay

We evaluated the antimutagenic effects kinds of bioactive phytochemicals and of 10 some phytochemical combinations against methotrexate （MTX）-induced genotoxicity by the umu test in Salmonella typhimurium TA1535/rpSK1002 combined with a micronucleus assay. We observed that allicin, proanthocyanidins, polyphenols, eleutherosides, and isoflavones had higher antimutagenic activities than the other five types of bioactive phytochemicals. At the highest dose tested, MTX-induced genotoxicity was inhibited by 25%-75%. Kunming mice treated by MTX along with bioactive phytochemical combinations showed significant reduction in micronucleus induction and sperm abnormality rate （P〈0.01）. These results indicate that bioactive phytochemical combinations can be potentially used as new cytoprotectors.

Coking wastewater increases micronucleus frequency in mouse in vivo via oxidative stress

Coking wastewater has caused serious health risk in coal-producing areas of China, however its toxic effects have not been well understood. The genotoxicity induced by coking wastewater on mice in vivo and its possible oxidative mechanisms were investigated via observing the induction of micronuclei in polychromatic erythrocytes of mouse bone marrow, and subsequently determining the antioxidative enzyme activities （superoxide dismutase Cu, Zn-SOD, Se-dependent glutathione peroxidase, and catalase）, thiobarbituric acid reactive substance contents and protein carbonyl levels in brains and livers of mice. Results showed that the tested coking wastewater caused a significant increase of micronucleus frequencies in a concentration-dependent manner. Also, the sample increased lipid peroxidation and protein oxidation levels, which was accompanied by changes in antioxidative status. Interestingly, pre-treatment with an antioxidant （vitamin C） led to a statistical reduction in the micronucleus frequency caused by coking wastewater. This implies that coking wastewater induces evident genetic damage in mammalian cells, and exposure to polluted areas might pose a potential genotoxic risk to human beings; in the process, oxidative stress played a crucial role.

Influence of cosmic radiation on lymphocyte micronucleus,serum lipid peroxide and antioxidation capacity in aircrew members

This study looks into the influence of cosmic radiation at high altitudes on human bodies. Results reveal that the cytokinesis-block micronuclei (CBMN) and conventional cultured micronuclei in peripheral blood lymphocytes, serum levels of lipid peroxide, superoxide dismutase, and the total antioxidation capacity by chemical colorimetry all increased significantly in aircrew members. There exists a linear relationship between the CBMN and the average annual effective doses of radiation received or the average annual flying hours. With both of them, a trend shows that the serum lipid peroxide levels increase as well. Either the lipid peroxide or CBMN can sensitively reflect the recent changes in flight load. These findings indicate that cosmic radiation impairs the stability of chromosomes and genome, and induces lipid oxidative damage in aircrews; Lymphocyte CBMN and serum lipid peroxide can be used as monitoring indicators in the cosmic radiation protection for aircrew members.

Comet assay and cytokinesis-blocked micronucleus test for monitoring the genotoxic effects of X-ray radiation in humans

Obejctive To assess the genotoxic effects of X ray radiation on human populations Methods The single cell gel electrophoresis (SCGE) and cytokinesis blocked micronucleus (CBMN) test were applied as biological dosimeters to detect DNA damage and abnormalities in human peripheral lymphocytes of subpopulation exposed to X ray radiation The subjects were divided into four groups: 12 radiation patients; 13 intervention radiation therapy doctors; 32 radiation diagnostians; 28 controls Results The average comet lengths of the four groups were 128 17±4 49?μm, 88 09±5 39?μm, 72 68±2 57?μm and 32 87±0 57?μm, respectively The difference in average comet length between any two groups was highly significant ( P <0 01) The average micronucleated cell (MNC) rates (‰) of the four groups were 12 33±0 85, 9 75±1 02, 8 48±0 66 and 3 18±0 36, respectively The difference of MNC rates of Group 1 vs 3, 1 vs 4, 2 vs 4 and 3 vs 4 was highly significant ( P <0 01), and the difference of Group 1 vs 2 was significant ( P <0 05), but there was no difference of MNC rate in Group 2 vs 3 ( P >0 05) Conclusions This study showed that both the comet assay and the CBMN test could be used to monitor populations exposed to X ray radiation, but the comet assay seems to be more sensitive than the CBMN test

Evaluation of drinking water treatment combined filter backwash water recycling technology based on comet and micronucleus assay

