The In Vitro and In Vivo Biological Activities of the Leaf of Cape Myrtle, Myrsine africana L.

The cape myrtle, Myrsine africana L., is a widely used medicinal plant, which has not been well investigated. We assessed the in vivo hepatoprotective and in vitro antiproliferative and antioxidant effects of leaf extracts of M. africana chemically profiled using high-performance liquid chromatography. Three flavonoids were quantified, and gas chromatography-mass spectrometry analysis revealed the presence of common fatty acids. The animal study was conducted on mice treated with CCl4, using three doses each of the methanol and chloroform extract (100, 200 and 300 mg/kg b.w.),with silymarin as a positive control. Hepatoprotective effects were determined by analyzing blood for liver marker and antioxidant enzymes, direct bilirubins and total proteins. The methanol extract (300 mg/kg b.w.) showed the strongest hepatoprotective effects against abnormalities produced by CCl4. The in vivo hepatoprotective effects correlated well with the in vitro antioxidant and antiproliferative activities and with high levels of flavonoids in the extracts. Finally, molecular docking studies of the constituent quercetin were undertaken in silico and several sites of binding to human estrogen receptor (ER) protein, linked with alkaline phosphatase, identified. Copyright © 2017 John Wiley & Sons, Ltd.

Biostimulation potential of biochar for remediating the crude oil contaminated soil and plant growth.

Crude oil contamination is a serious environmental threat to soil and plants growing in it. Biochar has the potential of biostimulation for remediation of crude oil-contaminated soil. Therefore, the current research was designed to analyze the bio-stimulatory impact of biochar for remediating the crude oil contaminated soil (10%, and 15%), and growth of maize under glasshouse conditions. Biochar was produced by pyrolysis of Australian pines at 350 °C. Soil incubations were done for 20 days. The results of soil analysis showed that the crude oil degradation efficiency of biochar was 34%. The soil enzymatic activities had shown 38.5% increase in fluorescein diacetate (FDA) hydrolysis and 55.6% increase in dehydrogenase activity in soil incubated with biochar in comparison to control. The soil microbial diversity was improved to 41% in biochar treated soil with respect to untreated one, while microbial respiration rate had shown a 33.67% increase in soil incubated with biochar with respect to control under oil stress. Gas Chromatography Mass spectrometry (GC-MS) analysis had shown the high content of low molecular weight hydrocarbons (C9-C13) in the soil incubated with biochar in comparison to untreated soil. Biochar showed a significant increase in fresh and dry biomass (25%, 14.61%), leaf area (10%), total chlorophyll (11%), water potential (21.6%), osmotic potential (21%), and membrane stability index (12.7%). Moreover, biochar treatment showed a higher increase in the contents of proline (29%), total amino acids (18%), soluble sugars (30.4%), and antioxidant enzymes like superoxide dismutase (16.5%), catalase (11%), and peroxidase (12%). Overall, the results of the present study suggest the bio-stimulating potential of biochar for degradation of hydrocarbons in crude oil contaminated soil and their growth-stimulating effects on maize.

Isolation of Oxyberberine and ß-Sitosterol from Berberis lycium Royle Root Bark Extract and In Vitro Cytotoxicity against Liver and Lung Cancer Cell Lines.

Berberis lycium Royle has been traditionally used to cure rheumatism, eye and ear diseases, malarial fever, diabetes, stomach disorders, and skin diseases. There is a least amount of data available on cytotoxic capacity of Berberis lycium from Pakistani origin, so on this basis, the present study was aimed to screen Berberis lycium root bark extracts for cytotoxicity against cancer cell lines and isolation of chemical constituents from the most cytotoxic extract. Initial screening of extracts was performed on HepG2 cells at 100 µg/mL for 72 hours of treatment by using an MTT assay. Active fractions were subjected to a series of column chromatographies for the isolation of cytotoxic compounds. Molecular structures were elucidated by using combined data from 1H-NMR, 13C-NMR, and ESI-MS graphs. Assessment of reduction in cell proliferation by isolated compounds was performed on three human cancer cell lines (SK-Hep-1, HepG2, and NCI-H1299). Both n-hexane and chloroform fractions were found active with percent cell viabilities of 8.41 ± 2.23 and 22.31 ± 9.11 in HepG2 cells compared with lupeol 35.43 ± 3.35 percent viability. A protoberberine alkaloid identified as oxyberberine was isolated from chloroform fraction while ß-sitosterol was isolated from n-hexane fraction. Oxyberberine inhibited SK-Hep-1 cell proliferation under a dose-dependent manner with an IC50 value of 34.26 ± 3.34 µM while HepG2 cells showed 50% inhibition at 62.96 ± 4.12 µM. ß-Sitosterol showed reduction in cell viability in SK-Hep-1 cells and HepG2 cells with IC50 values of 123.12 ± 3.51 µM and 140 ± 4.21 µM. This is the first report on the isolation of oxyberberine and ß-sitosterol from Berberis lycium root bark and their cytotoxic evaluation against SK-Hep-1 and NCI-H1299 cells. The cytotoxic potential of Berberis lycium Royle extracts and isolated compounds is suggesting that it is a promising candidate for anticancer drug discovery.

Antioxidant and Cytotoxic Activities and Phytochemical Analysis of Euphorbia wallichii Root Extract and its Fractions.

Euphorbia wallichii a perennial herb growing mainly in Himalayas has been widely used in folk medicines for its medicinal properties. In the present study, the crude methanolic root extract (CME) and its fractions; n-Hexane Fraction (NHF), n-Butanol Fraction (NBF), Chloroform Fraction (CHF), Ethyl acetate Fraction (EAF) and Aqueous Fraction (AQF) of this plant specie were investigated for antioxidant and cytotoxic activities and phytochemical analysis. Antioxidant activity was determined by using 2,2-diphenyl-1-picryl-hydrazyl free radical (DPPH) and DNA protection assay performed on pBR322 plasmid DNA. In both these assays, promising results were obtained for CME as well as other fractions. The IC50 values for DPPH assay were in a range of 7.89 to 63.35 µg/ml in which EAF showed the best anti-oxidant potential and almost all the tested samples showed certain level of DNA protection. The cytotoxic activity was assessed by using Sulforhodamine B (SRB) assay on human cell lines; H157 (Lung Carcinoma) and HT144 (Malignant Melanoma). The IC50 values of the tested samples ranged from 0.18 to 1.4 mg/mL against H157 cell line whereas against HT144 cell line the IC50 values ranged from 0.46 to 17.88 mg/mL with NBF fraction showing maximum potential for both. Furthermore, the phytochemical analysis of CME and its fractions showed the presences of flavonoids, saponins, tannins, terpenoides and cardiac glycosides with varying concentrations.