A comparative account of phototoxicity of anthracene and pyrene in the tadpoles of the anuran amphibian Fejervarya limnocharis using multiple toxicological end points.

Anthracene (Anth) and pyrene (Pyr), two of the priority polycyclic aromatic hydrocarbons (PAHs), being lipophilic in nature, not only accumulate in animals, but also settle in the sediment of water bodies leading to continuous exposure for animals. Anth and Pyr when exposed to sunlight can be photoactivated and have harmful effects on aquatic organisms. A comparative analysis was carried out to assess the acute, sub-chronic, genetic and biochemical toxicity of Anth and Pyr in F. limnocharis tadpoles following short exposures to sunlight on a daily basis. In the bioaccumulation studies, it was found that both Anth and Pyr accumulated in the tadpole tissues in a concentration and time dependent manner. The LC50 values for Anth (under 15 min of daily sunlight exposure) were found to be 2.87, 2.59, 2.28, 1.80 mg/L at 24, 48, 72 and 96 h of the exposures. The corresponding LC50 values for Pyr were 1.03, 0.80, 0.62, 0.42 mg/L. Sublethal exposure of Anth and Pyr affected the survivality, time to metamorphosis as well as morphometric parameters under sunlight exposure. In the genotoxicity assessment studies, particularly the micronucleus test and comet assay, it was found that Pyr led to a higher incidence of micronucleus formation and DNA damage in comparison to Anth. The exposure to PAHs resulted in significant changes in the activity of antioxidant-mediated protective response, specifically the SOD activity, which varied between the groups treated with Anth and Pyr. On the other hand, Pyr treated group showed a higher level of GSH as compared to Anth treated groups. Moreover, the elevation in MDA level in the Anth and Pyr treated groups suggests an increase in lipid peroxidation. Future research should focus on understanding the ecotoxicological risk faced by anuran amphibia due to PAHs that frequently occur in aquatic environments and developing strategies to mitigate these risks.

Aqueous Extract of Moringa oleifera Exhibit Potential Anticancer Activity and can be Used as a Possible Cancer Therapeutic Agent: A Study Involving In Vitro and In Vivo Approach.

OBJECTIVES: New cases of cancers are increasing at an alarming rate globally. It has been hypothesized that modern cancer treatment is associated with lots of side effects and thus evoking the need to develop safer treatment measures. Thus, the present study was undertaken to evaluate the anti-carcinogenic potential of a highly nutricious plant "Moringa oleifera" (MO) in vitro and in vivo. METHODS: GC-MS analysis of aqueous extract of Moringa oleifera (AEMO) was employed to identify the bioactive compound present. Anti-tumor activity of AEMO was assessed in EAC (Ehrlich acites carcinoma) induced solid tumor bearing mice by analyzing tumor weight (TW) and Tumor volume (TV). To assess AEMO induced cytotoxicity, EAC and HEp-2 (Human laryngeal carcinoma) cells were treated with AEMO (0.05, 0.1, 0.25, 0.5 and 1 mg/ml) for both 48 h and 72 h and trypan blue, MTT and LDH released assay was done. Further, cell cycle assay and apoptosis assay was done in EAC cells to understand the mechanism of AEMO induced tumor regression. RESULTS: GC-MS analysis revealed the presence of quinic acid, octadecanoic acid, hexadecanoic acid (palmitic acid), α-tocopherol (Vitamin-E) and É£-sitosterol as major bioactive compounds. AEMO administration reduced the TV and TW of tumor-bearing mice and increases the life span. Side effect analysis showed that AEMO treatment did not induce significant alterations of liver and kidney function and hematological parameters. Further, in vitro cytotoxicity assays revealed that AEMO treatment induced dose and time-dependent toxicity in both the cell lines tested. Flow cytometric analysis confirmed significant induction of apoptotic cells by changing the mitochondrial membrane potential in EAC cell line. CONCLUSION: The results of the present study suggest that AEMO has immense potential to inhibit the tumor progression without affecting the normal physiology and functioning of the body and thus can be used as a cancer therapeutic agent.

