Potential usage of biosynthesized zinc oxide nanoparticles from mangosteen peel ethanol extract to inhibit Xanthomonas oryzae and promote rice growth.

In recent decades, the biosynthesis of nanoparticles using biological agents, such as plant extracts, has grown in popularity due to their environmental and economic benefits. Therefore, this study investigated into utilizing ethanol crude extract sourced from mangosteen peel for the synthesis of zinc oxide nanoparticles (ZnO NPs) and assessing their efficacy against the rice blight pathogen (Xanthomonas oryzae pv. oryzae) through antibacterial evaluations. Additionally, the effects of the synthesized ZnO NPs on rice plant growth was investigated. The X-ray diffraction analysis revealed the production of wurtzite ZnO NPs under specific synthesis conditions, exhibiting a crystallite size of 38.71 nm (or 387.122 Å) without any contamination. Analysis of the ultraviolet-visible optical absorption spectrum indicated a characteristic absorption peak at 363 nm, suggesting a calculated band gap energy of 2.88 eV for the ZnO NPs. Furthermore, Fourier transform infrared spectroscopy analysis confirmed the presence of active compounds functional groups from mangosteen peel in the synthesized ZnO NPs. These biosynthesized ZnO NPs demonstrated significant inhibition of X. oryzae pv. oryzae growth, exhibiting an in vitro 50 % inhibitory concentration (IC50) value of 1.895 mg/mL and a minimum inhibitory concentration (MIC) value of 4 mg/mL. The ZnO NPs treatments at two-fold IC50 values significantly enhanced root length, dry biomass, and chlorophyll a content in rice plants. Consequently, the results demonstrated the potential application of biosynthesized ZnO NPs from mangosteen peel extract in green agriculture, as an alternative to excessive antibiotic use, for combating bacterial plant diseases, and for enhancing plant growth.

In vitro evaluation of the anti­breast cancer properties and gene expression profiles of Thai traditional formulary medicine extracts.

Breast cancer is a leading cause of cancer-related deaths worldwide. Moreover, standard treatments are limited, so new alternative treatments are required. Thai traditional formulary medicine (TTFM) utilizes certain herbs to treat different diseases due to their dominant properties including anti-fungal, anti-bacterial, antigenotoxic, anti-inflammatory and anti-cancer actions. However, very little is known about the anti-cancer properties of TTFM against breast cancer cells and the underlying molecular mechanism has not been elucidated. Therefore, the present study, evaluated the metabolite profiles of TTFM extracts, the anti-cancer activities of TTFM extracts, their effects on the apoptosis pathway and associated gene expression profiles. Liquid chromatography with tandem mass spectroscopy analysis identified a total of 226 compounds within the TTFM extracts. Several of these compounds have been previously shown to have an anti-cancer effect in certain cancer types. The MTT results demonstrated that the TTFM extracts significantly reduced the cell viability of the breast cancer 4T1 and MDA-MB-231 cell lines. Moreover, an apoptosis assay, demonstrated that the TTFM extracts significantly increased the proportion of apoptotic cells. Furthermore, the RNA-sequencing results demonstrated that 25 known genes were affected by TTFM treatment in 4T1 cells. TTFM treatment significantly up-regulated Slc5a8 and Arhgap9 expression compared with untreated cells. Moreover, Cybb, and Bach2os were significantly downregulated after TTFM treatment compared with untreated cells. Reverse transcription-quantitative PCR demonstrated that TTFM extract treatment significantly increased Slc5a8 and Arhgap9 mRNA expression levels and significantly decreased Cybb mRNA expression levels. Moreover, the mRNA expression levels of Bax and Casp9 were significantly increased after TTFM treatment in 4T1 cells compared with EpH4-Ev cells. These findings indicated anti-breast cancer activity via induction of the apoptotic process. However, further experiments are required to elucidate how TTFM specifically regulates genes and proteins. This study supports the potential usage of TTFM extracts for the development of anti-cancer drugs.

Mulberroside F from In Vitro Culture of Mulberry and the Potential Use of the Root Extracts in Cosmeceutical Applications.

