Biochemical and gene expression alterations due to individual exposure of atrazine, dichlorvos, and imidacloprid and their combination in zebrafish.

In environmental toxicology, combined toxicity has emerged as an important concern. Atrazine (ATZ), dichlorvos (DIC), and imidacloprid (IMD) are the major pesticides, extensively used to control insect, flies, mosquitoes, and weed. Here, we investigate whether the exposure to three different types of pesticides individually and in combination for 24 h alters antioxidant enzyme responses in zebrafish (Danio rerio). Oxidative stress parameters (biochemical and mRNA expression), acetylcholinesterase (AChE) activity, and Metallothionein-II (MT-II) mRNA expression levels were measured. Present work includes toxicological assessment of individual and combined (CMD) exposure of ATZ (185.4 µM), DIC (181 µM), IMD (97.8 µ), and CMD (ATZ 92.7 µM + DIC 90.5 µM + IMD 48.9 µM), in the liver, kidney, and brain of adult zebrafish. Lipid peroxidation (LPO), glutathione (GSH) content, AChE, superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activity along with mRNA expression of SOD, CAT, GPx, and MT-II were evaluated. Briefly, LPO, GSH content, the activity of AChE, and all antioxidant enzymes enhanced significantly in individual exposure, which was further altered in the CMD group. The mRNA expression of SOD, CAT, GPx, and MT-II in the liver and kidney showed significant down-regulation in all exposed groups. In the brain, significant upregulation in mRNA expression of SOD, CAT, GPx, and MT-II was observed in DIC and IMD groups, while ATZ and CMD showed significant downregulation except for GPx. Findings postulate that the CMD group exhibits synergistic toxic manifestation. The present study provides the baseline data on the combined toxic effects of pesticides and suggests regulating the use of pesticides.

How to Train Your Posture: Haptic Feedback Can be Used for Postural Adaptation of the Trunk During Upper-Limb Motor Training.

Poor trunk posture, especially during long periods of sitting, could lead to problems such as Low Back Pain (LBP) and Forward Head Posture (FHP). Typical solutions are based on visual or vibration-based feedback. However, these systems could lead to feedback being ignored by the user and phantom vibration syndrome, respectively. In this study, we propose using haptic feedback for postural adaptation. In this two-part study, twenty-four healthy participants (age 25.87 ± 2.17 years) adapted to three different postural targets in the anterior direction while performing a unimanual reaching task using a robotic device. Results suggest a strong adaptation to the desired postural targets. Mean anterior trunk bending after the intervention is significantly different compared to baseline measurements for all postural targets. Additional analysis of movement straightness and smoothness indicates an absence of any negative interference of posture-based feedback on the performance of reaching movement. Taken together, these results suggest that haptic feedback-based systems could be used for postural adaptation applications. Also, this type of postural adaptation system can be used during the rehabilitation of stroke patients to reduce trunk compensation in lieu of typical physical constraint-based methods.

Let the force guide you: a performance-based adaptive algorithm for postural training using haptic feedback.

Motor learning is an essential component of human behavior. Many different factors can influence the process of motor learning, such as the amount of practice and type of feedback. Changes in task difficulty during training can also considerably impact motor learning. Typical motor learning studies include a sequential variation of task difficulty, i.e., easy to challenging, irrespective of user performance. However, many studies have reported the importance of performance-based task difficulty variation for effective motor learning and skill transfer. A performance-based adaptive algorithm for task difficulty variation based on the challenge-point framework is proposed in this study. The algorithm is described for postural adaptation during simultaneous upper-limb training. Ten healthy participants (28 ± 2.44 years) were recruited to validate the algorithm. Participants adapted to a postural target of 20° in the anterior direction from the initial upright posture while performing a unimanual reaching task using a robotic device. Results suggest a significant decrease in postural error after training. The algorithm successfully adapted the task difficulty based on the performance of the user. The proposed algorithm could be modified for different motor skills and can be further evaluated for different applications in order to maximize the potential benefits of rehabilitation sessions.

Haptic feedback system for postural adaptation during robotic rehabilitation of upper limb.

Although trunk compensation during stroke rehabilitation is widely studied, the proposed solutions primarily include a trunk constraint, which has several disadvantages. In this study, we have proposed a haptic feedback-based system for postural training during upper-limb motor rehabilitation. We have tested the proposed system on six healthy people in this preliminary study. Participants performed a simple 1-dimensional reaching task while their posture was being monitored. They received haptic feedback based on their trunk posture. Preliminary results revealed a significant decline in postural error (p<0.05) after the haptic-based training. The reduction in error was maintained even after haptic feedback was turned off. This study shows that haptic feedback could be a viable alternative to the traditional constraint-based methods for postural adaptation. Additional studies need to be conducted to further evaluate the influence of using such feedback strategies.

