基于彗星实验的洛克沙胂对秀丽隐杆线虫胚胎细胞DNA损伤的研究

为评价洛克沙胂的遗传毒性,采用单细胞凝胶电泳(彗星实验),研究了不同浓度的洛克沙胂对秀丽隐杆线虫胚胎细胞脱氧核醣核酸(DNA)的损伤作用。提取秀丽隐杆线虫的胚胎细胞,分别暴露于0(空白对照)、50、250、500μg·L~(-1)含洛克沙胂的溶液染毒1 h。用彗尾DNA百分比含量(TDNA%)、彗星尾长(TL)和Olive尾矩(OTM)作为DNA损伤的指标。实验结果表明,与空白对照组比较,处理组中彗尾DNA百分比含量、彗星尾长以及Olive尾矩显著增加(P0.01)。随着洛克沙胂浓度的增加,彗尾DNA百分比含量、彗星尾长以及Olive尾矩逐渐增加,其相关系数r0.99,说明在实验浓度范围内,存在极显著的浓度-效应关系。洛克沙胂对秀丽隐杆线虫胚胎细胞DNA具有损伤作用,彗星实验操作简便、快速、灵敏度高,能够反映出洛克沙胂的遗传毒性。因此,通过彗星实验建立实验室检测洛克沙胂遗传毒性的方法具有可行性。     

羰基镍急性中毒对大鼠骨髓细胞的DNA损伤

通过动物实验观察不同剂量羰基镍对大鼠骨髓细胞DNA损伤程度。采用SD大鼠,以135 mg·m~(-3)和250 mg·m~(-3)羰基镍为染毒组,250 mg·m~(-3)氯气为阳性对照组,静态方式染毒30 min。未染毒组为正常对照组,大鼠染毒后1、2、3和7 d分别采集样本。采用单细胞凝胶电泳检测每组大鼠骨髓细胞DNA的损伤程度。彗星尾长和Olive尾矩2个指标的分析结果表明,大鼠骨髓细胞DNA损伤程度随着羰基镍染毒剂量的增加而增加,在4个时间点各剂量组间均有显著差异(P0.05)且随时间的变化有一定的规律,损伤程度在3 d时达到最大,而后缓慢下降。羰基镍急性中毒对大鼠骨髓细胞DNA有一定的损伤,且存在剂量-效应关系,各剂量组损伤程度有一定的时间效应规律。     

Relative yields of monomeric and dimeric adducts induced by sulphur mustard in isolated and cellular DNA as determined by HPLC/tandem mass spectrometry

Sulphur mustard (SM) is known as an efficient vesicating agent as well as a carcinogenic chemical. This warfare agent remains a threat for both civilians and militaries. DNA alkylation is one of the critical molecular pathways at the origin of the symptoms associated with SM exposure. SM forms monoadducts with guanine and adenine as well as a biadduct between two guanine bases. The aim of the present work is to determine the relative yields of these three lesions in DNA samples after SM exposure without using radiolabeled SM as in earlier works. For this purpose, we have developed a high performance liquid chromatography/tandem mass spectrometric method to simultaneously quantify the SM monoadducts and biadduct in the same DNA sample. We observed in isolated and cellular DNA that the guanine monoadduct was the predominant lesion, while the biadduct was present in twofold lower yield. The adenine monoadduct was generated in lowest amounts. The analytical approach was extended to 2?chloroethyl ethyl sulphide, a widely used SM analog. Again, the adenine adduct was much less frequent than the guanine derivative. The developed assay will allow performing studies involving large numbers of samples.     

Effect of electronic waste on E. coli genomic integrity: a possible role for metal induced carcinogenesis

Electronic waste (e-waste) poses a major public health threat for developing countries’ populations. The hazards of e-waste are exacerbated by crude recycling methods. In this study, the presence of metals in e-waste samples obtained from Lagos, Nigeria, was assessed using atomic absorption spectrometry. The effect of e-waste on Escherichia coli (E. coli) PQ-37 genomic integrity was evaluated using the SOS chromotest. The means of metal concentrations in the evaluated samples were 16 (cadmium), 7.3 (nickel), 11 (chromium), 20 (lead), 3100 (iron), 90 (zinc), and 2000 (copper) μg_L^?1. Damage to E. coli deoxyribonucleic acid (DNA) increased proportionally to the metal concentrations. Significant amounts of DNA damaging agents from inadequately processed e-waste are present in the studied environment, which will have implications for adverse effects on public and ecological health. Existing policies against dumping of toxic materials in susceptible communities should be enforced.     

