Pramod Kushwaha

Effects of sub-acute exposure to TiO2, ZnO and Al2O3 nanoparticles on oxidative stress and histological changes in mouse liver and brain

Abstract Nanomaterials are at the leading edge of the rapidly developing field of nanotechnology. However the information regarding toxicity of these nanoparticles on humans and environment is still deficient. The present study investigated the toxic effects of three metal oxide nanoparticles, TiO2, ZnO and Al2O3 on mouse erythrocytes, brain and liver. Male mice were administered a single oral dose of 500 mg/kg of each nanoparticles for 21 consecutive days. The results suggest that exposure to these nano metallic particles produced a significant oxidative stress in erythrocyte, liver and brain as evident from enhanced levels of Reactive Oxygen Species (ROS) and altered antioxidant enzymes activities. A significant increase in dopamine and norepinephrine levels in brain cerebral cortex and increased brain oxidative stress suggest neurotoxic potential of these nanoparticles. Transmission electron microscopic (TEM) analysis indicated the presence of these nanoparticles inside the cytoplasm and nucleus. These changes were also supported by the inhibition of CuZnSOD and MnSOD, considered as important biomarkers of oxidative stress. The toxic effects produced by these nanoparticles were more pronounced in the case of zinc oxide, followed by aluminum oxide and titanium dioxide, respectively. The present results further suggest the involvement of oxidative stress as one of the main mechanisms involved in nanoparticles induced toxic manifestations.

Alpha-lipoic acid protects oxidative stress, changes in cholinergic system and tissue histopathology during co-exposure to arsenic-dichlorvos in rats.

We investigated protective efficacy of α-lipoic acid (LA), an antioxidant against arsenic and DDVP co-exposed rats. Biochemical variables suggestive of oxidative stress, neurological dysfunction, and tissue histopathological alterations were determined. Male rats were exposed either to 50 ppm sodium arsenite in drinking water or in combination with DDVP (4 mg/kg, subcutaneously) for 10 weeks. α-Lipoic acid (50mg/kg, pos) was also co-administered in above groups. Arsenic exposure led to significant oxidative stress along, hepatotoxicity, hematotoxicity and altered brain biogenic amines levels accompanied by increased arsenic accumulation in blood and tissues. These altered biochemical variables were supported by histopathological examinations leading to oxidative stress and cell death. These biochemical alterations were significantly restored by co-administration of α-lipoic acid with arsenic and DDVP alone and concomitantly. The results indicate that arsenic and DDVP induced oxidative stress and cholinergic dysfunction can be significantly protected by the supplementation of α-lipoic acid.

Oxidative stress following exposure to silver and gold nanoparticles in mice

Silver (Ag) and gold nanoparticles (Au NPs) have wide applications. They are increasingly being used in the medical devices, biosensors, cancer cell imaging, and cosmetics. Increased applications of these NPs in the technological advances have also led to the risk of exposure to these particles. This study investigated the toxic effects of Ag and Au NPs (1 μM and 2 μM, oral) on mouse erythrocytes and tissues after 14 consecutive days’ exposure. Our results demonstrate significant increase in reactive oxygen species (ROS) and depletion of antioxidant enzyme status in erythrocytes and tissues. Hepatic and renal toxicity was evident from liver and kidney function tests. Inflammatory markers, interleukin-6 and nitric oxide synthase increased in plasma on administration following exposure to these NPs at both the doses. A more pronounced increase was noted in kidney metallothionein (MT) compared to liver MT on exposure to these NPs. Toxic potential of these NPs was further confirmed by increased 8-hydroxy-2′-deoxyguanosine levels in urine, a biomarker of DNA damage. Among the two NPs, Ag NP was more toxic at 2 μM dose compared to lower dose of 1 μM. The study suggests oxidative stress as the major mechanism responsible for the toxic manifestations induced by Ag and Au NPs.

Effects of combined exposure to dichlorvos and monocrotophos on blood and brain biochemical variables in rats

Dichlorvos (DDVP) and monocrotophos (MC) are systemic insecticides and known to produce cholinergic and non-cholinergic effects. Individual toxic effects of these chemicals are known but their combined effects have not been studied. We studied the effect of concomitant exposure to DDVP and MC on selected biochemical variables suggestive of liver damage, changes in whole brain biogenic amines levels, acetylcholinesterase (AchE) and monoamine oxidase (MAO) activities in rats. Female rats were exposed to DDVP (2.5 mg/kg subcutaneously) and MC (1.8 mg/kg oral) either individually or in combination for 4 weeks. We observed significant decrease in more pronounced depletion in norepinephrine (NE) and dopamine (DA) levels during co-exposure to DDVP and MC. Brain AChE activity increased and activity of MAO showed significant depletion on co-exposure to DDVP and MC. Brain glutathione (GSH) and oxidized glutathione (GSSG) ratio decreased significantly during exposure to DDVP or MC while co-exposure to these toxicants led to a more pronounced depletion of GSH: GSSG ratio. Serum aspartate amino transferase (AST) and alkaline phosphatase (ALP) activities increased significantly on exposure to MC suggesting liver injury, while DDVP alone had no effect on these variables. There were no effects of DDVP and MC exposure on haematological biochemical variables except for depletion in serum glucose level after MC exposure which was more pronounced DDVP + MC during co-exposure. It can be concluded that only moderate synergistic effects occur between MC and DDVP during co-exposure. A more detailed study with variable doses, prolonged exposure and alterations in different brain regions is recommended.

