Ultrastructural Effects Of Air Pollution on Lung Cells

The lungs of exposed mice taken during 2- to 3-hour heavy smog periods (over 0.4 ppm total oxidants) showed various degrees of cytoplasmic damage in the alveolar epithelial cells. The extent of damage was markedly age-dependent. Alveolar wall cells taken during heavy smog from 5-month-old animals contained slightly more lamellar inclusion bodies than corresponding animals kept in clean air. The cytoplasm of alveolar cells of 9-month-old animals sacrificed during heavy smog was severely disorganized; however, animals of this age showed a marked recovery 14 hours following the smog peak. In a group of older mice (21 months), similar cytoplasmic damage was obvious, and those sacrificed 24 hours after the heavy smog peak showed even more cellular disruption, suggesting irreversible damage in the older animals. The effect of synthetic photochemical smog showed a pattern of ultrastructural alterations similar to that of the heavy natural smog. Some permanent changes occurred in alveolar cells of 15-month-old mice. Partial recovery of lining cells took place, but few wall cells and phagocytes remained. If older lung tissue has relatively fewer wall cells as is indicated, recovery is decreased to the point of permanent damage. Coupled with extensive disruption of lining membranes, exposures of this nature may well cause the death of older animals.     

Selenium extractability from a contaminated fine soil fraction: implication on soil cleanup

Two batches of fine soil fraction of an acidic soil were deliberately contaminated with selenite (Se(IV)) and selenate (Se(VI)), respectively, and aged for more than 220 days. Speciation analysis using continuous flow-through hydride generation atomic absorption spectrometry (HGAAS) indicated that the species were predominant in their respective aged soils. A selective sequential extraction scheme was employed to fractionate the Se retained in the soils into six fractions of varying retentions. Abilities of various chemical reagents in extracting the Se in the two soil batches were then evaluated. The reagents investigated were sodium salts such as sodium chloride (NaCl), sodium sulfate (Na2SO4), sodium carbonate (Na2CO3), and sodium phosphate (Na3PO4), and two oxidants, namely, hydrogen peroxide (H2O2) and potassium permanganate (KMnO4). It was found that NaCl, Na2SO4, and Na2CO3 could only extract the exchangeable fraction of Se, while Na3PO4 could extract the exchangeable and strongly-bound fractions. Selenate was extracted more than Se(IV) by the salts. The kinetics of Se(IV) extraction by Na3PO4 could be best described by the Elovich model, while the Ritchie second-order model was the most appropriate to describe Se(VI) extraction. Efficiencies of the oxidants in Se(IV) extraction highly depended on their applied dosages. Both H2O2 and KMnO4 were able to extract greater than 93% of total Se, and therefore were significantly more effective than the salts in Se(IV) extraction.     

Atrazine promotes biochemical changes and DNA damage in a Neotropical fish species

The effects of Atrazine, an herbicide used worldwide and considered as a potential contaminant in aquatic environments, were assessed on the Neotropical fish Prochilodus lineatus acutely (24 and 48 h) exposed to 2 or 10 μg L⁻¹ of atrazine by using a set of biochemical and genetic biomarkers. The following parameters were measured in the liver: activity of the biotransformation enzymes ethoxyresorufin-O-deethylase (EROD) and glutathione S transferase (GST), antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), content of reduced glutathione (GSH), generation of reactive oxygen species (ROS) and occurrence of lipid peroxidation (LPO); in brain and muscle the activity of acetylcholinesterase (AChE) and DNA damage (comet assay) on erythrocytes, gills and liver cells. A general decreasing trend on the biotransformation and antioxidant enzymes was observed in the liver of P. lineatus exposed to atrazine; except for GR, all the other antioxidant enzymes (SOD, CAT and GPx) and biotransformation enzymes (EROD and GST) showed inhibited activity. Changes in muscle or brain AChE were not detected. DNA damage was observed in the different cell types of fish exposed to the herbicide, and it was probably not from oxidative origin, since no increase in ROS generation and LPO was detected in the liver. These results show that atrazine behaves as enzyme inhibitor, impairing hepatic metabolism, and produces genotoxic damage to different cell types of P. lineatus.     

