• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.778-783
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• 1999

Campylobacter is a pathogen for both humans and animals that can be transferred from animals to humans. Four isolates, which grew under 5-10% $CO_2$ and had small and translucent colonies, were obtained from swine gastric mucosa and characterized using various methods. These bacteria were gram negative, spirally shaped with round ends. One or two non-sheathed polar flagella were observed under electron microscopy. A PCR with species-specific protein (SSP) primers for 16S rRNA gene in Campylobacter produced a typical 462 bp fragment. The isolates had various biochemical and molecular characteristics which differentiated them from other Campylobacters. The isolates were catalase and oxidase positive, urease (rapid) negative, nitrate reduction positive, indoxyl acetate hydrolysis positive, y-glutamyl transpeptidase negative, and alkaline phosphatase negative. All four isolates showed growth at $37^{\circ}C{\;}and{\;}42^{\circ}C{\;}but{\;}not{\;}at{\;}25^{\circ}C$, were resistant to cephalotin and cefoperazone, and susceptible to carbenicillin. The isolates showed various results in the reduction of chloride to triphenyl tetrazolium (TTC) and a susceptibility to nalidixic acid. Western blot analysis of these isolates with antiserum raised against one isolate showed different patterns from those of reference strains. A dendrogram drawn with the RAPD results showed that these isolates belonged to a new Campylobacter spp. group different from those of C. jejuni, C. doylei, C. lari, and C. coli.

• Journal of Microbiology
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• v.40 no.3
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• pp.211-218
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• 2002

A 4.8-kb DNA fragment encoding the P-450 type hydroxylase and ferredoxin genes was cloned from Pseudonocardia autotrophica IFO 12743 that can convert vitamin D$\_$3/ into its hydroxylated active forms. In order to isolate the P-450 gene cluster in this organism, we designed PCR primers on the basis of the regions of an oxygen binding site and a heme ligand pocket that are general characteristics of the P-450 hydroxylase. Sequencing analysis of the BamHI fragment revealed the presence of four complete and one incomplete ORFs, named PauA, PauB, PauC, and PauD, respectively. As a result of computer-based analyses, PauA and PauB have homology with enoyl-CoA hydratase from several organisms and the positive regulators belonging to the tetR family, respectively. PauC and PauD show similarity with SuaB/C proteins and ferredoxins, respectively, which are composed of P-450 monooxygenase systems for metabolizing two sulfonylurea herbicides in Streptomyces griseolus PauC shows the highest similarity with another CytP-450$\_$Sca2/ protein that is responsible for production of a specific HMG-CoA reductase inhibitor, pravastatin, in S. carbophilus. Cultures of Steptomyces lividans transformant, containing the P-450 gene cluster on the pWHM3 plasmid, was unable to convert vitamin D$\_$3/ to its hydroxylated forms.

• Journal of Pharmacopuncture
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• v.17 no.1
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• pp.44-50
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• 2014

Objectives: This study was undertaken to isolate a fibrinolytic enzyme from the snake venom of Gloydius blomhoffii siniticus and to investigate its enzymatic characteristics and hemorrhagic activity as a potential pharmacopuncture agent. Methods: The fibrinolytic enzyme was isolated by using chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and fibrin plate assay. The characteristics of the enzyme were investigated using fibrin plate assay, protein hydrolysis analysis, and hemorrhage assay. Its amino acid composition was determined. Results: The fibrinolytic enzyme with the molecular weight of 32kDa (FE-32kDa) from Gloydius blomhoffii siniticus showed a fibrin hydrolysis zone at the concentration of 0.2 mg/mL in the fibrin plate assay. The fibrin hydrolysis activity of the enzyme was inhibited completely by ethylenediaminetetraacetic acid (EDTA), ethyleneglycoltetraacetic acid (EGTA), and 1, 10-phenanthroline, thiothreitol and cysteine, and partially by phenylmethanesulfonylfluoride (PMSF). Metal ions such as $Fe^{2+}$ and $Hg^{2+}$ inhibited the fibrin hydrolysis completely, but $Zn^{2+}$ enhanced it. FE-32kDa hydrolyzed ${\alpha}$-chain but did not hydrolyze ${\beta}$-chain and ${\gamma}$-chain of fibrinogen. High-molecular-weight polypeptides of gelatin were hydrolyzed partially into low-molecular-weight polypeptides, but the extent of hydrolysis was limited. FE-32kDa induced hemorrhage beneath back skin of mice at the dose of $2{\mu}g$. Conclusions: FE-32kDa is a ${\alpha}$-fibrin(ogen)olytic metalloprotease that requires $Zn^{2+}$ for fibrinolytic activity and causes hemorrhage, suggesting that the enzyme is not appropriate for use as a clinical pharmacopuncture.

• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.79-84
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• 2002

Suppression subtractive hybridization (SSH) was used to isolate wound-induced cDNAs from wounded soybean. One of wound-induced cDNA, gmwi33 showed high homology with genes encoding $\beta$-amyrin synthase. The full length cDNA of gmwi33, designated GmAMS1, is 2416 bp long and contains an open reading frame consisted of 739 amino acids. GmAMS1 protein showed 89% identity with licorice GgbAS1 and 86% identity with pea OSCPSY. In 5 day-old, dark-grown seedlings, the expression of GmAMS1 was most strongly induced by light and weakly induced by methyl jasmonate and by low temperature. However, GmAMS1 was not induced by elicitor or UV-B treatment. Such expression pattern might be closely related with the oxygen-radical scavenging activity of soyasaponin.

• Weed & Turfgrass Science
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• v.6 no.2
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• pp.157-164
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• 2017

Functional microorganisms decompose various organic matter by enzyme activity and suppress plant disease caused by pathogen. This study was conducted to isolate and select functional microorganisms with protein or carbohydrate degradation activities and antagonistic activity against turfgrass fungal pathogens for eco-friendly turfgrass management in golf course from compost containing livestock manure of poultry or swine. Totally 68 isolates collected from livestock manure compost strains were isolated and tested for their activities of amylase, protease and lipase and antagonistic activities against Rhizoctonia solani AG2-2, R. solani AG1-1, and Sclerotinia homoeocarpa. Among the isolates, 34 strains were selected as functional microbes showing higher activities of amylase and protease. Three isolates of ASC-14, ASC-18, and ASC-35 among the 34 strains were selected as antifungal bacterial strains repressing the above 3 turfgrass fungal pathogens. Analysis results of 16s rRNA gene sequence and phylogenic cluster indicated that ASC-14 and ASC-18 belonged to Bacillus amyloliquefaciens, while ASC-35 was B. subtilis, respectively.

• Food Science of Animal Resources
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• v.38 no.3
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• pp.580-592
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• 2018

The formulation of an oil/water (o/w) emulsion made up of a mixture of perilla oil and canola oil (30/70 w/w) was optimized using a response surface methodology to find a replacement for animal fat in an emulsion-type meat product. A 12 run Plackett-Burman design (PBD) was applied to screen the effect of potential ingredients in the (o/w) emulsion, including polyglycerol polyricinoleate (PGPR), fish gelatin, soy protein isolate (SPI), sodium caseinate, carrageenan (CR), inulin (IN) and sodium tripolyphosphate. The PBD showed that SPI, CR and IN showed promise but required further optimization, and other ingredients did not affect the technological properties of the (o/w) emulsion. The PBD also showed that PGPR played a critical role in inhibiting an emulsion break. The level of PGPR was then fixed at 3.2% (w/w total emulsion) for an optimization study. A central composite design (CCD) was applied to optimize the addition levels of SPI, CR or IN in an (o/w) emulsion and to observe their effects on emulsion stability, cooking loss and the textural properties of a cooked meat emulsion. Significant interactions between SPI and CR increased the cooking loss in the meat emulsion. In contrast, IN showed interactions with SPI leading to a reduction in cooking loss. Thus, CR was also removed from the formulation. After optimization, the level of SPI (4.48% w/w) and IN (14% w/w) was validated, leading to a perilla-canola oil (o/w) emulsion with the ability to replace animal fat in an emulsion-type meat products.

