School Of Basic And Applied Sciences
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Item Assessment of antioxidant potential of phytochemicals in human glioblastoma (U-87 MG) cells(Central University of Punjab, 2014) Kaur, Manpreet; Mantha, Anil K.Imbalance between production of reactive oxygen/nitrogen species (ROS/RNS) leads to oxidative stress and has been well documented for mitochondrial dysfunction, a prime cause towards pathogenesis of neurological diseases and cancer. Glioblastoma Multiforme (GBM) is a highly aggressive, invasive and primary brain tumor which shows resistance to chemotherapy and radiotherapy. Superoxide dismutase (SOD) is an antioxidant enzyme that scavenges the production of superoxide radicals and dismutases into H?O? which is further converted into H?O and O? by catalase (CAT) enzyme. Apurinic/Apyrimidinic endonuclease (APE1) is a central enzyme of base excision repair (BER) pathway with two important functions; DNA repair and redox regulation of transcription factors (TFs) responsible for cell survival. In this study, it was seen that oxidative stress induced by endogenously found oxidants H?O? and glucose oxidase (GO) enhanced the activities of both CuZn-SOD and MnSOD in U-87 MG cells. In addition, CuZn-SOD levels were found to be increased in H?O?-induced oxidative stress and MnSOD levels were found to be increased in both H?O? and GO- induced oxidative stress. Further, pretreatment with phytochemicals Curcumin and Quercetin modulated the activities and expression of both forms of SOD studied. The BER-pathway enzyme, APE1 level was found to be decreased in mitochondria of oxidative stress induced U-87 MG cells by H?O? and GO, and in contrast APE1 level was found to be increased in cytosol, which indicates that oxidative stress affects the expression level and sub-cellular localization of APE1. Taken together, these results indicate that in GBM it is more likely that activated SOD a key player of antioxidant system and APE1 a key player in BER-pathway might be facilitating cancer cells to survive in oxidative stress environment.Item Assessment of Extract of Syzygium cumini Against Doxorubicin Induced Cardiotoxicity(Central University of Punjab, 2018) Chayan, Mukherjee; Monisha DhimanFor the past four decades, doxorubicin (DOX) has been used to treat cancer, mainly solid tumours and haematological malignancies. However, clinical community is greatly concerned regarding the administration of this as DOX treatment is commonly associated with dose-dependent cardiotoxicity. Attempts at alleviating drug generated cardiac damage using an extract from different parts of plants with radical scavenging property are a promising area of research. Hydroalcoholic extract derived from fruit pulp of Syzygiumcumini which has a significant antiradical scavenging effect. This study aims to assess the effect of parallel administration of SC fruit pulp extract (SC) on mitigating or preventing DOX induced cardiotoxicity in vitro using H9c2 cardiomyoblast cell lines. Addition of SC fruit pulp extract and DOX were performed for both treatment and control sets on H9c2 cells. SC fruit pulp extract showed strong ABTS cation radical scavenging activity in a dose dependent manner. MTT assay was used to study the cytotoxic effect of SC fruit pulp extract and DOX. ROS levels were estimated using NBT assay and DHE assay. The results showed that DOX has significant cytotoxic effect in a dose dependent manner while SC fruit pulp extract did not display any significant cytotoxicity on H9c2 cells. The DOX induced ROS production was found to be significantly reduced in SC fruit pulp extract treated cells. Results of the current study also suggest that the treatment of SC fruit pulp extract along with DOX, displayed cardioprotective potential in H9c2 cells by: 1) reducing lipid peroxidation; 2) decreasing extracellular nitric oxide (NO); 3) decreasing the expression of the protein p47phox and iNOS/NOS-2. These results clearly suggest that treatment of SC fruit pulp extract along with DOX reduces the DOX induced toxicity and hence can be a promising therapeutic intervention in managing DOX mediated cardiotoxicity.Item Oxidative stress responses to sub-lethal dose of Cry toxin in the larvae of castor semilooper, Achaea janata(Central University of Punjab, 2018) Singh, Kanika; Chaitanya,R.K.Development of synthetic insecticides to reduce the level of infestation led to deleterious effects on environment and human health. This lead to the development of ecofriendly pest management alternatives including Bacillus thuringensis (Bt). Bt produce Crystal (Cry), Cytotoxic (Cyt) and Vegetative (Vip) proteins with insecticidal activity against different orders of lepidoptera. Of late, pest resistance against Bt is reported in countries.The reduced toxicity of Bt formulation from degradation by UV light, wash-off by rain, drying, temperature, and soil acidity as well as its chemistry. Further, insects sense pesticides through odorant receptors and move away quickly, there