Glucose-rich diet aggravates monocrotophos-induced dopaminergic neuronal dysfunction in Caenorhabditis elegans.

The present study aimed to obtain insights into the mechanism(s) by which glucose-rich diet aggravates monocrotophos (MCP)-induced dopaminergic neuronal dysfunction in Caenorhabditis elegans. In this study, we exposed three different strains of worms (wild-type N2, CB1112 (cat-2(e1112)II, tyrosine hydroxylase-deficient mutant, catecholamine absent) and the transgenic BZ555 (egls1-dat-1p::green fluorescent protein [GFP]) (in which bright GFP is tagged to the dopamine neuronal soma and processes) grown and maintained in normal nematode growth medium or 2% glucose enriched-nematode growth medium to MCP (0.75 mm) for 48 h. After the exposure, dopamine-mediated behaviors such as repulsion to nonanone, chemotaxis index and basal slowing response were determined in worms. Dopamine, 3,4-dihydroxy phenyl acetic acid and homovanillic acid content were quantified in N2 worms. The extent of neurodegeneration was visualized and quantified in dat-1::GFP worms. Basal slowing response study clearly indicated that cat-2 worms exposed to MCP and glucose were less affected compared to N2 of the same treatment. Learning and memory were affected by MCP and glucose. While MCP-treated worms showed lesser repulsion to nonanone compared to control worms, MCP-treated, glucose-fed worms showed a greater reduction in repulsion to nonanone. Further, MCP-treated, glucose-fed worms exhibited a marked reduction in dopamine content and an increase in 3,4-dihydroxy phenyl acetic acid and homovanillic acid levels compared to that in control. Dat-1::GFP showed a significant degeneration of dopaminergic neurons when exposed to glucose and MCP. Thus, our results clearly demonstrate that glucose-rich diet aggravates the dopaminergic neuronal dysfunction induced by MCP in C. elegans. Copyright © 2016 John Wiley & Sons, Ltd.

Microplate-based kinetic method for assay of mitochondrial NADH-- and succinate--cytochrome c reductase activities.

This article describes a microplate-based kinetic assay for mitochondrial NADH-- and succinate--cytochrome c reductase activities in rat brain mitochondria. The assay reported here is based on the conventional spectrophotometric method and involves substrate-driven reduction of exogenous cytochrome c. Conditions regarding linearity with respect to time and protein concentration have been standardized. Furthermore, the methods were tested for inhibition of respective activities by specific inhibitors. The microplate format described here can be employed for rapid and simultaneous measurements of mitochondrial NADH-- and succinate--cytochrome c reductase activities in a large number of samples.

Alpha-tocopherol ameliorates cypermethrin-induced toxicity and oxidative stress in the nematode Caenorhabdtis elegans.

Oxidative stress and other effects induced by cypermethrin (CYP, 15 mM) and their amelioration by alpha-tocopherol (400 microM) was studied in the nematode Caenorhabditis elegans. The worms exposed for 4 h to CYP showed increased levels of reactive oxygen species (46%), H2O2 (37%) and protein carbonyls (29%), accompanied by decreased lifespan and brood size. However, exposure to both CYP and alpha-tocopherol resulted in diminution of above alterations with the worms exhibiting relatively lower levels of ROS (30%), H2O2 (15%), protein carbonyls (14%), altered antioxidant enzyme activities and normal lifespan and brood size. The results suggest that CYP induces oxidative stress in C. elegans and the strategy of intervention with alpha-tocopherol could be exploited to offset this induced oxidative stress.

Cypermethrin elicited responses in heat shock protein and feeding in Caenorhabditis elegans.

Heat shock proteins (Hsp) are a family of stress proteins, which are elicited in response to a variety of stressors in organisms. The objective of this study was to explore the potential of cypermethrin, a synthetic pyrethroid insecticide, on induction of small heat shock proteins (Hsp16) and feeding response in transgenic Caenorhabditis elegans PC72 (Hsp16-LacZ). A concentration-dependent inhibition in feeding was evident in worms exposed to cypermethrin (31%, 46% and 56% at 5, 10, 15 mM, respectively) beyond 4h while marked induction of heat shock protein-16 expression was evident after 12h exposure (as evident from quantitative analysis). Maximum expression of Hsp16 was observed throughout the body of the worms 24h after exposure similar to that evident in the worms exposed to heat shock at 30 degrees C. These data suggest that cypermethrin possesses the potential to induce Hsp16 as well as inhibit feeding in C. elegans at non-lethal concentrations. C. elegans (PC72) thus could serve as a convenient model to study the early toxic effects of xenobiotics.

