纳米银和PVP包被纳米银对HepG2细胞遗传毒性的比较研究

为了探讨纳米银对HepG2细胞DNA损伤、染色体畸变等遗传毒性指标的影响,以期为纳米银体外遗传毒性评价提供参考依据,本文采用2种纳米银材料(20 nm-PVP包被纳米银、20 nm-无包被纳米银),分别以20μg·mL~(-1)、40μg·mL~(-1)、80μg·mL~(-1)、160μg·mL~(-1)的剂量对HepG2细胞染毒24 h,用Hoechst-33258染色法检测细胞凋亡,彗星实验检测DNA损伤,胞质分裂阻滞微核细胞组学试验法检测染色体畸变。结果表明,20 nm Ag NPs组在160μg·mL~(-1)时引起细胞凋亡数显著增多(P0.05);20 nm PVP-Ag NPs组在80μg·mL~(-1)和160μg·mL~(-1)剂量组中细胞凋亡数显著增多(P0.01)。2种纳米银引起HepG2细胞发生细胞凋亡,并呈剂量效应关系。彗星试验结果表明,20 nm Ag NPs和20 nm PVP-Ag NPs在40μg·mL~(-1)、80μg·mL~(-1)、160μg·mL~(-1)剂量组中,Olive尾矩、尾长和尾部DNA百分比与空白对照组相比均有显著差异(P0.05)。2种纳米银对HepG2细胞DNA损伤程度为:20 nm Ag NPs20 nm PVP-Ag NPs。胞质分裂阻滞微核细胞组学试验结果表明,2种纳米银均不会引起核质桥数发生明显改变(P0.05),20 nm Ag NPs在高染毒剂量下引起微核总数、I型微核、II型微核、核芽数明显升高(P0.05);20 nm PVP-Ag NPs在各染毒剂量下均会引起微核总数及I型微核数量升高(P0.01),II型微核数在160μg·mL~(-1)剂量下升高明显(P0.01),剂量大于20μg·mL~(-1)时核芽数升高(P0.01)。20 nm PVP-Ag NPs对细胞核的影响大于20 nm Ag NPs(P0.05)。总之,2种纳米银材料均会引起HepG2细胞DNA损伤及染色体畸变等遗传毒性效应的改变,无包被纳米银比PVP包被纳米银更容易引起DNA损伤,PVP包被纳米银比无包被纳米银更容易引起细胞染色体畸变相关效应;2种材料对HepG2细胞的损伤存在浓度-效应关系,浓度越高遗传毒性损伤越严重。     

Simple and rapid determination of dioxin in fish and sea food using a highly sensitive reporter cell line, CBG 2.8D

Food, especially animal origin food is the main source of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), and dioxin-like polychlorinated biphenyls (dl-PCBs) for human exposure. So, a simple, rapid and cheap bioassay method is needed for determination of dioxins in food samples. In this study, we used a new highly sensitive reporter cell line to determine the concentration of dioxins in 33 fish and seafood samples. The samples were extracted by shaking with water/isopropanol (1:1 v/v) and hexane and cleaned-up by a multi layered silica gel column and an alumina column, then analyzed using CBG 2.8D cell line. We compared the results obtained from the CBG 2.8D cell assay to those obtained from conventional High-Resolution Gas Chromatography–High Resolution Mass Spectrometry (HRGC–HRMS) analysis. Good correlations were observed between these two methods (r²=0.93). While the slope of regression line was 1.76, the bioanalytical equivalent (BEQ) values were 1.76 folds higher than WHO-TEQ values and the conversion coefficient was 0.568 (the reciprocal of 1.76). In conclusion, CBG 2.8D cell assay was an applicable method to determine dioxins levels in fish and sea food samples.     

Studies on Formation of Liquid and Gaseous Formaldehyde-Induced DNA-Protein Crosslinks in Rat Marrow Cells

甲醛是一种遗传毒物,流行病学研究表明甲醛可能具有诱导白血病的作用,然而甲醛诱导白血病的机制目前还不清楚. 以不同浓度液态和气态甲醛对大鼠骨髓细胞进行染毒,采用KCl-SDS 法检测了骨髓细胞 DNA-蛋白质交联程度,并采用单细胞凝胶电泳技术(彗星实验)检测了骨髓细胞 DNA 链断裂程度. 研究结果表明:与对照组相比,低浓度甲醛(液态甲醛浓度为:5μmol·L-1和25μmol·L-1;气态甲醛浓度为:0.5mg·m-3 和 1.0mg·m-3)可以引起 DNA断裂水平显著增高 (p<0.01);而高浓度甲醛 (液态甲醛浓度为:125μmol·L-1 和 625μmol·L-1;气态甲醛浓度为:3.0mg·m-3)则可以引起 DNA-蛋白质交联水平显著增高(p<0.01; p<0.05). 研究结果提示:甲醛染毒可以导致大鼠骨髓细胞DNA的损伤,暗示甲醛诱导白血病具有高度的可能性.     