Lead and Ethanol Co-Exposure Lead to Blood Oxidative Stress and Subsequent Neuronal Apoptosis in Rats

AIMS The present study was aimed at investigating chronic exposure to lead and ethanol, individually and in combination with blood oxidative stress leading to possible brain apoptosis in rats. METHODS Rats were exposed to lead (0.1% w/v in drinking water) or ethanol (1 and 10%) either individually or in combination for four months. Biochemical variables indicative of oxidative stress (blood and brain) and brain apoptosis were examined. Native polyacrylamide agarose gel electrophoresis was carried out in brain homogenates for glucose-6-phosphate dehydrogenase (G6PD) analysis, whereas western blot analysis was done for the determination of apoptotic markers like Bax, Bcl-2, caspase-3, cytochrome c and p53. RESULTS The results suggest that most pronounced increase in oxidative stress in red blood cells and brain of animals co-exposed to lead and 10% ethanol compared all the other groups. Decrease in G6PD activity followed the same trend. Upregulation of Bax, cytochrome c, caspase-3, p53 and down-regulation of Bcl-2 suggested apoptosis in the rat brain co-exposed to lead and ethanol (10%) compared with their individual exposures. Significantly high lead accumulation in blood and brain during co-exposure further support synergistic toxicity. CONCLUSION The present study thus suggests that higher consumption of ethanol during lead exposure may lead to brain apoptosis, which may be mediated through oxidative stress.

Effects of co-exposure to arsenic and dichlorvos on glutathione metabolism, neurological, hepatic variables and tissue histopathology in rats

The individual toxic effects of arsenic and organophosphate pesticides are known but there is a lack of data on their combined effects. The present study investigates the toxic effects following combined exposure to organophosphate and arsenic on: (i) alterations in brain biogenic amines, (ii) oxidative stress and its correlation with glutathione linked enzymes, cell death and histopathological observations, and (iii) arsenic concentration in soft tissues. Rats were exposed to arsenic (1 mg kg−1 body weight, orally) and dichlorvos (4 mg kg−1 body weight, subcutaneously) either individually or in combination for 16 weeks. Arsenic alone and in combination with DDVP led to a significant increase in reactive oxygen species (ROS), thiobarbituric acid reactive substance (TBARS), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) activities accompanied by a decreased glutathione reductase, reduced and oxidized glutathione (GSH and GSSG) levels in tissues. Arsenic and DDVP also produced a significant depletion in brain biogenic amines; however, acetylcholinesterase (AChE) activity increased moderately on arsenic exposure. Arsenic and DDVP co-exposure exhibited synergism in the case of ROS while no such effect was noted in the case of TBARS. Interestingly, combined exposure to arsenic and DDVP resulted in more pronounced toxic effects compared to their individual effects based on various biochemical variables, cell death (TUNEL assay) and histopathological observations. Interestingly, brain arsenic levels decreased on co-exposure to arsenic and DDVP accompanied with prominent elevation of liver arsenic content in co-exposed groups. The present study thus provides some interesting observations on the interaction between arsenic and DDVP including (i) co-exposure to arsenic and DDVP might lead to significant oxidative stress and (ii) their co-exposure produced synergistic effects on some liver variables but some antagonistic effects on the brain.

Sodium tungstate induced neurological alterations in rat brain regions and their response to antioxidants.

Tungsten, recognized recently as an environmental contaminant, is being used in arms and ammunitions as substitute to depleted uranium. We studied the effects of sodium tungstate on oxidative stress, few selected neurological variables like acetylcholinesterase, biogenic amines in rat brain regions (cerebral cortex, hippocampus and cerebellum) and their prevention following co-administration of N-acetylcysteine (NAC), naringenin and quercetin. Animals were sub-chronically exposed to sodium tungstate (100 ppm in drinking water) and orally co-supplemented with different antioxidants (0.30 mM) for three months. Sodium tungstate significantly decreased the activity of acetylcholinesterase, dopamine, nor-epinephrine and 5-hydroxytryptamine levels while it increased monoamine oxidase activity in different brain regions. Tungstate exposure produced a significant increase in biochemical variables indicative of oxidative stress while, neurological alterations were more pronounced in the cerebral cortex compared to other regions. Co-administration of NAC and flavonoids with sodium tungstate significantly restored glutathione, prevented changes in the brain biogenic amines, reactive oxygen species (ROS) and TBARS levels in the different brain regions. The protection was more prominent in the animals co-administered with NAC. We can thus conclude that sodium tungstate induced brain oxidative stress and the alterations in some neurological variables can effectively be reduced by co-supplementation of NAC.

