Effect of Extraction Methods on the Antioxidant Potential and Cytotoxicity of the Combined Ethanolic Extracts of Daucus carota L., Beta vulgaris L., Phyllanthus emblica L. and Lycopersicon esculentum against Gastric Adenocarcinoma Cells.

Frequent consumption of fruits and vegetables in the daily diet may alleviate the risk of developing chronic diseases. Daucus carota L. (carrot), Beta vulgaris L. (beetroot) Phyllanthus emblica L. (amla), and Lycopersicon esculentum M (tomatoes) are traditionally consumed functional foods that contain a high concentration of antioxidants, ascorbic acid, polyphenols, and numerous phytochemicals. This study assessed how three distinct preparation methods affect the phenolic, flavonoid, carotenoid, and ascorbic acid contents, antioxidant level, and cytotoxicity of the combined fruit extract. The fruit samples were taken in the ratio of carrot (6): beetroot (2): tomato (1.5): amla (0.5) and processed into a lyophilized slurry (LS) extract, lyophilized juice (LJ) extract, and hot-air oven-dried (HAO) extract samples. The sample extracts were assessed for their phytoconstituent concentrations and antioxidant and cytotoxic potential. The total phenolic content in LS, LJ, and HAO extracts was 171.20 ± 0.02, 120.73 ± 0.02, and 72.05 ± 0.01 mg gallic acid equivalent/100 g, respectively and the total flavonoid content was 23.635 ± 0.003, 20.754 ± 0.005, and 18.635 ± 0.005 mg quercetin equivalent/100 g, respectively. Similarly, total ascorbic acid content, carotenoids, and antioxidant potential were higher in the LS and LJ extracts than in HAO. Overall, the LS extract had a substantially higher concentration of phytochemicals and antioxidants, as well as higher cytotoxic potential, compared to the LJ and HAO extracts. The LS extract was tested in the MKN-45 human gastric cancer cell line to demonstrate its effective antioxidant potential and cytotoxicity. Hence, lyophilization (freezing) based techniques are more effective than heat-based techniques in preserving the phytoconstituents and their antioxidant and cytotoxic potential.

Death receptor pathway genes (Caspase and BID) expression analysis in cancerous cells in response to extracts of amla and turmeric.

Plant extracts antiproliferative effects were determined by using mammalian cells along the expression profile of Caspases 3, 8 and the BID gene of the death receptor-induced pathway. Two medicinal plants viz., Turmeric (Curcuma longa) and Amla (Emblica officinalis) extracts were examined for antiproliferative effect through Neutral Red-Dye uptake assay on Vero and MDA-MB 231 cell lines. A reverse transcriptase polymerase chain reaction was used to determine the expression of genes while GAPDH expression was used as an internal control. Expression of BID was up-regulated in methanolic turmeric extract-induced MDA-MB 231 cells while Caspases 3,8 expressions were the same in induced and uninduced MDA-MB 231 cells. Activated BID cleaved into tBID and activated the intrinsic pathway which caused death in methanolic turmeric extract-induced cancerous cells. Ethanolic extracts of turmeric exerted the strongest antiproliferative effects on Vero and methanolic extracts on MDA-MB 231 cells. The morphological studies of cell lines and gene expression analysis of turmeric methanolic extract-treated cells showed activation of apoptosis via converting BID into t-BID (intrinsic pathway) and activating Caspase-3 and Caspase-8 (extrinsic pathway). With the differential cytotoxicity and induction of apoptosis in induced cancer cells in comparison to uninduced cancerous cells, hence turmeric is a natural source of new anti-cancerous compounds.

Characterization and metabolomic profiling of two pigment producing fungi from infected fruits of Indian Gooseberry.