Based on the fact that recycling of combined filter backwash water（CFBW）directly to drinking water treatment plants（WTP）is considered to be a feasible method to enhance pollutant removal efficiency,we were motivated to evaluate the genotoxicity of water samples from two pilot-scale drinking water treatment systems,one with recycling of combined backwash water,the other one with a conventional process.An integrated approach of the comet and micronucleus（MN）assays was used with zebrafish（Danio rerio）to investigate the water genotoxicity in this study.The total organic carbon（TOC）,dissolved organic carbon（DOC）,and trihalomethane formation potential（THMFP）,of the recycling process were lower than that of the conventional process.All the results showed that there was no statistically significant difference（P〉0.05）between the conventional and recycling processes,and indicated that the genotoxicity of water samples from the recycling process did not accumulate in 15 day continuous recycling trial.It was worth noting that there was correlation between the concentrations of TOC,DOC,UV（254）,and THMFPs in water and the DNA damage score,with corresponding R^2 values of 0.68,0.63,0.28,and 0.64.Nevertheless,both DNA strand breaks and MN frequency of all water samples after disinfection were higher than that of water samples from the two treatment units,which meant that the disinfection by-products（DBPs）formed by disinfection could increase the DNA damage.Both the comet and MN tests suggest that the recycling process did not increase the genotoxicity risk,compared to the traditional process.

DNA damaging effects of carbofuran and its main metabolites on mice by micronucleus test and single cell gel electrophoresis

The DNA damaging effects of the carbamate pesticide carbofuran and its four metabolites (carbofuranphenol, 3-ketocarbofuran, 3-hydrocarbofuran and nitrosocarbofuran) on mice were evaluated by single cell gel electrophoresis (SCGE) assay and micronucleus test. KM mice were exposed to test compounds with different doses of 0.1, 0.2 and 0.4mg/kg through intraperitoneal injection two times with an internal of 24 h, and then killed by cervical dislocation 6 h after the second injection. In SCGE assay, isolated mice peripheral blood lymphocytes were employed to determine DNA damaging degree after a 1 h treatment by test compounds and a following electrophoresis. Carbofuran and carbofuranphenol showed negative results in both test and had no obvious toxicity. 3-hydrocarbofuran and nitrosocarbofuran were positive.3-ketocarbofuran could not induce micronucleus formation but caused significant DNA migration in SCGE test. These tests revealed that 3-ketocarbofuran, 3-hydrocarbofuran and nitrosocarbofuran are potential mutagesis and further research is needed.

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

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.

Chromosome aberration and micronucleus frequency in peripheral blood lymphocytes in radiation workers

Objective To investigate chromosome aberration and micronucleus frequency in peripheral blood lymphocytes in workers engaged in radiation for a long time,to reduce occupational hazard caused by ionizing radiation and to further strengthen health surveillance.Methods A total of 366 members of medical staff engaged in radiation work who underwent physical examinations in

Micronuclei and cell survival in human liver cancer cells irradiated by 25MeV/u^(40)Ar^(14+)

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Evaluation of antigenotoxic effects of carotenoids from green algae Chlorococcum humicola using human lymphocytes

Objective:To identify the available phytochemicals and carotenoids in the selected green algae and evaluate the potential genotoxic/antigenotoxic effect using lymphocytes.Methods:Organic,solvent extracts of Chlorococcum humicola(C.humicola)were used for the phytochemical analysis.The available carotenoids were assessed by HPLC,and LC-MS analysis.The genotoxicity was induced by the benzo(a)pyrene in the lymphocyte culture,the genotoxic and antigenotoxic effects of algal carotenoids with and without genotoxic inducer were evaluated by chromosomal aberration(CA),sister chromatid exchange(SCE)and micronucleus assay(MN).Results:The results of the analysis showed that the algae were rich in carotenoids and fatty acids.In the total carotenoids lutein,β-carotene andα-carotene were found to be present in higher concentration.The frequency of CA and SCE increased by benzo(a)pyrene were significantly decreased by the carotenoids(P<0.05 for CA,P<0.001 for SCE).The MN frequencies of the cells were significantly decreased by the treatment with carotenoids when compared with the positive controls(P<0.05).Conclusions:The findings of the present study demonstrate that,the green algae C.humicola is a rich source of bioactive compounds especially carotenoids which effectively fight against environmental genotoxic agents,the carotenoids itself is not a genotoxic substance and should be further considered for its beneficial effects.

ANEUPLOIDY INDUCTION BY THE WATER EXTRACT OF TRIPTERYGIUM HYPOGLAUCUM(LEVEL) HUTCH IN MOUSE BONE MARROW CELLS

The aneuploldy inducing activity of a Chinese medicinal herb, Tripterygium hypoglaucum (level) Hutch (THH) were investigated by means of thhree cytogenetic end points, namely C-mitotic (CM) effects, micronucleus (MN) and parallel chromosome structural aberration (CA) analyses in vivo. The experiments were performed on mouse bone marrow cells. THH showed similar gentoxic effects to colchicine (COL) in CM, MN and CA analyses; positive CM effects were observed accompanied with increases of mitotic index and frequencies of C-mltotic cells as well as decreased frequencies of anaphase in all of the THH-treated groups. The compound showed a positive MN response in bone marrow polychromatic erythrocytes but was negative in CA analyses. The preliminary results suggested that THH is an aneuploldy inducer in mouse bone marrow cells under present experiment conditions.

