Deltamethrin-induced oxidative stress and mitochondrial caspase-dependent signaling pathways in murine splenocytes.

Deltamethrin (DLM) is a well-known pyrethroid insecticide used extensively in pest control. Exposure to DLM has been demonstrated to cause apoptosis in various cells. However, the immunotoxic effects of DLM on mammalian system and its mechanism is still an open question to be explored. To explore these effects, this study has been designed to first observe the interactions of DLM to immune cell receptors and its effects on the immune system. The docking score revealed that DLM has strong binding affinity toward the CD45 and CD28 receptors. In vitro study revealed that DLM induces apoptosis in murine splenocytes in a concentration-dependent manner. The earliest markers of apoptosis such as enhanced reactive oxygen species and caspase 3 activation are evident as early as 1 h by 25 and 50 µM DLM. Western blot analysis demonstrated that p38 MAP kinase and Bax expression is increased in a concentration-dependent manner, whereas Bcl 2 expression is significantly reduced after 3 h of DLM treatment. Glutathione depletion has been also observed at 3 and 6 h by 25 and 50 µM concentration of DLM. Flow cytometry results imply that the fraction of hypodiploid cells has gradually increased with all the concentrations of DLM at 18 h. N-acetyl cysteine effectively reduces the percentage of apoptotic cells, which is increased by DLM. In contrast, buthionine sulfoxamine causes an elevation in the percentage of apoptotic cells. Phenotyping data imply the effect of DLM toxicity in murine splenocytes. In brief, the study demonstrates that DLM causes apoptosis through its interaction with CD45 and CD28 receptors, leading to oxidative stress and activation of the mitochondrial caspase-dependent pathways which ultimately affects the immune functions. This study provides mechanistic information by which DLM causes toxicity in murine splenocytes. © 2014 Wiley Periodicals, Inc. Environ Toxicol 31: 808-819, 2016.

Immunomodulatory role of piperine in deltamethrin induced thymic apoptosis and altered immune functions.

Deltamethrin (DLM), a well-known pyrethroid insecticide, is a potent immunotoxicant. In rodents, it is primarily characterized by marked thymic apoptosis. Mechanism of DLM induced thymic apoptosis in primary murine thymocytes has been recently explored. Oxidative stress and activation of caspase dependent pathways appear to be involved in the DLM induced thymic injury. Thus, for the amelioration of its effect, this study has been designed to first observe the binding affinity of piperine to immune cell receptors and its protective effects on the DLM induced immunotoxicity under in vitro condition. The docking results demonstrated that piperine has good binding affinity towards CD4 and CD8 receptors. In vitro study results have shown that piperine (1, 10 and 50 µg/ml) increased cell viability in a concentration dependent manner. The early activated markers of apoptosis such as enhanced reactive oxygen species (ROS) and caspase-3 activation by DLM was significantly reduced by piperine treatment. GSH depletion induced by DLM has been also restored by piperine treatment. At 18 h, all concentration of piperine (1, 10 and 50 µg/ml) significantly ameliorated the DLM induced apoptosis. Further, DLM induced phenotypic changes were mitigated by the piperine. In addition, piperine also restored the cytokine levels, which were suppressed by DLM treatment. These findings strongly indicate the anti-oxidative, anti-apoptotic and chemo-protective ability of piperine in the DLM induced thymic apoptosis.

Deltamethrin induced an apoptogenic signalling pathway in murine thymocytes : exploring the molecular mechanism.

Deltamethrin (DLM) is a well-known pyrethroid insecticide; however, the immunotoxic effects of DLM on the mammalian system and its mechanism is still unclear. This study has been designed to first observe the binding affinity of DLM to immune cell receptors and its effects on the immune system. The docking score revealed that DLM has a strong binding affinity towards the CD4 and CD8 receptors. DLM induces apoptosis in murine thymocytes in a concentration-dependent manner. The ear\ly markers of apoptosis such as enhanced reactive oxygen species (ROS) and caspase-3 activation are evident as early as 1 h by 25 and 50 µM DLM. Glutathione (GSH) depletion has also been observed at 1 h by 50 µM DLM concentration. In cell-cycle studies using flow cytometry, the fraction of hypodiploid cells has gradually increased with all the concentrations of DLM at 18 h. The Annexin V binding assay measures the effect of DLM on apoptotic and necrotic cells. The apoptotic cells raised from 18.6% to 35.21% (10-50 µM DLM) at 18 h. N-acetyl cysteine (NAC) effectively reduced the percentage of apoptotic cells which is increased by DLM. In contrast, buthionine sulfoximine (BSO) caused an elevation in the percentage of apoptotic cells. These results demonstrate that caspase activation, ROS activation and GSH act as critical mediators in a DLM-induced apoptogenic signalling pathway in murine thymocytes. In the presence of caspase inhibitor, the percentage of apoptotic cells is partially decreased. Thus, there may be the possibility of some other caspase-independent pathways in DLM-induced apoptosis.