Joint detection of claudin-1 and junctional adhesion molecule-A as a therapeutic target in oral epithelial dysplasia and oral squamous cell carcinoma.

Abnormal expression of claudin-1 (CLDN-1) and junctional adhesion molecule-A (JAM-A) has been described in certain malignancies but their clinical relevance is poorly understood. The present study aims to elucidate the role of CLDN-1 and JAM-A in oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC). Changes in the expression of these proteins were identified immunohistochemically on tissue sections from patients with OED and OSCC and compared with control. A correlation between the expression level of proteins and clinicopathological features was analyzed by Pearson's correlation χ2 test. The survival curve of the follow-up data was estimated by the Kaplan-Meier method followed by the log-rank test. CLDN-1 and JAM-A were highly expressed in OED and OSCC tissues when compared to control. Also, delocalization of CLDN-1 from the membrane to the cytoplasm to the nucleus was observed as the cell proceeds from normal to malignancy. Increased expression of CLDN-1 and JAM-A in both OED and OSCC were concomitant with histological grades. In addition, increased JAM-A was associated with perineural invasion of cancer cells. A positive correlation between the expression level of proteins was observed in OED (r = 0.733) and OSCC (r = 0.577). Kaplan-Meier analysis in patients with OSCC showed that the survival rate was lower in patients with high CLDN-1 and high JAM-A expression compared to low expressed patients. To conclude, the elevated level and delocalization of CLDN-1 and JAM-A suggest their use as tumor markers. A positive correlation between CLDN-1 and JAM-A suggests joint detection of these proteins as a future diagnostic tool in oral precancerous and cancerous conditions.

Systems Biology Approach Reveals a Calcium-Dependent Mechanism for Basal Toxicity in Daphnia magna.

The expanding diversity and ever increasing amounts of man-made chemicals discharged to the environment pose largely unknown hazards to ecosystem and human health. The concept of adverse outcome pathways (AOPs) emerged as a comprehensive framework for risk assessment. However, the limited mechanistic information available for most chemicals and a lack of biological pathway annotation in many species represent significant challenges to effective implementation of this approach. Here, a systems level, multistep modeling strategy demonstrates how to integrate information on chemical structure with mechanistic insight from genomic studies, and phenotypic effects to define a putative adverse outcome pathway. Results indicated that transcriptional changes indicative of intracellular calcium mobilization were significantly overrepresented in Daphnia magna (DM) exposed to sublethal doses of presumed narcotic chemicals with log Kow ≥ 1.8. Treatment of DM with a calcium ATPase pump inhibitor substantially recapitulated the common transcriptional changes. We hypothesize that calcium mobilization is a potential key molecular initiating event in DM basal (narcosis) toxicity. Heart beat rate analysis and metabolome analysis indicated sublethal effects consistent with perturbations of calcium preceding overt acute toxicity. Together, the results indicate that altered calcium homeostasis may be a key early event in basal toxicity or narcosis induced by lipophilic compounds.

Assessment of toxicity, genotoxicity and oxidative stress in Fejervarya limnocharis exposed to tributyltin.

