Mitochondrial dysfunction,impaired oxidative-reduction activity,degeneration, and death in human neuronal and fetal cells induced by low-level exposure to thimerosal and other metal compounds

Thimerosal (ethylmercurithiosalicylic acid), an ethylmercury (EtHg)-releasing compound (49.55% mercury (Hg)), was used in a range of medical products for more than 70 years. Of particular recent concern, routine administering of Thimerosal-containing biologics/childhood vaccines have become significant sources of Hg exposure for some fetuses/infants. This study was undertaken to investigate cellular damage among in vitro human neuronal (SH-SY-5Y neuroblastoma and 1321N1 astrocytoma) and fetal (nontransformed) model systems using cell vitality assays and microscope-based digital image capture techniques to assess potential damage induced by Thimerosal and other metal compounds (aluminum (Al) sulfate, lead (Pb)(II) acetate, methylmercury (MeHg) hydroxide, and mercury (Hg)(II) chloride) where the cation was reported to exert adverse effects on developing cells. Thimerosal-associated cellular damage was also evaluated for similarity to pathophysiological findings observed in patients diagnosed with autistic disorders (ADs). Thimerosal-induced cellular damage as evidenced by concentration- and time-dependent mitochondrial damage, reduced oxidative–reduction activity, cellular degeneration, and cell death in the in vitro human neuronal and fetal model systems studied. Thimerosal at low nanomolar (nM) concentrations induced significant cellular toxicity in human neuronal and fetal cells. Thimerosal-induced cytoxicity is similar to that observed in AD pathophysiologic studies. Thimerosal was found to be significantly more toxic than the other metal compounds examined. Future studies need to be conducted to evaluate additional mechanisms underlying Thimerosal-induced cellular damage and assess potential co-exposures to other compounds that may increase or decrease Thimerosal-mediated toxicity.     

Maternal transfer of mercury to the developing embryo/fetus: is there a safe level?

Mercury (Hg) exposure is ubiquitous in modern society via vaccines, fish/crustacea, dental amalgam, food, water, and the atmosphere. This article examines Hg exposure in the context of primary exposure to pregnant women and secondary exposure experienced by their unborn babies. Babies in utero are particularly at risk of higher Hg exposure than adults (on a dose/weight basis through maternal Hg transfer via the placenta), and are more susceptible to adverse effects from mercury and its biologically active compounds. It is, therefore, critical that regulatory advisories around maximum safe Hg exposures account for pregnant women and secondary exposure that children in utero experience. This study focused on standardized embryonic and fetal Hg exposures via primary exposure to the pregnant mother of two common Hg sources (dietary fish and parenteral vaccines). Data demonstrated that Hg exposures, particularly during the first trimester of pregnancy, at well-established dose/weight ratios produced severe damage to humans including death. In light of research suggestive of a mercuric risk factor for childhood conditions such as tic disorders, cerebral palsy, and autism, it is essential that Hg advisories account for secondary prenatal human exposures.     

Photosynthetic performance,oxidative damage and antioxidants in <Emphasis Type="Italic">Cylindrotheca closterium</Emphasis> in response to high irradiance,UVB radiation and salinity

Cylindrotheca closterium is a common marine diatom living in intertidal environments where it can be present both in the water column and on sediments, depending on the tidal regime. In the present work this diatom was employed to investigate the responses to desiccation and to increase in PAR and UVB intensity, as occurs during emersion. Under these circumstances, the production of active oxygen species (AOS) may be enhanced resulting in an oxidative stress. Stress responses in this species were measured by exposing it to normal (30) and double salinity (60), supplying light of low or high intensity for 12 h, in the latter case either without or with moderate dose rates of UVB. Pulse amplitude modulated fluorometry was used to measure Chl a autofluorescence (F ₀), an index of photosynthetic efficiency of PSII (F _v/F _m) and the relative electron transfer rate (rETR). The oxidative stress was evaluated by analysing GSH pools and SOD activity. It was observed that at double salinity and under low light, intracellular pools of reduced glutathione (GSH) were higher than under the two conditions of high light without and with UVB at both salinities. The antioxidative defence activity of superoxide dismutase (SOD) was far higher under hypersaline conditions. The oxidative damage was evaluated as protein and lipid damage. The results showed that it expressed itself mainly through protein peroxidation: at normal salinity relative protein carbonyl content was (a) twice as high as in cells grown at double salinity, and (b) three times as high under UVB. Total unsaturated lipid contents doubled under hypersalinity conditions. The lipid peroxidation marker malondialdehyde showed the strongest response to low light and UVB at salinity value of 60. Lipid peroxide content was significantly higher at salinity of 60 compared to normal salinity and was the highest under low light and high light with UVB. The simulated emersion condition of the diatom seems to lead to the establishment of a balance between damage and repair, expressed mainly as (a) oxidative protein damage at normal salinity, in particular due to UV radiation, (b) sufficient protection by SOD activity mainly under hypersaline conditions.     