CNS depressant and anticonvulsant activities of the alcoholic extract of leaves of Ziziyphus nummularia.

ETHNOPHARMACOLOGICAL RELEVANCE: Ziziyphus nummularia (family: Rhamnaceae) is a xerophyte, grows in the grazing lands of the Thar Desert of Rajasthan. Ziziyphus nummularia (ZN) is used as sedative in ethnomedicine. The objective of this study is to investigate the anticonvulsant, anxiolytic and sedative activities of the alcoholic extract of leaves of Ziziyphus nummularia (EZN). MATERIALS AND METHODS: The anticonvulsant effect of the EZN (100, 200 and 300 mg/kg) was evaluated in mice using the pentylenetetrazole and maximal electroshock induced seizure models. Its anxiolytic activity was evaluated using the elevated plus maze, hole board and open field models board methods, while the pentobarbital induced sleep was used to evaluate the sedative activity. The acute toxicity and effect on motor coordination were also assessed. RESULTS: EZN (100-300 mg/kg) protected the mice against the pentylenetetrazole induced convulsions; it causes a significant (P<0.05) dose dependent increase in latency of convulsion. Treatment with EZN reduced the duration of the tonic hind limb extension induced by electroshock. Mice treated with EZN preferred the open arm of the plus maze and were found to be devoid of open-arm avoidance. EZN potentiation the barbiturate induce sleep in mice, it causes a decrease in the sleep latency and increases the duration of sleep. CONCLUSION: The results obtained from the experiments indicate that the EZN has CNS depressant and anticonvulsant activities.

Analgesic and anti-inflammatory studies of cyclopeptide alkaloid fraction of leaves of Ziziyphus nummularia.

Ziziyphus nummularia (family: Rhamnaceae) is a thorny small bush, grows in abundance in the grazing lands of the arid areas of Rajasthan, India. It is an important ethnomedicinal plant of the Thar Desert; local inhabitants use every part of the plant as medicine. Kernels are prescribed in pregnancy as soporific, antiemetic and for relieving abdominal pain. The insect gall is powered and given orally with water to cure bone fracture. Crushed root is applied on the paining shoulder of the bullock. The decoction of leaves is used for the treatment of cough and cold; leaves are also regarded as diaphoretic and prescribed in typhoid. Paste of leaves is used for healing of cuts, boils and cutaneous disease. It is widely used in pain and inflammatory conditions. Z. nummularia contains a unique group of alkaloids known as cyclopeptide alkaloids, in continuation of our work carried out on the leaves of Z. nummularia , present study was initiated to explore antiinflammatory and analgesic potential of cyclopeptide alkaloids isolated from the leaves of Z. nummularia (IFZN). Anti-inflammatory activity was tested against rat paw oedema, mouse peritonitis and cotton pellet granuloma. For screening of analgesic activity, acetic acid induced writhing, tail flick and hot plate test were performed. IFZN 30 mg/kg shows the anti-oedematogenic effect against paw oedema induced by carrageenan, dextran, serotonin and histamine; IFZN 20 and 30 mg/kg were found to have highly significant anti-nociceptive effects. Result of pharmacological studies indicated that IFZN is a potent and efficacious analgesic agent. The analgesic activity of IFZN is mediated by the peripheral as well as central pathways.

Attenuating Aß1-42-induced toxicity by a novel acetylcholinesterase inhibitor.

We explored the attenuating effects of NP-9 on ß-amyloid (Aß) aggregation and amyloid-induced toxicity. NP-9 is a recently reported monoamine oxidase B (MAO-B), and acetylcholinesterase (AChE) inhibitor. In the present study, we found that NP-9 inhibited AChE activity in a dose-dependent manner with a maximal inhibition dose of 8 mg/kg, i.p. It inhibited Aß aggregation, observed through thioflavin-T assay (IC50=60 µM) and scanning electron microscopy (S.E.M.) (no fibril formation). NP-9 has shown marked protection against scopolamine and Aß1-42-induced memory impairments. It also minimized neuronal loss and amyloid plaque deposition in the brains of Aß1-42-induced mice model. Therefore, NP-9 could be a promising lead molecule for AD, with effects against MAO-B, AChE, Aß aggregation, and Aß1-42 induced toxicity.