Tributyltin (TBT) is widely used in various commercial applications due to its biocidal properties. Toxicological and genotoxicological data on TBT exposure to amphibians is insufficient. Our study aimed to determine the acute toxicity and genotoxic potential of TBT in Fejervarya limnocharis tadpoles. Furthermore, oxidative stress was also investigated in TBT-treated tadpoles. Tadpoles of Gosner stage (26-30) were screened and subjected to increasing concentrations of TBT (0, 3, 7, 11, 15, 19, 23 µg/L) for determining the LC50 values for 24 h, 48 h, 72 h, and 96 h. LC50 values of TBT for 24 h, 48 h, 72 h, and 96 h were found to be 19.45, 15.07, 13.12, and 11.84 µg/L respectively. Based on the 96 h LC50 value (11.84 µg/L), tadpoles were exposed to different sub-lethal concentrations of TBT for the evaluation of its genotoxic potential and effects on oxidative balance. The role of TBT on survivability, growth, and time to metamorphosis was also assessed. TBT exposure significantly altered the life history traits measured, increased mortality, and delayed the time taken to metamorphosis. Results indicated significant induction of micronucleus (MN, p < 0.001) and other erythrocytic nuclear aberrations (ENA, p < 0.01) in the TBT-treated groups. Significant alterations in comet parameters and oxidative balance were also observed in the treated groups. The present study findings might add to the cause of the gradual population decline seen in the amphibians. This study also demonstrates the alteration of the life-history traits, oxidative balance, and DNA damage upon TBT exposure which can have long-term consequences for the anuran amphibian F. limnocharis.

Evaluation of arsenic induced genotoxicity and its impact on life processes of Daphnia magna.

Heavy metals like arsenic is ubiquitously present in the environment. Geologic and anthropogenic activities are the root cause behind high concentration of arsenic in natural water bodies demanding strict monitoring of water quality prior to human consumption and utilization. In the present study, we have employed Daphnia magna for studying the biological effects of environmentally relevant high concentration of arsenic in water. In acute toxicity study, the LC50 value for 24hr exposure was found to be 2.504 mg/L, which gradually decreased with increase in time period (24hr- 96hr) to 2.011 mg/ L at 96hr. Sub-chronic toxicity was evaluated over 12 days using sub-lethal concentrations (5 %, 10 %, 15 %, and 20 % of the 24-hr LC50). Survivability in Daphnia showed a decreasing trend from 96 % to 91 % with increase in arsenic concentrations from 5 % of LC50 24 hr value to 20 % of LC 50 24hr value respectively. Alongside decreased survivability, there was a significant reduction in body size, with organisms exposed to the highest concentration of arsenic measuring 0.87±0.01 mm compared to 1.51±0.10 mm in the control group. Reproductive potential declined concentration dependently with exposure, with the highest reduction observed at 20 % of LC50 24hr value, where offspring numbers decreased to 7±1 from 23±5 in the control. Heart rate decreased in concentration and time-dependent manners, with the lowest rates observed at the highest arsenic concentration (279±16 bpm after 24hr and 277±27 bpm after 48hr). Comet assay and micronucleus assay conducted after 48 hrs of exposure revealed concentration-dependent genotoxic effects in Daphnia magna. Our results indicate negative impact on physiology and reproduction of Daphnia magna at environmentally existent concentration of arsenic. Also Daphnia magna could serve as a sensitive test system for investigating arsenic contamination in water bodies.

Toxic effects of fenvalerate in tadpoles of Fejervarya limnocharis: a multiparametric study.

Pyrethroids are among the most widely used insecticides. Fenvalerate (FEN), a synthetic pyrethroid, is frequently used in domestic and agricultural settings to control insects which ultimately find its way into the aquatic ecosystems. The larval stages of amphibians, which are experiencing a rapid population decline, are spent in aquatic habitats, thus making them vulnerable to FEN exposure. The potential toxic effects of pyrethoids in general and FEN in particular are not well understood. The present study was carried out to assess the toxicity of FEN in tadpoles of Fejervarya limnocharis. FEN at different concentrations (0, 4, 5, 6, 7, and 8 mg/L) induced substantial lethal effects. The estimated LC50 values were 8.54, 6.73, 5.44, and 4.44 mg/L at 24, 48, 72, and 96 h respectively. Exposure to environmentally relevant sub-lethal concentrations delayed metamorphosis and reduced survivality. FEN was found to be genotoxic in erythrocyte micronucleus and comet assay. Further, sub-lethal concentrations of FEN adversely affected the antioxidant defense mechanism of the exposed individuals with parallel increase oxidative damage to membrane lipids. The swimming behavior in the form of startle response, swirl response, and total movements was decreased with a concomitant decrease in AChE activity. In addition, FEN exhibited significant cardiotoxicity by decreasing the cardiac rate of the exposed individuals. The present findings clearly indicate that FEN can cause significant toxicity to the tadpoles of F. limnocharis affecting their survival and fitness in the natural environment.