对二甲苯对褐牙鲆(Paralichthys olivaceus)幼鱼的毒性效应研究

采用急性和28 d慢性暴露试验研究了海洋典型危险化学品对二甲苯对褐牙鲆幼鱼的致死效应和生长抑制效应,并测定了遗传毒性、神经毒性以及免疫毒性效应相关的毒理学评价指标。研究结果表明:对二甲苯对褐牙鲆幼鱼的96 h-LC50为45.7 mg·L~(-1),根据国家环保局水和废水监测分析方法编委会规定的化学物质对鱼类毒性分级标准,属于中等毒性,浓度高于2.3 mg·L~(-1)的暴露能显著抑制褐牙鲆幼鱼的生长;4.6和9.2 mg·L~(-1)浓度组观察到对二甲苯暴露28 d后褐牙鲆肝脏中的丙二醛含量与DNA损伤程度显著升高,脑组织中乙酰胆碱酯酶(Ach E)活力被明显抑制;9.2 mg·L~(-1)浓度组暴露28 d后,褐牙鲆幼鱼体内总血细胞数量和溶菌酶活力显著降低。这些结果表明较高浓度的对二甲苯长期作用可导致褐牙鲆幼鱼的氧化胁迫,造成其肝脏毒性损伤并产生神经毒性与免疫毒性。本实验结果为了解对二甲苯对鱼类的毒性作用机制以及对二甲苯的海洋生态风险评估提供了科学依据。     

Involvement of mitochondrial-mediated caspase-3 activation and lysosomal labilization in acrylamide-induced liver toxicity

Acrylamide (ACR) is a chemical frequently used in both industrial and synthetic processes and may be produced during food processing. ACR at very high concentrations is postulated to exert its toxicity through the stimulation of an oxidative stress. ACR in excessive doses induces the central nervous system, reproduction, and genetic toxicity. However, ACR effects on the liver, a major organ of drug metabolism, have not been adequately explored. In addition, the role of mitochondria in an ACR-mediated hepatotoxicity is still unclear. The aim of this study was to investigate the cytotoxic mechanisms attributed to ACR using isolated rat hepatocytes. Hepatocytes were isolated by the collagenase perfusion method and incubated with an EC50_2hr concentration of ACR for 3 hr. The EC50_{2 hr} of ACR on isolated rat hepatocytes was determined to be 1 mM. Based on our results, hepatocytes cytotoxicity of ACR (1 mM) was mediated by a reactive oxygen species formation and lipid peroxidation. Incubation of hepatocytes with ACR produced rapid hepatocyte glutathione depletion which is another marker of the cellular oxidative stress. ACR cytotoxicity was also associated with mitochondrial injury as evidenced by the decline of mitochondrial membrane potential and lysosomal membrane leakiness. Our results also showed that ACR induced caspase-3 activation, the final mediator of apoptosis signaling. These findings contribute to a better understanding underlying mechanisms involved in ACR hepatotoxicity originating from the oxidative stress and ending in mitochondrial/lysosomal damage and cell death signaling.     

Inorganic and methyl-mercury speciation in sediments of the Swarzędzkie Lake

Sediment samples were analysed for mercury and methylmercury content in different parts of the bottom sediment of Swarz ?dzkie Lake, which were influenced by different external pollution sources. The results of determination with two methods of mineralization using two separate media (HNO₃/H₂O₂ and HF) were compared. The accuracy of the studied methods was analysed using certified reference material IAEA 405 of river sediment. The recovery of mercury was satisfactory and ranged from 97.5 to 98.8%. Methylmercury compounds in the studied sediments were found in limited concentrations. Their concentrations ranged from 0.26 to 58.1 μg kg^?1, i.e. 1.0–7.4% of the total mercury content. The depth profile displayed high values of both total and methylmercury content at a depth of 10–20 cm, related to the heavy pollution of the lake in the 1980s. Canonical analysis displayed the relationship of both total mercury and methylmercury concentrations with organic matter and phosphorus content in bottom sediments.     

