Rajbir Kaur

Free radical scavenging potential of some Indian medicinal plants

Indian medicinal herbs and plants are used since ancient times to treat different diseases and ailments as these natural products exert broad-spectrum actions. The present study was aimed to explore the hydrogen donating and hydroxyl radical scavenging potential of methanol extract of 10 medicinal plants belonging to six families including mimosaceae, Apocynaceae, moraceae, sapindaceae, rutaceae and meliaceae using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and plasmid nicking assay, respectively. The total phenol and flavonoid content of these extracts was also estimated using Folin-Ciocalteu and colorimetric assay respectively. It was found that methanol leaf extract of Koelreutaria paniculata, Acacia catechu and Mimusops hexandra showed strong inhibitory activity whereas that of Hamelia patens exhibited moderate DPPH radical scavenging activity at concentration of 200 µg/ml. However, methanol extract of Swietenia mahogoni, Murraya exotica, Murraya koenigii, Alstonia scholaris, Ficus benjamina and Sapindus trifoliatus exhibited weak hydrogen donating potential in DPPH assay. The methanol extract of these plants was effective in plasmid nicking assay and the activity was found to be correlated to the phenolic and flavonoid content in these fractions. These results emphasized the benefit of the phenolic compounds rich plant extracts and thus augmented the urge of in vivo studies to further confirm the beneficial effect of these extracts.

Interactions of betulinic acid with xenobiotic metabolizing and antioxidative enzymes in DMBA-treated Sprague Dawley female rats

Cancer chemoprevention is related to classical epidemiology and involves the use of agents that inhibit, delay, or reverse the carcinogenesis that occurs as a result of accumulation of mutations and increased proliferation. Betulinic acid is known for its cytotoxic effects against a panel of cancer cell lines. In the present study, interactions of betulinic acid (BA) with xenobiotic metabolizing enzymes including mixed function oxidases (cytochrome b5, P420, P450, NADPH cytochrome P450 reductase, and NADH cytochrome b5 reductase), phase II enzymes (GST, DT-diaphorase, γ-glutamyl transpeptidase), LDH, antioxidative enzymes (glutathione reductase, SOD, catalase, ascorbate peroxidase, and guaiacol peroxidase), and lipid peroxidation are studied alone as well as in the presence of 7,12 dimethylbenzanthracene (DMBA)--a potent carcinogen using Sprague Dawley female rats. The effect of BA on reduced glutathione content and protein content is also taken into consideration. It has been found that administration of BA decreased the level of mixed function oxidases that are involved in the conversion of carcinogen to electrophile, elevated the level of phase II enzymes which participated in the removal of electrophiles by sulfation, conjugation etc. It has been found that BA effectively removed or neutralized the reactive species by the action of phase II enzymes and such an effect was reflected from the specific activities of antioxidative enzymes which were found to be lower as compared to positive control (DMBA-treated group) and in some cases even that of untreated control. BA was also found to have a pronounced effect in protecting the animals from lipid peroxidation as evident from the reduced levels of TBARS, conjugated diene, and lipid hydroperoxide formation. This study highlights the role of BA in modulating the activities of xenobiotic and antioxidative enzymes that have putative roles in cancer initiation and proliferation.

Diminution of Hepatic Response to 7, 12-dimethylbenz(α)anthracene by Ethyl Acetate Fraction of Acacia catechu Willd. through Modulation of Xenobiotic and Anti-Oxidative Enzymes in Rats