Fine atmospheric particles emitted by industrial,traffic and urban sources in France: characterization and genotoxicity

Health risks associated with inhalation of fine particulate matter of 2.5 µm in diameter or smaller depend on their atmospheric levels and physicochemical properties. The relationships between chemical compositions and genotoxic activities of particles emitted by mineral industries, traffic and urban sources during summer and winter in the region of Provence-Alpes-Côte d'Azur (France) were investigated.

The fine particles were separated in respect to water-soluble (13 minerals and metals) and organic-extractable (16 polycyclic aromatic hydrocarbons) components that were quantified. The chromosome damaging properties of the hydrophilic and lipophilic extracts were assessed using the centromeric micronucleus assay on a human lung fibroblast cell line.

The composition of the fine particulate matter was variable and depended upon the sources and seasons. Both the hydrophilic and lipophilic extracts induced chromosome damage: (1) in hydrophilic extracts, Ca and Zn affected chromosome losses induction; (2) acenapthylene affected chromosome damage (breakages and losses) induction and naphthalene affected chromosome damage and losses induction in lipophilic extracts without metabolic activation; and (3) benzo[a]pyrene affected chromosome losses induction in lipophilic extracts with metabolic activation. Fine particulate matter arising from coal-fired power station, road traffic, and other urban sources were the most efficient to induce chromosome breakage.     

Genotoxicity in the freshwater gastropod Lymnaea luteola L: assessment of cell type sensitivities to lead nitrate

The aquatic ecosystems are converting into the highly contaminated site due to environmental pollutants. The present study explores the oxidative stress and toxic potential of lead nitrate in freshwater snail Lymnaea luteola (L. luteola) L. The snails were exposed to an environmentally relevant concentration of lead nitrate for 24 and 96?h. Later exposure to lead nitrate (0, 10, 20 and 40?µg/mL) to the freshwater snail, the level of reactive oxygen species, malondialdehyde and nitric oxide (NO) were increased and glutathione, glutathione-S-transferase were decreased. Lead-nitrate-induced haemocyte cell death and it was observed by using Annexin-V FITC/PI through a flow cytometer. DNA damage in haemocyte cells was measured at above doses of lead-nitrate exposure for 24 and 96?h and it was compared to the untreated snail. Average tail DNA (%) and olive tail moment in single-cell gel test were increased dose and duration fashion and maximum DNA damage was measured at 96?h. These results indicate the potential toxicity and genotoxicity of lead nitrate in acute treatment to L. luteola and single-cell gel test are the assay for rapid detection of genetic effects.     

纳米SiO2与常规SiO2颗粒对Hela细胞的细胞毒性作用

为了探讨纳米SiO2和常规SiO2颗粒对Hela细胞的细胞毒性作用,采用不同浓度的纳米SiO2和常规SiO2颗粒(0.05、0.1、0.2、0.4、0.8、1.6μg·μL-1)对Hela细胞进行12h染毒,应用MTT法检测细胞毒性效应.研究发现,较低浓度(≤0.2μg·μL-1)的纳米SiO2和常规SiO2对Hela细胞无明显细胞毒性(p>0.05);较高浓度时,纳米SiO2(≥0.4μg·μL-1)和常规SiO2(≥0.8μg·μL-1)对Hela细胞具有明显细胞毒性作用(p<0.01),并且随浓度增大细胞毒性增强;当浓度≥0.4μg·μL-1时,纳米SiO2的细胞毒性明显高于相同浓度的常规SiO2(p<0.05).以上结果表明,纳米SiO2和常规SiO2颗粒均能对Hela细胞产生细胞毒性,且纳米SiO2的细胞毒性强于常规SiO2;低浓度(≤0.2μg·μL-1)的纳米SiO2和常规SiO2具有很好的生物相容性.     

Phenolic content,antioxidant and antibacterial activity of selected natural sweeteners available on the Polish market

Seventeen natural sweeteners available on the Polish market were screened for total phenolic content, by the Folin-Ciocalteu method, and for antioxidant activity, using the ferric reducing antioxidant power (FRAP) assay and the 2,2′-Azinobis (3-ethylbenzthiazoline-6-sulphonic acid) radical cation decolorization assay (ABTS^·+). In addition, we analyzed antibacterial activities against Staphylococcus aureus strains: both those susceptible and those resistant to methicillin (MRSA). The results of the study showed that total phenolic content, antioxidant activity and antibacterial activity differ widely among different samples of sweeteners. Phenolic content, expressed as a gallic acid equivalent, ranged from 0 mg kg^?1 in white, refined sugar, xylitol and wheat malt syrup to 11.4 g kg^?1 in sugarcane molasses. Antioxidant activity was lowest in refined white sugar, xylitol, brown beet sugar, liquid fructose, and rape honey; it was average in spelt syrup and corn syrup, and highest in sugar cane, beet molasses, date and barley syrups. Despite the great variety of sweeteners, a strong correlation was noted between the concentration of phenolics and antioxidant properties, as determined by the ABTS^·+ method (r = 0.97) and the FRAP assay (r = 0.77). The strongest antibacterial activity was observed in sugarcane molasses, which was lethal to S. aureus strains at 2 and 4% concentrations in medium for susceptible and MRSA strains respectively. Other sweeteners kill bacteria in 6–15% solutions, whereas some did not show any antibacterial activities against S. aureus strains, even at 20% concentrations. Due to their high antioxidant and antibacterial activities, some of the tested sweeteners have potential therapeutic value as supporting agents in antibiotic therapy.     

