The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform

H. C. Goel - One of the best experts on this subject based on the ideXlab platform.

  • Podophyllum hexandrum modulates gamma radiation induced immunosuppression in balb c mice implications in radioprotection
    Molecular and Cellular Biochemistry, 2007
    Co-Authors: H. C. Goel, Hridayesh Prakash, Arif Ali, Madhu Bala
    Abstract:

    Aqueous extract of Podophyllum hexandrum (RP-1), which has been reported to render more than 82% survival against whole body lethal (10 Gy) gamma-irradiation in mice, was further investigated for its immunomodulatory potential. In this study, no significant change could be scored in peritoneal macrophages survival up to 8th day after whole body irradiation. RP-1 treatment (200 mg/kg body weight, i.p.) alone or 2 h before whole body irradiation enhanced macrophage survival significantly (p < 0.05) as compared to irradiated control mice. In irradiated animals, there was significant (p < 0.01) reduction in splenocyte survival and proliferation as revealed by 3H-TdR method. RP-1 treatment (200 mg/kg) alone or 2 h before irradiation countered the decrease in survival of splenocytes and proliferation significantly (p < 0.05) as compared to irradiated control group. Whole body irradiation also significantly (p < 0.05) reduced the population of CD4+ and CD8+ T cells and bone marrow GM-CFU at 24 h and 72 h post-irradiation intervals, respectively, as compared to unirradiated control. RP-1 treatment 2 h before whole body irradiation countered the decrease in CD4+ and CD8+ T cells populations and CGM-CFU. Nitric oxide free radicals generation was enhanced significantly (p < 0.05) in the supernatant of peritoneal macrophage cultures exposed to 2 Gy gamma radiation ex vivo in comparison to unirradiated control, which was reduced by pre-irradiation (−2 h) administration of RP-1. Whole body irradiation (10 Gy) also reduced the serum titres of IL-3, IL-1 and various IgG isotypes observed at different post-irradiation time interval. RP-1 treatment alone or before whole body irradiation countered radiation induced decrease in the titre of IL-1, IL-3 and IgG’s in the serum of mice. These findings indicate immunostimulatory potential of RP-1.

  • anti inflammatory effects of Podophyllum hexandrum rp 1 against lipopolysaccharides induced inflammation in mice
    Journal of pharmacy & pharmaceutical sciences : a publication of the Canadian Society for Pharmaceutical Sciences, 2005
    Co-Authors: Hridayesh Prakash, Arif Ali, Madhu Bala, H. C. Goel
    Abstract:

    PURPOSE: Down-regulation of lipopolysaccharide (LPS) induced hyper-inflammatory response by non-toxic pharmacological agents acquires paramount importance for countering bacterial sepsis. Anti-inflammatory potential of aqueous extract of Podophyllum hexandrum, a plant well documented in Ayurvedic literature for various therapeutic purposes, was investigated. METHODS: In vivo studies were performed on Balb/c mice pre-treated with supra- lethal dose of LPS endotoxin (E.coli 055:B5) with or without treatment with P. hexandrum extract (RP-1). Mouse peritoneal macrophage cultures were used to understand ex vivo effects of RP-1 on LPS generated nitric oxide (NO), secretion of IFN-γ, IL-6 and TNF-α. Griess assay and sandwich ELISA method were used to quantify inducible NO and cytokines respectively. RESULTS: Minimal dose of LPS that rendered 100% mortality to mice was found to be 450 μg/kg b.w. Administration of RP-1 (200 mg/kg b.w., i.p.) one hour before lethal LPS treatment (0.5 mg/kg b.w.) ren- dered maximum (78%) survival. Ex vivo study revealed that RP-1 (50 μg/ml) treatment to peritoneal macroph- ages inhibited LPS (5 μg/ml) induced nitrite generation to 37%, IFN-γ secretion to 5%, IL-6 secretion to 50% and TNF-α secretion to 50 % of LPS treated control values. CONCLUSION: This study has demonstrated anti-inflammatory potential of aqueous extract of P. hexandrum.

  • radioprotective mechanism of Podophyllum hexandrum during spermatogenesis
    Molecular and Cellular Biochemistry, 2004
    Co-Authors: Namita Samanta, K Kannan, M Bala, H. C. Goel
    Abstract:

    RP-1, a herbal preparation of Podophyllum hexandrum has already been reported to provide protection against whole body lethal gamma irradiation (10 Gy). It has also been reported to render radioprotection to germ cells during spermatogenesis. Present study was undertaken to unravel the cellular and molecular mechanism of action of RP-1 on testicular system in strain 'A' mice. Various antioxidant parameters such as thiol content, glutathione peroxidase (GPx), glutathione reductase (GR), glutathione-S-transferase (GST) enzyme activity, lipid peroxidation (LPO) and total protein levels were investigated. Thiol content was seen to increase significantly (p < 0.05) in both RP-1 alone and RP-1 pretreated irradiated groups over the irradiated groups at 8, 16 and 24 h. Irradiation (10 Gy) significantly decreased GPx, GST and GR activity in comparison to untreated control but RP-1 treatment before irradiation significantly (p < 0.05) countered radiation-induced decrease in the activity of these enzymes. Radiation-induced LPO was also found to be reduced at all time intervals by RP-1 treatment before irradiation. As compared to irradiated group the protein content in testicular tissue was increased in RP-1 pretreated irradiated group at 4 and 16 h significantly (p < 0.05). Comets revealed by single-cell gel electrophoresis were significantly longer (p < 0.001) in irradiated mice than in unirradiated control. RP-1 treatment before irradiation, however, rendered significant increase (p < 0.05) in comet length over the corresponding control and irradiated group initially at 4 h but at later time points, this was reduced significantly (p < 0.01) as compared to the irradiated group. RP-1 treatment alone rendered shorter comets at 8, 16 and 24 h than irradiated groups (p < 0.001). This study implies that RP-1 offers radioprotection at biochemical and cytogenetic level by protecting antioxidant enzymes, reducing LPO and increasing thiol content.