Two pigment producing fungi, Talaromyces atroroseus and Penicillium choerospondiatis, were isolated and identified from infected fruits of Phyllanthus emblica L. based on amplification and sequencing of internal transcribed spacer region and beta-tubulin gene. This is the first occurrence report of these two fungi from fruits of P. emblica. Culture extract containing metabolites of T. atroroseus and P. choerospondiatis contained phenolics of 26.35 mg and 30.89 mg GAE/g dry extract respectively; whereas no significant amount of flavonoids and tannins were detected. P. choerospondiatis metabolites extract showed higher DPPH and ABTS activity with IC50 values of 21.94 mg/ml and 27.03 mg/ml respectively than T. atroroseus. LC-HRMS analysis of metabolites extract of T. atroroseus revealed presence of trimethyl-isopropyl-butanamide, perlolyrine, N-hexadecanoylpyrrolidine etc. whereas P. choerospondiatis displayed presence of tangeraxanthin, ugaxanthone, daphniphylline, etc. Therefore, fungal metabolites are rich natural sources of diversified compounds that can be utilized in dyeing industries, cosmetics and novel drug development.

Biological Activities of Phenolics from the Fruits of Phyllanthus emblica L. (Euphorbiaceae).

Seven phenolic compounds, 1 - 7, including a new organic acid gallate, mucic acid 1-ethyl 6-methyl ester 2-O-gallate (7), were isolated from the MeOH extract of the fruits of Phyllanthus emblica L. (Euphorbiaceae). The structures were elucidated on the basis of extensive spectroscopic analysis and comparison with literature data. Upon evaluated for their antioxidant abilities by 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) assays. The inhibitory activities against melanogenesis in B16 melanoma cells induced by α-MSH, as well as cytotoxic activities against four human cancer cell lines were also evaluated. All phenolic compounds, 1 - 7, exhibited potent antioxidant abilities (DPPH: IC50 5.6 - 12.9 µm; ABTS: 0.87 - 8.43 µm Trolox/µm; FRAP: 1.01 - 5.79 µm Fe2+ /µm, respectively). Besides, 5 - 7, also exhibited moderate inhibitory activities against melanogenesis (80.7 - 86.8% melanin content), even with no or low toxicity to the cells (93.5 - 101.6% cell viability) at a high concentration of 100 µm. Compounds 1 - 3 exhibited cytotoxic activity against one or more cell lines (IC50 13.9 - 68.4%), and compound 1 with high tumor selectivity for A549 (SI 3.2).

Photocatalytic degradation of nitrophenol using biologically active Phyllanthus emblica seed extract.

The present study deals about the vicinity of phytochemicals present in the Phyllanthus emblica (P. emblica) seed extract. The bio-active compounds present in the methanolic seed extract have been identified using Gas Chromatography Mass Spectroscopy (GC-MS)·The antioxidant activity of P. emblica seed extract was evaluated using assistance of DPPH (2,2-Diphenyl-1-picrylhydrazyl) assay. The determination of total phenol and flavonoid substance were contemplated. Further blood clot lysis activity was also done to check the percentage of clot lysis in methanolic seed extract. The result proved that seed extract has potential application. The GCMS results of P. emblica suggest that Octyl-ß-d-Glucopyranoside is present in major quantity. The work has been designed towards the degradation of 2-nitrophenol and 4-nitrophenol using P. emblica methanolic seed extract. The progress of nitrophenol degradation has been observed in UV-visible spectroscopy. At 5min duration, the 4-nitrophenol has been degraded up to 82.42%. This may be due to the presence of secondary metabolites such as alkaloids, carbohydrate and phenols in the P. emblica seed extract. The seed extract showed good scavenging activity which resulted in IC50 value of 85.92µg/mL. The total phenol and flavonoid content present in the extract were 48.242 and 12.72mg/mL. Also the seed extract showed good lysis when compared to the standard streptokinase.

Anticancer Properties of Phyllanthus emblica (Indian Gooseberry).