Effects of changed water regime on the toxicity of silver nanoparticles (AgNPs) in tadpoles of Fejervarya limnocharis.

Climate change is viewed as one of the important causes of the amphibian population decline. Aspects of climate change like increase in water temperature and drying up of habitats have been underrepresented. The expanding production and usage of metal nanoparticles like silver nanoparticles (AgNPs) make them likely to end up in aquatic ecosystems. To arrive at a realistic assessment of the impact of AgNPs in a warming world, we have investigated the effects of temperature on the acute toxicity of AgNPs in tadpoles of Fejervarya limnocharis at 24, 48, 72 and 96 h of exposure. The various aspects of sub-lethal toxicities of AgNPs with increase in temperature were also investigated. Besides, the effects of habitat desiccation on the sub-lethal toxicities of AgNPs in the tadpoles were analysed. The LC50 values of AgNPs at four different time points were found to be significantly different between the two different temperatures. Alterations in survival pattern, life history traits, amplifications in genotoxic potential and oxidative stress were observed with increased water temperature following AgNP exposure. The phenomenon of habitat desiccation was also found to significantly affect the toxicity of AgNPs with respect to alterations in mortality rate, time to metamorphosis and morphometric parameters of metamorphosed tadpoles. The findings suggest that changed water regime such as increased water temperature as well as reduction in water level accelerated the toxic effects of AgNPs in F. limnocharis tadpoles which is likely to affect their natural populations.

Effects of radiation and vitamin C treatment on metronidazole genotoxicity in mice.

The impact of exposure to low dose radiation (LDR) on human health is not clear. Besides, cross adaptation or sensitization with pharmaceutical agents may modify the risk of LDR. In the present study, we analyzed the interaction of radiation and metronidazole (MTZ) in inducing chromosome aberration (CA) and micronucleus (MN) in the bone marrow cells of Balb/C mice in vivo. Further, we evaluated the efficacy of vitamin C to reduce MTZ induced genotoxicity. We found that 10, 20 and 40mg/kg of MTZ induced dose dependent increase in the frequency of CA (r=0.9923, P<0.01) as well as MN (r=0.9823, P<0.05) in polychromatic erythrocytes. However, MTZ did not affect the ratio of polychromatic erythrocytes to normochromatic erythrocytes indicating lack of cytotoxicity. Supplementation with vitamin C prior to MTZ treatment significantly reduced the frequency of CA (P<0.001) as well as MN (P<0.001). Radiation (0.5Gy) exposure prior to MTZ treatment produced a less than additive (for CA) to additive (for MN) effects. However, radiation exposure following MTZ treatment produced additive (for CA) and synergistic (for MN) effects. Further, vitamin C pre-treatment also reduced the genotoxicity indices following the combined treatment of MTZ and radiation. Our findings suggest that MTZ may sensitize bone marrow cells to radiation exposure and enhances genotoxicity. We recommend more studies on the interaction of LDR and marketed pharmaceuticals to minimize possible harmful outcomes through appropriate precautionary measures.

Areca nut and smokeless tobacco exposure induces micronucleus, other nuclear abnormalities and cytotoxicity in early chick embryo.