Autometallographic tracing of mercury in frog liver

The distribution of mercury in the liver of the frog Rana ridibunda with the autometallographic method was investigated. The mercury specific autometallographic (HgS/SeAMG) technique is a sensitive histochemical approach for tracing mercury in tissues from mercury-exposed organisms. Mercury accumulates in vivo as mercury sulphur/mercury selenium nanocrystals that can be silver-enhanced. Thus, only a fraction of the Hg can be visualized. Six animals were exposed for one day and another group of six animals for 6 days in 1 ppm mercury (as HgCI2 ) dissolved in fresh water. A third group of six animals, served as controls, were sacrificed the day of arrival at the laboratory. First, mercury appears in the blood plasma and erythrocytes. Next, mercury moves to hepatocytes and in the apical part of the cells, that facing bile canaliculi. In a next step, mercury appears in the endothelial and Kupffer cells. It seems likely that, the mercury of hepatocytes moves through bile canaliculi to the gut, most probably bound to glutathione and/or other similar ligands. Most probably, the endothelial and Kupffer cells comprise the first line of defense against metal toxicity.     

Acid-base properties of humic substances from composted and thermally-dried sewage sludges and amended soils as determined by potentiometric titration and the NICA-Donnan model

The acid-base properties of humic acids (HAs) and fulvic acids (FAs) isolated from composted sewage sludge (CS), thermally-dried sewage sludge (TS), soils amended with either CS or TS at a rate of 80 t ha(-1)y(-1) for 3y and the corresponding unamended soil were investigated by use of potentiometric titrations. The non-ideal competitive adsorption (NICA)-Donnan model for a bimodal distribution of proton binding sites was fitted to titration data by use of a least-squares minimization method. The main fitting parameters of the NICA-Donnan model obtained for each HA and FA sample included site densities, median affinity constants and widths of affinity distributions for proton binding to low and high affinity sites, which were assumed to be, respectively, carboxylic- and phenolic-type groups. With respect to unamended soil HA and FA, the HAs and FAs from CS, and especially TS, were characterized by smaller acidic functional group contents, larger proton binding affinities of both carboxylic- and phenolic-type groups, and smaller heterogeneity of carboxylic and phenolic-type groups. Amendment with CS or TS led to a decrease of acidic functional group contents and a slight increase of proton binding affinities of carboxylic- and phenolic-type groups of soil HAs and FAs. These effects were more evident in the HA and FA fractions from CS-amended soil than in those from TS-amended soil.     

Interactions of forests with secondary air pollutants: some challenges for future research

The effects of ozone and other photochemical oxidants on individual trees have been studied for several decades, but there has been much less research on the potential effects on entire forest ecosystems. Given that ozone and other oxidants affect the production and subsequent fate of biogenic volatile organic compounds that act as signalling molecules, there is a need for more detailed study of the role of oxidants in modifying trophic interactions in forests. Deposition of fine particulates to forests may act as a source of nutrients, but also changes leaf surface properties, increasing the duration of surface wetness and modifying the habitat for epiphytic organisms, leading to increased risks from pathogens. Even where this pathway contributes a relatively small input of nutrients to forests, the indirect effects on canopy processes and subsequent deposition to the forest floor in throughfall and litter may play a more important role that has yet to be fully investigated.     

Surfactant production accompanying the modified Fenton oxidation of hydrocarbons in soil

A hydrocarbon-contaminated soil was treated in laboratory slurry reactors using two types of modified Fenton (MF) chemistry. Liquid hydrogen peroxide (HP) with Fe(3+) was compared with a calcium peroxide (CaO(2))-based oxidant (Cool-Oxtrade mark). Bioslurry treatment served as a control. During oxidation, samples of slurry filtrate were tested to quantify hydrocarbon concentrations and bulk surfactant concentrations, using the critical micelle dilution method. The results showed that both oxidants resulted in the temporary accumulation of surfactants to maximum levels of four times the critical micelle concentration, but that surfactants were completely removed by the end of treatment. Removal of surfactants was complete within 2h for liquid HP treatment versus 2 days for the CaO(2)-based oxidant. For both MF oxidants, hydrocarbon concentrations in filtrate were 3-4 orders of magnitude greater than in the biological control. Both MF oxidants also showed enhanced removal of the high molecular weight fractions of the petroleum hydrocarbons relative to biological treatment, though this effect was considerably greater with the CaO(2)-based oxidant. The chemical treatments did not considerably reduce numbers of culturable hydrocarbon-degrading microorganisms relative to the bioreactor, suggesting that chemical and biological oxidation may have occurred simultaneously in the slurry.     

