For the first time, different morphologies of zinc oxide (ZnO) superstructures are synthesized by a simple and environmental friendly route using Nerium oleander leaf extract as fuel. Powder X-ray diffraction, scanning electron microscopy, UV−visible spectroscopy, and photoluminescence studies are performed to ascertain the formation and characterization of ZnO. X-ray diffraction confirmed the crystalline nature of the compound with hexagonal Wurtzite structure. When the concentration of the leaf extract is varied, different morphologies are formed. ZnO are tested for antifungal using soybean seed-borne fungi by food-poison method and antibacterial activity against bacterial human pathogens by a broth microplate dilution method using 96-well plates. Among the screened soybean seed-borne fungi, Fusarium equisiti was found to be more susceptible, which was followed by Macrophomina phaseolina for ZnO nanoparticles (NPs) prepared using 0.2188 mol/dm 3 N. oleander leaf extract. It was observed that NPs exhibited pronounced antifungal activity in a dose-dependent manner with a relatively high percentage of mycelial inhibition. ZnO obtained with the concentration of 0.2188 mol/dm 3 leaf extract showed both minimum inhibitory concentration and minimum bactericidal concentration effectiveness compared to other synthesized compounds. It is observed that the samples with small crystallite size show greater antibacterial activity than those of larger crystallite size. Further, we found that crystallite size and morphology significantly affects the antibacterial activity of ZnO. Prepared compounds showed significant inhibition against Escherichia coli, Staphylococcus aureus, Bacillus subtilis, and Pseudomonas aeurginosa. Among the tested bacteria, P. aeurginosa is more susceptible and E. coli is the least effective against bacterial pathogens. The antibacterial activities of the as-formed ZnO are preliminarily studied against Gram-positive (B. subtilis and S. aureus) and Gram-negative (E. coli and P. aeruginosa) bacteria and are found to be dependent on the shape of the nanostructures.
The present study focuses on isolation and evaluation of the anti-cancer activity of compounds from the leaves of . The bioassay-directed strategy was adopted using chromatographic, gas chromatographic-mass spectrum analysis, nuclear magnetic resonance and X-ray crystallography techniques for purification and characterization of active cytotoxic compounds. Further, MDA-MB-231 breast cancer cell lines and 7,12-dimethylbenz (a) anthracene (DMBA) induced virgin female Sprague Dawley (SD) rats were used for in vitro and in vivo cytotoxicity evaluation. Stigmasterol hemihydrate and 9,12-Octadecadienoic acid (Z,Z)-2-hydroxy-1-(hydroxymethyl)ethyl ester or (β-monolinolein) were the two main cytotoxic constituents of leaf extract of, with an IC value of 74.2 and 13.2 µg/ml, respectively, in MDA-MB-231 cells. Additionally, the treatment with the stigmasterol and β-monolinolein as a combinatorial drug therapy in DMBA-induced female SD rats led to recovery of body weight, decreased tumor weight and volume, without any toxic side effects. Immunohistochemical examination showed extensive cell death and low proliferation in the treated tumor tissues that was confirmed by results from H and E staining, TUNEL assay and Ki-67 index as compared to control animal group. The reversion of glycoprotein, lysosomal and tumor marker enzyme levels back to near-normal levels after treatment with the plant compounds clearly demonstrated the reduction of tumor burden in these animals. This is the first report on isolation and characterization of the two active cytotoxic components from leaves of . Additionally, the profound cytotoxic and tumor-suppressive effect of these two compounds as a combinatorial therapy provide an alternative option for breast cancer treatment.
Die-back disease caused by Phomopsis (Diaporthe) azadirachtae is the devastating disease of Azadirachta indica. Accurate identification of P. azadirachtae is always problematic due to morphological plasticity and delayed appearance of conidia. A species-specific PCR-based assay was developed for rapid and reliable identification of P. azadirachtae by designing a species-specific primer-targeting ITS region of P. azadirachtae isolates. The assay was validated with DNA isolated from different Phomopsis species and other fungal isolates. The PCR assay amplified 313-bp product from all the isolates of P. azadirachtae and not from any other Phomopsis species or any genera indicating its specificity. The assay successfully detected the pathogen DNA in naturally and artificially infected neem seeds and twigs indicating its applicability in seed quarantine and seed health testing. The sensitivity of the assay was 100 fg when genomic DNA of all isolates was analysed. The PCR-based assay was 92% effective in comparison with seed plating technique in detecting the pathogen. This is the first report on the development of species-specific PCR assay for identification and detection of P. azadirachtae. Thus, PCR-based assay developed is very specific, rapid, confirmatory and sensitive tool for detection of pathogen P. azadirachtae at early stages.
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