Areca nut (AN) and smokeless tobacco (SLT) are indiscriminately consumed among the populations of Southeast and South Asian countries, even by women during the gestational period. This study aimed to investigate the genotoxic and cytotoxic potentials of AN and Sadagura (SG), a unique homemade SLT preparation, alone and in combination in early chick embryos. Fertile white leghorn chicken eggs were randomly divided into five treatment groups: vehicle control, positive control (Mitomycin C, 20 µg/egg), AN, SG, and AN+SG. AN, SG, and AN+SG were given at dosages of 0.125, 0.25, and 0.5 mg/egg. The hen's egg test for micronucleus induction (HET-MN) was performed in chick embryos to evaluate the genotoxic potential of the test agents. Furthermore, the cytotoxic potential was assessed by studying erythroblast cell populations and the polychromatic erythrocytes (PCEs) to normochromatic erythrocytes (NCEs) ratio. Our results indicated a significant increase (p < .001) in MN frequency and other nuclear abnormalities, suggesting the potential of AN and SG to cause genotoxicity. Also, AN and SG exposure alone and in combination considerably altered the erythroblast cell population (%) and the PCE to NCE ratio in all the treatment periods. Our findings established the genotoxic and cytotoxic potential of both AN and SG alone and in combination during early embryonic development in the chick embryo.

Role of quercetin on mitomycin C induced genotoxicity: analysis of micronucleus and chromosome aberrations in vivo.

Quercetin, a flavonol group of plant flavonoid, has generated immense interest because of its potential antioxidant, anti-proliferative, chemoprotective, anti-inflammatory and gene expression modulating properties. However, the pro-oxidant chemistry of quercetin is important as it is related to the generation of mutagenic quinone-type metabolites. In the present study, 25mg/kg, 50mg/kg and 100mg/kg of quercetin given through the intra peritoneal (i.p.) route induced 2.31 ± 0.27%, 4.72 ± 0.58% and 6.38 ± 0.68% (control value=0.67 ± 0.30%) respectively, of cells with micronucleus (MN) in polychromatic erythrocytes in bone marrow cells and 10.93 ± 0.98%, 10.00 ± 0.89% and 14.27 ± 3.94% (control 2.61 ± 0.48) of cells with chromosome aberrations (CA) following 24h of the treatments. Higher frequencies of MN and CA were also observed after 48h of the treatments. To verify the effect of route of treatment on the quercetin induced damage, 100mg/kg b.w. was given through oral route which declined frequency of MN (P<0.001) as well as CA (P<0.05) as compared to the i.p. route for the same dose. Quercetin also induced higher frequency of metaphases with sticky chromosomes and C-mitosis. Pre-treatment with quercetin significantly reduced the frequency of mitomycin C (MMC) induced MN as well as CA, but no clear correlation between the dose and effect could be observed. Further studies are required to elucidate the possible interaction of quercetin with DNA as well as with other DNA damaging agents like MMC in vivo. The protective action of quercetin was not enhanced when given orally. Our findings suggest that quercetin may result in genomic instability in the tested dose range and significant reduction in MMC induced genotoxicity in the highest dose tested. These effects of quercetin are to be taken into consideration while evaluating the possible use of quercetin as a therapeutic agent.

Consumption pattern and genotoxic potential of various smokeless tobacco products in Assam, India: A public health concern.

Smokeless tobacco (SLT) consumption is presumed to be one of the major causes of high incidence of oral cancer in India. The present study aimed to document various types of SLT products consumed and their potential impact on the genome instability on the population from Assam state in Northeast India. A cross-sectional study (n = 5000) showed that 60.56 % of the study population consumed at least one of the three forms (sadagura, zarda and khaini) of SLT of which 52.0 % were only sadagura users. Genotoxicity assessment using buccal cytome assay in 240 age and sex matched volunteers revealed that except for zarda, other forms of SLT induced significantly higher incidence micronuclei in the buccal epithelial cells compared to the control individuals. Similar effects were also observed in other cytome parameters related to cell proliferation, cytokinesis defects and cell death. Significantly higher incidence of micronucleus was observed among sadagura and khaini users in lymphocyte cytokinesis-blocked micronucleus assay. The addition of lime in sadagura increased the pH and anion levels which possibly result in higher absorption and may lead to the development of cellular anomalies.

Phenanthrene alters oxidative stress parameters in tadpoles of Euphlyctis cyanophlyctis (Anura, Dicroglossidae) and induces genotoxicity assessed by micronucleus and comet assay.