Volatile organic sulfur compounds in a stratified lake

Three volatile organic sulfur compounds (VOSCs), dimethyl sulfide (DMS), carbon disulfide (CS(2)), and dimethyl disulfide (DMDS), were detected in the stratified water column of a lake (Linsley Pond) in Connecticut. The compounds DMS and DMDS appeared in both the oxic and the anoxic portions of the water column, CS(2) was primarily found in anoxic hypolimnion. Algal metabolism and/or bacterial degradation of sulfur-containing amino acids or other organic materials are potential sources of VOSCs in the oxic lake water. Reactions of hydrogen sulfide with organic compounds and microbial degradation of organic matter may be responsible for the production of VOSCs in the anoxic lake water. The vertical distribution patterns of these three VOSCs varied from month to month in the summer, but the daily profiles obtained in one 5-day period in the summer displayed consistency. No clear diurnal pattern for any of the three VOSCs was observed. Based on observation that these VOSCs were not present in surface and near surface waters of Linsley Pond, freshwater inputs of reduced sulfur compounds to the atmosphere may be insignificant.     

An insight into the mechanism of biosorption of copper by Bacillus polymyxa

Our main objective was to isolate bacteria from soil contaminated with effluents from electroplating wastewater and assess their potential to remove copper from the aqueous phase. The strain identified as Bacillus polymyxa accumulated copper inside the cell, where it bound to the cell wall. The intracellular metal accumulation led to the inhibition of dehydrogenases, which is essential for the energy deriving reactions. Addition of thiol group containing compounds, like reduced glutathione, dithiothreitol and mercapto-ethanol, revived the enzyme activity, implicating the sulfhydryl groups as the target of metal action. However, when the cells were exposed to higher concentration of Cu(II), irreversible enzyme denaturation occurred. Desorption with DTT and citrate showed that 82% of the metal was accumulated on cell surface and the contribution from metabolism-dependent intracellular accumulation was only 10-12%. The surface characterization of B. polymyxa indicated the presence of carboxyl, imidazolium, amino and phenolic groups, which might be responsible for metal uptake. Electron paramagnetic resonance (EPR) spectroscopy studies indicated that the metal coordinating environment could be either CuN₂O₂ or CuNO₃ chromophores, which may be the result of binding of Cu(II) to oxygen atoms of carboxylic groups of cell wall peptidoglycan and nitrogen atoms of amino-sugars or structural proteins. This environment got slightly altered after several treatments of B. polymyxa, leading to the formation of CuO₄, suggesting the coordination of copper to four oxygen atoms derived from carboxyl groups of peptidoglycan.     

Understanding particulate matter formation in the California San Joaquin Valley: conceptual model and data needs

Quantitative information from the 1995 Integrated Monitoring Study (IMS95) is used to develop a conceptual model, which describes the chemical characteristics and the physical processes responsible for the accumulation of PM in the San Joaquin Valley of California. One significant finding of the conceptual model is the sensitivity of ammonium nitrate (46% of winter PM_2.5) and nitric acid to oxidants, which may be VOC-sensitive rather than NO_x-sensitive. Key gaps in current knowledge are identified using the conceptual model, e.g., the relative sensitivity of winter oxidants to VOC and NO_x, mechanistic details of secondary organic aerosol formation, mechanisms of dispersion under calm conditions, and the importance of dry deposition. Some recommendations are also provided for the formulation of air quality models suitable to address the accumulation of PM in the San Joaquin Valley.     

Effect of methamidophos and urea application on microbial communities in soils as determined by microbial biomass and community level physiological profiles