  • modification of radiation damage to mitochondrial system in vivo by Podophyllum hexandrum mechanistic aspects
    Molecular and Cellular Biochemistry, 2004
    Co-Authors: Damodar Gupta, Rajesh Arora, A P Garg, Madhu Bala, H. C. Goel
    Abstract:

    The present study was undertaken to investigate whether RP-1 treatment protected mitochondrial system against radiation damage and also to unravel the mechanism associated with this process. Radioprotection of mitochondrial system by Podophyllum hexandrum (RP-1) was investigated to understand its mechanism of action. Levels of superoxide anion (O2−), reduced or oxidized glutathione (GSH or GSSG), thiobarbituric acid reactive substance (TBARS), protein carbonyl (PC), ATP, NADH-ubiquinone oxidoreductase (complex-I), NADH-cytochrome c oxidoreductase (complex I/II), succinate-cytochrome c oxidoreductase (complex II/III) and mitochondrial membrane potential (MMP) were studied in mitochondria isolated from liver of mice belonging to various treatment groups. Whole body γ-irradiation (10 Gy) significantly (p < 0.01) increased the formation of O2−, PC, and TBARS, upto 24 h as compared to untreated control. RP-1 treatment (200 mg/kg b.w.) to mice 2 h before irradiation reduced the radiation-induced O2− generation within 4 h and formation of TBARS and PC upto 24 h significantly (p < 0.01). Singularly irradiation or RP-1 treatment significantly (p < 0.01) increased the levels of glutathione within an hour, as compared to untreated control. Pre-irradiation administration of RP-1 enhanced levels of GSH induced increase in complex I (upto 16 h), complex I/III (4 h) complex II/III activity (upto 24 h;p < 0.01) and inhibited the radiation-induced decrease in MMP significantly (24 h;p < 0.01). The present study indicates that RP-1 itself modulates several mitichondrial perameters due to its influence on the biochemical milieu within and outside the cells. However, RP-1 treatment before irradiation modulates radiation induced perturbations such as the increase in electron transport chain enzyme activity, formation of O2−, TBARS and PC to offer radioprotection (Mol Cell Biochem 266: 65–77, 2004)

  • radiation protection of hepg2 cells by Podophyllum hexandrum royale
    Molecular and Cellular Biochemistry, 2003
    Co-Authors: Damodar Gupta, Rajesh Arora, A P Garg, H. C. Goel
    Abstract:

    Radioprotection by an aqueous extract of Podophyllum hexandrum (RP-1) was investigated in HepG2 cells by evaluating colony forming efficacy (CFE), redox status of mitochondria, reactive oxygen species (ROS), generation of nitric oxide (NO), peroxidation of lipids and intracellular glutathione. Lower concentrations of RP-1 (0.1 and 1 microg/ml) rendered maximum radioprotection when administered 1 or 2 h before irradiation. Higher concentrations (5 and 10 microg/ml) however were less effective when administered 1 or 2 h before irradiation, but were more effective with increased time intervals (4 or 8 h) between RP-1 administration and irradiation. RP-1 pre-treatment also significantly inhibited radiation-induced MTT reduction in a concentration and time-dependent manner by decreasing gamma radiation-induced leakage of electrons from electron transport chain. Pre-irradiation administration of RP-1 significantly reduced both ROS and NO generation and enhanced glutathione levels, thereby inhibiting lipid peroxidation.

Manoharan Rajesh - One of the best experts on this subject based on the ideXlab platform.

  • establishment of somatic embryogenesis and podophyllotoxin production in liquid shake cultures of Podophyllum hexandrum royle
    Industrial Crops and Products, 2014
    Co-Authors: Manoharan Rajesh, Ganeshan Sivanandhan, Markandan Manickavasagam, Kondeti Subramanyam, Gnanajothi Kapildev, Balusamy Jaganath, S Kasthurirengan, A Ganapathi
    Abstract:

    Abstract The objective of this study was to establish a simple and efficient method for high frequency of somatic embryogenesis and plant regeneration of Podophyllum hexandrum and podophyllotoxin (PTOX) production. Embryogenic callus (100 mg FW) cultured on 0.75 strength MS medium with 4% sucrose and 3 g/l PVP produced maximum number of somatic embryos (1089) after five weeks of culture under dark condition. When these somatic embryos were subcultured to the same MS medium with 1 mg/l ABA, the embryos matured after 2 weeks of culture. The MS medium supplemented with1.0 mg/l GA 3 led to high frequency germination (91.1%). The accumulation of PTOX varied in somatic embryos, matured embryos and germinated embryos. Highest PTOX accumulation (2.8 mg/l) was recorded in somatic embryos cultured in 0.75 strength MS medium with 8% sucrose. Regenerated plantlets were successfully acclimatized in the growth chamber. The protocol established in this study will be helpful for large scale production of podophyllotoxin.