is always a possibility of a population of larvae to get exposed to sub-lethal doses of toxin which might exhibit variable effects and escape mortality and eventually generate resistance. Sub-lethal dose lead to the generation of oxidative stress in the insect and eventually scavenged by anti-oxidant enzymes. These stress responses would enhance our understanding of adaptations for survival and resistance development. The current study is an attempt to monitor the antioxidative responses at the transcriptional level upon sub-lethal exposure of Cry toxin in the larvae of an polyphagous pest castor semilooper, Achaea janata. prevalent in the Indian subcontinent.Item Pyramiding of tea Dihydroflavonol reductase and Anthocyanidin reductase increases flavan-3-ols and improves protective ability under stress conditions in tobacco(Springer Verlag, 2017) Kumar, Vinay; Yadav, Sudesh KumarTea (Camellia sinensis) is one of the richest sources of flavan-3-ols, an important class of flavonoids. The expression level of gene-encoded key regulatory enzymes of flavan-3-ol/anthocyanin biosynthetic pathway, dihydroflavonol 4-reductase (DFR) and anthocyanidin reductase (ANR), has been highly correlated with the flavan-3-ol contents and antioxidant activity in tea plant. In the present study, pyramiding of CsDFR and CsANR in tobacco was achieved. However, single transgenic tobacco overexpressing either CsDFR or CsANR was documented earlier. In continuation, pyramided transgenic lines were evaluated for the possible, either same or beyond, effect on flavan-3-ol accumulation and protective ability against biotic and abiotic stresses. The pyramided transgenic lines showed early flowering and improved seed yield. The transcript levels of flavan-3-ol/anthocyanin biosynthetic pathway and related genes in pyramided transgenic lines were upregulated as compared to control tobacco plants. The accumulations of flavan-3-ols were also found to be higher in pyramided transgenic lines than control tobacco plants. In contrast, anthocyanin content was observed to be decreased in pyramided transgenic lines, while DPPH activity was higher in pyramided transgenic lines. In pyramided transgenic lines, strong protective ability against feeding by Spodoptera litura was documented. The seeds of pyramided transgenic lines were also found to have better germination rate under aluminum toxicity as compared to control tobacco plants. Interestingly, the synergistic effect of these two selected genes are not beyond from transgenic lines expressing either CsDFR and CsANR alone as published earlier in terms of flavan-3-ols accumulation. However, the unique flower color and better seed germination rate are some interestingly comparable differences that were reported in pyramided lines in relation to individual transgenic plants. In conclusion, the present results reveal an interesting dynamic between CsDFR and CsANR in modulating flavan-3-ol/anthocyanin levels and functional analysis of stacked CsDFR and CsANR transgenic tobacco lines. ? 2017, Springer-Verlag GmbH Germany.Item Composition, bioactive compounds and antioxidant activity of common Indian fruits and vegetables(Springer India, 2016) Singh,Jatinder Pal; Kaur, Amritpal; Shevkani, Khetan; Singh, NarpinderThe present work was undertaken to evaluate the chemical composition (proximate, minerals and dietary fibre), colour parameters, antioxidant activity and polyphenol profiles of different fruits (pomegranate, kinnow, mango, banana, jambolan, grapes and sapodilla) and vegetables (beetroot, brinjal, orange carrot, bitter gourd, mentha and spinach). The amount of insoluble dietary fibre was higher than soluble dietary fibre for all fruits and vegetables. Vegetables showed superior mineral composition (higher amounts of K, Ca and Fe) as compared to fruits. Total phenolic content (TPC) and antioxidant activity (ABTS and DPPH) ranged from 354.9 to 1639.7?mg?GAE/100?g, 2.6 to 5.5 and 3.0 to 6.3?mM?TE/g, respectively for different fruits, while it ranged from 179.3 to 1028.6?mg?GAE/100?g, 2.1 to 4.7 and 2.0 to 5.0?mM?TE/g, respectively for different vegetables. Gallic acid, protocatechuic acid, catechin, caffeic acid, ferulic acid, sinapic acid, quercetin, resveratrol and kaempferol were detected and quantified in different fruits and vegetables. The results highlighted that fruit peels could be used as valuable sources of minerals and polyphenols having high antioxidant activity. ? 