Risperidone induced alterations in feeding and locomotion behavior of Caenorhabditis elegans.

Antipsychotic drugs (APDs) are prescribed for the treatment of psychiatric illness. However, these drugs can also contribute to several developmental and behavioral disorders. Contemporary studies to evaluate the toxic effects of numerous atypical antipsychotics are reported to cause behavioral alteration at variable doses in mammals and nematodes. Risperidone, the second most prescribed drug in India, requires more exploration of its adverse effects on humans. Here, we explore effects on feeding behavior and locomotion patterns due to risperidone exposure in C. elegans model. The study targets to work out the toxic effects of risperidone exposure on feeding and locomotion behavior in addition to the expected pharmacological effects. N2 wild type strain was exposed in liquid culture assay for 2, 4, 6, 8, 10, and 12 hours with fixed 50 µM concentration. Feeding behavior was depleted due to inhibition in pharyngeal pumping varying from 11.05% - 45.67% in a time-dependent manner. Results of locomotion assay also show time-varying increase in reversals (4.9%-34.03%) and omega bends (26.23%-62.17%) with reduction in turn counts (29.07%- 42.2%) and peristaltic speed (31.38%-42.22%) amongst exposed groups as to control. The present work shows behavioral alterations due to risperidone exposure (50 µM) in C. elegans is in a time-dependent manner. The study concludes that risperidone exposure in C. elegans produces toxic effects with time, possibly caused by antagonizing other receptors apart from serotonin (5-H2T) and dopamine (D2) adding to its expected pharmacological effects.

Impact of co-exposure of aldrin and titanium dioxide nanoparticles at biochemical and molecular levels in Zebrafish.

Aldrin (ALD), a persistent-organic-pollutant (POP), an organochlorine-cyclodiene-pesticide is highly toxic in nature. Titanium dioxide nanoparticles (TNP) are widely used for various industrial applications. Despite the remarkable research on pesticide toxicity, the work with impact of nanoparticles on POP has been dealt with marginally. Chemicals co-exist in the environment and exhibit interactive effects. An investigation was carried out to evaluate the individual and combined effects of ALD (6 ppm) and TNP (60 ppm) exposure at sub-lethal concentration for 24 h in zebrafish. Significant reversal of lipid peroxidation level in liver and brain tissues and restoration in enhanced catalase activity in all examined tissues were observed in combined group. For other parameters, combined exposure of ALD and TNP does not show significant reversal action on ALD toxicity. Further studies are inline to understand combined effects of both to achieve significant reversal of ALD toxicity by TNP nanoparticles with threshold concentration of aldrin.

Oxidative injury caused by individual and combined exposure of neonicotinoid, organophosphate and herbicide in zebrafish.

The greatest challenge in environmental toxicology is to understand the effects of mixture toxicity as environmental pollutants co-exist and exhibit combined effects. Thus, it is necessary to evaluate the mixture toxicity associated with two or more co-existing compounds. Pesticides are widely used to control pest, they are ubiquitous in nature and present in all environmental components. Pesticide residue can be detected in almost all components of environment and food samples. Imidacloprid (IMD) (neonicotinoid), dichlorvos (DIC) (organophosphate) and atrazine (ATZ) are three widely used pesticides for commercial uses. Present work includes the assessment of effects of individual exposure of IMD (27.5 mg/L), DIC (15 mg/L), and ATZ (03 mg/L) and in combination of three (CMD) (13.75 + 7.5 + 1.5 mg/L IMD, DIC & ATZ, respectively) in terms of LPO, GSH content and antioxidant enzymes activities (superoxide dismutase, catalase and glutathione peroxidase) in zebrafish (Danio rerio), exposed for 24 h. CMD group exhibits highest lipid peroxidation than other individually exposed groups. Similarly, the activities of antioxidant enzymes were highest in CMD group with reduced GSH content. Results indicate that exposure to mixture of pesticides develops synergistic effects which were more toxic in compare to individual exposure and also produce toxicity in all examined tissues rather than selective organ toxicity.

Small molecule non-peptide inhibitors of botulinum neurotoxin serotype E: Structure-activity relationship and a pharmacophore model.

