Mass production of rubusoside using a novel stevioside-specific β-glucosidase from Aspergillus aculeatus

Rubusoside (R) is a natural sweetener and a solubilizing agent with antiangiogenic and antiallergic properties. However, currently, its production is quite expensive, and therefore, we have investigated nine commercially available glycosidases to optimize an economically viable R-production method. A stevioside (ST)-specific β-glucosidase (SSGase) was selected and purified 7-fold from Aspergillus aculeatus Viscozyme L by a two-step column chromatography procedure. The 79 kDa protein was stable from pH 3.0 to pH 7.0 at 50-60 °C. Hydrolysis of ST by SSGase produced R and steviol monoglucosyl ester as determined by (1)H and (13)C nuclear magnetic resonance (NMR). Importantly, SSGase showed higher activity toward ST than other β-linked glucobioses. The optimal conditions for R production were 280 mM ST and 16.6 μL of SSGase at pH 5.1 and 63 °C. This is the first discussion detailing the production of R by enzymatic hydrolysis of ST and is useful for the food additive and pharmaceutical industries.     

Influenza virus neuraminidase inhibitory activity of phlorotannins from the edible brown alga Ecklonia cava

Influenza A virus infections continue to pose a major threat to humans and several animal species. Neuraminidase (NA) is one of the most promising targets for the development of drugs against influenza viruses because of its critical role in the viral life cycle. During the course of a search for NA inhibitors from edible natural sources, we found that the ethyl acetate layer of ethanol extracts of Ecklonia cava showed extremely high NA-inhibitory activity (72.1% inhibition at 30 μg/mL). Bioactivity-guided fractionation of the ethyl acetate layer yielded five phlorotannins, identified as phloroglucinol (1), eckol (2), 7-phloroeckol (3), phlorofucofuroeckol (4), and dieckol (5). The inhibitory activities of these compounds (1-5) against NAs from group-1 (A/Bervig_Mission/1/18 [H1N1], A/PR/8/34 [H1N1]) and group-2 (A/Hong Kong/8/68 [H3N2], A/Chicken/Korea/MS96/96 [H9N2]) influenza A were evaluated to determine potencies and kinetic behavior. Analyses using various in vitro influenza A virus NA assays showed that all five phlorotannin derivatives were selective NA inhibitors. Of the phlorotannins, phlorofucofuroeckol (4) exhibited the most potent inhibitory activities toward group-1 NAs (IC?? values, 4.5 and 14.7 μM), whereas dieckol (5) potently inhibited group-2 NAs. Kinetic analyses indicated that compounds 1-5 were all noncompetitive. Notably, these noncompetitive inhibitors synergized with oseltamivir to enhance the NA-inhibitory effects of oseltamivir.     

Detection and characterization of bovine-like coronaviruses from four species of zoo ruminants

Five coronaviruses (CoVs) were detected in diarrheal feces from four zoo ruminant species: one wisent (Bison bonasus), two Himalayan tahr (Hemitragus jemlahicus), one sitatunga (Tragelaphus spekii), and one nyala (Tragelaphus angasii). We sequenced and analyzed the spike (S) and hemagglutinin/esterase (HE) genes of these viruses and compared the nucleotide (nt) and deduced amino acid (aa) sequences with those of other bovine CoV (BcoV) strains. Comparison of the entire deduced aa sequences of the S and HE glycoproteins revealed no specific differences that would account for discrimination between bovine-like CoV strains from zoo ruminants and BcoVs strains. In addition, the 99.9% aa identity among the five CoV strains revealed that the ruminants were infected by the same strain. Phylogenetically, bovine-like CoVs belong to group 2a CoVs, which are related most closely to the BcoV strains recently isolated in Korea. These data suggest that cattle are potential reservoirs for CoVs that are capable of infecting zoo ruminants.     

Dyeing of cotton and polyester/cotton blend with disperse dyes using sodium 2-(2,3-dibromopropionylamino)-5-(4,6-dichloro-1,3,5-triazinylamino)-benzenesulfonate

The cotton fabrics were pretreated by sodium 2-(2,3-dibromopropionylamino)-5-(4,6-dichloro-1,3,5-triazinylamino)-benzenesulfonate (DBDCBS) at alkaline condition of room temperature and then dyed with four disperse dyes having amino groups (C.I. Disperse Yellow 9, C.I. Disperse Red 11, C.I. Disperse Blue 56 and C.I. Disperse Violet 1) at acidic condition of high temperature. A novel hetero-bifunctional bridge compound,DBDCBS, has two reactive groups such as dichloro-s-triazinyl group andα, β-dibromopropionylamido group. The first has reactivity towards hydroxy group of cellulosic fiber and the second shows reactivity towards amino groups of disperse dye containing amino groups. The results indicate that it is possible to dye polyester/cotton blend at one-bath dyeing using one kind of disperse dye containing amino groups. Therefore, two kinds of dyeing methods such as two-bath process one-bath dyeing and one-bath process one-bath dyeing were investigated and their dyeabilities were compared. The differences between these two methods were negligibly small so that perfect one-bath one-step dyeing of polyester/cotton blend by one kind of disperse dye was achieved.     