  • an efficient in vitro system for somatic embryogenesis and podophyllotoxin production in Podophyllum hexandrum royle
    Protoplasma, 2014
    Co-Authors: Manoharan Rajesh, Ganeshan Sivanandhan, Markandan Manickavasagam, Murugaraj Jeyaraj, Natesan Selvaraj, Rajan Chackravarthy, Andy Ganapathi
    Abstract:

    Podophyllum hexandrum Royle known as Indian mayapple is an important medicinal plant found only in higher altitudes (2,700 to 4,200 m) of the Himalayas. The highly valued anticancer drug Podophyllotoxin is obtained from the roots of this plant. Due to over exploitation, this endemic plant species is on the verge of extinction. In vitro culture for efficient regeneration and the production of podophyllotoxin is an important research priority for this plant. Hence, in the present study, an efficient plant regeneration system for mass multiplication through somatic embryogenesis was developed. We have screened P. hexandrum seeds collected from three different regions in the Himalayas to find their regenerative potentials. These variants showed variation in germination percentage as well as somatic embryogenic frequency. The seeds collected from the Milam area of Pithoragarh district showed better germination response (99.3 %) on Murashige and Skoog (MS) medium fortified with Gibberellic acid (GA3 [5 mg/l]) and higher direct somatic embryogenic frequency (89.6 %). Maximum production of embryogenic callus (1.2 g fresh weight [FW]) was obtained when cotyledons containing the direct somatic embryo clusters were cultured in MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D [1.5 mg/l]) after 4 week of culture in complete darkness. In the present investigation, somatic embryogenesis was accomplished either by direct organogenesis or callus mediated pathways. The latter method resulted in a higher frequency of somatic embryo induction in hormone-free MS medium yielding 47.7 embryos/50 mg of embryogenic callus and subsequent germination in MS medium supplemented with GA3 (5 mg/l). Seventy-nine percent of embryos attained complete maturity and germinated into normal plants with well-developed roots. Systematic histological analysis revealed the origin of somatic embryo and their ontogenesis. The higher level of podophyllotoxin (1.8 mg/g dry weight [DW]) was recorded in germinated somatic embryos when compared to field grown plants. The present system can be widely used for mass propagation, transgenic recovery, and podophyllotoxin production for commercial utilization.

  • factors influencing podophyllotoxin production in adventitious root culture of Podophyllum hexandrum royle
    Acta Physiologiae Plantarum, 2014
    Co-Authors: Manoharan Rajesh, Ganeshan Sivanandhan, Markandan Manickavasagam, Muthukrishnan Arun, Venkatachalam Vasudevan, Jeevaraj Theboral, Shanmugam Girija, Natesan Selvaraj, Andy Ganapathi
    Abstract:

    To determine the possibility of generating the Podophyllotoxin accumulation using Podophyllum hexandrum adventitious roots derived from root segments, several nutrient constituents (carbon sources, media strength, initial medium pH, ammonium and nitrate proportion and phosphate ratio) were evaluated in culture. The maximum biomass accumulation was recorded in 0.50 MS medium containing 3 mg/l indole-3-butyric acid and 2 % sucrose, and the maximum accumulation of Podophyllotoxin was documented in the same strength of MS medium with 6 % of sucrose. When the initial medium pH was on 6 in the optimized MS medium, the biomass and Podophyllotoxin accumulations were highest. The lower concentration of ammonium (10 mM) in combination with a moderate concentration of nitrate (20 mM) was found ideal for maximum accumulations of biomass and Podophyllotoxin. Maximum Podophyllotoxin accumulation (6.4 mg/g dry weight) was recorded at the higher concentration of phosphate (2.25 mM), and lower concentration of phosphate (1.25 mM) showed highest growth accumulation. The outcome of the present work will be helpful for the large-scale cultivation of adventitious root for the production of Podophyllotoxin.

  • agrobacterium mediated transformation of the medicinal plant Podophyllum hexandrum royle syn p emodi wall ex hook f thomas
    Plant Cell Tissue and Organ Culture, 2013
    Co-Authors: Manoharan Rajesh, Ganeshan Sivanandhan, Murugaraj Jeyaraj, Kondeti Subramanyam, Thankaraj Salammal Mariashibu, Subramanian Mayavan, Gnanajothi Kapil Dev, Vasudevan Ramesh Anbazhagan, Markandan Manickavasagam
    Abstract:

    An efficient Agrobacterium-mediated genetic transformation method has been developed for the medicinal plant Podophyllum hexandrum Royle, an important source of the anticancer agent podophyllotoxin. Highly proliferating embryogenic cells were infected with Agrobacterium tumefaciens harbouring pCAMBIA 2301, which contains npt II and gusA as selection marker and reporter genes, respectively. The transformed somatic embryos and plantlets were selected on Murashige and Skoog (MS) basal medium containing kanamycin and germination medium, respectively. GUS histochemical analysis, polymerase chain reaction and Southern blot hybridisation confirmed that gusA was successfully integrated and expressed in the P. hexandrum genome. Compared with cefotaxime, 200 mg l−1 timentin completely arrested Agrobacterium growth and favoured somatic embryo development from embryogenic cells. Among the different Agrobacterium strains, acetosyringone concentrations and co-cultivation durations tested, embryogenic callus infected with A. tumefaciens EHA 105 and co-cultivated for 3 days on MS basal medium containing 100 μM acetosyringone proved to be optimal and produced a transformation efficiency of 29.64 % with respect to germinated GUS-positive plantlets. The Agrobacterium-mediated genetic transformation method developed in the present study facilitates the transference of desirable genes into P. hexandrum to improve the podophyllotoxin content and to enhance other useful traits.