Effect of chronic exposure to monocrotophos on white adipose tissue in rats and its association with metabolic dyshomeostasis.

Earlier, we demonstrated that chronic exposure to monocrotophos (MCP) elicits insulin resistance in rats along with increased white adipose tissue (WAT) weights. This study was carried out to delineate the biochemical and molecular changes in adipose tissues of rats subjected to chronic exposure to MCP (0.9 and 1.8 mg/kg bw/d for 180 days). Pesticide-treated rats exhibited increased fasting glucose and hyperinsulinemia as well as dyslipidemia. Tumor necrosis factor-alpha and leptin levels were elevated, while adiponectin level was suppressed in plasma of treated rats. MCP treatment caused discernable increase in the weights of perirenal and epididymal WAT. Acetyl coenzyme A carboxylase, fatty acid synthase, glyceraldehyde-3-phosphate dehydrogenase, lipin-1, and lipolytic activities were elevated in the WAT of MCP-treated rats. Corroborative changes were observed in the expression profile of proteins that are involved in lipogenesis and adipose tissue differentiation. Our results clearly demonstrate that long-term exposure to organophosphorus insecticides (OPIs) such as MCP has far-reaching consequences on metabolic health as evidenced by the association of adipogenic outcomes with insulin resistance, hyperinsulinemia, endocrine dysregulations, and dyslipidemia. Taken together, our results suggest that long-term exposure to OPI may be a risk factor for metabolic dysregulations.

Reversible hyperglycemia in rats following acute exposure to acephate, an organophosphorus insecticide: role of gluconeogenesis.

The present study was undertaken to investigate the hyperglycemic potential of acute exposure to acephate and its etiology employing rat model system. Oral administration of acephate (140mg/kg b.w.) caused reversible hyperglycemia as evidenced by peak increase in blood glucose at 2h after the administration (87% over control) followed by trend of normalization. In further experiment carried out to understand the etiology of the induced hyperglycemia, we observed that 2h exposure to acephate caused significant increase in blood glucose, plasma corticosterone (78%) and activities of two gluconeogenesis enzymes in liver viz., glucose-6-phosphatase (91%) and tyrosine aminotransferase (84%) compared to that in control. When rats were exposed to acephate for 6h, decrement was observed in elevated levels of blood glucose, plasma corticosterone and the gluconeogenesis enzymes of the liver. Adrenal cholesterol levels in acephate-exposed rats were significantly depleted. While the glycogen content in liver of 2-h exposure group was comparable to control, a tremendous increase in liver glycogen content ( approximately 3.5 folds) was observed in rats of the 6-h exposure group. Our results demonstrate that acephate causes reversible hyperglycemia in rats probably by enhancing hepatic glucose output via gluconeogenesis. A role for hyperactivity of adrenal cortex is suggested in increased gluconeogenesis while significant attenuation in elevated levels of blood glucose and the activity the gluconeogenesis enzyme, glucose-6-phosphatase in liver with concomitant increase in liver glycogen are indicative of the onset of counter-regulatory responses such as hyperinsulinemia, to overcome the induced hyperglycemia.

Antioxidant-mediated protective effect of potato peel extract in erythrocytes against oxidative damage.

Potato peels are waste by-product of the potato processing industry. They are reportedly rich in polyphenols. Our earlier studies have shown that extracts derived from potato peel (PPE) possess strong antioxidant activity in chemical and biological model systems in vitro, attributable to its polyphenolic content. The main objective of this study was to investigate the ability of PPE to protect erythrocytes against oxidative damage, in vitro. The protection rendered by PPE in erythrocytes was studied in terms of resistance to oxidative damage, morphological alterations as well as membrane structural alterations. The total polyphenolic content in PPE was found to be 3.93 mg/g powder. The major phenolic acids present in PPE were predominantly: gallic acid, caffeic acid, chlorogenic acid and protocatechuic acid. We chose the experimental prooxidant system: FeSO(4) and ascorbic acid to induce lipid peroxidation in rat RBCs and human RBC membranes. PPE was found to inhibit lipid peroxidation with similar effectiveness in both the systems (about 80-85% inhibition by PPE at 2.5 mg/ml). While PPE per se did not cause any morphological alteration in the erythrocytes, under the experimental conditions, PPE significantly inhibited the H(2)O(2)-induced morphological alterations in rat RBCs as revealed by scanning electron microscopy. Further, PPE was found to offer significant protection to human erythrocyte membrane proteins from oxidative damage induced by ferrous-ascorbate. In conclusion, our results indicate that PPE is capable of protecting erythrocytes against oxidative damage probably by acting as a strong antioxidant.

