Protective role of epigallocatechin 3-gallate against lead-induced toxicity in human neuroblastoma cells

Lead (Pb) is a heavy metal, known to induce oxidative stress and produce damage to the antioxidant defence system ultimately leading to cell death. Antioxidants such as epigallocatechin 3-gallate (EGCG), a green tea polyphenol, was shown to play a protective role during Pb-exposure. In this study, human SH-SY5Y neuroblastoma cells were exposed to different concentrations (0.01–10?µM) of Pb for 48?h to determine effects on the viability of cells. It was observed that IC₅₀ was at 5?µM and at this concentration the cells exhibited a significant increase in caspase-3 activity, an indicator of apoptosis at least by 10-fold and the decrease of 59.4% in glutathione (GSH) content. The total cellular prostaglandin-E₂ (PGE₂) level was found to be elevated at least 10-fold upon Pb exposure. However, the effects of Pb on cells pre-incubated with 50?µM EGCG followed by 5?µM Pb showed 40% inhibition in cell viability, 17.3% decrease in caspase-3 activity, 23% increase in GSH content, and 11.4% fall in PGE₂ levels when compared with cells exposed to Pb only. Data suggest that EGCG exerted a significant protection to cell viability in preventing cell death and elevation in levels of GSH in cells exposed to Pb. However, EGCG did not elicit any significant effect on release of PGE₂ indicating the nature of EGCG as an effective anti-apoptotic, antioxidant, and anti-inflammatory agent.     

Halogenated hydrocarbons and aryl hydrocarbons identified in human tissues

New methods for the preparation and purification of nephrotoxic cysteine conjugates of chloro‐ and fluoro‐alkenes are described. The methods are applicable to a wide range of cysteine, N‐acetyl cysteine and glutathione conjugates and examples have been given of the synthesis and characterisation of two novel glutathione conjugates.     

Biochemical speciation in chromium genotoxicity

The biochemical speciation of chromium compounds in mammalian cells is discussed with respect to uptake, metabolism, DNA binding and damaging. Whereas soluble hexavalent chromium is taken up rapidly and accumulated intracellularly after its reduction, compounds of trivalent chromium penetrate biomembranes about three orders of magnitude slower. Cr(VI) after its uptake is metabolised by electron donating compounds via Cr(V) to Cr(III) compounds. Chromium from various Cr(III) compounds, but not chromate, binds to chromatin in isolated cell nuclei. The DNA‐protein crosslinks and DNA strand breaks observed in rat liver and kidney after chromate administration are also found in vitro, when Cr(III) compounds (but not chromate) interacts with isolated nuclei. In the Chinese Hamster cell HGPRT mutation assay, three out of four tested Cr(III) complexes were found to be mutagenic. In a direct DNA strand break assay with supercoiled bacteriophage PM 2 DNA, neither chromate nor the four Cr(III) compounds tested caused nicks. However, the combined action of chromate plus glutathione as well as the isolated complex of pentavalent chromium, Na₄Cr(glutathione)₄, did cause DNA breaks. Reactive oxygen species are inferred to be the ultimate DNA nicking agents in this assay. In conclusion there appear to be two mechanisms of chromate genotoxicity; one with direct DNA damage caused by Cr(V) species and one via DNA‐protein crosslinks formed with Cr(III), the final reduction state of chromate.     

Sublethal concentrations of azinphos-methyl induce biochemical and morphological alterations in Rhinella arenarum embryos

Considering that amphibians are good sentinels of environmental conditions, Rhinella arenarum embryos were used to investigate the effects of sublethal concentrations of the organophosphorus insecticide azinphos-methyl, focusing on its anticholinesterasic or pro-oxidant actions and its possible connection with the appearance of morphological alterations. Early amphibian embryos exposed to azinphos-methyl displayed a protective response through glutathione S-transferase induction, along with superoxide dismutase inhibition. At intermediate embryonic stages, embryos exposed to azinphos-methyl displayed superoxide dismutase inhibition and morphological alterations, although cholinesterase activity was not altered, suggesting that molecular targets other than cholinesterase were involved in the development of morphological alterations. At the end of embryonic development, decreases in reduced glutathione and cholinesterase inhibition were observed, along with a significant increase in the number of malformed embryos. The connection between biochemical alterations and the appearance of malformations was not evident in R. arenarum embryos. However, increased glutathione S-transferase and decreased superoxide dismutase activities could be considered as early markers of exposure to azinphos-methyl. The results obtained demonstrate that sublethal concentrations of azinphos-methyl are a serious threat to toad embryos in their natural habitats because biochemical and morphological alterations could impair their ability to deal with environmental stresses.     