Mulberry (Morus spp.) is primarily used in sericulture, and its uses also extend to the food, pharmaceutical, and cosmetic industries. Mulberry extracts are rich in many bioactive compounds that exhibit a wide range of biological properties. Mulberroside F (Moracin M-6, 3'-di-O-ß-D-glucopyranoside), one of the bioactive compounds found in mulberry, has previously been reported as a whitening agent by inhibiting melanin synthesis and exhibiting antioxidant effects. However, there is still limited information on the presence of this compound in plants cultured in vitro. In this study, the mulberroside F content, biochemical, and cytotoxic properties of the extracts from mulberry cultured in vitro were determined. The results revealed that both root and callus were found to be a potential source of mulberroside F. Furthermore, the mulberroside F content was positively correlated with the inhibitory effects on tyrosinase activity. Cell viability assay also revealed that crude extract of the mulberry root has no cytotoxicity in both human keratinocyte cell line (HaCaT) and Vero cells. Taken together, mulberry tissue culture represents a possible alternative and continuous production of mulberroside F, which could be further utilized in cosmeceutical applications.

Utilization of Cratoxylum formosum crude extract for synthesis of ZnO nanosheets: Characterization, biological activities and effects on gene expression of nonmelanoma skin cancer cell.

Cratoxylum formosum Dyer is a medicinal plant widely found in Asia and commonly consumed for food and folk medicine. It is rich in phenolic compounds. The present study utilized water crude extract of C. formosum leaves to synthesize zinc oxide nanoparticles (ZnO NPs) by green synthesis. The synthesized ZnO NPs with the average electronic band gap ∼3  eV were obtained and found to either have spherical shape or sheet-like structures depending on synthesis process and concentration of crude extract. Higher concentration of C. formosum extract also eliminates impurity of Zn(OH)2 during the synthesis. Results from an agar disk diffusion assay demonstrated that all synthesized ZnO samples inhibited growth of Gram-positive bacteria, Bacillus subtilis and Staphylococcus epidermidis and Gram-negative bacterium, Escherichia coli. Furthermore, all synthesized ZnO demonstrated potent anti-cancer activity against non-melanoma skin cancer cells (A431) and the intermediary of cancerous keratinocytes (HaCaT) without affecting normal cell lines (Vero). In addition, we observed that the ZnO nanosheet offered stronger cytotoxicity effects against A431 than spherical shaped ZnO particles. Analysis of RNA-sequencing data revealed that synthesized ZnO nanosheets altered the number of genes in pathways involved in cancer and MAPK signaling pathways in A431 cells. Several isoforms of metallothionein transcripts were upregulated including transcripts involved in inflammatory responses whereas transcripts promoted cell proliferation and apoptosis were downregulated. Therefore, these studies firstly reported potential usage of the green-synthesized ZnO nanosheets from C. formosum extract for development of antibacterial substances or anticancer drugs.

In vitro evaluation of anti-epidermoid cancer activity of Acanthus ebracteatus protein hydrolysate and their effects on apoptosis and cellular proteins.

Acanthus ebracteatus Vahl. is commonly consumed with the aim of curing cancer, inflammatory conditions and skin diseases in traditional Thai medicine. It is known to contain various phytochemicals; however, very little is known about the effects of A. ebracteatus protein hydrolysate on cancer cells, including its molecular mechanisms. The present study therefore investigated the anti-cancer activity of A. ebracteatus protein hydrolysates against epidermoid cancer of the skin cell line A431. Their effects on the apoptosis pathway and expression of proteins involved in the regulation of apoptosis, cell proliferation or cell cycle were also investigated. Crude extract of protein hydrolysate, partially purified peptides and purified peptides extracted from the aerial part of A. ebracteatus were administered to the A431 cells. The cytotoxicity effects were then determined using an MTT assay. As a result, A. ebracteatus protein hydrolysate significantly inhibited A431 cells with half inhibitory concentration equals to 425.9 ng protein/ml. By performing Annexin V assay, the partially purified peptides of A. ebracteatus were demonstrated to enhance the apoptosis pathway. Furthermore, western blot analysis revealed that the partially purified peptides of A. ebracteatus increased protein expression levels of RelA (p65) and Cyclin D1 proteins. However, A. ebracteatus did not increase the expression levels of p53-serine 15 phosphorylation (Ser15P).