Assessment of DNA damage and cytotoxicity of palmatine on human skin epithelial carcinoma cells

The present investigation was carried out to examine the cytotoxic and genotoxic effects of a Tinospora cordifolia crude methanolic extract (palmatine) on human skin epithelial carcinoma cells (A431). T. cordifolia is one of the indispensable medicinal plants used in Ayurvedic medicine for treatment of various diseases and recommended for improving the immune system. Cytotoxicity and genotoxicity evaluation was carried out using A431 cells treated with different concentrations of palmatine. The duration of the treatment was 24 and 48 hr. A cellular proliferative capacity test showed that palmatine produced cytotoxicity in concentration- and time-dependent manner. Further, palmatine induced significant intracellular reactive oxygen species generation and elevated lipid peroxidation, as well as activities of catalase and superoxide dismutase. DNA fragmentation analysis using the comet assay showed that palmatine induced genotoxicity in a concentration- and time-dependent manner. Evidence indicates palmatine is capable of induction of oxidative stress resulting in cell death and genomic instability.     

Cytotoxic activity of spark‐decomposed sulfur hexafluoride and analysis of cytotoxic contributions of individual spark decomposition products†

Sulfur hexafluoride decomposed by electrical sparks has been found to by cytotoxic to hamster cells when tested in an in vitro cell survival assay, while SF₆ shows no cytotoxic activity. Chemical analysis of spark‐decomposed SF₆ has identified and quantified the following compounds: SOF₂, SO₂F₂, SF₄, SOF₄, SiF₄, SO₂ and HF. Each of these gases, at concentration ranges expected in spark‐decomposed SF₆, were tested for cytotoxic activity toward hamster cells. Of the gases showing cytotoxic activity, SO₂F₂ and SOF₄ were similar in activity, as were SOF₂ and SF₄, while the behavior of SiF₄ was different from the rest. None of these individual gases, at concentrations expected in spark‐decomposed SF₆, has sufficient cytotoxic activity to account for the cytotoxic effect of spark‐decomposed SF₆ observed in our assay system. A four‐component mixture of some of the gases enumerated above (at concentrations overestimating their abundance in spark‐decomposed SF₆) was much less cytotoxic than the spark‐decomposed SF₆ gas. A mathematical simulation of the cytotoxic activity of a mixture of gases at concentrations found in spark‐decomposed SF₆ was made, assuming independent cytotoxic effects from each component. The simulated cytotoxic effect thus computed was less than that seen in spark‐decomposed SF₆. Since individual components or mixtures of the major decomposition products do not account for the observed biological activity of spark‐decomposed SF₆, this suggests there may be one or more components, present in the spark‐decomposed gas at very low concentrations, which may have a very strong cytotoxic activity.     

Voltammetric study on the interaction of the heavy metals Cd(II) and Mn(II) with transfer‐RNA,DNA and other polynucleotides

Small amounts of bivalent cations, usually provided by Mg²⁺, are in the living cell necessary for the biological activity of t‐RNA as these bivalent cations influence the tertiary and secondary structure of this globular polynucleotide.

In context with the discussed possibility of carcinogenic actions of ingested Cd it is of particular interest to check whether there exist specific strong interactions of this toxic heavy metal with nucleic acids.

