Enhancing Rubisco gene expression and metabolites accumulation for better plant growth in Ficus deltoidea under drought stress using hydrogen peroxide.

Growth improvement of the medicinal plant, Ficus deltoidea (Mas Cotek) under drought conditions is a vital issue in Malaysia since it is a slow-growing plant and disposed to leaf damage under the stresses of drought. Therefore, investigation was done to examine the outcomes of hydrogen peroxide (H2O2) application on Rubisco gene expression and metabolites accumulation of stressed F. deltoidea plants, and thereby to record the changes in leaf histology, photosynthesis, biochemical properties, and the growth of the plant. H2O2 at the rates of 0, 5, 10, 15, and 20 mM were foliar sprayed biweekly on the drought stressed plants using a hand sprayer. The application of 20 mM H2O2 amplified leaf number, tallness, stomatal conductance, and photosynthetic yield by 143, 24, 88, and 18%, respectively, over the control plant. A reduced transpiration rate and improved chlorophyll fluorescence were also noted in H2O2-treated plants. The treatment produced a greater amount of chlorophyll a, total phenols, total flavonoids, sugar content, and antioxidant activities by 1.61-, 1.30-, 1.98-, 1.92-, and 1.53-fold, respectively. Application of 15 mM H2O2 enhanced net photosynthetic rate and internal CO2 concentrations by 1.05- and 1.25-fold, respectively. Additionally, H2O2 treatments promoted stomatal closure, increased stomata size, the number of stomata, improved vein structure, and reduced the damage of the leaf margin and mesophyll cells of drought stressed plants. The application of H2O2 also accumulated significantly higher contents of sodium (Na+), calcium (Ca2+), potassium (K+), magnesium (Mg+), and iron (Fe2+) in stressed plants. Although the amount of Arsenic (As+) and Antimony (Sb3+) increased to some extent, the increases were not at a toxic level. The use of H2O2 enhanced the Rubisco gene expression to a greater level and the ratio of Rubisco expression increased up to 16-fold. Finally, thirteen (13) identified and five (5) unmatched volatile compounds with a quality score above 70% were identified by gas chromatography-mass spectrometry (GCMS). The GCMS analysis showed that the foliar application of H2O2 accumulates a higher percentage of volatile components in plants which helps to mitigate the negative effects of drought stress. It is concluded that under drought stressed conditions the F. deltoidea plants should be treated with 10-15 mM of H2O2 twice a week to improve leaf histology, photosynthesis, the level of Rubisco gene expression and volatile compounds accumulation, and plant growth and development.

Anti-Uterine Fibroid Effect of Standardized Labisia Pumila Var. Alata Extracts In Vitro and in Human Uterine Fibroid Cancer Xenograft Model.

BACKGROUND: Uterine fibroids are a common type of solid tumor presenting in women of reproductive age. There are very few alternative treatment available from conventional treatment involving surgeries. Labisia pumila var. alata or locally known as 'Kacip Fatimah' was widely used as traditional medicine in Malaysia. This plant has been used to maintain a healthy female reproductive system. The present study aimed to evaluate anti fibroid potential of L. pumila extracts through in vitro apoptosis activity against uterine leiomyoma cells (SK-UT-1) and in uterine leiomyoma xenograft model. Evaluation of bioactive markers content were also carried out. METHODS: Apoptotic induction of the extracts was determined by morphological examination of AO/PI dual staining assay by flourescent microscopy and flow cytometry analysis on Annexin V-FITC/PI stained cells. In vivo study was done in immune-compromised mouse xenograft model. HPLC analysis was employed to quantify marker compounds. RESULTS: Morphological analysis showed L. pumila induced apoptosis in a dose dependent manner against SK-UT-1 cells. In vivo study indicated that L. pumila significantly suppressed the growth of uterine fibroid tumor. All tested extracts contain bioactive marker of gallic acid and cafeic acid. CONCLUSION: This work provide significant data of the potential of L. pumila in management of uterine fibroids.
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Organic matter and moringa leaf extract's effects on the physiology and fruit quality of red seedless watermelon (Citrullus lanatus) / Efeitos da matéria orgânica e do extrato da folha de moringa sobre a fisiologia e qualidade do fruto da melancia vermelha sem sementes ( Citrullus lanatus )

This study was designed to assess the effects of different types of organic matter on the quality and biochemical properties of red seedlesswatermelon.The research was a single factor experiment, which involved six (6) treatments and four (4) replications. Watermelon grown and harvested from five (5) different organic matter sources, namely; vermicompost (VC), poultry manure (PM), cow dung (CD), goatdung (GD), and moringa leaf extract (MLE), at the rates of 10 ton, 20 ton, 30 ton, 30 ton, and 3000 L ha-1, respectively, were used in this study. The results revealed that the treatment of vermicompost improved the chlorophyll content, chlorophyll fluorescence, stomatal conductance, internal carbon dioxide, net photosynthetic rate, fruit weight and mineral nutrient content of watermelon. The application of cow dung reduced the rind thickness and increased the TSS and antioxidant activity of the watermelon. The goat dung treatment significantly affected fruit size, juice content and the pH content of the watermelon's fruit juice. The poultry manure (PM) and moringa extract (MLE) treatments resulted in the highest anthocyanin content of all the watermelon. Although all the organic matter, in their unique ways, improved the quality of red seedless watermelon, the study shows that the vermicompost application had better growth, yield and improvement in the quality of watermelon.

