Molecular and morphological discrimination of Chrysanthemum indicum using allele-specific PCR and T-shaped trichome.

Chrysanthemum indicum L. is a traditional oriental medicinal herb prepared as a tea from flowers that have been used in China and South Korea since ancient times. It has a long history in the treatment of hypertension, inflammation, and respiratory diseases. Among Chrysanthemum species, C. indicum has more active chemical components as well as better therapeutic effects, and C. indicum is mostly used for medicinal purposes in South Korea. However, the usage of C. indicum has become problematic over the years due to the abundance of adulterated Chrysanthemum and confusion with morphologically related species such as C. morifolium, C. boreale, and Aster spathulifolius. Thus, here we developed a method for molecular authentication using chloroplast universal region rpoC2 and morphological authentication based on T-shaped trichomes of the adaxial leaf surface. By using a species-specific primer derived from the rpoC2 region, we established a multiplex allele-specific PCR for the discrimination of C. indicum. Amplicons of 675 bp for C. indicum and 1026 bp for other Chrysanthemum species were produced using both rpoC2-specific and common primers. These primers can be used to analyze dried samples of Chrysanthemum. Morphological discrimination was performed using T-shaped trichomes present only on the adaxial leaf surface of C. indicum species, and then molecular markers were utilized to authenticate C. indicum products from adulterant samples available in the market. Our results indicate that these molecular markers in combination with morphological differentiation can serve as an effective tool for identifying C. indicum.

Rhizobium panacihumi sp. nov., an isolate from ginseng-cultivated soil, as a potential plant growth promoting bacterium.

A novel bacterial strain designated DCY116T was isolated from ginseng-cultivated soil in Gochang-gun, Republic of Korea. Strain DCY116T, belongs to the genus Rhizobium, and is closely related to Rhizobium yantingense H66T (98.3%), Neorhizobium huautlense S02T (98.2%), Rhizobium soli DS-42T (98.1%), Rhizobium smilacinae PTYR-5T (97.9%), and Neorhizobium alkalisoli CCBAU 01393T (97.9%) based on 16S rRNA gene sequence analysis. Analysis of the housekeeping genes atpD, recA, and glnII showed low levels of sequence similarity (96.8%) between strain DCY116T and other closely related species. Strain DCY116T was Gram-stain negative, motile by peritrichous flagella, rod-shaped, strictly aerobic, catalase- and oxidase-positive. Q-10 was the predominant ubiquinone. The major cellular fatty acids were identified as C16:0 and summed feature 8 (C18:1 ω7c and/or C18:1 ω6c). The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, and an unknown lipid (L1-3). Genomic DNA G + C content of strain DCY116T was determined to be 57.2 mol%. DNA-DNA homology values between strain DCY116T and closely related species of the genus Rhizobium were lower than 40%. Strain DCY116T produced indole-3-acetic acid, siderophores, and was able to solubilize phosphate as a potential plant growth promoting bacterium. In conclusion, the results of this study support strain DCY116T as a novel species of the genus Rhizobium, for which the name Rhizobium panacihumi is proposed. The type strain is DCY116T (= KCTC 62017T = JCM 32251T).

Gold Nanoparticles Synthesized with Fresh Panax ginseng Leaf Extract Suppress Adipogenesis by Downregulating PPARγ/CEBPα Signaling in 3T3-L1 Mature Adipocytes.

The obesity rate has been increasing worldwide, which is important because obesity has been linked to the development of various metabolic disorders, such as type 2 diabetes, hypertension, cancer, and stroke. Nanomedicine offers a new approach for treating many diseases, including metabolic disorders such as obesity. In this study, we explored the anti-adipogenic effects of spherical gold nanoparticles synthesized with fresh Panax ginseng leaves (P.g AuNPS) in vitro using 3T3-L1 mature adipocytes. Cell viability was assessed by quantitating preadipocyte growth at different time points. Furthermore, to assess the anti-adipogenic effects of P.g AuNPS, intracellular lipid accumulation was investigated in mature adipocytes. To this end, cells were observed under a microscope and OD measurements were taken after Oil Red O staining. In addition, transcriptional gene regulation was examined by performing real time PCR to assess the levels of adipogenic genes such as PPARγ, CEBPα, CEBPß, Jak2, STAT3, FAS, SREBP-1, and ap2. Moreover, protein levels were evaluated by immunoblotting. Altogether, these results confirm that P.g AuNPS exhibit anti-adipogenic effects at a concentration of 100 µg/ml and that these effects are mediated by the downregulation of PPARγ/CEBPα (major transcription factors) signaling in 3T3-L1 mature adipocytes.

Fermented Aloe arborescens Miller Leaf Extract Suppresses Acute Alcoholic Liver Injury via Antioxidant and Anti-Inflammatory Effects in C57BL/6J Mice.