Phenanthrene (PHE), a tricyclic polycyclic aromatic hydrocarbon (PAH), is ubiquitously found in aquatic environments. It is one of the major components in PAH mixtures. It has been identified as one of the 16 priority PAHs for toxicological evaluations. PHE is reported to induce lethal and sub-lethal toxicity in various aquatic indicator organisms. However, no toxicological data of PHE in anuran amphibians could be found. Amphibian larvae (tadpoles) develop in aquatic habitats. Therefore, exposure to PHE could negatively impact their development and fitness in later periods as they move in to the terrestrial habitat following metamorphosis. In the present study, we have analyzed the effects of PHE in Euphlyctis cyanophlyctis tadpoles. PHE induced concentration-dependent lethal effects in the tadpoles. The estimated LC50 values were 16.52, 15.29, 13.69, and 12.28 mg/L at 24, 48, 72, and 96 h of exposure respectively. These LC50 values are significantly higher than the reported environmental concentration of PHE. However, the strong negative correlation (R2 = 0.997, p < 0.001) between the LC50 value and exposure time indicates that longer exposure to lower concentration may cause significant lethal effects. Besides, PHE at environmentally relevant concentrations induced significant sub-lethal toxicities. Exposure to sub-lethal concentrations was found to be genotoxic in erythrocyte micronucleus as well as comet assays. Sub-lethal concentrations of PHE significantly increased superoxide dismutase activity and tissue glutathione level as well as induced lipid peroxidation. The present findings clearly indicate that PHE is a potential threat to the early life stages of amphibians. Further investigations are necessary to ascertain the implications of these early effects during adult life stages.

Arsenic and smokeless tobacco exposure induces DNA damage and oxidative stress in reproductive organs of female Swiss albino mice.

Arsenic contamination in the groundwater of Southern Assam, India is well-documented. A specific type of smokeless tobacco (sadagura, SG) is highly prevalent among the local population. Thus, the present study is aimed to evaluate the toxicological implications of arsenic and smokeless tobacco co-exposure on the reproductive health of female mice. The estrous cycle of experimental animals was monitored for 30 days. Histopathological studies and comet assay of ovarian and uterine tissues were performed after 30 days of exposure to SG and arsenic (sodium arsenite, SA). Oxidative stress was estimated biochemically by taking tissue glutathione, lipid peroxidation (LPO), and superoxide dismutase activity as endpoints. Our findings indicated a prolonged diestrus phase in the SG + L + SA group (p < 0.001). Histopathological study revealed abnormal tissue architecture in treated groups. Comet assay study showed that SG + SA exposure significantly induced DNA damage in test animals. The elevated LPO level in the SG + SA group indicated oxidative stress generation in the reproductive tissues. The present study suggests that female reproductive organs are vulnerable to SA and SG and oxidative stress generation may be the possible mechanism behind DNA damage, impaired follicular growth, atresia, and altered estrous cycle in the mice test system.

Micronucleus and other nuclear abnormalities among betel quid chewers with or without sadagura, a unique smokeless tobacco preparation, in a population from North-East India.

Genotoxicity is one of the important endpoints for risk assessment of various lifestyle factors. The study is the first report on the genotoxic effect associated with sadagura, a unique smokeless tobacco prepared in southern Assam province of North-East India. Sadagura is consumed with or without betel quid and/or smoking. In the present cytogenetic monitoring study, analysis of micronuceus (MN), nuclear bud, binucleated, karyorrhectic, karyolytic and pyknotic cells tests were performed in the exfoliated buccal cells of 75 habituates and compared to controls matched for gender, age, and habit. Significant increase in the frequency of MN was found in sadagura chewers (0.48%, P < 0.001), smokers (0.46%, P < 0.01), betel quid with sadagura chewers (0.91%, P < 0.001) and smokers chewing betel quid with sadagura (0.53%, P < 0.001) as compared to the unexposed control group (0.07%). Betel quid chewers showed significant increase (1.65%, P < 0.05) in the frequency of binucleated cells as compared to the control group (0.16%). Results of this study demonstrated that sadagura consumed as a single agent or in combination with betel quid, leads to a significant induction of cytogenetic damage in the buccal epithelial cells of habituates. We suggest that analysis of other degenerative nuclear changes in addition to MN can provide valuable information while evaluating potential genotoxic agents.