Wound healing activity of latex of Euphorbia caducifolia.

ETHNOPHARMACOLOGICAL RELEVANCE: [corrected] Euphorbia caducifolia is a Euphorbiaceae species native to Thar Desert of India, where latex of E. caducifolia (ECL) is used by the local inhabitants for treatment of bleeding wound, cutaneous eruption and other skin diseases. Looking to the medicinal importance of the plant, it was considered worthwhile to scientifically evaluate the ECL for the wound healing activity. MATERIAL AND METHODS: In vitro methods, clotting of platelet free plasma and angiogenesis in chick chorioallantoic membrane (CCM) were used for studying effect of ECL on clotting and angiogenesis. Excision and incision wounds model were used to study effect of ECL on wound contraction, tensile strength and hydroxyproline and DNA content. RESULTS: ECL treatment stimulates the blood vessel formation in CCM, at the all dose levels. 56.77, 74.48 and 78.09% increase in area of angiogenesis was observed in ECL 2.5, 5.0 and 10mg/ml treated groups respectively. The excised skin of ECL 5.0 and 10mg/ml treated animals found to have higher content of hydroxyproline and DNA, when compared with control group. An acceleration of the wound closure was observed with an enhanced number of fibroblasts and collagen content in ECL treated mice. CONCLUSION: Result of in vitro and in vivo experiments indicate that ECL exerts significant wound healing activity, demonstrating its relevant therapeutic potential.

Development of selective and reversible pyrazoline based MAO-B inhibitors: virtual screening, synthesis and biological evaluation.

In an effort to develop selective MAO (monoamine oxidase) B inhibitors, structure based virtual screening was initiated on an in-house library. Top 10 HITS were synthesized and evaluated for MAO (A and B) inhibitory activity, both against human and rat enzymes. All the compounds were found selective, reversible and active in nM range (100 times more potent than selegeline) towards MAO-B. Outstanding co-relation between predicted and experimental K(i) values were observed.

Aerva lanata: A review on phytochemistry and pharmacological aspects.

Aerva lanata (L.) A. L. Juss. ex Schultes. (Amaranthaceae) locally known as 'bui' is an erect or prostrate undershrub with a long tap-root and many wolly-tomentose branches, found in the wild, throughout India. In traditional medicine the plant is used in cough, strangury (slow to be and painful discharge of urine), headache and urolithiasis. The photochemical constituents present in the plant include alkaloids (ervine, methylervine, ervoside, aervine, methylaervine, aervoside, ervolanine, and aervolanine), flavanoids (kaempferol, quercetin, isorhamnetin, persinol, persinosides A and B), methyl grevillate, lupeol, lupeol acetate benzoic acid, ß-sitosteryl acetate and tannic acid. Pharmacological studies reported diuretic, anti-inflammatory, hypoglycemic, anti-diabetic, antiparasitic, antimicrobial, hepoprotective, anti-urolithiasis, antiasthmatic, antifertility and hypolipidemic properties of Aerva lanata. This review article includes the detailed exploration of the morphology, phytochemistry, and pharmacological aspects of Aerva lanata in an attempt to provide a direction for further research.

The antihyperglycemic effect of aerial parts of Salvia splendens (scarlet sage) in streptozotocin-induced diabetic-rats.

Salvia splendens (Labiatae) is widely used in Indian traditional medicine for the control of diabetes mellitus. In this study, the hypoglycemic effects produced by the acute and subacute administration of various extracts of S. splendens were investigated. Both the aqueous extract (SSAE) and the methanolic extract (SSME) from the aerial parts resulted in significant reductions of glycemia in streptozotocin (STZ)-induced diabetic rats after oral administration at a dose of 100 and 200 mg/kg, respectively. On oral administration, aqueous and methanolic extracts showed statistically significant (P < 0.001) effect by reducing the effect of glycemia in STZ-induced diabetic rats. These findings suggest the significant antihyperglycemic potential of the S. splendens extracts in ameliorating the diabetic conditions in diabetic rats. No significant effects were found in the normal rats.

Sedative and anticonvulsant effects of an alcoholic extract of Capparis decidua.