There is a wealth of information emanating from both in vitro and in vivo studies indicating fruit extract of the Phyllanthus emblica tree, commonly referred to as Indian Gooseberries, has potent anticancer properties. The bioactivity in this extract is thought to be principally mediated by polyphenols, especially tannins and flavonoids. It remains unclear how polyphenols from Phyllanthus emblica can incorporate both cancer-preventative and antitumor properties. The antioxidant function of Phyllanthus emblica can account for some of the anticancer activity, but clearly other mechanisms are equally important. Herein, we provide a brief overview of the evidence supporting anticancer activity of Indian Gooseberry extracts, suggest possible mechanisms for these actions, and provide future directions that might be taken to translate these findings clinically.

Antibacterial effects of biosynthesized MgO nanoparticles using ethanolic fruit extract of Emblica officinalis.

Magnesium oxides nanoparticles were successfully synthesized from Mg(NO3)(2)·6H2O through a simple greener route using fruit extract (Emblica officinalis). The synthesized samples were characterized by different techniques such as X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy and Scanning electron microscopy (SEM) with Energy dispersive X-ray spectroscopy (EDX) analysis. The XRD pattern shows the face centered cubic structure with 27 nm of crystalline size of MgO nanoparticles was confirmed by the Debye-Scherrer's Formula. The spherical in shape of MgO nanoparticles is confirmed by SEM analysis. MgO nanoparticles treated cotton fabric produced stronger antibacterial activity. These types of treated fabrics are used in medical application.

Type I pro-collagen promoting and anti-collagenase activities of Phyllanthus emblica extract in mouse fibroblasts.

As part of an ongoing search for the novel pharmacological activities of Phyllanthus emblica, the present study has shown its type I collagen promoting and anti-collagenase effects on primary mouse fibroblast cells. At a concentration of 0.1 mg/ml, emblica extract significantly increased the type I pro-collagen level up to 1.65-fold, and 6.78-fold greater than that of an untreated control, determined by immunocytochemistry and Western blot analysis, respectively. Emblica extract caused an approximately 7.75-fold greater type I pro-collagen induction compared to the known herbal collagen enhancer asiaticoside at the same treatment concentration (0.1 mg/ml). Moreover, emblica extract inhibited collagenase activity in a dose-dependent manner. Maximal inhibition was observed (78.67 +/- 3.51%) at a concentration of 1 mg/ml. In summary, emblica extract has a promising pharmacological effect that benefits collagen synthesis and protects against its degradation and could be used as a natural anti-aging ingredient.

Emblica officinalis exerts wound healing action through up-regulation of collagen and extracellular signal-regulated kinases (ERK1/2).

During wound healing, the wound site is rich in oxidants, such as hydrogen peroxide, mostly contributed by neutrophils and macrophages. Ascorbic acid and tannins of low molecular weight, namely emblicanin A (2,3-di-O-galloyl-4,6-(S)-hexahydroxydiphenoyl-2-keto-glucono-delta-lactone) and emblicanin B (2,3,4,6-bis-(S)-hexahydroxydiphenoyl-2-keto-glucono-delta-lactone) present in Emblica officinalis (emblica), have been shown to exhibit a very strong antioxidant action. We proposed that addition of these antioxidants to the wound microenvironment would support the repair process. The present investigation was undertaken to determine the efficacy of emblica on dermal wound healing in vivo. Full-thickness excision wounds were made on the back of the rat and topical application of emblica accelerated wound contraction and closure. Emblica increased cellular proliferation and cross-linking of collagen at the wound site, as evidenced by an increase in the activity of extracellular signal-regulated kinase 1/2, along with an increase in DNA, type III collagen, acid-soluble collagen, aldehyde content, shrinkage temperature and tensile strength. Higher levels of tissue ascorbic acid, alpha-tocopherol, reduced glutathione, superoxide dismutase, catalase, and glutathione peroxidase support the fact that emblica application promotes antioxidant activity at the wound site. In summary, this study provides firm evidence to support that topical application of emblica represents a feasible and productive approach to support dermal wound healing.

Reversal of hepatotoxin-induced pre-fibrogenic events by Emblica officinalis--a histological study.