Biochemical biomarkers and overall health status of mussels Mytilus galloprovincialis exposed to nickel and chromium

This study examined the effects of nickel and chromium exposure on two biochemical biomarkers, glutathione peroxidase (GPX) and acetylcholinesterase (ACHE), in parallel with an indicator of overall health, Scope for Growth (SFG), in the mussel Mytilus galloprovincialis to evaluate their ecological relevance and to assess their potential use as biomarkers of Ni and Cr pollution. SFG integrates major physiological responses into an index that represents the energy status of the animals and is considered an ecologically relevant biomarker. Following exposure to a range of Ni and Cr concentrations effects of Ni and Cr were evident on GPX and SFG but not on ACHE. Accumulation in the mussels’ tissues increased according to exposure concentrations. Adverse effects on the energy status and induction of GPX were found at the higher exposure concentrations while possible hormetic effects on the energy status and moderate GPX suppression were associated with lower exposure concentrations. Nevertheless GPX was negatively correlated with SFG suggesting that GPX responses reflect the overall health of the organisms and thus show potential as an early warning signal of population level effects of Ni and Cr. On the other hand, our results indicate that ACHE is not a suitable biomarker for Ni and Cr pollution.     

O3对水稻叶片氮代谢、脯氨酸和谷胱甘肽含量的影响

臭氧(O3)被认为是重要的气污染物之一,水稻又是主要的粮食作物,因而准确地评估O3浓度升高对水稻生长发育的影响具有十分重要的意义。采用开顶式气室法模拟研究了O3对水稻叶片可见伤害症状、氮代谢、脯氨酸和谷胱甘肽含量的影响。结果显示,O3污染胁迫会导致水稻叶片产生明显的伤害症状,具体表现为:老叶叶鞘褪绿,有褐斑,直至完全干枯;稻穗小且黄化,籽粒不饱满;水稻成熟期提前等。O3浓度升高对水稻叶片的硝酸还原酶活性有显著影响。当O3浓度为40、80和120nL.L-1时,水稻叶片硝酸还原酶活性与对照组相比均降低,其中,分蘖期分别降低了25.3%、67.4%和86.3%;拔节期分别降低了57.4%、75.7%和97.8%;抽穗期分别降低了91.0%、97.2%和99.3%;乳熟期分别降低了89.5%、89.5%和96.7%。水稻叶片铵态氮和硝态氮含量随着O3浓度的升高而显著地降低,例如当O3浓度为40、80和120nL.L-1时,与对照相比,水稻叶片硝态氮含量分别降低46.3%、52.7%和65.7%,铵态氮含量分别降低6.5%、12.9%和43.4%。O3污染胁迫下水稻叶片脯氨酸含量在不同生长期变化不同,分蘖期、拔节期和抽穗期脯氨酸含量在40nL.L-1浓度O3熏蒸下急剧地提高,但是随着O3浓度的增加,脯氨酸含量又不断地降低。在水稻乳熟期,脯氨酸含量均随着O3浓度的增加而显著地下降。O3污染胁迫导致水稻叶片还原型谷胱甘肽(GSH)含量显著低于对照组,而氧化型谷胱甘肽(GSSG)含量显著高于对照组。当O3浓度为40、80和120nL.L-1时,乳熟期水稻叶片GSH含量分别比对照组降低68.7%、80.2%和78.2%,GSSG含量分别比对照提高494.4%、527.2%和439.8%。研究表明,O3污染胁迫对水稻叶片氮代谢和抗氧化系统产生了极显著的影响。     

O3浓度升高对不同生长期冬小麦叶片抗氧化系统的影响

由于人口的快速增长,人类活动导致近地层O3浓度不断提高,O3浓度升高将对植物、动物和人体健康产生极大的危害.采用开顶式气室(OTC)原位实验方法,研究O3浓度升高对不同生长期冬小麦叶片抗氧化系统的影响,进而分析O3对植物的伤害机制.结果表明,O3浓度升高可导致冬小麦拔节期和抽穗期叶片超氧化物歧化酶(SOD)、过氧化物酶...     