Botulinum neurotoxins (BoNTs) are the most poisonous biological substance known to humans. They cause flaccid paralysis by blocking the release of acetylcholine at the neuromuscular junction. Here, we report a number of small molecule non-peptide inhibitors of BoNT serotype E. The structure-activity relationship and a pharmacophore model are presented. Although non-peptidic in nature, these inhibitors mimic key features of the uncleavable substrate peptide Arg-Ile-Met-Glu (RIME) of the SNAP-25 protein. Among the compounds tested, most of the potent inhibitors bear a zinc-chelating moiety connected to a hydrophobic and aromatic moiety through a carboxyl or amide linker. All of them show low micromolar IC50 values.

Acoustic Injectors for Drop-On-Demand Serial Femtosecond Crystallography.

X-ray free-electron lasers (XFELs) provide very intense X-ray pulses suitable for macromolecular crystallography. Each X-ray pulse typically lasts for tens of femtoseconds and the interval between pulses is many orders of magnitude longer. Here we describe two novel acoustic injection systems that use focused sound waves to eject picoliter to nanoliter crystal-containing droplets out of microplates and into the X-ray pulse from which diffraction data are collected. The on-demand droplet delivery is synchronized to the XFEL pulse scheme, resulting in X-ray pulses intersecting up to 88% of the droplets. We tested several types of samples in a range of crystallization conditions, wherein the overall crystal hit ratio (e.g., fraction of images with observable diffraction patterns) is a function of the microcrystal slurry concentration. We report crystal structures from lysozyme, thermolysin, and stachydrine demethylase (Stc2). Additional samples were screened to demonstrate that these methods can be applied to rare samples.

Acoustic transfer of protein crystals from agarose pedestals to micromeshes for high-throughput screening.

Acoustic droplet ejection (ADE) is an emerging technology with broad applications in serial crystallography such as growing, improving and manipulating protein crystals. One application of this technology is to gently transfer crystals onto MiTeGen micromeshes with minimal solvent. Once mounted on a micromesh, each crystal can be combined with different chemicals such as crystal-improving additives or a fragment library. Acoustic crystal mounting is fast (2.33 transfers s(-1)) and all transfers occur in a sealed environment that is in vapor equilibrium with the mother liquor. Here, a system is presented to retain crystals near the ejection point and away from the inaccessible dead volume at the bottom of the well by placing the crystals on a concave agarose pedestal (CAP) with the same chemical composition as the crystal mother liquor. The bowl-shaped CAP is impenetrable to crystals. Consequently, gravity will gently move the crystals into the optimal location for acoustic ejection. It is demonstrated that an agarose pedestal of this type is compatible with most commercially available crystallization conditions and that protein crystals are readily transferred from the agarose pedestal onto micromeshes with no loss in diffraction quality. It is also shown that crystals can be grown directly on CAPs, which avoids the need to transfer the crystals from the hanging drop to a CAP. This technology has been used to combine thermolysin and lysozyme crystals with an assortment of anomalously scattering heavy atoms. The results point towards a fast nanolitre method for crystal mounting and high-throughput screening.

Loss of quaternary structure is associated with rapid sequence divergence in the OSBS family.

The rate of protein evolution is determined by a combination of selective pressure on protein function and biophysical constraints on protein folding and structure. Determining the relative contributions of these properties is an unsolved problem in molecular evolution with broad implications for protein engineering and function prediction. As a case study, we examined the structural divergence of the rapidly evolving o-succinylbenzoate synthase (OSBS) family, which catalyzes a step in menaquinone synthesis in diverse microorganisms and plants. On average, the OSBS family is much more divergent than other protein families from the same set of species, with the most divergent family members sharing <15% sequence identity. Comparing 11 representative structures revealed that loss of quaternary structure and large deletions or insertions are associated with the family's rapid evolution. Neither of these properties has been investigated in previous studies to identify factors that affect the rate of protein evolution. Intriguingly, one subfamily retained a multimeric quaternary structure and has small insertions and deletions compared with related enzymes that catalyze diverse reactions. Many proteins in this subfamily catalyze both OSBS and N-succinylamino acid racemization (NSAR). Retention of ancestral structural characteristics in the NSAR/OSBS subfamily suggests that the rate of protein evolution is not proportional to the capacity to evolve new protein functions. Instead, structural features that are conserved among proteins with diverse functions might contribute to the evolution of new functions.