In this study, we evaluated the effect of the application by two agrochemicals, methamidophos (O,S-dimethyl phosphoroamidothioate) and urea, on microbial diversity in soil, using the combined approaches of soil microbial biomass analysis and community level physiological profiles (CLPPs). The results showed that both a low and a high level of methamidophos application (CS2 and CS3) and urea application (CS4) significantly decreased microbial biomass C (Cmic) by 41-83% compared with the control (CS1). The soil organic C (Corg) values of CS3 and CS4 were significantly higher and lower by 24% and 14%, respectively, than that of CS1. Similarly to Cmic, the values of Cmic/Corg of the three applied soils which decreased were lower by 31-84% than that of CS1. In contrast, the respiration activity of the three applied soils were significantly higher than the control. Agrochemical application also significantly increased the soil total of N and P (Ntol and Ptol) and decreased the Corg/Ntol and Corg/Ptol values. The CLPPs results showed that the AWCD (average well color development) of the three applied soils were significantly higher than that of CS1 during the incubation period. Substrate richness, Shannon and Simpson indices of microbial communities under chemical stresses, increased significantly. In addition, the CFU (colony-forming unit) numbers of methamidophos metabolized bacteria in CS2 and CS3 also increased significantly by 86.1% and 188.9% compared with that of CS1. The combined results suggest that agrochemicals reduce microbial biomass and enhance functional diversities of soil microbial communities; meanwhile, some species of bacteria may be enriched in soils under methamidophos stress.     

Occurrence and determination of organic oxidants in rivers and wastewaters

Two iodometric methods for the concentration and analysis of water samples containing organic oxidants were applied to rivers and wastewaters in Illinois, Delaware, and Pennsylvania. Oxidant concentrations detected were as high as 2.5 § 10⁻⁵ M; the methods were capable of detecting as little as 10⁻⁸ M. Compounds possibly responsible for the observed oxidizing activity include N-chloro compounds, organic peroxides, or quinones.     

Disruption of thyroid hormone binding to sea bream recombinant transthyretin by ioxinyl and polybrominated diphenyl ethers

A number of chemicals released into the environment share structural similarity to the thyroid hormones (THs), thyroxine (T(4)) and triiodothyronine (T(3)) and it is thought that they may interfere with the thyroid axis and behave as endocrine disruptors (EDs). One of the ways by which such environmental contaminants may disrupt the TH axis is by binding to TH transporter proteins. Transthyretin (TTR) is one of the thyroid hormone binding proteins responsible for TH transport in the blood. TTR forms a stable tetramer that binds both T(4) and T(3) and in fish it is principally synthesized in the liver but is also produced by the brain and intestine. In the present study, we investigate the ability of some chemicals arising from pharmaceutical, industrial or agricultural production and classified as EDs, to compete with [I(125)]-T(3) for sea bream recombinant TTR (sbrTTR). Ioxinyl, a common herbicide and several polybrominated diphenyl ethers were strong inhibitors of [I(125)]-T(3) binding to TTR and some showed even greater affinity than the natural ligand T(3). The TTR competitive binding assay developed offers a quick and effective tool for preliminary risk assessment of chemicals which may disrupt the thyroid axis in teleost fish inhabiting vulnerable aquatic environments.     

Fluorescence spectroscopy of aromatic species produced in rich premixed ethylene flames

The fluorescence spectra of the condensed species (CS) collected in the soot inception region of a rich premixed laminar ethylene/oxygen flame have been measured by excitation in the UV at 266 and 355 nm excitation wavelength. The contribution of the most abundant polycyclic aromatic hydrocarbons (PAH) to the CS fluorescence has been evaluated in order to attribute the CS fluorescence at different emission wavelengths to specific aromatic structures. The fluorescence peaks detected in the UV region of the CS fluorescence spectrum was found to be mainly due to a typical PAH like fluorene, that is, the most fluorescent one among the PAH analyzed in the CS by chromatographic analysis. The CS exhibited the larger emission in the visible where the PAH contribution has been shown to be negligible and other fluorescing aromatic species, not identified by chromatographic analysis of the CS, have to be considered responsible for the visible fluorescence. Laser induced fluorescence (LIF) flame measurements excited at 266 nm and detected at two selected wavelengths (310 and 410 nm) have been performed along the flame axis and compared with the CS fluorescence intensity. The LIF and CS fluorescence signals show quite similar axial trends demonstrating that the LIF signals are related to CS fluorescence. In particular, the LIF fluorescence signals detected in the UV could be attributed to the PAH fluorescence whereas the unidentified species contained in the CS can be followed by LIF detection in the visible region.     