Effect of latex material on antioxidant enzymes,lipid peroxidation,DNA damage,and chromosomal aberration

Cell integrity is affected by oxidative stress when the production of active oxidants overwhelms antioxidant defense mechanisms. Latex, a natural polymer obtained from Hevea brasiliensis, is used in medical industry for manufacturing surgical gloves, urinary catheters, and dental dams. The aim of this study was to evaluate the effects of latex material on oxidative stress by in vivo and in vitro methods. In addition, the material was screened for its ability to induce any chromosomal aberrations (CAs) by in vitro method. In vivo studies were carried out with implanted latex material onto subcutaneous tissue of various batches of experimental Wistar rats. At the end of experimental period, animals were anesthetized, blood was collected for serum analysis, and sacrificed. Liver was excised for the determination of antioxidant enzymes and lipid peroxidation (LPO). Subcutaneous tissues were obtained for the extraction of genomic DNA from implanted animals and checked for the presence of 8-hydroxy-2-deoxyguanosine (8-OHdG), considered an indicator of DNA damage. Simultaneously, in vitro studies were carried out using fresh liver and subcutaneous tissue obtained from Swiss albino mice treated with physiological saline extract of latex material. For the estimation of both in vitro and in vivo oxidative stress, 10% liver homogenate was assessed for stress indicators like reduced glutathione, glutathione reductase, glutathione peroxidase, LPO and protein content. The results of both in vivo and in vitro studies indicated that the chemical leachents from the latex material did not significantly affect LPO and the levels of antioxidant enzymes. There was also no significant increase in 8-OHdG content due to the presence of implanted latex material. Finally, the results of in vitro CA test and G banding indicated that extracts of test material did not induce any chromosomal abnormalities.     

Ameliorating effects of hormone and Cu on salt stressed Brassica nigra (L.) Koch with reference to growth and biochemical characteristics

The ameliorating effects of exogenous hormone and calcium were investigated in salt-stressed black mustard seedlings (Brassica nigra L.). The seedlings were subjected to various treatments: control (nutrient solution), salt stress, kinetin/abscisic acid (ABA)/calcium chloride (CaCl₂). Sodium chloride (NaCl) inhibited shoot length (45–55%) and root (<75%). Further, alterations in superoxide dismutase and catalase activities were consistent with changes in levels of reactive oxygen species and malondialdehyde (MDA). The beneficial effects of ABA and CaCl₂ in ameliorating salt stress may be attributed to the reduction of membrane peroxidation and increased glutathione (GSH) levels. These observations suggest that oxidative stress resulting from salt stress in B. nigra may result in production of antioxidative enzymes to counteract oxidative damage, and the enzymes may contribute to the ability of B. nigra to survive adverse conditions. Data indicate that hormone and ion in the order of kinetin > ABA > calcium alleviate the adverse impact of salt on B. nigra.     

Structural and molecular alterations in arsenic-induced hepatic oxidative stress in rats: a FTIR study

Arsenic (As) is an ubiquitously distributed environmental toxicant predominantly contaminating drinking water. A number of studies indicated that oral exposure of humans to inorganic As produced damage to various body tissues including liver. Oxidative stress is thought to play a major role in As-induced hepatotoxicity. In this study, Fourier transforms infrared (FTIR) spectroscopy approach was applied to determine whether chronic As exposure at 25 ppm, intragastrically for 12 weeks, affected oxidative stress status in rat liver. Data demonstrated that chronic As administration exacerbated oxidative stress as was evidenced by suppressed antioxidant defense system and increased lipid peroxidation and protein oxidation. The FTIR study showed that peak area value of amide A decreased significantly followed by reduced amide I and amide II peak area in an As-treated rat liver suggesting altered protein profile. The change of Olefinic?CH stretching band and C?O stretching of triglycerides band indicated the altered lipid levels due to metallic exposure. The fall in the peak area of PO₂^? asymmetric stretching in the As-treated group might be due to compositional changes of nucleic acids. Hence, the results of this study indicate that As-induced oxidative stress was associated with structural and molecular modifications in proteins, lipids, and glycogen in a rat liver that may help to elucidate molecular mechanisms underlying metal-mediated hepatic damage.     

