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    • "The bacterial cells were subsequently washed using de-ionized water to remove free ions or copper ions adsorbed on the cell wall, if any, and then resuspended in 100 µl deionized water. Differential staining of these bacterial cells was carried out using Acridine Orange (AO) and Ethidium Bromide (EB) (Sigma-Aldrich, India) (Kasibhatla et al., 2006). "
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    ABSTRACT: Atomic Force Microscopy (AFM) has been used to elucidate the morphostructural changes occurring as a result of copper internalization in E. coli. The study documents a gradual aqueous leaching and built-up of copper ions within the cell using Atomic Absorption Spectroscopy (AAS). It further correlates the same to the morphometric changes studied using AFM and the cell viability studied using confocal microscopy. The cell perturbations take place within one hour of exposure to copper ions in water. Cell elongation takes place with the increase in exposure time and therefore copper ion concentration. Partially perturbed structure of the cell and accumulation of the cytosolic contents towards the apical ends of the cell reveal the inside-out mode of cell damage caused by copper ions.
    World Journal of Pharmaceutical Research 09/2015; 4(10):837-852.
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    • "Cells treated with H 2 O 2 were used as apoptotic death control [35] and for the necrotic control PC3 cells were treated with boiling water. Then, each cell culture was washed with 100 mL of PBS and stained with 100 í µí¼‡L AO/EB solution (100 í µí¼‡g/mL AO, 100 í µí¼‡g/mL EB), according to reported procedures [36]. The cells were observed using a fluorescence microscope (Olympus Co., Tokyo, Japan, with emission at 521 nm). "
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    ABSTRACT: The cytotoxic activity and the chemical composition of the dichloromethane/methanol root extract of Linum scabrellum Planchon (Linaceae) were analyzed. Using NMR spectra and mass spectrometry analyses of the extract we identified eight main constituents: oleic acid (1), octadecenoic acid (2), stigmasterol (3), α-amyrin (4), pinoresinol (5), 6 methoxypodophyllotoxin (6), coniferin (7), and 6-methoxypodophyllotoxin-7-O-β-D-glucopyranoside (8). By using the sulforhodamine B assay, an important cytotoxic activity against four human cancer cell lines, HF6 colon (IC50 = 0.57 μg/mL), MCF7 breast (IC50 = 0.56 μg/mL), PC3 prostate (IC50 = 1.60 μg/mL), and SiHa cervical (IC50 = 1.54 μg/mL), as well as toward the normal fibroblasts line HFS-30 IC50 = 1.02 μg/mL was demonstrated. Compound 6 (6-methoxypodophyllotoxin) was responsible for the cytotoxic activity exhibiting an IC50 value range of 0.0632 to 2.7433 µg/mL against the tested cell lines. Cell cycle studies with compound 6 exhibited a cell arrest in G2/M of the prostate PC3 cancer cell line. Microtubule disruption studies demonstrated that compound 6 inhibited the polymerization of tubulin through its binding to the colchicine site (binding constant K b = 7.6 × 10(6) M(-1)). A dose-response apoptotic effect was also observed. This work constitutes the first investigation reporting the chemical composition of L. scabrellum and the first study determining the mechanism of action of compound 6.
    Evidence-based Complementary and Alternative Medicine 08/2015; 2015(1):298463. DOI:10.1155/2015/298463 · 1.88 Impact Factor
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    • "Sulforhodamine-B and Trypan blue exclusion assays [12, 13] were performed using HCT-116, MCF-7, MDA-MB-231, and EAC cell lines. Similarly, AO/EB and Hoechst 33342 staining [14, 15] were done to examine the nuclear morphological changes within the cells. SRB assay which measures total protein content of the cells is directly proportional to viable cell count. "
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    ABSTRACT: The aim of the present study was to evaluate the antitumor potential of iminoflavones in in vitro and in vivo anticancer models. Preliminary screening in various cancer cell lines revealed four potential iminoflavones out of which IMF-8 was taken based on its activity against colon cancer cells. This was further confirmed by observing the nuclear changes in the cells by AO/EB and Hoechst 33342 staining studies. In vivo activity was assessed by dimethyl hydrazine-(DMH-) induced colon cancer model in rats. Animals were administered DMH (20 mg/kg, b.w. for 10 weeks and 30 mg/kg b.w., i.p. for 10 weeks) and were supplemented with (IMF-8) iminoflavone-8 (200 mg/kg, p.o. for 14 days). Results showed that DMH induced 100% aberrant crypt foci (ACF) and polyps which were significantly reduced in the IMF-8 treated group. IMF-8 significantly increased the catalase and GSH levels whereas it reduced the TNF-α and IL-6 levels markedly which suggests the antioxidative and anti-inflammatory actions of flavonoids present in IMF-8. The histopathological images of the IMF-8 treated colon showed no signs of mucosal crypt abscess. These findings suggest that the semi-synthetic iminoflavones, IMF-8, effectively inhibit DMH-induced ACFs and colonic crypts by alleviating the oxidative stress and suppressing the inflammation.
    BioMed Research International 06/2014; 2014(Article ID 569130):1-7. DOI:10.1155/2014/569130 · 3.17 Impact Factor
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