Capparis decidua (frock) Edgew (family Capparidaceae) is a xerophytic shrub, commonly known as karrel or ker, whose bark and shoot are used as analgesic, anti-inflammatory, hypolipidemic, and antidiabetic agents. The plant contains generous quantities of alkaloids. An alcoholic extract of aerial parts of C. decidua, including flowers and fruits, was screened for central nervous system (CNS) activity using conventional behavioral animal models. In the open field test all doses of C. decidua extract tested decreased the number of rearings, grooming, and fecal bolus (P < 0.001) when compared with control. In the barbiturate-induced sleeping test a significant (P < 0.001) a decrease in latency of sleeping and increase in sleeping time were observed at all doses (100, 200, and 300 mg/kg). C. decidua extract increased the percentage of animals exhibiting motor deficit in the rotarod test. In the pentylenetetrazole-induced seizures test the C. decidua extract dose-dependently decreased (P < 0.05) the number of animals with convulsions and increased convulsion latency (P < 0.001); none of the animals treated with extract died in the test. C. decidua extract decreased the duration of tonic hind leg extension in maximal electroshock-induced seizures (P < 0.001) when compared with control. The findings of the present animal study suggested that C. decidua has CNS depressant and anticonvulsant activities.

3D-QSAR study of 8-azabicyclo[3.2.1] octane analogs antagonists of cholinergic receptor.

3D-QSAR models of Comparative of Molecular Field Analysis (CoMFA) and Comparative of Molecular Similarities Indices Analysis (CoMSIA) of 20 8-azabicyclo[3.2.1] octane (potent muscarinic receptor blocker) was performed. These benztropine analogs were optimized using ligand based alignment method. The conventional ligand-based 3D-QSAR studies were performed based on the low energy conformations employing database alignment rule. The ligand-based model gave q(2) value 0.819 and 0.810 and r(2) value 0.991 and 0.988 for CoMFA and CoMSIA, respectively, and the predictive ability of the model was validated. Results indicate that the CoMFA and CoMSIA models could be reliable model which may be used in the design of novel muscarinic antagonists as leads.

A serological survey of antibodies against bovine herpesvirus-1 in yak (Poephagus grunniens) in Arunachal Pradesh in India.

Serum samples were collected from 254 yak (Poephagus grunniens, presently Bos grunniens) in different yak tracts of India. These samples were then screened by virus neutralisation test (VNT) and avidin-biotin enzyme-linked immunosorbent assay (AB-ELISA) to study the seroprevalence of antibodies against bovine herpesvirus type 1 (BHV-1). The overall seroprevalence in yak was found to be 41% (105) by VNT and AB-ELISA. The sex of the animal, whether it was on a farm or free-ranging and the location of the different yak tracts did not seem to have any effect on seroprevalence. However, seroprevalence was found to increase with the age of the animals, being highest in yak older than three years of age (49%). Yak generally share feeding, watering and grazing areas with other domestic and wild animals and this common ecological niche is thought to be a possible avenue of infection. This is the first time that the seroprevalence of antibodies against BHV-1 has been studied in yak in India.

Serological evidence of antibodies against Chlamydophila abortus in free-ranging yak (Poephagus grunniens) in Arunachal Pradesh, India.

Serum samples were randomly collected from 172 free-ranging yak (Poephagus grunniens, presently Bos grunniens) from six different yak tracts of Arunachal Pradesh, India, and subjected to enzyme-linked immunosorbent assay to detect the presence of specific antibodies against Chlamydophila abortus. The overall prevalence of this disease in yak was 35%. The prevalence of Cp. abortus-specific antibodies was significantly higher in yak cows (41%) than among bulls (25%). The highest prevalence (39%: 95% confidence interval [Cl] = 27, 55) was found in yak between one and three years of age, while the lowest prevalence (20%: 95% CI = 10, 41) was reported in yak below one year of age.

Evaluation of CNS Depressant Activity of Different Plant parts of Nyctanthes arbortristis Linn.

The present study was carried out with the water-soluble portion of the ethanol extracts of flowers, barks, seeds and leaves of Nyctanthes arbortristis Linn. to confirm their CNS depressant activity. The ethanol extracts of the plant parts were obtained by soxhlet extraction. After performing the gross behavioral study, the CNS depressant activity was evaluated by observing the prolongation of sleeping time induced by pentobarbital sodium in mice. Attempts have been made to explore the possible mechanism behind this activity by determining their effect on brain monoamine neurotransmitters like dopamine and serotonin. The gross behavioral study showed that ethanol extracts of the leaves, flowers and seeds possess significant CNS depressant activity. The leaves, flowers, seeds and barks (600 mg/kg) showed significant and dose-dependent prolongation of onset and duration of sleep and so found to cause decrease dopamine and increase serotonin level. From which it can be concluded that the CNS depressant activity of the ethanol extracts of seeds, leaves and flowers may be due to the decrease in dopamine and increase in serotonin level.