Ultrastructural responses and oxidative stress induced by cypermethrin in the liver of Labeo rohita

The present study was conducted to establish the relationship between selected oxidative stress parameters and ultrastructural responses in liver tissue of Labeo rohita fingerlings exposed to cypermethrin. Fish were exposed to lethal (4.0 μg L^?1) and sublethal (0.4 μg L^?1) concentrations of cypermethrin for a period of 24, 48, 72 and 96 h for acute studies and 1, 5, 10 and 15 days for subacute studies, respectively. Results showed increased catalase (CAT) and protease activity, hydrogen peroxide (H₂O₂), malondialdehyde (MDA), protein carbonyls and free amino acid (FAA) levels at both concentrations. This suggests participation of free-radical-induced oxidative cell injury in mediating the hepatotoxicity of cypermethrin. In corroboration of this, ultrastructural lesions witnessed a reduction in the number of cell organelles, swollen, vacuolated and condensed mitochondria, dilated rough endoplasmic reticulum, and reduced numbers of smooth enodplasmic reticulum, peroxisomes and lysosomes at the lethal (4.0 μg L^?1) concentration. At the sublethal (0.4 μg L^?1) concentration, cytoplasmic vacuolation, condensed, vacuolated and swollen mitochondria, dilated rough endoplasmic reticulum and an absence of hepatocyte microvilli were prominent. Ultrastructural changes were exhibited as subcellular responses due to the imbalance in cellular oxidative status by means of oxidative damage.     

GO通过氧化损伤激活秀丽线虫未折叠蛋白反应

以秀丽线虫为受试生物,探讨氧化石墨烯(GO)对秀丽线虫未折叠蛋白应答的激活及其与氧化应激的关系。将L4幼虫暴露于GO中24 h,通过检测活性氧(ROS)水平、乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)等指标评估GO对秀丽线虫所致氧化损伤;以hsp-4和hsp-6分别作为秀丽线虫内质网和线粒体未折叠蛋白反应(UPR)的报告基因评价GO对秀丽线虫UPR应答的激活;同时以谷胱甘肽(GSH)作为抗氧化剂探讨GO所致UPR应答与氧化应激的关系。研究结果显示,GO暴露后秀丽线虫体内ROS、LDH和MDA水平升高,抗氧化酶活性上升;内质网和线粒体UPR反应增强,GSH预处理可以降低内质网和线粒体UPR应答。研究表明,GO短期暴露可以诱导秀丽线虫机体氧化应激进而激活UPR应答,抗氧化保护可以降低UPR应答反应。     

Electron microscopy documenting the cellular metabolic fate of Zn IONS∗

In vertebrates and invertebrates Zn exist as complexed compounds with metallothioneins. However, its cellular level effects and metabolic fates are scantly documented. In the elucidation of this fact, EM results on hepatic cellular alterations in fish under lethal dose exposure at 4.0 ppm over a week is illustrated.

A large number of lysosomes in hepatic cells prevailed on exposure to Zn in its sulfate form. Evidently, due to metal compound stress and cellular damage lysosomal activity is augmented. The lysosomes harboured digestible material, presumably the aforementioned substrates. Contrary to it, fat droplets prevailed while glycogen depletion is noticeable. Unlike the effects of Hg, the nuclei were normal with granular chromatin and prominent nucleoli. However, the mitochondria contained some small intramitochondrial bodies. Similar to the effects of Hg, the cell membrane remained intact.

In vivo enzymatic studies indicated augmentation in catalase, acid‐ and alkaline‐phosphatases, while glucose‐6‐phosphatase is inhibited. However, only alkaline‐ and glucose‐6‐phosphatases are inhibited under in vitro conditions.

Thus, it is evident that Zn enhances cellular bioenergetic requirements culminating in glycogen depletion owing to stress, concomitantly envisaging inhibition of oxidative phosphorylation in the electron transport system.     