Efficacy of a herbal product of E. officinalis (fruit) (EO) has been evaluated against carbon tetrachloride (CCl4) and thioacetamide (TAA) induced changes in rat liver. Chronic treatment of CCl4 and TAA revealed abnormal histopathology indicative of pre-fibrogenic events. EO reversed such alterations with significant regenerative changes suggestive of its preventive role in prefibrogenesis of liver.

Separation and evaluation of free radical-scavenging activity of phenol components of Emblica officinalis extract by using an HPTLC-DPPH* method.

A new procedure has been developed to separate and quantify the free radical-scavenging activity of individual compounds from an Emblica officinalis extract based on the combination of HPTLC with a diode array detector (DAD) and postchromatographic DPPH* radical derivatization. Free gallic and ellagic acids and emblicanins A and B in the E. officinalis extract were separated by TLC and identified. All the compounds of the extract were capable of scavenging of DPPH* radicals. It was established that the DPPH* scavenging activity of emblicanins A and B was 7.86 and 11.20 times more than that of ascorbic acid and 1.25 and 1.78 times more than gallic acid, respectively. From the estimated ID50 values, it can be seen that the increasing order of activity was emblicanin B > emblicanin A > gallic acid > ellagic acid > ascorbic acid. Probably, the antioxidant activity of E. officinalis extract is associated with the presence of hydrolyzable tannins having ascorbic acid-like action.

Biosynthesis of gold and silver nanoparticles using Emblica Officinalis fruit extract, their phase transfer and transmetallation in an organic solution.

The design, synthesis and characterization of biologically synthesized nanomaterials have become an area of significant interest. In this paper, we report the extracellular synthesis of gold and silver nanoparticles using Emblica Officinalis (amla, Indian Gooseberry) fruit extract as the reducing agent to synthesize Ag and Au nanoparticles, their subsequent phase transfer to an organic solution and the transmetallation reaction of hydrophobized silver nanoparticles with hydrophobized chloroaurate ions. On treating aqueous silver sulfate and chloroauric acid solutions with Emblica Officinalis fruit extract, rapid reduction of the silver and chloroaurate ions is observed leading to the formation of highly stable silver and gold nanoparticles in solution. Transmission Electron Microscopy analysis of the silver and gold nanoparticles indicated that they ranged in size from 10 to 20 nm and 15 to 25 nm respectively. Ag and Au nanoparticles thus synthesized were then phase transferred into an organic solution using a cationic surfactant octadecylamine. Transmetallation reaction between hydrophobized silver nanoparticles and hydrophobized chloroaurate ions in chloroform resulted in the formation of gold nanoparticles.

Modulation of haematopoetic system and antioxidant enzymes by Emblica officinalis gaertn and its protective role against gamma-radiation induced damages in mice.

The radio protective effect of the fruit pulp of Emblica officinalis Gaertn (Emblica) was studied in adult Swiss albino mice. Mice were treated with 2.5 g/kg b.wt of Emblica for 10 consecutive days before irradiation and exposed to a single dose of 700 rads (7Gy) of radiation after the last dose. One group was given Emblica continuously for another 15 days after irradiation. Changes in the total leukocyte count, bone marrow viability and hemoglobin were studied after whole body irradiation. Administration of Emblica significantly increased these levels, which were lowered by irradiation. Animals were sacrificed at various time points after irradiation and the activities of the antioxidant enzymes catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPX), and glutathione-S-transferase (GST), and levels of glutathione were assayed in the blood. The damage to the cell membrane after whole body irradiation was studied by measuring the tissue lipid peroxides levels. Administration of Emblica significantly enhanced the activity of the various antioxidant enzymes and GST as well as glutathione system in the blood. Treatment with Emblica also lowered the elevated levels of lipid peroxides in the serum. The data clearly indicated that the extract significantly reduced the bioeffects of radiation. Emblica extract may be useful in reducing the side effects produced during therapeutic radiation.