  • An investigation on the cytotoxicity and caspase-mediated apoptotic effect of biologically synthesized silver nanoparticles using Podophyllum hexandrum on human cervical carcinoma cells
    Colloids and Surfaces B: Biointerfaces, 2013
    Co-Authors: Murugaraj Jeyaraj, Ganeshan Sivanandhan, Markandan Manickavasagam, Manoharan Rajesh, Gnanajothi Kapil Dev, Renganathan Arun, Davoodbasha Mubarakali, Gnanasekar Sathishkumar, Kumpati Premkumar, Nooruddin Thajuddin
    Abstract:

    Now-a-days synthesis and characterization of silver nanoparticles (AgNPs) through biological entity is quite interesting to employ AgNPs for various biomedical applications in general and treatment of cancer in particular. This paper presents the green synthesis of AgNPs using leaf extract of Podophyllum hexandrum Royle and optimized with various parameters such as pH, temperature, reaction time, volume of extract and metal ion concentration for synthesis of AgNPs. TEM, XRD and FTIR were adopted for characterization. The synthesized nanoparticles were found to be spherical shaped with average size of 14. nm. Effects of AgNPs were analyzed against human cervical carcinoma cells by MTT Assay, quantification of ROS, RT-PCR and western blotting techniques. The overall result indicates that AgNPs can selectively inhibit the cellular mechanism of HeLa by DNA damage and caspase mediated cell death. This biological procedure for synthesis of AgNPs and selective inhibition of cancerous cells gives an alternative avenue to treat human cancer effectively. © 2012 Elsevier B.V.

Markandan Manickavasagam - One of the best experts on this subject based on the ideXlab platform.

  • establishment of somatic embryogenesis and podophyllotoxin production in liquid shake cultures of Podophyllum hexandrum royle
    Industrial Crops and Products, 2014
    Co-Authors: Manoharan Rajesh, Ganeshan Sivanandhan, Markandan Manickavasagam, Kondeti Subramanyam, Gnanajothi Kapildev, Balusamy Jaganath, S Kasthurirengan, A Ganapathi
    Abstract:

    Abstract The objective of this study was to establish a simple and efficient method for high frequency of somatic embryogenesis and plant regeneration of Podophyllum hexandrum and podophyllotoxin (PTOX) production. Embryogenic callus (100 mg FW) cultured on 0.75 strength MS medium with 4% sucrose and 3 g/l PVP produced maximum number of somatic embryos (1089) after five weeks of culture under dark condition. When these somatic embryos were subcultured to the same MS medium with 1 mg/l ABA, the embryos matured after 2 weeks of culture. The MS medium supplemented with1.0 mg/l GA 3 led to high frequency germination (91.1%). The accumulation of PTOX varied in somatic embryos, matured embryos and germinated embryos. Highest PTOX accumulation (2.8 mg/l) was recorded in somatic embryos cultured in 0.75 strength MS medium with 8% sucrose. Regenerated plantlets were successfully acclimatized in the growth chamber. The protocol established in this study will be helpful for large scale production of podophyllotoxin.

  • an efficient in vitro system for somatic embryogenesis and podophyllotoxin production in Podophyllum hexandrum royle
    Protoplasma, 2014
    Co-Authors: Manoharan Rajesh, Ganeshan Sivanandhan, Markandan Manickavasagam, Murugaraj Jeyaraj, Natesan Selvaraj, Rajan Chackravarthy, Andy Ganapathi
    Abstract:

    Podophyllum hexandrum Royle known as Indian mayapple is an important medicinal plant found only in higher altitudes (2,700 to 4,200 m) of the Himalayas. The highly valued anticancer drug Podophyllotoxin is obtained from the roots of this plant. Due to over exploitation, this endemic plant species is on the verge of extinction. In vitro culture for efficient regeneration and the production of podophyllotoxin is an important research priority for this plant. Hence, in the present study, an efficient plant regeneration system for mass multiplication through somatic embryogenesis was developed. We have screened P. hexandrum seeds collected from three different regions in the Himalayas to find their regenerative potentials. These variants showed variation in germination percentage as well as somatic embryogenic frequency. The seeds collected from the Milam area of Pithoragarh district showed better germination response (99.3 %) on Murashige and Skoog (MS) medium fortified with Gibberellic acid (GA3 [5 mg/l]) and higher direct somatic embryogenic frequency (89.6 %). Maximum production of embryogenic callus (1.2 g fresh weight [FW]) was obtained when cotyledons containing the direct somatic embryo clusters were cultured in MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D [1.5 mg/l]) after 4 week of culture in complete darkness. In the present investigation, somatic embryogenesis was accomplished either by direct organogenesis or callus mediated pathways. The latter method resulted in a higher frequency of somatic embryo induction in hormone-free MS medium yielding 47.7 embryos/50 mg of embryogenic callus and subsequent germination in MS medium supplemented with GA3 (5 mg/l). Seventy-nine percent of embryos attained complete maturity and germinated into normal plants with well-developed roots. Systematic histological analysis revealed the origin of somatic embryo and their ontogenesis. The higher level of podophyllotoxin (1.8 mg/g dry weight [DW]) was recorded in germinated somatic embryos when compared to field grown plants. The present system can be widely used for mass propagation, transgenic recovery, and podophyllotoxin production for commercial utilization.

