Qualitative and Quantitative Studies on DNA and RNA Synthesis in Loxodes striatus Nuclei*

SYNOPSIS. In this study the characteristics of the synthesis of DNA and RNA in the nuclei of Loxodes were investigated. Loxodes striatus is a primitive ciliate with 2 pairs of structurally differentiated diploid nuclei, the macro- and micronuclei. The macronuclei are differentiated morphologically into a clearly recognizable central core and an outer zone. To determine DNA and RNA synthesis, individual organisms were analyzed by autoradiography after incubating groups of cells with a ³H-labeled precursor ([³H]thymidine for DNA and [³H]uridine for RNA). The following observations were made: (A) All portions of macro- and micronuclei appeared to contain DNA as judged by the localizations of incorporated [³H]thymidine. (B) The macro- and micronuclei did not synthesize DNA at the same time; moreover, the duration of DNA synthesis in the former was much longer than of the latter nucleus. (C) Replication of DNA in the inner core and outer zone of the macronucleus occurred at separate times with little if any overlap. (D) All of the detectable [³H]uridine incorporation was found in the macronucleus and none in the micronucleus. Within the macro-nucleus the central core was more heavily labeled. (E) The quantitative differences in the label of the different components of the nuclear complex were investigated. (F) Contrary to the previously reported information our results suggest that DNA synthesis can occur in adult macronuclei. The possible explanation of these results is discussed in the context of the nuclear evolution of ciliates and of recent information on nuclear differentiation.     

甲硝唑诱导蚕豆根尖细胞微核的研究

本实验研究了甲硝唑诱导蚕豆根尖细胞微核的效应。实验表明：(1)浓度为0.1、6、12、40和500 mg/L甲硝唑均能使蚕豆根尖细胞微核率显著增加,且蚕豆根尖细胞微核率和甲硝唑浓度之间存在明显的剂量-效应关系;(2)甲硝唑能引起DNA损伤,具有分子诱变剂的性能。     

微核技术监测广州灵岗河水质污染的研究

水污染已成为全球普遍关注的一个重大问题。为探讨洗水印染企业废水和生活污水排放对环境的污染程度以及对周边生物的遗传损伤,研究利用蚕豆根尖细胞微核监测技术对位于广州番禺区灵岗河段淤泥浸出液和污水引发的环境情况进行了测定。数据显示用灵岗河段污水处理后的蚕豆根尖细胞微核发生率显著大于阴性对照组的微核率,水质污染指数为2．912,初步测定该河段属中度污染。     

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 in-traperitoneal 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 pote     

Impact of radiofrequency radiation on DNA damage and antioxidants in peripheral blood lymphocytes of humans residing in the vicinity of mobile phone base stations

Radiofrequency radiations (RFRs) emitted by mobile phone base stations have raised concerns on its adverse impact on humans residing in the vicinity of mobile phone base stations. Therefore, the present study was envisaged to evaluate the effect of RFR on the DNA damage and antioxidant status in cultured human peripheral blood lymphocytes (HPBLs) of individuals residing in the vicinity of mobile phone base stations and comparing it with healthy controls. The study groups matched for various demographic data including age, gender, dietary pattern, smoking habit, alcohol consumption, duration of mobile phone use and average daily mobile phone use. The RF power density of the exposed individuals was significantly higher (p < 0.0001) when compared to the control group. The HPBLs were cultured and the DNA damage was assessed by cytokinesis blocked micronucleus (MN) assay in the binucleate lymphocytes. The analyses of data from the exposed group (n = 40), residing within a perimeter of 80 m of mobile base stations, showed significantly (p < 0.0001) higher frequency of micronuclei when compared to the control group, residing 300 m away from the mobile base station/s. The analysis of various antioxidants in the plasma of exposed individuals revealed a significant attrition in glutathione (GSH) concentration (p < 0.01), activities of catalase (CAT) (p < 0.001) and superoxide dismutase (SOD) (p < 0.001) and rise in lipid peroxidation (LOO) when compared to controls. Multiple linear regression analyses revealed a significant association among reduced GSH concentration (p < 0.05), CAT (p < 0.001) and SOD (p < 0.001) activities and elevated MN frequency (p < 0.001) and LOO (p < 0.001) with increasing RF power density.     