Anti-diarrheal activity of methanol extract of Litsea polyantha bark in mice.

The anti-diarrheal activity of methanol extract of dried bark and aerial parts of Litsea polyantha (MELP) has been evaluated in mice using different models (castor oil-induced diarrhea and propulsive gut motility in mice). MELP (50, 75, 100 mg/kg, p.o.) significantly (P<0.01) reduced the onset of diarrhea, fecal excretion and also showed a significant (P<0.001) reduction in gastrointestinal motility on charcoal meal test in mice. The results of the study support the folklore use of the plant for diarrheal remedies.

Diuretic activity of Nyctanthes arbortristis Linn.

Nyctanthes arbortristis Linn. is a well documented plant. The present study is done to establish the diuretic activity of the water-soluble portions of the ethanolic extracts of its flowers, barks, seeds and leaves. In toxicity study, the extracts were seen to be safe up to the dose of 2.0 gm/kg. For the estimation of diuretic activity, the parameters studied were total urine volume and urine concentration of Na(+), K(+) and Cl(-). The ethanolic extracts of different plant parts of Nyctanthes arbortristis L. possess significant diuretic activity as reflected by rise in urine volume with cation excretion. The ethanolic extracts of the seeds and leaves at their higher doses exhibited higher electrolyte excretion.

N-acetyl-D-glucosamine specific hemagglutinin receptor of Vibrio cholerae O1 in chicken erythrocyte membranes.

N-Acetyl-D-glucosamine specific cell-associated hemagglutinin (HA)/lectin, previously purified from a strain of Vibrio cholerae O1, had been established as an adhesin molecule of V. cholerae O1 cells. This communication records the isolation and purification of the glycoprotein receptor of the N-acetyl-D-glucosamine specific HA of the V. cholerae O1 strain from chicken erythrocyte membranes. The most salient feature of this study is that the pretreatment of partially purified glycoprotein with purified HA could completely inhibit the hemagglutinating activity of the V. cholerae O1 strain with chicken erythrocytes.

Antimicrobial activity of the mycotoxin citrinin obtained from the fungus penicillium citrinum.

The mycotoxin Citrinin was obtained from the fungus Penicillium citrinum. It was tested for it's Minimum Inhibitory Concentration (MIC) against some gram positive strains viz. Staphylococcus aureus, Bacillus pumilus, Bacillus subtillis, Bacillus cereus, Klebsiella pneumoniae, Streptococcus pneumoniae, Lactobacillus arabinosus and gram negative strains E.Coli, Shigella dysenteriae, shigella sonnei, shigella boydii, Salmonella typhimurium, Proteus mirabilis and Vibrio cholerae. Further the zones of inhibition produced by the fungal extract against the bacterial strains were assayed and compared with those produced by the standard antibiotic ciprofloxacin.

Mycotoxins - limits and regulations.

Since early years, a need has always been felt for some control on the quality of foodstuffs. With the discovery of aflatoxins in the early sixties, health authorities in man countries have become active in establishing regulations to protect their citizens and livestock fro t potential harm caused by mycotoxins. FDA mycotox-ins-in-foods sampling program is continuing with an objective to remove those foods from interstate commerce that contain Aflatoxins "at levels judged to be of regulator significance" Aflatoxins, Fumonisin B1 and B2, Deoxynivalenol (DON) Ochratoxin A and Patulin occur in a number of food products. FDA workers were instructed to sample and analyze all products for different types of mycotoxins. All baby foods should always be analyzed for all type of mycotoxins. The limits of Aflatoxins B1,B2,! < G2, and M1 in foods and feed stuffs varies from (0-40) ppb for foods & 0-1000ppb for food); for Ochratoxin A(0-50 ppb in food and 0-1000ppb in feed); for Don (500-2000ppb in food & 5-10,000 ppb in feed); for Zearalenone (0-1000 ppb in food); for Patulin (0-50 ppb in foods), for Diacetoxyscirpenol (0-100 ppd in feed); for chetomin (0ppb I feed); for stachybotryotoxin (0ppb in feeds and for Fumonisins (0-1000 ppb in food 5000-50,000 ppb in feedstuffs).