Response of Lymphocytes to Radiation in Untreated Breast Cancer Patients as Detected with Three Different Genetic Assays

Objective To detect the response of lymphocytes to radiation in untreated breast cancer patients with three different genetic assays. Methods Blood samples were collected from 25 untreated patients and 25 controls. Each blood sample was divided into two parts： one was irradiated by 3-Gy X-ray （irradiated sample）, the other was not irradiated （non-irradiated sample）. The radiosensitivity of lymphocytes was assessed by comet assay, cytokinesis-block micronucleus （CBMN） assay and 6-TG-resistant cells scored （TG） assay. Results The baseline values of micronucleated cell frequency （MCF） and micronucleus frequency （MNF） in the patients were significantly higher than those in the controls （P〈0.01）, and 3-Gy X-ray induced genetic damage to lymphocytes in the patients increased significantly as compared with that in the controls as detected with the three genetic assays （P〈0.01）. The proportion of radiosensitive cases in the patient group was 48% for the mean tail length （MTL）, 40% for the mean tall moment （MTM）, 40% for MCF, 44% for MNF, and 48% for mutation frequencies of the hprt gene （Mfs-hprt）, respectively, whereas the proportion of radiosensitive cases in the control group was only 8% for all the parameters. Conclusion The difference in the lymphocyte radiosensitivity between the breast cancer patients and the controls is significant. Moreover, there are wide individual variations in lymphocyte radiosensitivity of patients with breast cancer. In some cases, the radiosensitivity of the same patient may be different as detected with the different assays. It is suggested that multiple assays should be used to assess the radiosensitivity of patients with breast cancer before therapy.

Inhibitory effects of the ferment liquids on sarcoma

Objective： To study the anti-tumor effects of the ferment liquid （FL） and to explore the anti-mutagenic effects induced by cyclophosphamide （CP）. Methods： Anti-tumor effects and Anti-mutagenic effects induced by cyclophosphamide （CP） of the ferment liquid were observed in tumor-bearing mice in vivo. Spleen lymphocytes proliferation of mice was stimulated by ConA and NK cell activity was analyzed by the lactate dehydrogenase （LDH） release assay. Results： FL Ⅰ,FL Ⅱ and FL Ⅲ of 16.7, 33.3 and 66.6 ml/kg inhibited growth of Sarcoma 180 sighificanth by 11.6%, 24.2%, 28.0%, 4.8%, 15.2%, 2.2% and 2.1%, 8.5%, 5.2% respectively. Polychromatic erythrocytes （PCE） at all doses of FL Ⅰ and FL Ⅱ were greatly decreased compared with control group （ P 〈 0.05）, but FL Ⅲ has no inhibition effents（ P 〉 0.05）. Production of IL-2 showed no significant difference between all treatment groups. The NK cell killing activity in all treated mice increased by FL Ⅰ and FL Ⅱ was significantly potentiated compared to controls （ P 〈 0.05）. Lymphocytes proliferation of ICR mice were stimulated by ConA 2.5 ml/L and there were no differences between groups. Conclusion： The present data suggested that three kinds of ferment liquid used as health drink have anfi-mutagenesis, immune enhancement effects and showed certain inhibitory effects on S180 tumor growth in vivo.

Radiation signature on exposed cells: Relevance in dose estimation

The radiation is considered as a double edged sword, as its beneficial and detrimental effects have been demonstrated. The potential benefits are being exploited to its maximum by adopting safe handling of radionuclide stipulated by the regulatory agencies. While the occupational workers are monitored by personnel monitoring devices, for general publics, it is not a regular practice. However, it can be achieved by using biomarkers with a potential for the radiation triage and medical management. An ideal biomarker to adopt in those situations should be rapid, specific, sensitive, reproducible, and able to categorize the nature of exposure and could provide a reliable dose estimation irrespective of the time of the exposures. Since cytogenetic markers shown to have many advantages relatively than other markers, the origins of various chromosomal abnormalities induced by ionizing radiations along with dose-response curves generated in the laboratory are presented. Current status of the gold standard dicentric chromosome assay, micronucleus assay, translocation measurement by fluorescence insitu hybridization and an emerging protein marker the g-H2 AX assay are discussed with our laboratory data. With the wide choice of methods, an appropriate assay can be employed based on the net.