Hitting the target: fragment screening with acoustic in situ co-crystallization of proteins plus fragment libraries on pin-mounted data-collection micromeshes.

Acoustic droplet ejection (ADE) is a powerful technology that supports crystallographic applications such as growing, improving and manipulating protein crystals. A fragment-screening strategy is described that uses ADE to co-crystallize proteins with fragment libraries directly on MiTeGen MicroMeshes. Co-crystallization trials can be prepared rapidly and economically. The high speed of specimen preparation and the low consumption of fragment and protein allow the use of individual rather than pooled fragments. The Echo 550 liquid-handling instrument (Labcyte Inc., Sunnyvale, California, USA) generates droplets with accurate trajectories, which allows multiple co-crystallization experiments to be discretely positioned on a single data-collection micromesh. This accuracy also allows all components to be transferred through small apertures. Consequently, the crystallization tray is in equilibrium with the reservoir before, during and after the transfer of protein, precipitant and fragment to the micromesh on which crystallization will occur. This strict control of the specimen environment means that the crystallography experiments remain identical as the working volumes are decreased from the few microlitres level to the few nanolitres level. Using this system, lysozyme, thermolysin, trypsin and stachydrine demethylase crystals were co-crystallized with a small 33-compound mini-library to search for fragment hits. This technology pushes towards a much faster, more automated and more flexible strategy for structure-based drug discovery using as little as 2.5 nl of each major component.

Discovery of a fluorene class of compounds as inhibitors of botulinum neurotoxin serotype E by virtual screening.

Botulinum neurotoxins are one of the most poisonous biological substances known to humans and present a potential bioterrorism threat. There are no therapeutic interventions developed so far. Here, we report the first small molecule non-peptide inhibitor for botulinum neurotoxin serotype E discovered by structure-based virtual screening and propose a mechanism for its inhibitory activity.

Structural insight into mechanism and diverse substrate selection strategy of L-ribulokinase.

The araBAD operon encodes three different enzymes required for catabolism of L-arabinose, which is one of the most abundant monosaccharides in nature. L-ribulokinase, encoded by the araB gene, catalyzes conversion of L-ribulose to L-ribulose-5-phosphate, the second step in the catabolic pathway. Unlike other kinases, ribulokinase exhibits diversity in substrate selectivity and catalyzes phosphorylation of all four 2-ketopentose sugars with comparable k(cat) values. To understand ribulokinase recognition and phosphorylation of a diverse set of substrates, we have determined the X-ray structure of ribulokinase from Bacillus halodurans bound to L-ribulose and investigated its substrate and ATP co-factor binding properties. The polypeptide chain is folded into two domains, one small and the other large, with a deep cleft in between. By analogy with related sugar kinases, we identified (447)GGLPQK(452) as the ATP-binding motif within the smaller domain. L-ribulose binds in the cleft between the two domains via hydrogen bonds with the side chains of highly conserved Trp126, Lys208, Asp274, and Glu329 and the main chain nitrogen of Ala96. The interaction of L-ribulokinase with L-ribulose reveals versatile structural features that help explain recognition of various 2-ketopentose substrates and competitive inhibition by L-erythrulose. Comparison of our structure to that of the structures of other sugar kinases revealed conformational variations that suggest domain-domain closure movements are responsible for establishing the observed active site environment.

Lucanthone and its derivative hycanthone inhibit apurinic endonuclease-1 (APE1) by direct protein binding.

Lucanthone and hycanthone are thioxanthenone DNA intercalators used in the 1980s as antitumor agents. Lucanthone is in Phase I clinical trial, whereas hycanthone was pulled out of Phase II trials. Their potential mechanism of action includes DNA intercalation, inhibition of nucleic acid biosyntheses, and inhibition of enzymes like topoisomerases and the dual function base excision repair enzyme apurinic endonuclease 1 (APE1). Lucanthone inhibits the endonuclease activity of APE1, without affecting its redox activity. Our goal was to decipher the precise mechanism of APE1 inhibition as a prerequisite towards development of improved therapeutics that can counteract higher APE1 activity often seen in tumors. The IC(50) values for inhibition of APE1 incision of depurinated plasmid DNA by lucanthone and hycanthone were 5 µM and 80 nM, respectively. The K(D) values (affinity constants) for APE1, as determined by BIACORE binding studies, were 89 nM for lucanthone/10 nM for hycanthone. APE1 structures reveal a hydrophobic pocket where hydrophobic small molecules like thioxanthenones can bind, and our modeling studies confirmed such docking. Circular dichroism spectra uncovered change in the helical structure of APE1 in the presence of lucanthone/hycanthone, and notably, this effect was decreased (Phe266Ala or Phe266Cys or Trp280Leu) or abolished (Phe266Ala/Trp280Ala) when hydrophobic site mutants were employed. Reduced inhibition by lucanthone of the diminished endonuclease activity of hydrophobic mutant proteins (as compared to wild type APE1) supports that binding of lucanthone to the hydrophobic pocket dictates APE1 inhibition. The DNA binding capacity of APE1 was marginally inhibited by lucanthone, and not at all by hycanthone, supporting our hypothesis that thioxanthenones inhibit APE1, predominantly, by direct interaction. Finally, lucanthone-induced degradation was drastically reduced in the presence of short and long lived free radical scavengers, e.g., TRIS and DMSO, suggesting that the mechanism of APE1 breakdown may involve free radical-induced peptide bond cleavage.