Partially Purified Gloriosa superba Peptides Inhibit Colon Cancer Cell Viability by Inducing Apoptosis Through p53 Upregulation.

BACKGROUND: Colon cancer is a major health problem worldwide. Available treatments such as surgery, chemotherapy, radiation and anticancer drugs are limited due to stage of cancer, side effects and altered biodistribution. The use of peptides extracted from natural products has appeared as a potential therapy. Gloriosa superba is known to contain colchicine and other alkaloids with anticancer activity. However, these peptides contained within the extracts have not been studied. This study, therefore, focuses on an investigation of anti-colon cancer activity from a partially purified protein hydrolysate of G superba rhizome. METHODS: Dried G superba rhizome was extracted using 0.5% sodium dodecyl sulfate and digested with pepsin. The protein hydrolysates with molecular weight lesser than 3kDa were collected and subjected for cell viability assay. Then, the partial purification of the protein hydrolysate was performed using reverse-phase high-performance liquid chromatography. Fractions containing anticancer peptides were investigated, and their effects on apoptosis and protein expression using apoptosis test and Western blot, respectively. RESULTS: Partially purified peptides of G superba rhizome demonstrated anticolon activity in SW620 cells by inducing apoptosis through upregulation of p53 and downregulation of nuclear factor kappa B (NF-κB). CONCLUSIONS: Consequently, G superba peptides showed high potential for further purification and development of anticolon therapeutics.

Human miR-5193 Triggers Gene Silencing in Multiple Genotypes of Hepatitis B Virus.

Hepatitis B virus (HBV) infection can lead to various disease states including asymptomatic, acute hepatitis, chronic hepatitis, liver cirrhosis and hepatocellular carcinoma (HCC), which remain a major health problem worldwide. Previous studies demonstrated that microRNA (miRNAs) plays an important role in viral replication. This study aimed to predict and evaluate human miRNAs targeting multiple genotypes of HBV. Candidate human miRNAs were analyzed by data obtained from miRBase and RNAhybrid. Then miRNAs were selected based on hybridization patterns and minimum free energy (MFE). The silencing effect of miRNA was evaluated by real-time PCR, the luciferase reporter assay and the ELISA assay. Five human miRNAs including miR- 142-5p, miR-384, miR-500b, miR-4731-5p and miR-5193 were found to target several HBV genotypes. Interestingly, miR-5193 was found to be the most potent miRNA that could target against all HBV transcripts in almost all HBV genotypes with a highly stable hybridization pattern (5' canonical with MFE lower than -35 kcal/mol). Moreover, miR-5193 caused significant silencing in luciferase activity (53% reduction), luciferase transcript (60% reduction) and HBV surface antigen (HBsAg) production (20-40% reduction depending on genotypes). Therefore, miR-5193 might be useful and have a vital role for inhibition of HBV replication in the future.

A rational study for identification of highly effective siRNAs against hepatitis B virus.

RNA interference (RNAi) is a powerful gene knockdown technique used for study gene function. It also potentially provides effective agents for inhibiting infectious and genetic diseases. Most of RNAi studies employ a single siRNA designing program and then require large-scale screening experiments to identify functional siRNAs. In this study, we demonstrate that an assembly of results generated from different siRNA designing programs could provide clusters of predicting sites that aided selection of potent siRNAs. Based on the clusters, three siRNA target sites were selected on a conserved RNA region of hepatitis B virus (HBV), known as HBV post-transcriptional regulatory element (HBV PRE) at nucleotide positions 1317-1337, 1357-1377 and 1644-1664. All three chosen siRNAs driven by H1 promoter were highly effective and could drastically decrease expression of HBV transcripts (core, surface and X) and surface protein without induction of interferon response and cell cytotoxicity in liver cancer cell line (HepG2). Based on prediction of secondary structures, the silencing effects of siRNAs were less effective against a loop sequence of the mRNA target with hairpin structure. In summary, we demonstrate an effectual approach for identification of functional siRNAs. Moreover, highly potent siRNAs identified here may serve as novel agents for development of nucleic acid-based HBV therapy.