  • factors influencing podophyllotoxin production in adventitious root culture of Podophyllum hexandrum royle
    Acta Physiologiae Plantarum, 2014
    Co-Authors: Manoharan Rajesh, Ganeshan Sivanandhan, Markandan Manickavasagam, Muthukrishnan Arun, Venkatachalam Vasudevan, Jeevaraj Theboral, Shanmugam Girija, Natesan Selvaraj, Andy Ganapathi
    Abstract:

    To determine the possibility of generating the Podophyllotoxin accumulation using Podophyllum hexandrum adventitious roots derived from root segments, several nutrient constituents (carbon sources, media strength, initial medium pH, ammonium and nitrate proportion and phosphate ratio) were evaluated in culture. The maximum biomass accumulation was recorded in 0.50 MS medium containing 3 mg/l indole-3-butyric acid and 2 % sucrose, and the maximum accumulation of Podophyllotoxin was documented in the same strength of MS medium with 6 % of sucrose. When the initial medium pH was on 6 in the optimized MS medium, the biomass and Podophyllotoxin accumulations were highest. The lower concentration of ammonium (10 mM) in combination with a moderate concentration of nitrate (20 mM) was found ideal for maximum accumulations of biomass and Podophyllotoxin. Maximum Podophyllotoxin accumulation (6.4 mg/g dry weight) was recorded at the higher concentration of phosphate (2.25 mM), and lower concentration of phosphate (1.25 mM) showed highest growth accumulation. The outcome of the present work will be helpful for the large-scale cultivation of adventitious root for the production of Podophyllotoxin.

  • agrobacterium mediated transformation of the medicinal plant Podophyllum hexandrum royle syn p emodi wall ex hook f thomas
    Plant Cell Tissue and Organ Culture, 2013
    Co-Authors: Manoharan Rajesh, Ganeshan Sivanandhan, Murugaraj Jeyaraj, Kondeti Subramanyam, Thankaraj Salammal Mariashibu, Subramanian Mayavan, Gnanajothi Kapil Dev, Vasudevan Ramesh Anbazhagan, Markandan Manickavasagam
    Abstract:

    An efficient Agrobacterium-mediated genetic transformation method has been developed for the medicinal plant Podophyllum hexandrum Royle, an important source of the anticancer agent podophyllotoxin. Highly proliferating embryogenic cells were infected with Agrobacterium tumefaciens harbouring pCAMBIA 2301, which contains npt II and gusA as selection marker and reporter genes, respectively. The transformed somatic embryos and plantlets were selected on Murashige and Skoog (MS) basal medium containing kanamycin and germination medium, respectively. GUS histochemical analysis, polymerase chain reaction and Southern blot hybridisation confirmed that gusA was successfully integrated and expressed in the P. hexandrum genome. Compared with cefotaxime, 200 mg l−1 timentin completely arrested Agrobacterium growth and favoured somatic embryo development from embryogenic cells. Among the different Agrobacterium strains, acetosyringone concentrations and co-cultivation durations tested, embryogenic callus infected with A. tumefaciens EHA 105 and co-cultivated for 3 days on MS basal medium containing 100 μM acetosyringone proved to be optimal and produced a transformation efficiency of 29.64 % with respect to germinated GUS-positive plantlets. The Agrobacterium-mediated genetic transformation method developed in the present study facilitates the transference of desirable genes into P. hexandrum to improve the podophyllotoxin content and to enhance other useful traits.

  • An investigation on the cytotoxicity and caspase-mediated apoptotic effect of biologically synthesized silver nanoparticles using Podophyllum hexandrum on human cervical carcinoma cells
    Colloids and Surfaces B: Biointerfaces, 2013
    Co-Authors: Murugaraj Jeyaraj, Ganeshan Sivanandhan, Markandan Manickavasagam, Manoharan Rajesh, Gnanajothi Kapil Dev, Renganathan Arun, Davoodbasha Mubarakali, Gnanasekar Sathishkumar, Kumpati Premkumar, Nooruddin Thajuddin
    Abstract:

    Now-a-days synthesis and characterization of silver nanoparticles (AgNPs) through biological entity is quite interesting to employ AgNPs for various biomedical applications in general and treatment of cancer in particular. This paper presents the green synthesis of AgNPs using leaf extract of Podophyllum hexandrum Royle and optimized with various parameters such as pH, temperature, reaction time, volume of extract and metal ion concentration for synthesis of AgNPs. TEM, XRD and FTIR were adopted for characterization. The synthesized nanoparticles were found to be spherical shaped with average size of 14. nm. Effects of AgNPs were analyzed against human cervical carcinoma cells by MTT Assay, quantification of ROS, RT-PCR and western blotting techniques. The overall result indicates that AgNPs can selectively inhibit the cellular mechanism of HeLa by DNA damage and caspase mediated cell death. This biological procedure for synthesis of AgNPs and selective inhibition of cancerous cells gives an alternative avenue to treat human cancer effectively. © 2012 Elsevier B.V.

Sharmila Chattopadhyay - One of the best experts on this subject based on the ideXlab platform.