A comparative study on the relationship between various toxicological endpoints in Caenorhabditis elegans exposed to organophosphorus insecticides.

The toxicity of 10 organophophorus (OP) insecticides-acephate, dimethoate, dichlorvos, dicrotophos, monocrotophos, methamidophos, phosphamidon, omethoate, phosdrin, and trichlorfon-was evaluated in Caenorhabditis elegans using lethality, movement, and acetylcholinesterase (AChE) activity as the endpoints after a 4-hr- exposure period. The OP insecticides tested showed LC50 values ranging from 0.039 mM (for dichlorovs) to 472.8 mM (for methamidophos). The order of toxicity for lethality and movement was not significantly different when tested using the rank order correlation coefficient. AChE activity was markedly affected by all the OP insecticide exposures that caused significant inhibition in movement, indicating that the mechanism of toxicity of OP insecticides in C. elegans is the same as in higher animals. All OP insecticides induced greater than 50% inhibition of AChE at the lowest tested OP insecticide concentration resulting in inhibition in movement. While a significant correlation was evident between LC50 values in C. elegans and the LD50 values in rats for the 10 OP insecticides studied, a correlation was not evident between EC50 values in C. elegans and LD50 values in rats. Overall, the two endpoints, LC50 and movement, were more reliable and easier to perform than measurement of AChE activity in C. elegans for determining the toxicity of OP insecticides. Further, ranking of these endpoints with respect to the OP insecticides studied indicates that these parameters in C. elegans are predictive of OP insecticides mammalian neurotoxicity.

Protective effect of potato peel extract against carbon tetrachloride-induced liver injury in rats.

Our earlier studies have shown that extracts derived from potato peel (PPE) are rich in polyphenols and possess strong antioxidant activity both in vitro and in vivo. The objective of the present study was to investigate its potential to offer protection against acute liver injury in rats. Rats pretreated with PPE (oral, 100mg/kgb.w./day for 7 days) were administered a single oral dose carbon tetrachloride (CCl(4), 3ml/kg b.w., 1:1 in groundnut oil) and sacrificed 8h of post-treatment. Hepatic damage was assessed by employing biochemical parameters (transaminase enzyme levels in plasma and liver [AST-aspartate transaminase; ALT-alanine transaminase, LDH-lactate dehydrogenase]). Further, markers of hepatic oxidative damage were measured in terms of malondialdehyde (MDA), enzymic antioxidants (CAT, SOT, GST, GPX) and GSH (reduced glutathione) levels. In addition, the CCl(4)-induced pathological changes in liver were evaluated by histopathological studies. Our results demonstrated that pretreatment of rats with PPE significantly prevented the increased activities of AST and ALT in serum, prevented the elevation of hepatic MDA formation as well as protected the liver from GSH depletion. PPE pretreatment also restored CCl(4)-induced altered antioxidant enzyme activities to control levels. The protective effect of PPE was further evident through the decreased histological alterations in liver. Our findings provide evidences to demonstrate that PPE pretreatment significantly offsets CCl(4)-induced liver injury in rats, which may be attributable to its strong antioxidant propensity.

Insights on the age dependent neurodegeneration induced by Monocrotophos, (an organophosphorous insecticide) in Caenorhabditis elegans fed high glucose: Evidence in wild and transgenic strains.