Surface chemistry of a pine-oil cleaner and other terpene mixtures with ozone on vinyl flooring tiles

Indoor environments are dynamic reactors where consumer products (such as cleaning agents, deodorants, and air fresheners) emit volatile organic compounds (VOCs) that can subsequently interact with indoor oxidants such as ozone (O₃), hydroxyl radicals, and nitrate radicals. Typically, consumer products consist of mixtures of VOCs and semi-VOCs which can react in the gas-phase or on surfaces with these oxidants to generate a variety of oxygenated products. In this study, the reaction of a pine-oil cleaner (POC) with O₃ (100 ppb) on a urethane-coated vinyl flooring tile was investigated at 5% and 50% relative humidity. These results were compared to previous α-terpineol + O₃ reactions on glass and vinyl surfaces. Additionally, other terpene and terpene alcohol mixtures were formulated to understand the emission profiles as seen in the POC data. Results showed that the α-terpineol + O₃ reaction products were the prominent species that were also observed in the POC/O₃ surface experiments. Furthermore, α-terpineol + O₃ reactions generate the largest fraction of oxygenated products even in equal mixtures of other terpene alcohols. This finding suggests that the judicial choice of terpene alcohols for inclusion in product formulations may be useful in reducing oxidation product emissions.     

Partitioning of Cd and Pb in the blood of European blackbirds (Turdus merula) from a smelter contaminated site and use for biomonitoring

Blood composition is commonly measured to assess exposure to and effects of metals on birds. In most of passerine species, only small volumes of blood may be sampled safely (<500 μL), which limit the measure of several markers of health status and chemical residues. Here, we documented the partitioning of Cd and Pb in the blood of European Blackbirds Turdus merula in order to propose usable relationships between whole blood concentrations (for which toxicological benchmarks exist) and those measured in the erythrocytes. Sixty-two blackbirds were trapped along a pollution gradient (smelter of Metaleurop Nord, Northern France). Blood was sampled and Cd and Pb concentrations were measured both in whole blood and in the erythrocytes only. Birds coming from the most contaminated sites exhibited high residues for both Cd and Pb. We assessed that 73% and 99% of Cd and Pb, respectively, were associated to erythrocytes. Strong linear relationships, that were not influenced by neither the age nor the sex, were established between whole blood residues and those of erythrocytes for Cd and Pb (adj-R² = 0.78 and 0.93, respectively). Present findings are promising to optimize the use of small blood samples in order to investigate several responses relative to wildlife health status.     

工业废气中氮氧化物的治理研究

对碱液吸收和活性炭吸附两级联合治理氮氧化物废气进行了实验研究,结果表明:对于氮氧化物进口浓度为7000～10 000 mg/m3的氮氧化物废气,氮氧化物的平均脱除率可以达到99%,出口浓度低于99 mg/m3;还进行了以硝酸溶液、双氧水溶液和高锰酸钾溶液为氧化剂湿法氧化碱吸收的实验研究,结果显示3种氧化剂都能显著提高氮氧化物的脱除率,其中以高锰酸钾溶液作氧化剂的脱除效果最好,其次是双氧水溶液.     

How Plants Cope with Foreign Compounds. Translocation of xenobiotic glutathione conjugates in roots of barley (Hordeum vulgare) (9 pp)