有机磷酸酯对斑马鱼的早期神经毒性作用研究

国内外研究已证实,有机磷酸酯广泛分布于多种环境介质中,但目前仍缺乏足够的数据阐明有机磷酸酯具有早期神经毒性效应及其可能的毒性作用机制。本研究采用模式动物斑马鱼(Danio rerio)作为研究对象,选择了环境中3种典型的有机磷酸酯类化合物包括磷酸三苯酯(TPP)、2-乙基己基二苯基磷酸酯(EHDPP)和磷酸三(2-氯)乙酯(TCEP),从斑马鱼运动行为、氧化应激和神经发育关键基因的转录等方面阐述有机磷酸酯的早期神经毒性作用及可能的作用机制。研究发现,TPP(0.1和1 mg·L~(-1))、EHDPP(0.2和2 mg·L~(-1))和TCEP(0.5和5 mg·L~(-1))可能通过诱导氧化应激并下调神经发育关键基因(mbp和syn2a)的转录从而显著抑制斑马鱼的运动行为。本研究可以为有机磷酸酯类阻燃剂及其替代产品的生产、使用和危险度评估提供直接依据。     

Impaired behavior and changes in some biochemical markers of bivalve (Ruditapes decussatus) due to zinc toxicity

Susceptibility and response of bivalves, the clam Ruditapes decussatus, to zinc (Zn) were studied by monitoring behavior using valve movement and some biochemical parameters. The LC₅₀ was 4.1 mg L^?1 at 7 days. Depuration of Zn from their tissues was also followed. Accumulation of Zn increased with concentration and duration of exposure. After 20 days, Zn caused impairment of valve movement as well as the antioxidant system, decreasing glutathione and protein levels and increasing the lipid peroxidation which is indicative of oxidative damage. Results suggested that behavior and biochemical parameters of clam R. decussatus were sensitive and suitable responses for assessing the effects of Zn on the aquatic ecosystems. It could be concluded that there is a time–dose–response relationship of Zn with behavior and oxidative stress of R. decussatus.     

全氟辛烷磺酸(PFOS)对三角帆蚌肝胰腺的氧化性损伤

PFOS是典型的持久性有机污染物,迁移能力强,具有较高的生物可利用性和蓄积能力,且具有广泛的生物毒性。为探究PFOS对淡水底栖生物的毒性作用机制,以三角帆蚌为研究对象,进行了不同剂量(0.1、1.0、5.0 mg·L-1)的PFOS胁迫和净水恢复实验,期间对受试生物肝胰腺中的谷胱甘肽(GSH)含量、谷胱甘肽-S转移酶(GST)活性、超氧化物歧化酶(SOD)活性,以及谷丙转氨酶(ALT)和谷草转氨酶(AST)的活性进行了连续测定。结果发现,低浓度胁迫(0.1 mg·L-1)对各项指标均有不同程度的诱导作用,且持续时间较长;而在中高浓度PFOS胁迫下,则呈现出明显的诱导向抑制过渡的时间效应。GSH含量和GST活性具有较高的相关性(P0.05)。恢复实验中,所测指标普遍未恢复到对照组水平,说明P FOS胁迫损伤的恢复需要更长的时间。研究表明,PFOS对三角帆蚌肝胰腺的氧化胁迫显著,并能快速地激活肝胰腺细胞的解毒代谢;但长期的PFOS胁迫则会造成肝胰腺细胞的实质性损伤。     

BDE-47对斑马鱼胚胎-幼鱼的急性毒性及氧化应激作用

为研究2,2',4,4'-四溴联苯醚(BDE-47)对斑马鱼胚胎-幼鱼急性毒性、氧化应激及细胞凋亡的影响,以受精后3 h的斑马鱼胚胎为染毒对象,用概率单位法计算BDE-47对斑马鱼胚胎-幼鱼的96 h-LC₅₀;再参照96 h-LC₅₀按一定比例级差设置0.25、0.5、1.0、2.0 mg·L^-1 4个浓度组和1个对照组(0 mg·L^-1)进行96 h半静水式毒性试验,检测斑马鱼超氧化物歧化酶(SOD)及过氧化氢酶(CAT)活性、丙二醛(MDA)含量和细胞凋亡情况。结果表明,BDE-47对斑马鱼96 h的急性毒性LC₅₀为3.77 mg·L^-1(95%可信区间1.93～10.27 mg·L^-1);2.0 mg·L^-1剂量组与对照组相比,SOD活性和MDA含量显著增加,且CAT活性与BDE-47染毒浓度之间存在明显的剂量-效应关系;0.5 mg·L^-1 BDE-47染毒96 h后,即观察到斑马鱼幼鱼出现明显细胞凋亡,主要集中于神经管和脑部。研究表明,BDE-47可以影响斑马鱼体内抗氧化防御系统,并能诱导细胞凋亡;BDE-47导致神经组织的氧化损伤可能在动物神经毒性中起重要作用。     