Low pH-induced pore formation by the T domain of botulinum toxin type A is dependent upon NaCl concentration.

Botulinum neurotoxins (BoNTs) undergo low pH-triggered membrane insertion, resulting in the translocation of their light (catalytic) chains into the cytoplasm. The T (translocation) domain of the BoNT heavy chain is believed to carry out translocation. Here, the behavior of isolated T domain from BoNT type A has been characterized, both in solution and when associated with model membranes. When BoNT T domain prepared in the detergent dodecylmaltoside was diluted into aqueous solution, it exhibited a low pH-dependent conformational change below pH 6. At low pH the T domain associated with, and formed pores within, model membrane vesicles composed of 30 mol% dioleoylphosphatidylglycerol/70 mol% dioleoylphosphatidylcholine. Although T domain interacted with vesicles at low (50 mM) and high (400 mM) NaCl concentrations, the interaction required much less lipid at low salt. However, even at high lipid concentrations pore formation was much more pronounced at low NaCl concentrations than at high NaCl concentration. Increasing salt concentration after insertion in the presence of 50 mM NaCl did not decrease pore formation. A similar effect of NaCl concentration upon pore formation was observed in vesicles composed solely of dioleoylphosphatidylcholine, showing that the effect of NaCl did not solely involve modulation of electrostatic interactions between protein and anionic lipids. These results indicate that some feature of membrane-bound T domain tertiary structure critical for pore formation is highly dependent upon salt concentration.

Effects of sub-chronic low-level lead exposure on the homeostasis of copper and zinc in rat tissues.

Information about the health risks or the subtle adverse health effects that might be associated with low-level lead exposure on micronutrient metabolism are scarce in the literature. The present work investigated the subtle adverse health effects of exposure to progressively low levels of lead on the metabolism of two micronutrients, copper and zinc in different tissues of the rat. Rats were exposed to 200, 300 and 400 ppm lead in their drinking water for 12 weeks. Lead, copper and zinc concentrations were determined in blood, liver, kidney, heart, spleen and brain of the animals. While the imbalance in zinc metabolism was characterized by a deposition of zinc in the kidney and to a lesser extent in the heart of the animals, imbalance in copper metabolism was characterized by a depletion of blood and splenic copper concentrations as well as renal and cardiac accumulation of copper. Hepatic and brain copper and zinc contents, together with blood zinc were not affected by the 12-week lead exposure. A linear relationship was observed between lead dose and lead accumulation in the spleen, whereas a non-linear relationship was observed between lead dose and lead accumulation in blood, liver, kidney and heart. Our findings indicate that exposure to progressively low-level lead concentrations results in imbalance in copper and zinc in the organism and this might be a factor in propensity toward behavioral disorders observed in lead exposure.

Discovery and structure determination of the orphan enzyme isoxanthopterin deaminase .

