Molecular and morphological evaluation of transgenic Persian walnut plants harboring Fld gene under osmotic stress condition.

BACKGROUND: Soil drought stress is a limiting factor of productivity in walnut (Juglans regia L). Ferredoxin (Fd) level decreases under adverse environmental stress. Functional replacement of decreased Fd by Fld (Flavodoxin) had been shown to have protective effect under abiotic stress condition. This study aimed to evaluate four transgenic lines (L3, L4, L13 and L17) along with non-transgenic line under three osmotic stresses levels (0, 10 and 12% PEG). METHODS AND RESULTS: This experiment carried out based on a completely randomized design with four replications. To confirm that the Fld gene is successfully integrated into the walnut genome, PCR and dot blot analysis were carried out. The transgenic lines of walnut expressing Fld displayed increased tolerance to osmotic stress at 10 and 12% PEG condition. Lines expressing Fld exhibited increasing tolerance to drought stress and maintained health of plants under osmotic conditions. Results of real time PCR showed that expression level of Fld gene in L4 was higher than the others. Among transgenic lines, L4 was more tolerant than other lines under osmotic stress. CONCLUSIONS: These findings indicate that expression of Fld gene can increase tolerance to osmotic stress in Persian walnut and is useful tool for walnut production in arid and semi-arid regions.

Anticancer effect of Psidium guajava (Guava) leaf extracts against colorectal cancer through inhibition of angiogenesis

Objective: To evaluate the anti-angiogenic and anticancer activities of Psidium guajava leaf extracts against angiogenesis-dependent colorectal cancer. Methods: Three extracts were produced using distilled water, ethanol, and n-hexane as solvents. The extracts were physically characterised through gas chromatography-mass spectrometry, ultraviolet-visible spectroscopy, and Fourier transform infrared spectroscopy. Their antioxidant activity was evaluated using the 2,2-diphenyl-1-picrylhydrazyl, total phenolic content, and total flavonoid content assays. To assess their anti-angiogenic activity, cell viability and rat aortic ring assays were conducted, while cell migration, tube formation, colony formation, and VEGF ELISA assays were conducted to elucidate their effects on different aspects of angiogenesis. Molecular docking was used to assess the anti-angiogenic potential of some possible compounds in the extracts. Tumour spheroid assay was used to assess the extracts' potential as a treatment for colorectal cancer. Results: The ethanol extract showed the best antioxidant activity. The distilled water and ethanol extracts exhibited more inhibitory activity against EA.hy926 cell viability and aortic ring microvessel growth. In addition, the ethanol extract performed significantly better than the distilled water extract against cell migration and colony formation, and VEGF expression of the cells was suppressed by the ethanol extract. Both the distilled water and ethanol extracts showed significant inhibitory effect on EA.hy926 tube formation and tumour spheroids consisting of EA.hy926 and HCT116 cells. The ethanol extract contained β-caryophyllene and β-elemene by phytochemical analysis and subsequent docking studies, which may contribute to its anti-angiogenic activity. Conclusions: The ethanol extract of Psidium guajava has potential in the treatment of colorectal cancer through the inhibition of angiogenesis.

Antiangiogenic activity of sophorolipids extracted from refined bleached deodorized palm olein.

OBJECTIVES: Sophorolipids (SLs) are a group of surface-active glycolipids produced by a type of nonpathogenic yeast Candida bombicola in the presence of vegetable oil through fermentation technology. SLs have shown antitumor activity; however, the mechanism of action underlying the anticancer activity of SLs is poorly understood. This work evaluated the anticancer activity of SLs fermented from palm oil by exploring its antiangiogenic activity. MATERIALS AND METHODS: The SLs that were fermented and further characterized for their biochemical activities. Cytotoxicity study was performed to assess cytostatic properties. A series of in vitro and ex vivo angiogenesis assay was also carried out. The relative fold change in the expression of p53 mRNA by SLs was also studied. RESULTS: Altogether, the data show that SLs derived from palm oil fermentation process inhibited neovascularization in the ex vivo tissue segments and also the endothelial cell proliferation between 50% and 65% inhibition as a whole. The palm oil derived SLs also caused downregulation of the suppression level of vascular endothelial growth factor and also upregulate the p53 mRNA level. The analytical studies revealed the presence of high amount of phenolic compounds but with relatively weak antioxidant activity. The gas chromatography-mass spectrometry studies revealed abundant amount of palmitic and oleic acid, the latter an established antiangiogenic agent, and the former being proangiogenic. CONCLUSION: Therefore, it can be concluded from this study that SLs derived from fermented palm oil have potent antiangiogenic activity which may be attributed by its oleic acid component.