  • Transcriptome-wide identification and characterization of CAD isoforms specific for podophyllotoxin biosynthesis from Podophyllum hexandrum
    Plant Molecular Biology, 2016
    Co-Authors: Dipto Bhattacharyya, Saptarshi Hazra, Anindyajit Banerjee, Riddhi Datta, Deepak Kumar, Saikat Chakrabarti, Sharmila Chattopadhyay
    Abstract:

    Podophyllotoxin (ptox) is a therapeutically important lignan derived from Podophyllum hexandrum and is used as a precursor for the synthesis of anticancer drugs etoposide, teniposide and etopophose. In spite of its enormous economic significance, genomic information on this endangered medicinal herb is scarce. We have performed de novo transcriptome analysis of methyl jasmonate (MeJA)-treated P. hexandrum cell cultures exhibiting enhanced ptox accumulation. The results revealed the maximum up-regulation of several isoforms of cinnamyl alcohol dehydrogenase (CAD). CAD catalyzes the synthesis of coniferyl alcohol and sinapyl alcohol from coniferaldehyde (CAld) and sinapaldehyde respectively. Coniferyl alcohol can produce both lignin and lignan while sinapyl alcohol produces only lignin. To isolate the CAD isoforms favoring ptox, we deduced full length cDNA sequences of four CAD isoforms: PhCAD1, PhCAD2, PhCAD3 and PhCAD4 from the contigs of the transcriptome data. In vitro enzyme assays indicated a higher affinity for CAld over sinapaldehyde for each isoform. In silico molecular docking analyses also suggested that PhCAD3 has a higher binding preference with CAld over sinapaldehyde, followed by PhCAD4, PhCAD2, and PhCAD1, respectively. The transgenic cell cultures overexpressing these isoforms independently revealed that PhCAD3 favored the maximum accumulation of ptox as compared to lignin followed by PhCAD4 and PhCAD2 , whereas, PhCAD1 favored both equally. Together, our study reveals transcriptome-wide identification and characterization of ptox specific CAD isoforms from P. hexandrum . It provides a useful resource for future research not only on the ptox biosynthetic pathway but on overall P. hexandrum , an endangered medicinal herb with immense therapeutic importance.

  • identification of conserved mirnas and their putative target genes in Podophyllum hexandrum himalayan mayapple
    Plant Gene, 2016
    Co-Authors: Soumi Biswas, Saptarshi Hazra, Sharmila Chattopadhyay
    Abstract:

    MicroRNAs (miRNAs) are a class of naturally occurring, endogenous, small non-coding RNA molecules that play a crucial role in post-transcriptional gene regulation either by directing mRNAs for cleavage or translational repression. Plant miRNAs show near-perfect complementarity to their target mRNAs. Podophyllum hexandrum is an endangered medicinal plant. The rhizome and roots of P. hexandrum contain podophyllotoxin that possess antitumor activity. Several miRNAs have been identified in different plant species to date but, miRNAs and their target genes are still unknown in P. hexandrum. In the present study, a total of 60 mature miRNAs and 6 pre-miRNAs were identified in P. hexandrum by 454 pyrosequencing. Quantitative real-time PCR was used to validate the expression levels of some of the identified miRNAs. The P. hexandrum miRNAs targeted a variety of mRNAs. Gene ontology (GO) analysis of these target genes revealed 13 different biological processes. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that the identified miRNAs regulate various metabolic pathways, especially biosynthesis of secondary metabolites. Our research work provides some elementary information about miRNA mediated regulation of secondary metabolite biosynthesis in P.hexandrum.

  • de novo transcriptome analysis using 454 pyrosequencing of the himalayan mayapple Podophyllum hexandrum
    BMC Genomics, 2013
    Co-Authors: Dipto Bhattacharyya, Saptarshi Hazra, Riddhi Datta, Ragini Sinha, Sharmila Chattopadhyay
    Abstract:

    The Himalayan or Indian Mayapple (Podophyllum hexandrum Royle) produces podophyllotoxin, which is used in the production of semisynthetic anticancer drugs. High throughput transcriptome sequences or genomic sequence data from the Indian Mayapple are essential for further understanding of the podophyllotoxin biosynthetic pathway. 454 pyrosequencing of a P. hexandrum cell culture normalized cDNA library generated 2,667,207 raw reads and 1,503,232 high quality reads, with an average read length of 138 bp. The denovo assembly was performed by Newbler using default and optimized parameters. The optimized parameter generated 40, 380 assembled sequences, comprising 12,940 contigs and 27,440 singlets which resulted in better assembly as compared to default parameters. BLASTX analysis resulted in the annotation of 40,380 contigs/singlet using a cut-off value of ≤1E-03. High similarity to Medicago truncatula using optimized parameters and to Populus trichocarpa using default parameters was noted. The Kyoto encyclopedia of genes and genomes (KEGG) analysis using KEGG Automatic Annotation Server (KAAS) combined with domain analysis of the assembled transcripts revealed putative members of secondary metabolism pathways that may be involved in podophyllotoxin biosynthesis. A proposed schematic pathway for phenylpropanoids and podophyllotoxin biosynthesis was generated. Expression profiling was carried out based on fragments per kilobase of exon per million fragments (FPKM). 1036 simple sequence repeats were predicted in the P. hexandrum sequences. Sixty-nine transcripts were mapped to 99 mature and precursor microRNAs from the plant microRNA database. Around 961 transcripts containing transcription factor domains were noted. High performance liquid chromatography analysis showed the peak accumulation of podophyllotoxin in 12-day cell suspension cultures. A comparative qRT-PCR analysis of phenylpropanoid pathway genes identified in the present data was performed to analyze their expression patterns in 12-day cell culture, callus and rhizome. The present data will help the identification of the potential genes and transcription factors involved in podophyllotoxin biosynthesis in P. hexandrum. The assembled transcripts could serve as potential candidates for marker discovery and conservation, which should form the foundations for future endeavors.