Cholesterol contributes to dopamine-neuronal loss in MPTP mouse model of Parkinson's disease: Involvement of mitochondrial dysfunctions and oxidative stress.

Hypercholesterolemia is a known contributor to the pathogenesis of Alzheimer's disease while its role in the occurrence of Parkinson's disease (PD) is only conjecture and far from conclusive. Altered antioxidant homeostasis and mitochondrial functions are the key mechanisms in loss of dopaminergic neurons in the substantia nigra (SN) region of the midbrain in PD. Hypercholesterolemia is reported to cause oxidative stress and mitochondrial dysfunctions in the cortex and hippocampus regions of the brain in rodents. However, the impact of hypercholesterolemia on the midbrain dopaminergic neurons in animal models of PD remains elusive. We tested the hypothesis that hypercholesterolemia in MPTP model of PD would potentiate dopaminergic neuron loss in SN by disrupting mitochondrial functions and antioxidant homeostasis. It is evident from the present study that hypercholesterolemia in naïve animals caused dopamine neuronal loss in SN with subsequent reduction in striatal dopamine levels producing motor impairment. Moreover, in the MPTP model of PD, hypercholesterolemia exacerbated MPTP-induced reduction of striatal dopamine as well as dopaminergic neurons in SN with motor behavioral depreciation. Activity of mitochondrial complexes, mainly complex-I and III, was impaired severely in the nigrostriatal pathway of hypercholesterolemic animals treated with MPTP. Hypercholesterolemia caused oxidative stress in the nigrostriatal pathway with increased generation of hydroxyl radicals and enhanced activity of antioxidant enzymes, which were further aggravated in the hypercholesterolemic mice with Parkinsonism. In conclusion, our findings provide evidence of increased vulnerability of the midbrain dopaminergic neurons in PD with hypercholesterolemia.

A highly reproducible mice model of chronic kidney disease: Evidences of behavioural abnormalities and blood-brain barrier disruption.

AIMS: In the present study, a novel mice model of chronic kidney disease (CKD) was developed, and psycho-motor behavioural abnormalities, blood-brain barrier (BBB) integrity and brain histology were studied. MAIN METHODS: Swiss albino female mice were given high adenine diet (0.3% w/w mixed with feed) for 4weeks. Serum urea and creatinine levels and renal histological studies were performed to validate the model. Psycho-motor behavioural abnormalities and neurological severity were studied. BBB integrity was assessed using Evans blue extravasation method. Nissl staining was performed to see possible morphological aberrations in brain. KEY FINDINGS: There was a significant increase in serum urea and creatinine levels in mice given high adenine diet, and the mice had abnormal kidney morphology. Deposition of adenine and 2,8-dihydroxyadenine crystals, and increased collagen deposits in the renal tissues were found, which validate induction of CKD in the mice. Motor behavioural abnormalities, depression-like and anxiolytic behaviour and increase in neurological severity were prevalent in mice with CKD. Evans Blue dye extravasation was found to occur in the brain, which signifies disruption of BBB. However, Nissl staining did not reveal any morphological aberration in brain tissue. SIGNIFICANCE: The present study puts forward a highly reproducible mice model of CKD validated with serum parameters and renal histopathological changes. The mice showed psycho-motor behavioural abnormalities and BBB disruption. It is a convenient model to study the disease pathology, and understanding the associated disorders, and their therapeutic interventions.

Chronic exposure of homocysteine in mice contributes to dopamine loss by enhancing oxidative stress in nigrostriatum and produces behavioral phenotypes of Parkinson's disease.