Evaluation of new metallized direct dyes for mutagenicity using the Salmonella mammalian mutagenicity assay

A series of new metallized direct dyes based on benzidine congeners, 2,2′-dimethyl-5,5′-dipropoxybenzidine and 5,5′-dipropoxybenzidine, were evaluated for mutagenicity in Salmonella typhimurium strains TA98 and TA100. All of the dyes examined were judged to be non-mutagenic with and without metabolic activation while toxicity was seen in some dyes at high doses. The study also suggested that the standard Salmonella mutagenicity plate-incorporated assay was an excellent method for evaluation of dyes for mutagenicity.     

Antiproliferative Activity of Korean Wild Vegetables on Different Human Tumor Cell Lines

This study was conducted to determine the antiproliferative activity of 24 Korean wild vegetables. The methanol extracts of these wild vegetables were used against lung, breast, colon and gastric cancer cells, and the results were assessed by MTT assay. It was found that at the extract concentration of 400 mgL⁻¹ 14 plants exercised antiproliferative activity over 80% against the lung cancer cells, one plant among six—against breast cancer cells, and two plants among six—against colon cancer cells, respectively. Eighteen wild vegetables had the hyperplasia inhibition activity against gastric cancer cells over 23.6% at all extract concentrations, however, only six plants had the antiproliferative activity over 80% in 600 mgL⁻¹. It was found that the extracts from Youngia sonchifolia, Synurus deltoides, Syneilesis palmata, and Cephalonoplos segetum, in concentration of 400 mgL⁻¹ inhibited the hyperplasia of lung cancer cells over 95% and Angelica gigas—both lung and colon cancer cells over 95%. In conclusion, the studied wild vegetables’ methanol extracts possess dose dependent antiproliferative properties, based on their bioactive compounds, mainly polyphenols, but some of them as Hypericum ascyron against lung cancer are not effective and even course harm.     

Application of the rpoS gene for the detection of Vibrio anguillarum in flounder and prawn by polymerase chain reaction

Vibrio anguillarum , an opportunistic fish pathogen, is the main species responsible for vibriosis, a disease that affects feral and farmed fish and shellfish, and causes considerable economic losses in marine aquaculture. In this study, we used polymerase chain reaction (PCR) to detect V. anguillarum . PCR specificity was evaluated by amplifying the rpoS gene, a general stress regulator, in six strains of V. anguillarum and 36 other bacterial species. PCR amplified a species-specific fragment (689 bp) from V. anguillarum . Furthermore, the PCR assay was sensitive enough to detect rpoS expression from 3 pg of genomic DNA , or from six colony-forming units (CFU) mL⁻¹ of cultured V. anguillarum . However, the assay was less sensitive when genomic DNA from the infected flounder and prawn was used (limit of detection, 50 ng and 10 ng g⁻¹ tissue, respectively). These data demonstrate that PCR amplification of the rpoS gene is a sensitive and species-specific method to detect V. anguillarum in practical situations.     

Fast neutron irradiation deteriorates hippocampus-related memory ability in adult mice

Object recognition memory and contextual fear conditioning task performance in adult C57BL/6 mice exposed to cranial fast neutron irradiation (0.8 Gy) were examined to evaluate hippocampus-related behavioral dysfunction following acute exposure to relatively low doses of fast neutrons. In addition, hippocampal neurogenesis changes in adult murine brain after cranial irradiation were analyzed using the neurogenesis immunohistochemical markers Ki-67 and doublecortin (DCX). In the object recognition memory test and contextual fear conditioning, mice trained 1 and 7 days after irradiation displayed significant memory deficits compared to the sham-irradiated controls. The number of Ki-67- and DCX-positive cells decreased significantly 24 h post-irradiation. These results indicate that acute exposure of the adult mouse brain to a relatively low dose of fast neutrons interrupts hippocampal functions, including learning and memory, possibly by inhibiting neurogenesis.