Draft Genome Sequence of Aspergillus oryzae BP2-1, Isolated from Traditional Malted Rice in South Korea.

The fungus Aspergillus oryzae strain BP2-1 was isolated from the traditional malted starter culture nuruk. We report here the draft whole-genome sequence of A. oryzae BP2-1, which is comprised of 14 scaffolds with a total length of 39,455,382 bp and a GC content of 47.13%.

The enhancing effect of Acanthopanax sessiliflorus fruit extract on the antibacterial activity of porcine alveolar 3D4/31 macrophages via NF-κB1 and lipid metabolism regulation.

OBJECTIVE: The demand for measures to improve disease resistance and productivity of livestock are increasing, as most countries prohibit the addition of antibiotics to feed. This study therefore aimed to uncover functional feed additives to help enhance livestock immunity and disease resistance, using Acanthopanax sessiliflorus fruit extract (ASF). METHODS: ASF was extracted with 70% EtOH, and total polyphenolic and catechin contents were measured by the Folin-Ciocalteu and vanillin assay, respectively. The 3D4/31 porcine macrophage cells (MΦ) were activated by Phorbol 12-Myristate 13-Acetate (PMA), and cell survival and growth rate were measured with or without ASF treatment. Flow-cytometric analysis determined the lysosomal activity, reactive oxygen species levels (ROS), and cell cycle distribution. Nuclear factor kappa B (NF-κB) and superoxide dismutase (SOD) protein expression levels were quantified by western blotting and densitometry analysis. Quantitative PCR was applied to measure the lipid metabolism-related genes expression level. Lastly, the antibacterial activity of 3D4/31 MΦ cells was evaluated by the colony forming unit assay. RESULTS: ASF upregulated the cell viability and growth rate of 3D4/31 MΦ, with or without PMA activation. Moreover, lysosomal activity and intracellular ROS levels were increased after ASF exposure. In addition, the antioxidant enzyme SOD2 expression levels were proportionately increased with ROS levels. Both ASF and PMA treatment resulted in upregulation of NF-κB protein, tumor necrosis factor (TNF) α mRNA expression levels, lipid synthesis, and fatty acid oxidation metabolism. Interestingly, co-treatment of ASF with PMA resulted in recovery of NF-κB, TNFα, and lipid metabolism levels. Finally, ASF pretreatment enhanced the in vitro bactericidal activity of 3D4/31 MΦ against Escherichia coli. CONCLUSION: s: This study provides a novel insight into the regulation of NF-κB activity and lipid metabolism in MΦ, and we anticipate that ASF has the potential to be effective as a feed additive to enhance livestock immunity.

Diversity analysis of Saccharomyces cerevisiae isolated from natural sources by multilocus sequence typing (MLST).

We used multilocus sequence typing (MLST) to analyze the diversity of natural isolates of Saccharomyces cerevisiae, the most important microorganism in alcoholic fermentation. Six loci, ADP1, RPN2, GLN4, ACC1, MET4, and NUP116, in S. cerevisiae genome were selected as MLST markers. To investigate genetic diversity within S. cerevisiae, 42 S. cerevisiae isolated from natural sources in Korea as well as six S. cerevisiae obtained from Genbank and four industrial S. cerevisiae were examined using MLST. Twenty-six polymorphic sites were found in the six loci. Among them, ACC1 had the most genetic variation with eight polymorphic sites. MLST differentiated the 52 strains into three clades. Alcohol fermentation results revealed that S. cerevisiae in Clade III produced less alcohol than those in Clades I and II. These results suggested that MLST is a powerful tool to differentiate S. cerevisiae and can potentially be used to select S. cerevisiae suitable for industrial use.

Extract of Makgeolli suldeot downregulates NF-κB-mediated MMP-9 expression, thereby inhibiting invasive activity in a human breast cancer cell line, MDA-MB-231.

Makgeolli is a Korean traditional alcoholic beverage prepared by fermenting rice starch using nuruk. It is known for its health benefits, but the mechanisms underlying its inhibitory effects on cancer remain unclear. Here a 70% ethanol extract of Makgeolli suldeot (70EMS) was tested for its inhibitory activities against cancer metastasis. This extract interrupted the formation of the neovasculature and reduced the migratory and invasive activity of breast cancer cells. Furthermore, it dose-dependently suppressed the activity of matrix metalloproteinase (MMP)-9 and phosphorylation of ERK1/2, JNK, and Akt. Moreover, 70EMS strongly inhibited the translocation of nuclear factor-kappa B (NF-κB) into the nucleus and expression of activator protein-1 (AP-1). These data indicate that the anti-invasive effect of 70EMS may be related to the inhibition of ERK1/2, phosphorylation of JNK, and reduced expression of NF-κB and AP-1, thereby attenuating MMP-9 in human breast cancer cells.

Development of titanium oxide layer containing nanocrystalline zirconia particles with tetragonal structure: Structural and biological characteristics.

This study investigated the microstructural, mechanical and biological properties of oxide layers containing tetragonal zirconia (t-ZrO2) particles on pure titanium produced by plasma electrolytic oxidation (PEO) process. For this purpose, PEO processes were carried out at an AC current density of 200mA/cm(2) for 180s in potassium pyrophosphate (K4P2O7) electrolytes with and without t-ZrO2 powder. Structural investigations using transmission electron microscopy exhibited that the present nanocrystalline oxide layer evidenced the successful incorporation of a myriad of t-ZrO2 particles working as an intermediate medium to reinforce the adhesion strength between the substrate and oxide layer. Regarding biomimetic apatite formation, the t-ZrO2 particles uniformly spread were of considerable importance in triggering the nucleation and growth of biomimetic apatite on the surface of the oxide layer immersed in a simulated body fluid solution. The growth and proliferation rates of the osteoblasts (MC3T3-E1) cultured on the oxide layer with t-ZrO2 particles were higher than that without t-ZrO2 particles due to the higher roughness providing the better sites for the filopodia extension and interlocking.