The higher susceptibility of high glucose fed C. elegans to Monocrotophos (MCP, an organophosphorus insecticide) - induced dopaminergic (DA) neuronal degeneration was recently demonstrated. Employing this acute exposure model, the impact of MCP on DA degeneration among worms of two age groups (8 and 13 d old) fed control (CO) and high glucose (GF) diet with specific focus on phenotypic alterations, oxidative impairments and associated molecular perturbations employing both wild (N2) and transgenic strains(BZ555 and NL5901) was investigated. In general, 13 d worms exhibited higher susceptibility to MCP intoxication compared to 8 d old worms. Further, MCP-exposure caused an enhanced degree of DA degeneration among glucose fed (GF) worms as evidenced by lower chemotaxis index, reduced long-term memory and increased nonanone repulsion. Biochemical analysis of 13 d GF worms also revealed a significant increase in ROS, protein carbonyls and reduced ADP/ATP ratio. Interestingly, marked increase in degeneration of dopaminergic neurons and increased in α-synuclein content was evident among 13 d GF worms exposed to MCP. Significant alterations in the mRNA expression levels of daf-2, age-1, sir 2.1 and aak-2 among 13 d GF worms was evident. Collectively these findings suggest that high intake of glucose diet aggravates MCP associated dopaminergic neuronal degeneration and the impact of increasing age under such a condition. Moreover it provides an experimental paradigm to explore the molecular targets and mechanism/s underlying the possible relationship between insecticide exposure-associated dopaminergic degeneration in humans under hyperglycemic conditions.

Neuroprotective effect of Decalepis hamiltonii aqueous root extract and purified 2-hydroxy-4-methoxy benzaldehyde on 6-OHDA induced neurotoxicity in Caenorhabditis elegans.

In this study, we investigated the possible neuroprotective efficacy of Decalepis hamiltonii tuber extract against 6-Hydroxy dopamine (6-OHDA) induced neurotoxicity and associated effects in Caenorhabditis elegans. The major component of flavour rich extract from D. hamiltonii is 2-hydroxy-4-methoxy benzaldehyde (2H4MB) which is an isomer of vanillin. We have conducted preliminary experiments with different types of extracts and subsequently DHFE (D. hamiltonii Fresh Tuber Extract) and DHPF (D. hamiltonii purified 2H4MB fraction) were used for further studies. Here we attempted to enumerate the neuroprotective efficacy of the above compounds in worms by evaluating behavioural and mitochondrial function, dopamine content and selective degeneration of dopaminergic neurons in BZ555 strains in comparison with control and 6-OHDA treated organisms. The relative expression levels of selected antioxidant genes involved in defence mechanism like SOD-3, GST-2 and GST-4 were evaluated along with those of CAT-2 and DOP-2 at mRNA level. We observed that both DHPF and DHFE exhibited significant levels of neuroprotective property against 6-OHDA induced neurotoxicity, which was evident in mitochondrial/dopaminergic function and antioxidant defence mechanism.

Altered glucose homeostasis and oxidative impairment in pancreas of rats subjected to dimethoate intoxication.

The primary objective of this study was to investigate the effect of repeated sublethal doses of dimethoate (DM), an organophosphorus insecticide on glucose homeostasis, oxidative stress induction in pancreas and pancreatic damage in adult rats. Daily oral administration of DM (20 and 40 mg/kg b.w.) for 30 days induced a significant increase in blood glucose levels which was associated with impaired glucose tolerance. DM treatment resulted in elevated levels of pancreatic tissue specific markers such as activities of amylase and lipase in serum and pancreatic tissue indicating pancreatic dysfunction. Further, the activities of DT-diaphorase and NADPH-diaphorase in pancreas of DM treated rats were also found to be elevated. Interestingly, these biochemical dysfunctions were accompanied by a marked dose-related enhancement of lipid peroxidation and ROS levels in the pancreatic tissue indicating significant induction of oxidative damage. Additional evidence such as depletion in reduced glutathione levels and significant alterations in enzymic antioxidant defenses in pancreas among DM treated rats suggested induction of oxidative stress. Taken together, these findings provide experimental evidence that dimethoate at subchronic oral doses has the propensity to impair glucose homeostasis, induce significant pancreatic damage and also provide an account of the associated oxidative damage to pancreatic tissue in adult rats.

Angiotensin converting enzyme inhibitory activity of amino acid esters of carbohydrates.