2016, Association of Food Scientists & Technologists (India).Item Bioactive constituents in pulses and their health benefits(Springer India, 2017) Singh,Balwinder; Singh, Jatinder Pal; Shevkani, Khetan; Singh, Narpinder; Kaur, AmritpalPulses are good sources of bioactive compounds such as polyphenols, phytosterols and non-digestible carbohydrates that play important physiological as well as metabolic roles. These compounds vary in concentration amongst different pulse species and varieties. Pulse seed coats are rich in water-insoluble fibres and polyphenols (having high antioxidant activities), while cotyledons contain higher soluble fibres, oligosaccharides, slowly digestible and resistant starch content. Ferulic acid is the most abundant phenolic acid present in pulses, while flavonol glycosides, anthocyanins and tannins are responsible for the seed coat colour. Sitosterol (most abundant), stigmasterol, and campesterol are the major phytosterols present in pulses. Pulse fibres, resistant starch and oligosaccharides function as probiotics and possess several other health benefits such as anti-inflammatory, anti-tumour, and reduce glucose as well as lipid levels. Beans and peas contain higher amounts of oligosaccharides than other pulses. Processing methods affect resistant starch, polyphenol composition and generally increase antioxidant activities of different pulses. In this review, the current information on pulse polyphenols, phytosterols, resistant starch, dietary fibre, oligosaccharides, antioxidant and associated health benefits are discussed. ? 2016, Association of Food Scientists & Technologists (India).Item Anthocyanins enriched purple tea exhibits antioxidant, immunostimulatory and anticancer activities(Springer India, 2017) Joshi, Robin; Rana, Ajay; Kumar, Vinay; Kumar, Dharmesh; Padwad, Yogendra S.; Yadav, Sudesh Kumar; Gulati, AshuPurple coloured tea shoot clones have gained interest due to high content of anthocyanins in addition to catechins. Transcript expression of genes encoding anthocyanidin reductase (ANR), dihydroflavonol-4-reductase (DFR), anthocyanidin synthase (ANS), flavonol synthase (FLS) and leucoantho cyanidin reductase (LAR) enzymes in three new purple shoot tea clones compared with normal tea clone showed higher expression of CsDFR, CsANR, CsANS and lower expression of CsFLS and CsLAR in purple shoot clones compared to normal clone. Expression pattern supported high content of anthocyanins in purple tea. Four anthocyanins (AN1?4) were isolated and characterized by UPLC-ESI-QToF-MS/MS from IHBT 269 clone which recorded highest total anthocyanins content. Cyanidin-3-O-?-d-(6-(E)-coumaroyl) glucopyranoside (AN2) showed highest in vitro antioxidant activity (IC50?DPPH?=?25.27???0.02??g/mL and IC50?ABTS?=?10.71???0.01??g/mL). Anticancer and immunostimulatory activities of cyanidin-3-glucoside (AN1), cyanidin-3-O-?-d-(6-(E)-coumaroyl) glucopyranoside (AN2), delphinidin-3-O-?-d-(6-(E)-coumaroyl) glucopyranoside (AN3), cyanidin-3-O-(2-O-?-xylopyranosyl-6-O-acetyl)-?-glucopyranoside (AN4) and crude anthocyanin extract (AN5) showed high therapeutic perspective. Anthocyanins AN1?4 and crude extract AN5 showed cytotoxicity on C-6 cancer cells and high relative fluorescence units (RFU) at 200??g/mL suggesting promising apoptosis induction activity as well as influential immunostimulatory potential. Observations demonstrate potential of purple anthocyanins enriched tea clone for exploitation as a nutraceutical product. ? 2017, Association of Food Scientists & Technologists (India).Item Phytochemical Ginkgolide B Attenuates Amyloid- 1 - 42 Induced Oxidative Damage and Altered Cellular Responses in Human Neuroblastoma SH-SY5Y Cells(IOS Press, 2017) Gill, Iqbal; Kaur, Sukhchain; Kaur, Navrattan; Dhiman, Monisha; Mantha, Anil K.Oxidative stress is an upsurge in reactive oxygen/nitrogen species (ROS/RNS), which aggravates damage to cellular components viz. lipids, proteins, and nucleic acids resulting in impaired cellular functions and neurological pathologies including Alzheimer's disease (AD). In the present study, we have examined amyloid-β (Aβ)-induced oxidative stress responses, a major cause for AD, in the undifferentiated and differentiated human neuroblastoma SH-SY5Y cells. Aβ1-42-induced oxidative damage was evaluated on lipids by lipid peroxidation; proteins by protein carbonyls; antioxidant status by SOD and GSH enzyme activities; and DNA and RNA damage levels by evaluating the number of AP sites and 8-OHG base damages produced. In addition, the neuro-protective role of the phytochemical ginkgolide B (GB) in countering Aβ1-42-induced oxidative stress was assessed. We report that the differentiated cells are highly vulnerable to Aβ1-42-induced oxidative stress events as exerted by the deposition of Aβ in AD. Results of the current study suggest that the pre-treatment of GB, followed by Aβ1-42 treatment for 24 h, displayed neuro-protective potential, which countered Aβ1-42-induced oxidative stress responses in both undifferentiated and differentiated SH-SY5Y neuronal cells by: 1) hampering production of ROS and RNS; 2) reducing lipid peroxidation; 3) decreasing protein carbonyl content; 4) restoring antioxidant activities of SOD and GSH enzymes; and 5) maintaining genome integrity by reducing the oxidative DNA and RNA base damages. In conclusion, Aβ1-42 induces oxidative damage to the cellular biomolecules, which are associated with AD pathology, and are protected by the pre-treatment of GB against Aβ-toxicity. Taken together, this study advocates for phytochemical-based therapeutic interventions against AD.