Background Liver is the primary metabolizing site of body and is prone to damage by exogenous as well as endogenous intoxicants. Polycyclic aromatic hydrocarbons such as 7, 12- dimethylbenz(α)anthracene (DMBA) is an exogenous hepatotoxin, which is well known for modulating phase I, II and anti-oxidative enzymes of liver. Plants contain plethora of polyphenolic compounds which can reverse the damaging effect of various xenobiotics. The present study investigated protective role of the ethyl acetate fraction of Acacia catechu Willd. (EAF) against DMBA induced alteration in hepatic metabolizing and anti-oxidative enzymes in rats. Methodology and Principal Findings The rats were subjected to hepatic damage by treating with DMBA for 7 weeks on alternative days and treatment schedule was terminated at the end of 14 weeks. The rats were euthanized at the end of protocol and livers were homogenized. The liver homogenates were used to analyse phase I (NADPH-cytochrome P450 reducatse, NADH-cytochrome b5 reductase, cytochrome P420, cytochrome b5), phase II (glutathione-S-transferase, DT diaphorase and γ-Glutamyl transpeptidase) and antioxidative enzymes (catalase, superoxide dismutase, ascorbate peroxidase, glutathione reductase, guiacol peroxidase and lactate dehydrogenase). Furthermore, other oxidative stress parameters (thiobarbituric acid reactive substances, lipid hydroperoxides and conjugated dienes and reduced glutathione) and liver marker enzymes (serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase and alkaline phosphatase) were also studied. The DMBA induced significant changes in activity of hepatic enzymes that was reversed by treatment with three dose levels of EAF. Conclusion It is concluded that EAF affords hepato-protection against DMBA in rats through modulation of phase I, II and anti-oxidative enzymes.

Inhibition of lipid peroxidation by extracts/subfractions of Chickrassy (Chukrasia tabularis A. Juss.)

Polyphenols and polyphenol-rich fractions of plants have been reported to have protective effects against lipid peroxidation, most probably by serving as scavengers of free radicals and/or by chelating metal ions. In the present study, the effect of different extracts/subfractions of Chickrassy (Chukrasia tabularis) on peroxyl radical mediated damage to the polyunsaturated fatty acids was investigated. Liver homogenate was used as experimental material. The production of malondialdehyde served as a marker of lipid peroxidation and oxidative stress. It was observed that polyphenol-rich fractions, particularly the ethyl acetate fractions of bark and leaves, showed the highest protective activity of 83.02% and 88.62% inhibition, respectively. This study will help in knowing the scientific validation of this plant, for its use in ayurvedic formulations.

Assessment of mutagenic, genotoxic, and cytotoxic potential of water samples of Harike wetland: a Ramsar site in India using different ex vivo biological systems

Harike is a wetland of international importance under the Ramsar Convention. The present study entails the investigation of mutagenic, genotoxic and cytotoxic effect of surface water samples collected from five different areas of the Harike wetland by using the histidine reversion point mutation assay in Salmonella typhimurium (TA98 and TA100) strain with or without S9, bioluminescence mutagenicity assay using Vibrio harveyi (A16) strain, plasmid-nicking assay using pBR322 and 3(4,5-dimethyl-thiazol-2-yl)2,5-diphenyl-tetrazolium bromide assay as well as confocal imaging studies using Chinese hamster ovarian cell line, respectively. It was observed that although, the water sample of all the areas of wetland demonstrated mutagenic, genotoxic as well as cytotoxic activity, the effect was quite significant with the water samples from River Satluj and Khatan area (i.e. reservoir mainly contains Satluj water). The metal analysis of water samples was also conducted with atomic absorption spectrophotometer. The mutagenicity, genotoxicity and cytotoxicity of water samples emerged to be correlated with metal concentration. The source of toxic components seems to be associated with various industrial effluents and agricultural run-off. The results of the present study carry great importance in documenting the water quality monitoring data of the wetland.

Antimutagenic and Antioxidant Characteristics of Chukrasia tabularis A Juss Extracts

The current study aims to evaluate the antioxidative and antimutagenic activities of methanol extract and different fractions of Chukrasia tabularis leaves. The antioxidative potential was evaluated using 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) cation and superoxide anion radical-scavenging assay. The antimutagenic potential was evaluated against direct-acting mutagens, 4-nitro-o-phenylenediamine and sodium azide; and S9-dependent mutagen, 2-aminofluorene in TA98 and TA100 strains of Salmonella typhimurium using Ames assay. It has been found that methanol extract and its fractions were more efficient against S9-dependent mutagen in pre-incubation mode of treatment as compared to direct-acting mutagens in both the strains. Methanol extract and its fractions also exhibited strong radical-scavenging potential. High-performance liquid chromatography (HPLC) analysis of methanol extract showed the presence of gallic acid, epicatechin, 7-hydroxycoumarin, and rutin. From the study, it could be concluded that antioxidative and antimutagenic activity of methanol extract and its fractions was related to the synergistic interactions among different chemical compounds.