L-alanyl-D-glucose, L-valyl-D-glucose, L-phenylalanyl-D-glucose and L-phenylalanyl-lactose esters were synthesized enzymatically using two lipases viz., Rhizomucor miehei lipase (RML) and porcine pancreas lipase (PPL) and tested for their potential as inhibitors of angiotensin converting enzyme (ACE) in vitro. The esters exhibited concentration related ACE inhibitory activity. The potency of the various esters measured in terms of IC50 values were as follows: L-phenylalanyl-D-glucose, IC50-0.121 mM (mixture of five diastereomeric esters: 6-O-24.1%; 3-O-23.3%; 2-O-19.2%; 2,6-di-O-16.6% and 3,6-di-O-16.8% from the total yield of 92.4%); L-phenylalanyl-lactose, IC50-0.229 mM (mixture of three diastereomeric esters: 6-O-42.1%; 6'-O-30.9%; and 6,6'-di-O-27.0% from the total yield of 50.58%); alanyl-D-glucose, IC50-0.23 mM (mixture of five diastereomeric esters: 6-O-46.7%; 3-O-11.5%; 2-O-19.9%; 2,6-di-O-6.6% and 3,6-di-O-15.3% from the total yield of 26.5%) and L-valyl-D-glucose, IC50-0.396 mM (mixture of five diastereomeric esters: 6-O-32.4%; 3-O-26.5%; 2-O-26.4%; 2,6-di-O-8.8% and 3,6-di-O-5.9% from the total yield of 68.2%). These in vitro data suggest a potential therapeutic role for the aminoesters of carbohydrates as inhibibitors of ACE.

Antioxidant and neuroprotective potential of chitooligomers in Caenorhabditis elegans exposed to Monocrotophos.

The objectives of this investigation were to establish the propensity of the chitooligomers (COS) to ameliorate neurodegeneration and oxidative stress in Caenorhabditis elegans induced by an organophosphorus insecticide, Monocrotophos (MCP). COS was prepared from α-chitosan by the enzymatic method using chitosanase and characterized by HPLC and electron spray ionization-TOF-(ESI-TOF)-MS. We exposed age synchronized L4 C. elegans worms (both wild type N2 and transgenic strain BZ555 (Pdat-1:GFP) to sublethal concentration of MCP (0.75mM) for 24h in the presence or absence of COS (0.2mM). The neuroprotective effect of COS was examined in N2 worms in terms of brood size, lifespan, egg laying, dopamine content, acetylcholinesterase and carboxylesterase activity and by direct visualization and quantification of degeneration of dopaminergic neurons in BZ555. Exposure to COS extended lifespan, normalized egg laying, increased brood size, decreased the dopaminergic neurodegeneration, increased the dopamine content and increased AChE and carboxylesterase activity in C. elegans treated with MCP. COS induced a significant decrease in reactive oxygen species and increased the reduced glutathione level as well as increased superoxide dismutase and catalase activity. Our findings demonstrate that COS significantly inhibits the dopaminergic neurodegeneration and associated physiological alterations induced by MCP in C. elegans by attenuating the oxidative stress as well.

Attenuation of hyperglycemia and associated biochemical parameters in STZ-induced diabetic rats by dietary supplementation of potato peel powder.

BACKGROUND: Both dietary fiber and polyphenols have been reported to exert antihyperglycemic effect. Potato peel (PP), a waste by product of potato processing, is found to be a good source of both dietary fiber and polyphenols. The current study examined the attenuating influence of dietary potato peel (PP) powder on hyperglycemia and various oxidative stress-associated biochemical parameters in diabetic rats. METHODS: Streptozotocin (STZ)-induced diabetic male Wistar rats were used as experimental models. The rats were divided into nondiabetic (control), diabetic, potato-peel-supplemented (5% and 10%) and diabetic-PP (5% and 10%)-supplemented groups and were maintained for 4 weeks on the experimental regime. The modulatory role of PP was assessed by determining its effect on blood glucose, urine output, body weight gain, lipid peroxidation, reduced glutathione, serum aminotransferases, lipid profiles, selected antioxidant enzymes in liver/kidney and selected eye lens parameters. RESULTS: Diabetic rats fed with PP-powder-supplemented diet for 4 weeks showed a significant decrease in blood glucose levels. Incorporation of PP powder reduced significantly the hypertrophy of both liver and kidney of STZ-diabetic rats and also normalized the activities of serum ALT and AST, hepatic and renal MDA and GSH, as well as activities of various antioxidant enzymes in liver and kidney of diabetic rats. Furthermore, PP powder in the diet also appeared to attenuate the eye lens damage associated with the diabetic condition. CONCLUSION: Potato peel powder supplementation in diet was found to effectively attenuate diabetic alterations in rats.