Testosterone concentrations and male genital organ morphology in Greenland sledge dogs (Canis familiaris) dietary exposed to organohalogen contaminants

This study investigated whether low-level, long-term in utero and post-natal exposure to organohalogen pollutants disrupts male reproductive organ morphology and testosterone production in Greenland sledge dogs (Canis familiaris), as a model of Arctic top predators feeding on marine mammals. Six male dogs were followed for 1 year and testosterone concentrations, testes/baculum morphology and baculum bone mineral density (BMD) was determined. Three males were exposed to organohalogenated contaminants (OHCs) in utero through maternal dietary intake of minke whale blubber (Balaenoptera acutorostrata), with a post-weaning ∑OHC intake of 10.4–11.7 µg kg^?1 day^?1 resulting in an adipose tissue range of ∑OHC 4518–5729 ng (g lw)^?1 after 1 year. Three control males were exposed to very low concentrations of OHCs through pork fat. No significant differences were seen in plasma testosterone concentrations, baculum weight, BMD, and testicular length in the six male dogs (control, n = 3 and exposed, n = 3) measured at 3, 5, 7, 9, and 12 months of age. Testicular weights were significantly lower in the exposed group (p = 0.015, n = 2). Although this study had a limited number of animals, it was observed that in utero and the following 12 months of chronic exposure to a complex mixture of contaminants in the form of naturally accumulated OHCs does not affects testosterone levels, but possibly affects testicular weights in sledge dogs.     

铅与纳米SiO2联合染毒对A549细胞氧化损伤的影响

为了研究铅与纳米SiO2联合染毒所致的细胞损伤特征,并从氧化应激方面探讨其可能的作用机制。用铅和SiO2处理A549细胞,采用四唑盐(MTT)比色法检测细胞存活率,评价铅和SiO2联合染毒所致的细胞损伤特征;采用硫代巴比妥酸(TBA)比色法检测细胞内丙二醛(MDA)含量,评价铅与SiO2联合染毒所致细胞的氧化应激状态;检测了细胞内抗氧化物还原型谷胱甘肽(GSH)含量以及细胞内抗氧化酶的活性,以评价铅与SiO2联合染毒对细胞抗氧化系统的影响。将实验数据进行ANOVA分析。结果表明,铅、SiO2单独染毒组各指标没有明显改变;而联合染毒能造成细胞氧化损伤,表现为细胞存活率、GSH水平、超氧化物歧化酶(SOD)及谷胱甘肽过氧化物酶(GSH-Px)活性显著低于对照组及2个单独染毒组(P<0.05),细胞内MDA含量显著高于对照组及各单独染毒组(P<0.05)。可见,联合染毒可引起明显的细胞毒性,氧化损伤可能是铅与SiO联合染毒致肺细胞毒性损伤的作用机制之一。     

Toxicity of 4-methylimidazole on isolated brain mitochondria: using both in vivo and in vitro methods

4-methylimidazole (4MI) is a compound widely used in various industrial and consumer applications. The most important sources of exposure include chemical caramel coloring, ammoniated molasses, dyes and pigments, rube, cleaning and agricultural chemicals. Toxicity attributed to 4MI in foods has recently become a focus of research. Recent studies showed that 4MI induced adverse changes in various target tissues. Brain is known to be a target organ for 4MI-induced toxicity but its cytotoxic mechanisms have not yet been elucidated. In this study, experiments were divided into two parts: (1) using in vivo methodology, doses of 4MI at 100, 200, or 300 mg/kg were administered orally to mice daily for 14 to obtain brain mitochondria; and (2) utilizing in vitro methodology, brain mitochondria were incubated with 4MI at 400, 800, or 1600 μM concentrations. Subsequently, the neurotoxicity of 4MI was assessed using mitochondrial dysfunction tests, including reactive oxygen species (ROS) formation, mitochondrial membrane potential (MMP) collapse, mitochondrial swelling, and cytochrome c release. Our results from both in vivo and in vitro experiments on isolated brain mitochondria showed a significant decrease in complex II activity and also marked elevation in the ROS formation, MMP collapse, mitochondrial swelling, and enhanced release of cytochrome c. Data indicated that 4MI induced neurotoxicity through the impairment of electron transfer chain especially at complex II and elevated ROS formation leading to subsequent oxidative stress events including mitochondrial membrane depolarization, mitochondrial swelling, and release of cytochrome c, which is the starting point of mitochondrial-mediated apoptosis signaling and neurodegeneration.