Therefore, the binding of the toxic heavy metal ion Cd²⁺ and the essential heavy metal ion Mn²⁺ to t‐RNA and for comparison to DNA and the polynucleotides poly‐U, poly‐A and poly‐A‐poly‐U has been studied. Free metal ion concentrations have been determined by differential pulse polararography. Association constants and the number of binding sites have been evaluated by the Scatchard method and alternatively according to a simple electrostatic model of the polyelectrolytes. With the Scatchard method for t‐RNA and all polynucleotides with helical structure two different binding sites of different strength are observed. Those with higher association constants are assigned to the helical parts of t‐RNA. Interaction sites with low association constants correspond to the parts with no ordered tertiary structure, as their exclusive occurrence for poly‐U, having a completely stochastic coil structure, reflects. The values of the association constants for the stronger and weaker association sites are in the respective polynucleotides for both investigated bivalent metal ions of comparable magnitude. This emphasizes that the interaction is essentially of electrostatic nature and depends primarily on the charge of the interacting species.

Thus the specific strong interaction of Cd by the intercalation into the tertiary structure of nucleic acids or by chelation of their base units can be ruled out as one possibility for carcinogenity of Cd.

Moreover, under physiological conditions the high excess of competitive Mg²⁺ will suppress the interaction of Cd based on electrostatic forces.     

Recent advances in arsenic mutagenesis and carcinogenesis†

Arsenic is ubiquitously distributed in nature and is released into the environment through non‐ferrous smelting operations, generation of power from coal, and agriculture.

Epidemiological studies have shown that the incidences of epidermoid carcinomas of the skin and lungs, and of pre‐cancerous dermal keratoses are significantly increased in human subjects exposed to arsenic compounds by oral or respiratory routes. The negative results obtained on animals treated with arsenicals suggest strongly that this metal is probably a cocarcinogen.

The results of the short‐term tests suggest that the cocarcinogenic properties of arsenic could be related to its ability to inhibit DNA repair.     

Fluorescence studies of dye displacement from DNA by chromium(III) complexes: Evidence for cation induced DNA condensations

The interactions of 10 different chromium(III) complexes with isolated calf thymus DNA have been analysed by studying the electronic and fluoresence spectra of intercalated ethidiumbromide. Triply charged cationic complexes including: [Cr(urea)₆]Cl₃.3H₂O, [Cr(1,10‐phenanthroline)₃](ClO₄)₃.2H₂O, [Cr(2,2'‐bipyridyl)₃] (ClO₄)₃.2H₂O, [Cr(ethylendiamine)₃]Cl₃.3.5H₂O and [Cr(NH₃)₆](NO₃)₃ displaced the dye from DNA. Similar effects were observed in experiments using the non‐intercalating dye bisbenzimidazole ("Hoechst 33258"). However, singly charged cationic, anionic and uncharged chromium(III) complexes such as: cis‐[Cr(1,10‐phenanthroline)₂Cl₂]Cl.2H₂O, cis‐[Cr(2,2'‐bipyridyl)₂Cl₂]Cl.2H₂O, [Cr(glutathione)₂]Na₂, [Cr(cysteine)₂]Na.2H₂O and [Cr(glycine)₃] were unable to displace both ethidiumbromide and bisbenzimidazole from DNA. There was no evidence for the formation of co‐ordinate bonds between chromium(III) and DNA for any of the above complexes. The charge and type of ligand are important in controlling the interaction of chromium(III) with isolated DNA in vitro. Our findings indicate that the outer sphere interaction of a chromium(III) complex with DNA is weak and unlikely to be the mechanism by which chromate causes DNA impairments in vivo and in vitro.     

Organ differences in microsomes and cytosol metabolism of Aflatoxin B1 in piglets

In order to delineate the features of aflatoxin B1 (AFB1) metabolism in various organs of piglets, in vitro metabolism of AFB1 by microsomes and cytosol of the various piglet organs was studied. The AFB1 was converted efficiently to AFP1 by the kidney microsomes. A less efficient metabolism was noted from the AFB1 to AFQ1, AFM1, and aflatoxicol (AFL) in the various organs. The microsomal ability to form AFB1-DNA adduct was higher in liver when compared to the other organs. The cytosolic glutathione-S-transferase activity to convert AFB1-epoxide to AFB1-glutathione conjugate product was relatively higher in the liver and the small intestine. The reductase activity to convert AFB1-dialdehyde to AFB1-dialcohol was similar in all the organs. The results suggest that the variation in susceptibility to the aflatoxin among different organs is attributable mainly to the organ differences in cytochrome P450 activity to form AFB1-epoxide.