Background, Aim and Scope Numerous herbicides and xenobiotic organic pollutants are detoxified in plants to glutathione conjugates. Following this enzyme catalyzed reaction, xenobiotic GS-conjugates are thought to be compartmentalized in the vacuole of plant cells. In the present study, evidence is presented for long range transport of these conjugates in plants, rather than storage in the vacuole. To our knowledge this is the first report about the unidirectional long range transport of xenobiotic conjugates in plants and the exudation of a glutathione conjugate from the root tips. This could mean that plants possess an excretion system for unwanted compounds which give them similar advantages as animals. Materials and Methods: Barley plants (Hordeum vulgare L. cv. Cherie) were grown in Petri dishes soaked with tap water in the greenhouse. - Fluorescence Microscopy. Monobromo- and Monochlorobimane, two model xenobiotics that are conjugated rapidly in plant cells with glutathione, hereby forming fluorescent metabolites, were used as markers for our experiments. Their transport in the root could be followed sensitively with very good temporal and spatial resolution. Roots of barley seedlings were cut under water and the end at which xenobiotics were applied was fixed in an aperture with a thin latex foil and transferred into a drop of water on a cover slide. The cover slide was fixed in a measuring chamber on the stage of an inverse fluorescence microscope (Zeiss Axiovert 100). - Spectrometric enzyme assay. Glutathione S-transferase (GST) activity was determined in the protein extracts following established methods. Aliquots of the enzyme extract were incubated with 1-chloro-2,4-dinitrobenzene (CDNB), or monochlorobimane. Controls lacking enzyme or GSH were measured. - Pitman chamber experiments. Ten days old barley plants or detached roots were inserted into special incubation chambers, either complete with tips or decapitated, as well as 10 days old barley plants without root tips. Compartment A was filled with a transport medium and GSH conjugate or L-cysteine conjugate. Compartments B and C contained sugar free media. Samples were taken from the root tip containing compartment C and the amount of conjugate transported was determined spectro-photometrically. Results: The transport in roots is unidirectional towards the root tips and leads to exsudation of the conjugates at rates between 20 and 200 nmol min-1. The microscopic studies have been complemented by transport studies in small root chambers and spectroscopic quantification of dinitrobenzene-conjugates. The latter experiments confirm the microscopic studies. Furthermore it was shown that glutathione conjugates are transported at higher rates than cysteine conjugates, despite of their higher molecular weights. This observation points to the existence of glutathione specific carriers and a specific role of glutathione in the root. Discussion: It can be assumed that long distance transport of glutathione conjugates within the plant proceeds like GSH or amino acid transport in both, phloem and xylem. The high velocity of this translocation of the GS-X is indicative of an active transport. For free glutathione, a rapid transport-system is essential because an accumulation of GSH in the root tip inhibits further uptake of sulfur. Taking into account that all described MRP transporters and also the GSH plasmalemma ATPases have side activities for glutathione derivatives and conjugates, co-transport of these xenobiotic metabolites seems credible. - On the other hand, when GS-B was applied to the root tips from the outside, no significant uptake was observed. Thus it can be concluded that only those conjugates can be transported in the xylem which are formed inside the root apex. Having left the root once, there seems to be no return into the root vessels, probably because of a lack of inward directed transporters. Conclusions: Plants seem to possess the capability to store glutathione conjugates in the vacuole, but under certain conditions, these metabolites might also undergo long range transport, predominantly into the plant root. The transport seems dependent on specific carriers and is unidirectional, this means that xenobiotic conjugates from the rhizosphere are not taken up again. The exudation of xenobiotic metabolites offers an opportunity to avoid the accumulation of such compounds in the plant. Recommendations and Perspectives: The role of glutathione and glutathione related metabolites in the rhizosphere has not been studied in any detail, and only scattered data are available on interactions between the plant root and rhizosphere bacteria that encounter such conjugates. The final fate of these compounds in the root zone has also not been addressed so far. It will be interesting to study effects of the exuded metabolites on the biology of rhizosphere bacteria and fungi.     

Antioxidant effect of Sargassum polycystum (Phaeophyceae) against acetaminophen induced changes in hepatic mitochondrial enzymes during toxic hepatitis

The present study was aimed to examine the protective effects of Sargassum polycystum (Phaeophyceae) alcoholic extract on changes in liver mitochondrial enzymes against acetaminophen induced toxic hepatitis in experimental rats. The levels of protein, lipid peroxide, glutathione (GSH) in mitochondrial fraction, superoxide dismutase (SOD) and catalase (CAT) were also determined. The activities of tricarboxylic acid enzymes such as isocitrate dehydrogenase (ICD), alpha-ketoglutarate dehydrogenase (alpha-KGD), succinate dehydrogenase (SD), malate dehydrogenase (MD), NADH dehydrogenase, and cytochrome-c-oxidase were determined in mitochondrial fraction. The rats intoxicated with acetaminophen showed significant elevation in the levels of lipid peroxides with decreased levels of protein, GSH, SOD, CAT and impaired tricarboxylic acid cycle enzyme activities. The prior oral administration of S. polycystum alcoholic extract showed significant diminution in the severity of toxic hepatitis in acetaminophen-induced rats by maintaining the activities of tricarboxylic acid enzymes with concomitant improvement in the hepatic mitochondrial antiperoxidative status when compared with intoxicated animals. The results obtained in the present study indicate that the protective effects of S. polycystum extract may be due to the presence of some active compounds that are inhibitory against the free radicals generated during lipid peroxidation in acetaminophen induced toxic hepatitis.