Effects of sodium tungstate on oxidative stress enzymes in rats

Abstract Tungsten, due to its distinguished physical properties, has wide industrial and military applications. Environmental exposure to tungsten, which mainly occurs through various sources like food, water, soil, etc., is of growing concern as various toxic effects have recently been reported. In this study, we investigated the effects of oral and intraperitoneal (i.p.) administration of sodium tungstate on various biochemical variables indicative of oxidative stress in erythrocytes and soft tissue damage in rats. Male rats were administered to 119 mg, 238 mg/kg of sodium tungstate orally or 20 mg and 41 mg/kg through i.p. route, for 14 consecutive days. The results demonstrated a significant increase in Reactive Oxygen Species (ROS) and an increase in catalase and glutathione peroxidase antioxidant enzymes activities in erythrocytes. Erythrocyte glutathione-S-transferase (GST) activity showed significant inhibition, while tissue ROS and thiobarbituric acid reactive substance levels increased accompanied by a decreased reduced glutathione, oxidized glutathione (GSH:GSSG) ratio. These changes were supported by an increase in plasma transaminases activities, creatinine, and urea levels, suggesting hepatic and renal injury. These biochemical alterations were prominent in rats intraperitoneally administrated with sodium tungstate than oral administration, suggesting more pronounced toxicity. The study also suggests oxidative stress as one of the major mechanism involved in the toxic manifestations of sodium tungstate.

Monensin potentiates lead chelation efficacy of MiADMSA in rat brain post chronic lead exposure.

The present study evaluates combination therapy with a chelating agent, MiADMSA and a Na(+) ionophore, monensin against sub-chronic lead toxicity in rats. Animals were exposed to 0.1% lead in drinking water for 16 weeks and then treated with either MiADMSA at 50mg/kg body weight, or monensin at 10mg/kg, or both in combination for a period of 5 days was administered. Biomarkers indicative of oxidative stress like ROS, GSH, GSSG and TBARS demonstrated lead-induced toxic manifestations in blood, kidney and brain. Antioxidants like SOD, catalase and glutathione peroxidase along with specific lead biomarker, blood ALAD were also severely depleted in lead intoxicated animals. Serum parameters and histopathological findings supported the said results. MiADMSA treatment during both mono- and combination therapy with monensin, restored the antioxidant status and recovered biochemical and haematological variables due to lead. However, monensin alone was not found to be effective in the given scenario. Interestingly, combination therapy in its ability to revert lead-induced overall systemic toxicity was only found at par with the MiADMSA monotherapy except for its chelation potential. Monensin given in combination with MiADMSA potentiated its lead chelation ability especially from brain, along with maintaining the normal copper concentrations in the organ unlike MiADMSA monotherapy.

Comparative safety evaluation of riot control agents of synthetic and natural origin

Abstract Riot control agents (RCA) are lachrymatory, irritating compounds which temporarily incapacitate the uncontainable crowd. Ortho-Chlorobenzylidene-malononitrile (CS), 2-chloroacetophenone (CN), dibenz[b,f]1:4-oxazepine (CR), and nonivamide (PAVA) are synthetic RCAs, while oleoresin extract of chili known as oleoresin capsicum (OC) a natural irritant has been in use by various law enforcement agencies. Though efficacy of these agents is beyond doubt, they suffer from certain drawbacks including toxicity, production cost, and ecological compatibility. Presently, we have evaluated the safety of CR, OC, and PAVA on inhalation variables along with oral lethality. Additionally, the liver function test (LFT) in serum and lungs function was evaluated in broncho-alveolar-lavage fluid (BALF), both collected on the 14th day after RCA exposure. Animals then sacrificed and histopathology of liver and lungs was carried out. Results showed OC and PAVA to be more toxic than CR with an oral LD50 of 150 and 200 mg/kg body weight, respectively, while CR was safe at >3 g/kg body weight. All three agents caused severe impairment of respiratory variables bringing down normal respiration by >80% with rise in sensory irritation. Recovery from the irritating effect of CR was more rapid than OC and PAVA. LFT and BALF variables were not significantly different from that of control. There were no remarkable histopathological changes in liver and lungs. Hence, as per results, CR is safest among all synthetic and natural origin RCAs and can be safely used for effective dispersion of disobedient mob.