Cyanidin-3-O-sambubioside from Acanthopanax sessiliflorus fruit inhibits metastasis by downregulating MMP-9 in breast cancer cells MDA-MB-231.

Anthocyanins play an important role in physiological functions related to human health. This study aimed to investigate the inhibitory effect of cyanidin-3-O-sambubioside isolated from Acanthopanax sessiliflorus fruit on cancer cell metastasis. The inhibitory effect of cyanidin-3-O-sambubioside on angiogenesis was assessed by chorioallantoic membrane and tube formation assays. Cyanidin-3-O-sambubioside interrupted the formation of neovasculature and tube-like structures in a dose-dependent manner. It inhibited the wound healing migration and invasion of MDA-MB-231 cells through reconstituted extracellular matrix (Matrigel). Gelatin zymography revealed that cyanidin-3-O-sambubioside significantly decreased the secretion of matrix metalloproteinase-9 and inhibited matrix metalloproteinase-9 expression. In additional, cyanidin-3-O-sambubioside exerted an inhibitory effect on Akt phosphorylation. These results suggest that cyanidin-3-O-sambubioside inhibits metastasis processes, such as angiogenesis and invasion, in breast cancer cells through regulation of matrix metalloproteinase-9 activity.

Bioactive 3,4-seco-Triterpenoids from the Fruits of Acanthopanax sessiliflorus.

Eight new 3,4-seco-lupane triterpenes and glycosides, acanthosessiligenins I and II (1, 3) and acanthosessiliosides A-F (2, 4-8), as well as six known 3,4-seco-lupane triterpenes (9-14) were isolated from an ethanolic extract of Acanthopanax sessiliflorus fruits. The chemical structures of 1-8 were determined by spectroscopic data interpretation. All isolated compounds were tested for their cytotoxicity against six human cancer cell lines and their ability to inhibit LPS-induced nitric oxide production in RAW 264.7 macrophages.

Anti-inflammatory lignans from the fruits of Acanthopanax sessiliflorus.

A new lignan, named acanthosessilin A (1), as well as eight known lignan and lignan glycosides 2-9 were isolated from an ethanolic extract of Acanthopanax sessiliflorus fruits. The chemical structures were determined by spectroscopic methods, including HR-EIMS, 1D NMR (1H, 13C, DEPT), 2D NMR (gCOSY, gHSQC, gHMBC, NOESY), and IR spectroscopy. All isolated compounds were tested for the ability to inhibit LPS-induced nitric oxide production in RAW264.7 macrophages.

Sterols isolated from Nuruk (Rhizopus oryzae KSD-815) inhibit the migration of cancer cells.

An activity-guided fractionation method was used to isolate anticancer components from Nuruk (Rhizopus oryzae KSD-815:KSD-815). Dried powder of KSD-815 was extracted with 80% methanol and partitioned successively using nhexane, ethyl acetate, n-butanol, and water. The n-hexane and n-butanol fractions showed a strong antimigratory effect on human cancer cells. Both of these fractions were subjected to separation and purification procedures using silica gel, octadecyl silica gel, and Sephadex LH-20 column chromatographies to afford four purified compounds. These were identified as ergosterol peroxide (1), stigmast- 5-en-3beta,7beta-diol (2), ergosta-7,22-dien-3beta,5alpha,6beta,9alpha-tetraol (3), and daucosterol (4), respectively, by spectroscopic methods such as nuclear magnetic resonance spectrometry, mass spectrometry, and infrared spectroscopy, and comparison with those in the literature. Compounds 1-4 were isolated from KSD-815 for the first time. Compounds 1 and 4 inhibited the migration of MDA-MB-231 cells at concentrations lower than 20 micronM.

Nuruk extract inhibits lipopolysaccharide-induced production of nitrite and interleukin-6 in RAW 264.7 cells through blocking activation of p38 mitogen-activated protein kinase.

Nuruk, which is a natural inoculator and source of amylolytic enzymes, is used in Korean traditional rice wine. A methanol extract of nuruk (NE) attenuated lipopolysaccharide (LPS)- induced nitrite and interleukin (IL)-6 in RAW 264.7 cells. Both the n-hexane and water fractions from NE (MEH and MW, respectively) inhibited the production of nitrite and IL-6 in RAW 264.7 cells. MEH and MW also inhibited the LPS-induced inducible nitric oxide synthase expression. Further, and MEH protected against the LPS-induced activation of p38 mitogen-activated protein kinase. Together, these results indicate that nuruk may contribute to the anti-inflammatory and cancer-preventive effects of Korean traditional rice wine.

Stability of new carbapenem DA-1131 to renal dipeptidase (dehydropeptidase I).

The stability of DA-1131 to renal dipeptidase (RDPase) (EC 3.4.13.19) was compared with that of imipenem and meropenem by V(max)/K(m) ratios as an index of the enzyme's preference for substrates. Our results showed a decreasing order of imipenem (6.24), meropenem (2.41), and DA-1131 (1.39). The biochemical evaluation of DA-1131 as the least preferred substrate of RDPase suggests its potential use as a novel beta-lactam antibiotic which may be usable without coadministration of RDPase inhibitors once its clinical suitability is proven.