Este estudo foi desenhado para avaliar os efeitos de diferentes tipos de matéria orgânica sobre a qualidade e propriedades bioquímicas da melancia vermelha sem sementes. A pesquisa foi um experimento de fator único, que envolveu seis (6) tratamentos e quatro (4) repetições. Melancia cultivada e colhida de cinco (5) diferentes fontes de matéria orgânica, nomeadamente; vermicomposto (VC), esterco de galinha (PM), estrume de vaca (CD), esterco de cabra (GD) e extrato de folhas de moringa (MLE), nas doses de 10 ton, 20 ton, 30 ton, 30 ton e 3000 L ha-1, respectivamente, foram utilizados neste estudo. Os resultados revelaram que o tratamento do vermicomposto melhorou o teor de clorofila, a fluorescência da clorofila, a condutância estomática, o dióxido de carbono interno, a taxa fotossintética líquida, o peso do fruto e o teor de nutrientes minerais da melancia. A aplicação de esterco bovino reduziu a espessura da casca e aumentou o TSS e a atividade antioxidante da melancia. O tratamento com esterco de cabra afetou significativamente o tamanho do fruto, o teor de suco e o teor de pH do suco de fruta da melancia. Os tratamentos com esterco de galinha (PM) e extrato de moringa (MLE) resultaram no maior teor de antocianina de todas as melancias. Apesar de toda a matéria orgânica, em seus meios únicos, melhorar a qualidade da melancia vermelha sem sementes, o estudo mostra que a aplicação do vermicomposto apresentou melhor crescimento, rendimento e melhora na qualidade da melancia.

Phosphorylation of importin-α1 by CDK1-cyclin B1 controls mitotic spindle assembly.

Importin-α serves as an adaptor linking importin-ß to proteins carrying a nuclear localization sequence (NLS). During interphase, this interaction enables nuclear protein import, while in mitosis it regulates spindle assembly factors (SAFs) and controls microtubule nucleation, stabilization and spindle function. Here, we show that human importin-α1 is regulated during the cell cycle and is phosphorylated at two sites (threonine 9 and serine 62) during mitosis by the major mitotic protein kinase CDK1-cyclin B. Mutational analysis indicates that the mitotic phosphorylation of importin-α1 inhibits its binding to importin-ß and promotes the release of TPX2 and KIFC1, which are then targeted like importin-ß to the spindle. Loss of importin-α1 or expression of a non-phosphorylated mutant of importin-α1 results in the formation of shortened spindles with reduced microtubule density and induces a prolonged metaphase, whereas phosphorylation-mimicking mutants are functional in mitosis. We propose that phosphorylation of importin-α1 is a general mechanism for the spatial and temporal control of mitotic spindle assembly by CDK1-cyclin B1 that acts through the release of SAFs such as TPX2 and KIFC1 from inhibitory complexes that restrict spindle assembly.

Orthosiphon stamineus Extracts Inhibits Proliferation and Induces Apoptosis in Uterine Fibroid Cells

Objectives: The effects of water and 50% ethanolic-water extracts of Orthosiphon stamineus Benth (OS) on cell proliferation and apoptotic activity against uterine leiomyosarcoma (SK-UT-1) cells were investigated. Methods: Anti-proliferation effect was evaluated through cell cycle analysis whereas apoptotic activity was determined via screening and quantifying using fluorescence microscopy and flow cytometric analysis, respectively. The effect of extracts on molecular mechanism was studied using real-time reverse transcription polymerase chain reaction and Western blotting. Results: Cell cycle flow cytometric analysis showed the induction of cell cycle arrests were behaves in a p53-independent manner. The examination using fluorescence microscopy and Annexin V flow cytometry revealed the presence of morphological features of apoptotic bodies. Downregulation of anti-apoptotic gene (Bcl-2) supports the apoptotic activity of OS extracts although poorly induce PARP-1 cleavage in Western blot analysis. The extracts also inhibit the SK-UT-1 growth by suppressing VEGF-A, TGF-ß1 and PCNA genes, which involved in angiogenesis and cell proliferation. Conclusion: This study demonstrates that O. stamineus extracts are able to inhibit proliferation and induced apoptosis of uterine fibroid cells and is worth further investigation.

Selected metabolites profiling of Orthosiphon stamineus Benth leaves extracts combined with chemometrics analysis and correlation with biological activities.

BACKGROUND: Studies on selected metabolites profiling of Orthosiphon stamineus extracts using chromatographic and spectroscopic techniques combined with chemometric tools have not been fully elucidated. Thus present study was performed to profile selected metabolites in O. stamineus leaves extracts using HPLC and FTIR combined with chemometric tools and correlated with biological activities. METHODS: Five different extracts were prepared using three methods; maceration, soxhlet and reflux. The extracts were analyzed using UV-Vis, HPLC and FTIR techniques. Analysis of selected primary and secondary metabolites was also evaluated. The antioxidant and cytotoxic activities of the extracts were evaluated. Chemometric tools were employed to classify the extracts based on HPLC analysis and FTIR fingerprints. RESULTS: The ethanolic extract using maceration characterized high content of phenolics and flavonoids, (rosmarinic acid and eupatorin) with high antioxidant activity. Ethanolic (50%) and methanolic extracts using soxhlet showed high proteins and glycosaponins. Water extracts using reflux and maceration showed high polysaccharides. Methanolic extract (50%) using soxhlet and methanolic extract using maceration showed strong cytotoxic effect against MCF7 and HCT116 cell lines, respectively. Antioxidant and cytotoxic activities showed significant correlation with selected primary and secondary metabolites. HPLC fingerprints combined with chemometrics showed the extracts have been clustered based on selected major peaks profile. FTIR fingerprints combined with chemometrics showed that the extracts have been clustered based on protein and polysaccharide contents. CONCLUSION: Ten different extracts of O. stamineus have showed significant differences in the content of selected primary and secondary metabolites as well as the biological activities. Chemometric tools were able to classify and discriminate the distinctive features of extracts thus can be correlated with the biological activities.