This study confirmed the change in functional composition and alcohol-induced acute liver injury in Aloe arborescens after fermentation. An acute liver injury was induced by administration of ethanol (3 g/kg/day) to C57BL/6J mice for 5 days. A fermented A. arborescens Miller leaf (FAAL) extract was orally administered 30 minutes before ethanol treatment. After fermentation, the emodin content was approximately 13 times higher than that of the raw material. FAAL extract significantly attenuated ethanol-induced aspartate aminotransferase, alanine aminotransferase, and triglyceride increases in serum and liver tissue. Histological analysis revealed that FAAL extract inhibits inflammatory cell infiltration and fat accumulation in liver tissues. The cytochrome P450 2E1, superoxide dismutase, and glutathione (GSH), which involved in alcohol-induced oxidative stress, were effectively regulated by FAAL extract in serum and liver tissues, except for GSH. FAAL also maintained the antioxidant defense system by upregulating heme oxygenase 1 and nuclear factor erythroid 2-related factor 2 protein expression. In addition, FAAL extract inhibited the decrease in alcohol dehydrogenase and aldehyde dehydrogenase activity, which promoted alcohol metabolism and prevented the activation of inflammatory response. Our results suggest that FAAL could be used as a potential therapeutic agent for ethanol-induced acute liver injury.

Lysobacter panacihumi sp. nov., isolated from ginseng cultivated soil.

A Gram-negative, non-motile, aerobic, catalase-, and oxidasepositive bacterial strain, designated DCY117T, was isolated from ginseng cultivated soil in Gochang-gun, Republic of Korea, and was characterized taxonomically using a multifaceted approach. 16S rRNA gene sequence analysis revealed that strain DCY117T showed highest similarity to Lysobacter ruishenii CTN-1T (95.3%). Phylogenetic analysis revealed that closely related relatives of strain DCY117T were L. aestuarii S2-CT (95.1%), L. daejeonensis GH1-9T (95.0%), and L. caeni BUT-8T (94.9%). Diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), and phosphatidylethanolamine (PE) were the major polar lipids of strain DCY117T. The major isoprenoid quinone was Q-8. The major cellular fatty acids of strain DCY117T were iso-C15:0, iso-C16:0, and summed feature 9 (comprising iso-C17:1ω9c and/or 10-methyl-C16:0). Genomic DNA G + C content was 61.8 mol%. On the basis of our findings, strain DCY117T is a novel species in the genus Lysobacter. We propose the name Lysobacter panacihumi sp. nov., and the type strain is DCY117T (= KCTC 62019T = JCM 32168T).

Caspase-3/MAPK pathways as main regulators of the apoptotic effect of the phyto-mediated synthesized silver nanoparticle from dried stem of Eleutherococcus senticosus in human cancer cells.

Siberian ginseng (Eleutherococcus senticosus) was used for the synthesis of an ecofriendly silver nanoparticle (Sg-AgNP), which has exhibited antibacterial, antioxidant effect and lower cytotoxicity to normal cells in comparison to human cancer cells. Although, the potential anticancer activity of Sg-AgNP has not been determined. In this study, two cancer cell lines were used to evaluate the cytotoxicity and apoptotic effect of Sg-AgNP along with the determination of the role of the Caspase-3 / p38 MAPK pathways. Results shown that Sg-AgNP reduced the cell viability of colon cancer cells HT29 and lung cancer cells A549. The cytotoxic effect was higher than the effect exhibited by a commercial silver nanoparticle and Cisplatin. Reactive oxygen species were observed to be superior in both cell lines in the presence of Sg-AgNPs than c-AgNPs and Cisplatin. It was observed an activation of MAPK14 gene and phosphorylation of p38 MAPK protein in both cell lines induced by Sg-AgNPs treatment. Furthermore, induction of morphological changes in the nucleus was done by Sg-AgNPs at 10 µg/mL in both cell lines. On the other hands, the activation of CASP3 gene and Caspase-3 protein was observed in HT29 cells but only at protein level in A549 cells. These results, suggest that Sg-AgNPs anticancer potential activity might be linked to the induction of apoptosis though the generation of ROS by activation of the Caspase-3/p38 MAPK pathway.

Intracellular synthesis of gold nanoparticles with antioxidant activity by probiotic Lactobacillus kimchicus DCY51T isolated from Korean kimchi.

A straightforward synthesis of gold nanoparticles (AuNps) is achieved by novel probiotic Lactobacillus kimchicus DCY51T isolated from Korean kimchi via an intracellular membrane-bound mechanism. The bioreduction of HAuCl4 into AuNps was verified by ultraviolet-visible spectrophotometry at ∼540nm. AuNps were spherical with varying sizes of 5-30nm. AuNps maintained an average crystallite size of 13nm and demonstrated long-term stability in physiological buffer and biological media. Furthermore, the protective capping layer consisted of amino acid residues and surface-bound proteins rendered them non-toxic to murine macrophage (RAW264.7) and human colorectal adenocarcinoma (HT29) cell lines. Finally, biosynthesized AuNps served as superior free radical scavengers against 2,2-diphenyl-1-picrylhydrazyl (DPPH) in contrast to their corresponding gold salt. In short, this green synthesis is cost-effective and advantageous for the development of a new class of probiotics mediated and non-toxic carriers in targeted drug delivery systems, cancer diagnostic, photothermal therapy, biosensing, and medical imaging.