Colour changes in Labeo rohita (Ham.) due to pigment translocation in melanophores, on exposure to municipal wastewater of Tung Dhab drain, Amritsar, India

The present study was aimed to evaluate the effect of municipal wastewater of Tung Dhab drain on morphology of scale melanophores in freshwater fish Labeo rohita (Ham.). Chronic, non-renewal toxicity tests were performed with sub lethal concentrations (17.7, 26.6 and 35.4%) of wastewater for exposure durations of 15, 30 and 60 days. Recovery experiments were also performed for duration of 60 days. The scales were removed, processed and diameters of melanophores were measured using an ocular micrometer. The results showed concentration and duration dependent changes in melanophore morphology (size and dendricity) in experimental fish as compared to control fish. The values of mean melanophore size index (MMSI) varied from 5.37 ± 0.49 (17.7%) to 12.12 ± 0.81 (35.5%) in comparison to control values of 4.32 ± 0.32 and 4.55 ± 0.29 for 15 and 60 days respectively. The recovery experiments suggested that observed dark colouration due to pigment translocation is reversible, even after chronic exposure for 60 days.

Municipal wastewater-induced histopathological lesions in gills of freshwater fish Labeo rohita – A SEM study

The fingerlings of fish Labeo rohita were subjected to chronic (15, 30, and 60 days) and recovery (60 days) experiments for histopathological studies using sublethal concentrations (35.4%, 26.6%, and 17.7% v/v) of municipal wastewater of Tung Dhab drain, Amritsar, India. Control fish were maintained for same exposure periods using tap water. Scanning electron microscopic studies exhibited dose- and duration-dependent alterations on gill surface ultra-structure. Control fish exhibited functionally normal gills with normal mean degree of tissue change. Fifteen different types of lesions were identified out of which 9, 5, and 1 were of the stage I, II, and III, respectively. The mean degree of change values in the treated groups ranged from 14.8 ± 0.5 (17.7%, 15 days) to 116. ± 17 (35.4%, 60 days) demonstrated gill damages of slight to irreparable lesions. The recorded alterations were epithelial lifting and intraepithelial edema, reduction of water spaces, lamellar fusion, lamellar telangiectasia, swelling and fusion of filaments, sloughing of lamellar epithelium, hypertrophy and hyperplasia of mucous and chloride cells, and necrotic lamellae. The recovery experiments showed positive improvement as the intensity of damage was less than recorded for 60 days of treatment. This study clearly indicated toxicity of drain water to fish at cellular and organ level. The study recommends environmental agencies to make and enforce decisions regarding improvement of water quality of Tung Dhab drain, and further incorporating histopathological studies as an analytical tool.

Heavy ion ranges in glass detectors

Glass detectors, viz. sodalime, phosphate and quartz, were irradiated at GSI heavy ion facility at Darmstadt, Germany and at heavy ion Cyclotron facility at JINR, Dubna, USSR using 238U(5.9 and 15.0 MeV/u), 131 Xe (5.9,11.56 and 14.5 MeV/u) ,197 Au (11.4,11.67 and 15.96 MeV/u) , 56 Fe (4.0 MeV/u) and 48 Ti(4.0 MeV/u) beams.Heavy ion ranges are measured in glass detectors using appropriate conditions. The calibration curves are obtained for all these glass detectors using different ion beams. The present study may be used for particle identification.

Assessment of Water Quality of Tung Dhab Drain—An International Water Channel—Using Multivariate Statistical Techniques

Different multivariate statistical techniques were applied to interpret the temporal variations in water quality of Tung Dhab drain, Amritsar, India and further to identify water pollution sources. Data was collected seasonally for a period of two years (2012-2013) using 34 water quality parameters. The recorded values for variables like turbidity, total suspended solids, biochemical oxygen demand, chemical oxygen demand, oil & grease, nitrate as N, lead, chromium, nickel and zinc were much higher than the recommended permissible discharge limits into inland waters. Significant correlations were found in between different physicochemical parameters ( p ≤ 0.05; p ≤ 0.01). Cluster Analysis (CA) grouped four sampling seasons into two clusters. CA confirmed that the water quality of rainy season was different from other three seasons in terms of similarity and distance indices. Principal Component Analysis/Factor Analysis (PCA/FA) explained minerals, organic, agricultural and industrial pollutants responsible for deterioration of drain water quality. The present study will help environmental agencies to make and enforce decisions regarding improvement of water quality of Tung Dhab drain.