Morphological and biochemical perturbations in rat erythrocytes following in vitro exposure to Fenvalerate and its metabolite.

Erythrocytes are a convenient model to understand the membrane oxidative damage induced by various xenobiotic-prooxidants. In this investigation, we have examined the potency of Fenvalerate (FEN) and its metabolite, p-chlorophenyl isovaleric acid (p-CPIA) to induce oxidative stress response in rat erythrocytes in vitro in terms of lipid peroxidation and effects on selected antioxidant enzymes. Susceptibility of erythrocytes to FEN exposure was further investigated in terms of morphological alterations by scanning electron microscopy and protein damage by gel electrophoresis of erythrocyte ghosts. Following in vitro exposure, FEN caused a significant induction of oxidative damage in erythrocytes at concentrations beyond 0.1 mM as evidenced by increased thiobarbituric acid reactive substances (TBARS) levels. The response was both concentration and time dependent. At higher concentrations, significant decreases in the activities of vital antioxidant enzymes viz., catalase, superoxide dismutase, glutathione transferase and glutathione reductase were also discernible clearly suggesting the potency of both, parent compound and its metabolite to induce oxidative stress in erythrocytes. Scanning electron micrographs of erythrocytes following FEN exposure at higher concentrations revealed various degrees of distortion in shape and ruptured membranes. Furthermore, gel electrophoresis studies revealed consistent and significant aggregation of only band 3 protein in erythrocyte membranes exposed to either FEN or p-CPIA at higher concentrations. These in vitro findings show that FEN and its metabolite have the propensity to cause significant oxidative damage in rat erythrocytes, which is associated with marked damage to membrane proteins. These data suggest that both structural and functional perturbations may ensue in erythrocytes following exposure to FEN at higher concentrations under in vivo situations.

Fenvalerate-induced oxidative damage in rat tissues and its attenuation by dietary sesame oil.

The primary objective of this study was to investigate the propensity of Fenvalerate (FEN), a synthetic pyrethroid to induce oxidative stress (OS) in various tissues of growing male rats following a short-term (28 days) dietary regimen and its possible attenuation by dietary (10%) sesame oil (SO). FEN incorporated diet was fed to weanling male rats at the dosages of 0, 250, 500 and 1000 ppm. Terminally, significant induction of OS in liver, thymus, spleen and erythrocytes was noticed at higher doses as evidenced by the elevated levels of lipid peroxidation (LPO). Significant dose-dependent depletion of GSH levels, perturbations in antioxidant enzymes, and enhanced protein carbonyls further confirmed the potential of FEN to induce OS in hepatic tissue. In addition, FEN also caused significant increases in activities of hepatic transaminases, ALP and LDH. Interestingly, dietary SO significantly attenuated FEN-induced oxidative damage in liver and other tissues. The degree of protection was remarkably high, since LPO and GSH status, protein carbonyl content and antioxidant defenses in liver and other tissues were brought down to basal levels in the SO + FEN1ooo group. These results clearly indicate the potential of FEN to induce oxidative damage in vivo and also suggest the ability of SO, a dietary fat to significantly offset the oxidative damage which may related to the presence of antioxidant compounds in the oil.

Alpha-linolenic acid suppresses dopaminergic neurodegeneration induced by 6-OHDA in C. elegans.