  • proteins differentially expressed in elicited cell suspension culture of Podophyllum hexandrum with enhanced podophyllotoxin content
    Proteome Science, 2012
    Co-Authors: Dipto Bhattacharyya, Saptarshi Hazra, Amrita Chakraborty, Srijani Ghanta, Ragini Sinha, Sharmila Chattopadhyay
    Abstract:

    Background Podophyllotoxin (PTOX), the precursor for semi-synthesis of cancer therapeutics like etoposide, teniposide and etophos, is primarily obtained from an endangered medicinal herb, Podophyllum hexandrum Royle. PTOX, a lignan is biosynthetically derived from the phenylpropanoid pathway. The aim of this study is to investigate changes in the P. hexandrum cell proteome potentially related to PTOX accumulation in response to methyl jasmonate (MeJA) elicitation. High-resolution two-dimensional gel electrophoresis (2-DE) followed by colloidal Coomassie staining and mass spectrometric analysis was used to detect statistically significant changes in cell’s proteome.

  • proteins differentially expressed in elicited cell suspension culture of Podophyllum hexandrum with enhanced podophyllotoxin content
    Proteome Science, 2012
    Co-Authors: Dipto Bhattacharyya, Saptarshi Hazra, Amrita Chakraborty, Srijani Ghanta, Ragini Sinha, Sharmila Chattopadhyay
    Abstract:

    Podophyllotoxin (PTOX), the precursor for semi-synthesis of cancer therapeutics like etoposide, teniposide and etophos, is primarily obtained from an endangered medicinal herb, Podophyllum hexandrum Royle. PTOX, a lignan is biosynthetically derived from the phenylpropanoid pathway. The aim of this study is to investigate changes in the P. hexandrum cell proteome potentially related to PTOX accumulation in response to methyl jasmonate (MeJA) elicitation. High-resolution two-dimensional gel electrophoresis (2-DE) followed by colloidal Coomassie staining and mass spectrometric analysis was used to detect statistically significant changes in cell’s proteome. The HPLC analysis showed approximately 7–8 fold change in accumulation of PTOX, in the 12day old cell suspension culture (i.e. after 9days of elicitation) elicited with 100 μM MeJA as compared to the control. Using 2-DE a total of 233 spots was detected, out of which 105 spots were identified by MALDI TOF-TOF MS/MS. Data were subjected to functional annotation from a biological point of view through KEGG. The phenylpropanoid and monolignol pathway enzymes were identified, amongst these, chalcone synthase, polyphenol oxidase, caffeoyl CoA 3-O-methyltransferase, S-adenosyl-L-methionine-dependent methyltransferases, caffeic acid-O-methyl transferase etc. are noted as important. The relation of other differentially accumulated proteins with varied effects caused by elicitors on P. hexandrum cells namely stress and defense related protein, transcription and DNA replication and signaling are also discussed. Elicitor-induced PTOX accumulation in P. hexandrum cell cultures provides a responsive model system to profile modulations in proteins related to phenylpropanoid/monolignol biosynthesis and other defense responses. Present findings form a baseline for future investigation on a non-sequenced medicinal herb P. hexandrum at molecular level.

Virendra S. Bisaria - One of the best experts on this subject based on the ideXlab platform.

  • enhanced production of podophyllotoxin by fed batch cultivation of Podophyllum hexandrum
    Canadian Journal of Chemical Engineering, 2008
    Co-Authors: Saurabh Chattopadhyay, Sant S. Bhojwani, Virendra S. Bisaria, Ashok K. Srivastava
    Abstract:

    Podophyllum hexandrum (Indian May Apple) was successfully cultivated in a 3 L stirred tank bioreactor under low shear conditions in batch and fed-batch modes of operation. A statistically optimized culture medium was used for the batch cultivation of Podophyllum hexandrum. Under optimum culture conditions of P. hexandrum, the batch culture showed a growth-associated product formation with a maximum biomass of 21.4 g/L dry cell weight (DCW) basis and a podophyllotoxin production of 13.8 mg/L in 26 d. A mathematical model was developed to design the nutrient feeding strategies for a fed-batch cultivation to prolong the productive log phase of cultivation. The fed-batch cultivation was able to enhance the biomass and podophyllotoxin accumulation to 48 g/L (DCW basis) and 43.2 mg/L, respectively, in 60 d. The volumetric productivity of podophyllotoxin in fed-batch cultivation was found to be 0.72 mg/(L. d) as opposed to 0.53 mg/(L. d) in batch cultivation under optimized culture conditions. Podophyllum hexandrum (« Indian May Apple ») a ete cultive avec succes dans un bioreacteur agite de 3 L dans des conditions de faible cisaillement en mode de fonctionnement discontinu et semi-discontinu. On a utilise un milieu de culture optimise statistiquement pour la culture discontinue de Podophyllum hexandrum. Dans des conditions de culture optimales de P. hexandrum, la culture discontinue montre une formation de produit associee a la croissance avec une biomasse maximale sur la base de la masse seche des cellules (DCW) de 21,4 g/L et une production de podophyllotoxine de 13,8 mg/L en 26 jours. On a mis au point un modele mathematique qui a concevoir les strategies d'alimentation des nutriments pour une culture semi-discontinue pour prolonger la phase productive de la culture. La culture semi-discontinue est capable d'ameliorer l'accumulation de podophyllotoxine a 48 g/L (base DCW) et a 43,2 mg/L, respectivement, en 60 jours. La productivite volumetrique de podophyllotoxine dans la culture a alimentation discontinue s'avere de 0,72 mg/(L.j) comparativement a 0,53 mg/(L.j) dans la culture discontinue dans des conditions de culture optimales.