Increased homocysteine (Hcy) level has been implicated as an independent risk factor for various neurological disorders, including Parkinson's disease (PD). Hcy has been reported to cause dopaminergic neuronal loss in rodents and causes the behavioral abnormalities. This study is an attempt to investigate molecular mechanisms underlying Hcy-induced dopaminergic neurotoxicity after its chronic systemic administration. Male Swiss albino mice were injected with different doses of Hcy (100 and 250 mg/kg; intraperitoneal) for 60 days. Animals subjected to higher doses of Hcy, but not the lower dose, produces motor behavioral abnormalities with significant dopamine depletion in the striatum. Significant inhibition of mitochondrial complex-I activity in nigra with enhanced activity of antioxidant enzymes in the nigrostriatum have highlighted the involvement of Hcy-induced oxidative stress. While, chronic exposure to Hcy neither significantly alters the nigrostriatal glutathione level nor it causes any visible change in tyrosine hydroxylase-immunoreactivity of dopaminergic neurons. The finding set us to hypothesize that the mild oxidative stress due to prolonged Hcy exposure to mice is conducive to striatal dopamine depletion leading to behavioral abnormalities similar to that observed in PD.

Cadmium pollution and amphibians--Studies in tadpoles of Rana limnocharis.

Cadmium is released into the environment in increasing amounts from different natural and anthropogenic activities contaminating the aquatic habitats. Amphibian tadpoles develop in water and hence are likely to be adversely affected by cadmium present in the aquatic environment. We have studied the toxic and genotoxic effects of CdCl2 on the tadpoles of Rana limnocharis. CdCl2 in the concentration range between 0.1 and 0.4 mg/L induced significant mortality in R. limnocharis tadpoles in a dose and time dependent manner. The 10-day LC50 which has more ecological relevance was far less than the 24-h LC50. Tadpoles exposed to CdCl2 metamorphosed at an early age possibly as a survival strategy to move out of the stressful environment. The body weight of the CdCl2 exposed animals at metamorphosis was lower compared to the control individuals which may affect survival and reproductive fitness in adult life. Besides, the average body length of the metamorphosed individuals in the CdCl2 exposed group was higher than the control group. CdCl2 was found to be genotoxic in micronucleus test and comet assay. The ambient concentration of Cd could reach up to 60 µg/L or more. Exposure to 18.5 µg/L of CdCl2 (1% of 24-h LC50) induced significant increase in DNA strand breaks as compared to the control. The present findings demonstrate that presence of cadmium in the aquatic environment can significantly alter the life history traits and cause DNA damage in amphibians and hence, could contribute towards their population decline.

The antimalarial agent artesunate causes sperm DNA damage and hepatic antioxidant defense in mice.

Artesunate is an artemisinin derivative effective against multidrug resistant malaria. We analyzed the effects of artesunate 40 mg/kg b.w. as a single dose (ART1) or 13.3mg/kg b.w. for 3 days at 24h intervals (ART2) on mice spermatozoa at morphological and molecular level, and hepatic antioxidant status following 24h and 35 days following exposures in vivo. Artesunate significantly reduced epididymal sperm count and increased the frequency of sperms with abnormal head morphology following 24h of exposure. Comet assay analysis revealed significant increase in DNA strand breaks in spermatozoa evidenced by about 3-fold increase in comet tail DNA and up to 10-fold increase in Olive tail moment following 35 days of artesunate treatment. The damage index was significantly higher in the treated groups (40.27 ± 6.62 and 37.07 ± 5.35 for ART1 and ART2 respectively) as compared to the control group (16.13 ± 3.21) indicating the genotoxic effect of artesunate. The significant reduction in GSH, SOD and increase in lipid peroxidation indicate involvement of oxidative mechanisms in artesunate induced toxicity in mice. The present study suggests that artesunate has the potential to breach the testis-blood barrier and cause toxicity to male germ cells which may have implications in male reproductive toxicity.