Effect of in vitro administered 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on DNA-binding activities of nuclear transcription factors in NIH-3T3 mouse fibroblasts

Abstract

Most modern pesticides are expensive. Application of excessive dosage rates is likely to cause undesirable biological side‐effects and is economically wasteful. Non‐uniform distribution of the spray cloud, or application at the wrong time, may result in failure to control the pest. It is the responsibility of the field operator to acquire sufficient knowledge and skill to ensure proper use of the control agents, to increase efficiency of their usage and to reduce unwanted side‐effects. To achieve this goal, he must take into consideration the various physical factors that govern field performance of pesticides.

A simple relationship exists between the spray volume and emission rate used, and droplet size produced. The use of extremely low spray volumes (i.e., those less than 2.0 litre per ha) for forest insect control in Canada, as opposed to higher volumes used in agriculture, necessitates the release of fine droplets (ranging from 20 to 70 μm in diameter) to obtain adequate coverage of the target area. These droplets take a long time to sediment downwards, evaporate in‐flight, become smaller in size and/or form powdery residues, thus contributing to off‐target drift and impaired droplet adhesion to target surfaces. Physical factors such as rain washing, degradation by sunlight and erosion by wind also influence the longevity of pesticide deposits on foliage which is crucial during the critical period of pest control.

Factors affecting the mode of entry into insects are related to the type of ingredients used in formulation. If a pesticide acts via crawling contact, formulations which would provide surface deposits would be more beneficial than emulsions or oil‐based mixes which tend to undergo penetration into foliar cuticle. Physical factors that affect field performance of a pesticide tank mix are related to phase separation and ‘breakdown of emulsions’ in the application equipment; ‘agglomeration and caking’ of wettable powder dispersions at the bottom of the tank; impaired flow behaviour of highly viscous formulations; and coarse atomization of high‐viscosity tank mixes leading to poor target cover.     

Genotoxicity monitoring of freshwater environments using caged crayfish (Astacus leptodactylus)

Genotoxicity of freshwater pollution was assessed by measuring DNA damage in haemocytes of caged freshwater crayfish Astacus leptodactylus by the means of Comet assay and micronucleus test, integrated with the measurements of physiological (total protein concentration) and immunological (total haemocyte count) haemolymph parameters as biomarkers of undergone stress. Crayfish were collected at the reference site (River Mre?nica) and exposed in cages for 1 week at three polluted sites along the Sava River (Zagreb, Sisak, Krapje). The long term pollution status of these locations was confirmed by chemical analyses of sediments. Statistically significant increase in DNA damage measured by the Comet assay was observed at all three polluted sites comparing to the crayfish from reference site. In addition, native crayfish from the mildly polluted site (Krapje) cage-exposed on another polluted site (Zagreb) showed lower DNA damage than crayfish from the reference site exposed at the same location indicating adaptation and acclimatisation of crayfish to lower levels of pollution. Micronuclei induction showed similar gradient of DNA damage as Comet assay, but did not reach the statistical significance. Observed increase in total haemocyte count and total protein content in crayfish from polluted environments in the Sava River also confirmed stress caused by exposure to pollution. The results of this study have proved the applicability of caging exposure of freshwater crayfish A. leptodactylus in environmental genotoxicity monitoring using Comet assay and micronucleus test.     

Recombinant human AhR-mediated GUS reporter gene assays for PCB congeners in transgenic tobacco plants in comparison with recombinant mouse and guinea pig AhRs

Four expression plasmids for recombinant human aryl hydrocarbon receptor (hAhR) consisting of a ligand binding domain of hAhR, a DNA-binding domain of LexA and a transactivation domain of VP16 as well as β-glucuronidase (GUS) reporter genes were constructed. All the expression plasmids were transformed into tobacco plants. The selected transgenic tobacco plants were used to assay. PCB congeners showed GUS activity in a TEF-dependent manner. The selected transgenic tobacco plant XhD4V17 was compared with the transgenic tobacco plants XmD4V26 and XgD2V23 containing recombinant mouse (m) AhR-mediated GUS reporter gene expression cassette and recombinant guinea pig (g) AhR-mediated GUS reporter gene expression cassette for PCB congener-inducible GUS activity. The data revealed that the tobacco plant XgD2V23 was the most active in PCB congener-inducible GUS activity. In a 1:1 mixture of PCB126 and PCB80 a reduced PCB126-induced GUS activity was observed in plant XgD2V23, which could possibly be due to interaction between PCB126 and PCB80.