  • cytotoxicity of in vitro produced podophyllotoxin from Podophyllum hexandrum on human cancer cell line
    Natural Product Research, 2004
    Co-Authors: Saurabh Chattopadhyay, Virendra S. Bisaria, A K Panda, Ashok K. Srivastava
    Abstract:

    Podophyllotoxin was produced by cell culture of Podophyllum hexandrum under in vitro culture conditions. A maximum of 4.26 mg/L of podophyllotoxin was produced when P. hexandrum was cultivated in 3 L stirred tank bioreactor. The compound extracted from the cell culture was applied to the human breast cancer cell line (MCF-7) and 1 nM podophyllotoxin was able to inhibit the growth of the cancer cells by 50%. The most profound inhibitory effect of podophyllotoxin was observed when it was applied in the beginning of cell growth.

  • Enhanced production of podophyllotoxin by Podophyllum hexandrum using in situ cell retention bioreactor.
    Biotechnology progress, 2003
    Co-Authors: Saurabh Chattopadhyay, Virendra S. Bisaria, Ashok K. Srivastava
    Abstract:

    The rhizomes of the rare plant Podophyllum hexandrum contain podophyllotoxin, which is a precursor of the anticancer drugs etoposide and teniposide. Batch cultivation of Podophyllum hexandrum was conducted using optimized medium in a 3 L bioreactor, which resulted in biomass and podophyllotoxin concentrations of 21.4 g/L and 13.8 mg/L in 24 and 26 days, respectively. The batch kinetics was used to identify the mathematical model. The model was extrapolated to identify the nutrient feeding rate (150 mL/d) and substrate concentration (105 g/L) in the incoming feed for nonlimiting and noninhibitory glucose concentration in the cell retention bioreactor. An improvement in cell growth to 53 g/L and intracellular podophyllotoxin accumulation of 48.8 mg/L was achieved in 60 days, when the bioreactor was operated in continuous cell retention cultivation mode.

  • effect of major nutrients on podophyllotoxin production in Podophyllum hexandrum suspension cultures
    Applied Microbiology and Biotechnology, 2003
    Co-Authors: Saurabh Chattopadhyay, Ashok K. Srivastava, Sant S. Bhojwani, R S Mehra, Virendra S. Bisaria
    Abstract:

    The effect of major medium ingredients (sugar, nitrogen source and phosphate) in Podophyllum hexandrum suspension cultures was investigated in order to increase the production of podophyllotoxin, the raw material in the synthesis of anticancer drugs. Amongst B5, Eriksson, MS, Nitsch, Street and White's medium, MS medium resulted in high growth and podophyllotoxin accumulation. The optimum level of nitrogen was found to be 60 mM, with a combination of ammonium salts and nitrate in the ratio of 1:2. The highest level of podophyllotoxin was obtained at 60 g glucose/l and at 1.25 mM phosphate after 30 days. Statistical design was adopted to determine the optimum levels of the parameters for cell growth and podophyllotoxin production.

  • optimization of culture parameters for production of podophyllotoxin in suspension culture of Podophyllum hexandrum
    Applied Biochemistry and Biotechnology, 2002
    Co-Authors: Saurabh Chattopadhyay, Ashok K. Srivastava, Virendra S. Bisaria
    Abstract:

    The root explants of the germinated seedlings of Podophyllum hexandrum were grown in MS medium supplemented with indole acetic acid (IAA) (2 mg/L) and activated charcoal (0.5%), and healthy callus culture was obtained after incubation for 3 wk at 20°C. The cultivation of plant cells in shake flask was associated with problems such as clumping of cells and browning of media, which were solved by the addition of pectinase and polyvinylpyrrolidone. The effect of major media components and carbon source was studied on the growth and podophyllotoxin production in suspension culture. It was found that glucose was a better carbon source than sucrose and that NH4+:NO3− ratio (total nitrogen concentration of 60 mM) and PO43− did not have much effect on the growth and product formation. The relative effect of culture parameters (inoculum level, pH, IAA, glucose, NH4+:NO3− ratio, and PO43−) on the overall growth and product response of the plant cell suspension culture was further investigated by Plackett-Burman design. This indicated that inoculum level, glucose, IAA, and pH had significant effects on growth and production of podophyllotoxin. To identify the exact optimum concentrations of these parameters on culture growth and podophyllotoxin production, central composite design experiments were formulated. The overall response equations with respect to growth and podophyllotoxin production as a function of these culture parameters were developed and used to determine the optimum concentrations of these parameters, which were pH 6.0, 1.25 mg/L of IAA, 72 g/L of glucose, and inoculum level of 8 g/L.