一株白腐菌的食用安全性初步评价

本研究选取了一株白腐菌模式菌株进行了小鼠急性毒性试验、小鼠骨髓嗜多染红细胞微核试验以及小鼠精子畸形试验,以分析该白腐菌的食用安全性,为进一步应用白腐菌开发功能性食品提供数据支持。结果显示,小鼠经灌胃白腐菌,其LD50为6.76 g/kg(以白腐菌菌丝干重计)。微核试验和精子畸形试验结果均为阴性(P0.05)。试验结果表明,本次实验条件下,白腐菌对小鼠表现出一定的急性毒性,但未见遗传毒性,由此推断,在一定剂量范围内使用白腐菌作为食用材料是相对安全的,安全的剂量范围需要进一步扩大浓度梯度来确定。     

重铬酸钾对蚕豆根尖细胞致畸效应的研究

以蚕豆根尖为材料,研究重铬酸钾对蚕豆根尖细胞的致畸效应。采用蚕豆根尖细胞的微核试验和染色体畸变试验方法,以不同浓度的重铬酸钾为诱变剂,测定蚕豆根尖细胞的微核率和染色体畸变率。结果表明:重铬酸钾能诱发较高频率的微核率,即在一定浓度范围内,其微核率随重铬酸钾处理浓度的升高而增加,但高于一定浓度后反而呈下降趋势;不同浓度的重铬酸钾均使蚕豆根尖细胞有丝分裂指数增大;重铬酸钾还能诱导蚕豆根尖细胞产生较高频率的染色体畸变,且产生多种类型的染色体畸变。结论是重铬酸钾对蚕豆根尖细胞具有明显的致畸效应。     

SO2体内衍生物诱发蚕豆根尖细胞微核

以蚕豆为材料 ,研究 SO2 体内衍生物 -亚硫酸钠与亚硫酸氢钠混合液 (3:1)对根尖细胞的遗传毒效应。结果表明 :0 .0 5～2 .0 0 mmol/ L的 SO2 衍生物处理可诱发两品种蚕豆根尖中的微核细胞明显增加 ;不同浓度 (0 .0 5～ 15 .0 0 mmol/ L ) SO2 衍生物处理 12、2 4、36 h后 ,根尖微核细胞率是对照组的 2 .5～ 5 .0倍 ,显著高于对照组。在一定浓度范围内 ,蚕豆根尖细胞微核率与SO2 衍生物浓度之间具有正的线性相关。研究结果表明 ,SO2 衍生物能够破坏蚕豆根尖细胞的遗传稳定性。     

Evaluation of genotoxic effects in human leukocytes after in vitro exposure to 1950 MHz UMTS radiofrequency field

In the present study the third generation wireless technology of the Universal Mobile Telecommunication System (UMTS) signal was investigated for the induction of genotoxic effects in human leukocytes. Peripheral blood from six healthy donors was used and, for each donor, intermittent exposures (6 min RF on, 2 h RF off) at the frequency of 1950 MHz were conducted at a specific absorption rate of 2.2 W/kg. The exposures were performed in a transverse electro magnetic (TEM) cell hosted in an incubator under strictly controlled conditions of temperature and dosimetry. Following long duration intermittent RF exposures (from 24 to 68 h) in different stages of the cell cycle, micronucleus formation was evaluated by applying the cytokinesis block micronucleus assay, which also provides information on cell division kinetics. Primary DNA damage (strand breaks/alkali labile sites) was also investigated following 24 h of intermittent RF exposures, by applying the alkaline single cell gel electrophoresis (SCG)/comet assay. Positive controls were included by treating cell cultures with Mitomycin-C and methylmethanesulfonate for micronucleus and comet assays, respectively. The results obtained indicate that intermittent exposures of human lymphocytes in different stages of cell cycle do not induce either an increase in micronucleated cells, or change in cell cycle kinetics; moreover, 24 h intermittent exposures also fail to affect DNA structure of human leukocytes soon after the exposures, likely indicating that repairable DNA damage was not induced.     

Coffee-mediated protective effects against directly acting genotoxins and gamma-radiation in mouse lymphoma cells

The cytokinesis-block micronucleus test was performed using L5178Y mouse lymphoma cells to ascertain whether or not standard (caffeinated) instant coffee, the commonly consumed polyphenolic beverage with antioxidant activity can protect against chromosomal damage induced by the directly acting agents N-methyl-N-nitro-N-nitrosoguanidine (MNNG), mitomycin C (MMC), methyl methanesulfonate (MMS) and gamma radiation. Our results demonstrated significant reductions in the in vitro genotoxic effects of MNNG, MMC, and MMS following co-treatment of mouse lymphoma cells with standard instant coffee. Subsequently, the comet assay was carried out to assess the effect of coffee co-treatment on the level of DNA damage induced by MMS in mouse lymphoma cells. The results demonstrated a significant reduction in MMS-induced DNA damage following co-treatment with standard instant coffee. Protective effects were observed in mouse lymphoma cells which were treated with coffee immediately after exposure to gamma radiation (1 and 2 Gy). Another experiment showed protection when the mammalian cells were irradiated (0.5 and 1 Gy) midway (at 2 h) during a 4 h coffee treatment. However, the protective effect against the lower dose (0.5 Gy) was not significant. In addition we assessed the modulatory effect of coffee on MNNG-induced apoptotic frequency by flow cytometry. The results revealed only a minor influence of coffee on the frequency of apoptotic cells induced by the test compounds, rendering an increase in sensitivity for apoptosis as a reason for the reduced genomic damage an unlikely or at least incomplete explanation.