沙尘暴细颗粒物对大鼠肺泡巨噬细胞钙水平和脂质过氧化的影响

2004年3月采集甘肃省武威市和内蒙古包头市正常良好天气、局地扬沙天气和沙尘暴天气的大气细颗粒物(PM2．5)样品,以不同浓度的细颗粒物悬液体外处理大鼠肺泡巨噬细胞后测定细胞内丙二醛(MDA)、谷胱甘肽(IgSH)及胞质游离Ca^2 含量,并观察不同处理时间的细胞存活率．研究结果表明：(1)沙尘暴与非沙尘暴PM2．5均使细胞内MDA和Ca^2 含量升高、GSH含量下降,且对细胞毒性存在时间．效应和剂量．效应关系,(2)PM2．5对各指标的影响仅与处理剂量有关,而与样品种类无关．沙尘暴期间大气PM2．5浓度很高,因而沙尘暴PM2.5的危害作用不可忽视。     

Effect of herbicides on glutathione S-transferases in the earthworm, <Emphasis Type="Italic">Eisenia fetida</Emphasis>

AIM AND BACKGROUND: Earthworms have been studied as a readily available, easily maintainable and cheap test species for assessing chemical pollution, and may be an alternative to in vivo rodent bioassays. The current investigation aims to characterize detoxification enzymes in Eisenia fetida and stress response against two herbicides with different modes of action, namely, fenoxaprop and metolachlor. METHODS: Herbicides were applied to soil containing earthworms. Animals were then collected, sacrificed and shock-frozen. Extracted protein was analyzed for glutathione S-transferase (GST) activity using CDNB (1-chloro-2,4-dinitrobenzene), DCNB (1,2-dichloro-4-nitrobenzene), pNBC (p-nitrobenzylchloride), PNOBC (p-nitrobenz-o-ylchloride) and selected herbicides. GST isoenzymes were partially purified by affinity chromatography and molecular weights were estimated by SDS-PAGE. RESULTS: In E. fetida protein extracts, GST activity towards model compounds ranked as CDNB>DCNB>PNBOC>PNBC. Fluorodifen was not conjugated at all, but fenoxaprop and metolachlor were conjugated at low rates. Furthermore, the GST isoenzyme pattern changed during the incubation with herbicides, either due to stress or as a defense reaction. After incubation with monochlorobimane, a strong fluorescence of the intestinal tract and the intersegments was observed, indicating organ-specific GST induction. DISCUSSION: According to the author's knowledge, here, for the first time, evidence is presented that E. fetida GST are also capable of conjugating a wider range of xenobiotic substrates. Different forms of GST were observed and changes in GST isoforms due to the herbicide treatment were also noticed. GST conjugation rates varied between different herbicides used in this experiment. It might be assumed that herbicides may well be detoxified by earthworms, to a certain extent, but that they are also potent stress factors influencing the detoxification system of the animal. High doses or long exposure might lead to deleterious effects on earthworms and limit their survival rate. The use of the animals as bioindicators for herbicides and herbicide residues seems very promising, but is surely influenced by the lack of detoxification for some compounds. CONCLUSIONS: Conjugation of several xenobiotics with model substances and herbicides is proven in the earthworm E. fetida. However, E. fetida has only limited capabilities of detoxifying herbicidal compounds. Different isoforms of GST were involved and altered in their activity after treatment. RECOMMENDATIONS AND PERSPECTIVES: The accumulation of GS-conjugates and their determination via fluorescence microscopy is a quick and secure, additional marker for exposure that should be further developed to complement existing biotests. The described methods and endpoints might help to understand the complex reaction of earthworms towards herbicides and lead to an adapted test methodology.     

Changes in Lipid Peroxidation in the Gill and Muscle of the Marine Bivalve (Perna Viridis) During Exposure to Cadmium and Copper

The marine bivalve, Perna viridis was exposed to sublethal concentration (one third of 24 h LC₅₀) of either cadmium or copper for 1, 3 and 7 days and alterations in lipid peroxidation in the gill and muscle were investigated. Significant increase of the level of malondialdehyde, which is indicative of the peroxidative process, and a decrease of activity levels of glutathione peroxidase and glutathione transferase were observed in both the tissues of copper exposed mussels. the exposure of mussels to cadmium did not elicit any of the above changes.