Two previously uncharacterized proteins have been identified that efficiently catalyze the deamination of isoxanthopterin and pterin 6-carboxylate. The genes encoding these two enzymes, NYSGXRC-9339a ( gi|44585104 ) and NYSGXRC-9236b ( gi|44611670 ), were first identified from DNA isolated from the Sargasso Sea as part of the Global Ocean Sampling Project. The genes were synthesized, and the proteins were subsequently expressed and purified. The X-ray structure of Sgx9339a was determined at 2.7 A resolution (Protein Data Bank entry 2PAJ ). This protein folds as a distorted (beta/alpha)(8) barrel and contains a single zinc ion in the active site. These enzymes are members of the amidohydrolase superfamily and belong to cog0402 within the clusters of orthologous groups (COG). Enzymes in cog0402 have previously been shown to catalyze the deamination of guanine, cytosine, S-adenosylhomocysteine, and 8-oxoguanine. A small compound library of pteridines, purines, and pyrimidines was used to probe catalytic activity. The only substrates identified in this search were isoxanthopterin and pterin 6-carboxylate. The kinetic constants for the deamination of isoxanthopterin with Sgx9339a were determined to be 1.0 s(-1), 8.0 muM, and 1.3 x 10(5) M(-1) s(-1) (k(cat), K(m), and k(cat)/K(m), respectively). The active site of Sgx9339a most closely resembles the active site for 8-oxoguanine deaminase (Protein Data Bank entry 2UZ9 ). A model for substrate recognition of isoxanthopterin by Sgx9339a was proposed on the basis of the binding of guanine and xanthine in the active site of guanine deaminase. Residues critical for substrate binding appear to be conserved glutamine and tyrosine residues that form hydrogen bonds with the carbonyl oxygen at C4, a conserved threonine residue that forms hydrogen bonds with N5, and another conserved threonine residue that forms hydrogen bonds with the carbonyl group at C7. These conserved active site residues were used to identify 24 other genes which are predicted to deaminate isoxanthopterin.

Detoxification and antioxidant effects of curcumin in rats experimentally exposed to mercury.

Curcumin, a safe nutritional component and a highly promising natural antioxidant with a wide spectrum of biological functions, has been examined in several metal toxicity studies, but its role in protection against mercury toxicity has not been investigated. Therefore, the detoxification and antioxidant effects of curcumin were examined to determine its prophylactic/therapeutic role in rats experimentally exposed to mercury (in the from of mercuric chloride-HgCl(2), 12 micromol kg(-1) b.w. single intraperitoneal injection). Curcumin treatment (80 mg kg(-1) b.w. daily for 3 days, orally) was found to have a protective effect on mercury-induced oxidative stress parameters, namely, lipid peroxidation and glutathione levels and superoxide dismutase, glutathione peroxidase and catalase activities in the liver, kidney and brain. Curcumin treatment was also effective for reversing mercury-induced serum biochemical changes, which are the markers of liver and kidney injury. Mercury concentration in the tissues was also decreased by the pre/post-treatment with curcumin. However, histopathological alterations in the liver and kidney were not reversed by curcumin treatment. Mercury exposure resulted in the induction of metallothionein (MT) mRNA expressions in the liver and kidney. Metallothionein mRNA expression levels were found to decrease after the pre-treatment with curcumin, whereas post-treatment with curcumin further increased MT mRNA expression levels. Our findings suggest that curcumin pretreatment has a protective effect and that curcumin can be used as a therapeutic agent in mercury intoxication. The study indicates that curcumin, an effective antioxidant, may have a protective effect through its routine dietary intake against mercury exposure.

Evaluation of comparative effect of pre- and posttreatment of selenium on mercury-induced oxidative stress, histological alterations, and metallothionein mRNA expression in rats.

To evaluate the effect of pre- or posttreatment of selenium (6 micromol/kg b.w., single intraperitoneal injection) in mercury intoxication, rats were exposed to mercury (12 micromol/kg b.w., single intraperitoneal injection). Exposure to mercury resulted in induced oxidative stress in liver, kidney, and brain tissues. Marked changes in serum biochemical parameters together with alterations in histopathology and an induction in metallothionein-I and metallothionein-II mRNA expression in the liver and kidney were observed. Pretreatment with selenium to mercury-exposed animals had protective effect on the liver, whereas posttreatment had partial protection on restoration of altered oxidative stress parameters. In the kidney, pretreatment with selenium showed partial protection on restoration of altered biochemical parameters, whereas no protection was observed in posttreatment. The pretreatment with selenium resulted in restoration of mercury-induced metallothionein-I and metallothionein-II mRNA expression, which was completely restored in the liver whereas partial restoration was observed in the kidney. Posttreatment with selenium resulted in further induction in metallothionein-I and metallothionein-II mRNA expression in the liver and kidney. In the brain, selenium showed partial protection on alerted biochemical parameters. Results indicate that pretreatment with selenium is beneficial in comparison to posttreatment in mercury intoxication. Thus, dietary intake of selenium within safe limit may, therefore, enable us in combating any foreseen effects due to mercury exposure.