Parkinson's disease (PD) is the second most common neurodegenerative disorder characterized by the specific and massive loss of dopamine (DA) containing neurons in the substantia nigra pars compacta (SNpc) and aggregation of protein α-synuclein. There are a few animal studies, which indirectly implicate the neuroprotective action of omega-3 polyunsaturated fatty acids in neurodegenerative diseases. In this study, we exposed Caenorhabditis elegans (both wild type N2, and transgenic strain, UA44) to 6-hydroxydopamine (6-OHDA, the model neurotoxicant) and evaluated the extent of protection offered by alpha-linolenic acid (ALA). Larval stage worms (L1/L2) of N2 and transgenic strains were exposed to 6-OHDA (25 mM) with or without ALA (10, 50 and 100 µM) for 48 h at 20 °C. After 48 h, while the N2 worms were assessed for their responses in terms of locomotion, pharyngeal pumping, lifespan and AChE activity, the transgenic worms were monitored for dopaminergic neuronal degeneration. Worms exposed to 6-OHDA exhibited a significant reduction (48%) in the locomotion rate. Interestingly, supplementation with ALA increased the locomotion rate in 6-OHDA treated worms. A marked decrease (45%) in thrashing was evident in worms exposed to 6-OHDA while thrashing was slightly improved in worms co-exposed to 6-OHDA and higher concentrations of ALA. Interestingly, worms co-exposed to 6-OHDA with ALA (100 µM) exhibited a significant increase in thrashing (66 ± 1.80 thrashes/30s). The pharyngeal pumping rate declined significantly in the case of worms exposed to 6-OHDA (35%). However, the worms co-treated with ALA exhibited significant recovery in pharyngeal pumping. The mean survival for the control worms was 26 days, while the worms exposed to 6-OHDA, showed a marked reduction in survival (21 days). Worms co-exposed to 6-OHDA and ALA showed a concentration-dependent increase in lifespan compared to those exposed to 6-OHDA alone (23, 25 and 26 days respectively). Transgenic worms treated with 6-OHDA showed significant loss of processes of CEP and ADE neurons as evident from visibly marked reduction in GFP expression. Worms co-exposed to 6-OHDA and ALA showed visibly significant reduction in neuronal degeneration in both CEP and ADE. However, worms exposed to 6-OHDA together with ALA showed increased GFP expression within processes of CEP and ADE neurons. Overall, our results demonstrate that ALA significantly suppresses the dopaminergic neurodegeneration and movement disorder induced by 6-OHDA in C. elegans.

Effect of L-ascorbic acid supplementation on the toxicity of pirimiphos-methyl to albino rats.

The effect of L-ascorbic acid supplementation on Pirimiphos-methyl induced toxicity was studied in albino rats. Biochemical estimations were made in rats administered orally the insecticide at 100 and 200 mg/kg body weight with or without oral supplementation of L-ascorbic acid at 200 mg/kg b.w. The biochemical assessments included estimations of brain and plasma cholinesterases, levels of ascorbic acid in liver, kidney and adrenals, urinary levels of ascorbic acid and glucuronic acid. A lower degree of inhibition of the cholinesterases were evident in ascorbic acid supplemented rats. Marked elevation in urinary levels of ascorbic acid and glucuronic acid was observed in the insecticide treated rats. Results of this study suggests that L-ascorbic acid supplementation partially offsets Pirimiphos-methyl induced toxicity.

Oral exposure to the organophosphorus insecticide, Monocrotophos induces intestinal dysfunction in rats.

There is limited experimental evidence to imply the role of organophosphorus insecticides on intestinal dysfunctions. Residues of Monocrotophos (MCP), above maximum residue limits (MRL), have been reported in fruits and vegetables from various parts of India. Hence, in this study, we investigated the potential of MCP to induce intestinal dysfunction in rats. MCP was administered orally to rats at sublethal doses (0.45, 0.9 and 1.8 mg/kgb.w/d) for 30 days. MCP at the highest dose significantly increased the unit weight of the small intestine. MCP increased the activities of intestinal brush border disaccharidases, intestinal alkaline phosphatase, glycyl-glycine dipeptidase, and Na(+)/K(+)-ATPase while it decreased cholesterol: phospholipid ratio. Histology and scanning electron microscopy of small intestine of MCP treated rats revealed disruption in terms of congestion, increased length of villi, goblet cell hyperplasia, infiltration of inflammatory cells and necrotic villi tip. Further, the intestinal transit rate was found to be increased in MCP treated rats. Collectively, our findings provide evidence that repeated oral intake of MCP has the propensity to alter small intestinal structure and functions, which might lead to intestinal dysfunctions and abnormal nutrient uptake and thereby affect the human health. Although we have employed doses, which are higher than those likely to be encountered as residues, we speculate that further studies should be performed to determine whether MCP residues in foods in the long-term will interfere with the digestive capacity of the small intestine and thus exert adverse effects on the health of human.