• The Plant Pathology Journal
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• v.36 no.1
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• pp.76-86
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• 2020

Cucumber mosaic virus (CMV) is damaging to the growth and quality of lettuce crops in Lanzhou, China. Recently, however, for the first time an isolate of lettuce necrotic yellows virus (LNYV) has been detected in lettuce crops in China, and there is concern that this virus may also pose a threat to lettuce production in China. Consequently, there is a need to develop a rapid and efficient detection method to accurately identify LNYV and CMV infections and help limit their spread. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays were developed to detect the nucleoprotein (N) and coat protein (CP) genes of LNYV and CMV, respectively. RT-LAMP amplification products were visually assessed in reaction tubes separately using green fluorescence and gel electrophoresis. The assays successfully detected both viruses in infected plants without cross reactivity recorded from either CMV or LNYV or four other related plant viruses. Optimum LAMP reactions were conducted in betaine-free media with 6 mM Mg²⁺ at 65℃ for LNYV and 60℃ for 60 min for CMV, respectively. The detection limit was 3.5 pg/ml and 20 fg/ml using RT-LAMP for LNYV and CMV plasmids, respectively. Detection sensitivity for both RT-LAMP assays was greater by a factor of 100 compared to the conventional reverse transcription polymerase chain reaction assays. This rapid, specific, and sensitive technique should be more widely applied due to its low cost and minimal equipment requirements.

• Food Science of Animal Resources
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• v.28 no.1
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• pp.99-104
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• 2008

A lactic acid bacterium showing growth inhibitory activity against Enterobacter sakazakii was isolated from bovine intestinal tracts. From biochemical and molecular biological studies, the isolate was identified and named as Enterococcus faecium JH95. This strain was resistant to kanamycin and streptomycin at a concentration of $100{\mu}g/mL$. E. faecium JH95 had high antimicrobial activity against food-borne pathogens such as Escherichia coli O157:H7, Listeria monocytogenes, Salmonella typhimurium, Staphylococcus aureus, and Clostridium perfrigens. The culture supernatant of this strain did not have antimicrobial activity. The culture broth of this strain failed to show the antimicrobial activity by heat treatment at $100^{\circ}C$ for 5 min or by pretense treatments for 2 hr. This result suggested that the putative antimicrobial substance produced by E. faecium JH95 is likely a protein which is not secreted into culture medium.

• Korean Journal of Food Science and Technology
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• v.43 no.3
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• pp.321-328
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• 2011

Edible films were developed from jujube (Zizyphus jujube Miller) using depolymerization processes of ultrasound and high-pressure homogenization. A 4.6% (w/w) jujube hydrocolloid was treated by ultrasound (600W, 20 min) or homogenized at high pressure (172 MPa, 6 s) and mixed with whey protein isolate, glycerol, xanthan, and sucrose esters of fatty acids to form film-forming solutions from which films were formed by drying. The film prepared by highpressure homogenization (HPH film) produced more homogeneous films without particles than those prepared without depolymerization or with the ultrasound treatment. HPH films possessed the highest tensile strength (4.7MPa), the lowest water vapor permeability ($2.9g{\cdot}mm/kPa{\cdot}h{\cdot}m^2$), and the most uniform and dense microstructures among the films. Flavor profiles of jujube powder and the films were distinguishable. Heat seal strength and oxygen permeability of the HPH films were 44.4 N/m and $0.025mL{\cdot}{\mu}m/m^2$/day/Pa, respectively. Antioxidant activities of jujube power and HPH films were not significantly different.

• Clinical and Experimental Reproductive Medicine
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• v.45 no.1
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• pp.1-9
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• 2018

Objective: To determine the localization, expression, and function of Toll-like receptors (TLRs) in fallopian tube epithelial cells. Methods: The localization of TLRs in fallopian tube epithelial cells was investigated by immunostaining. Surprisingly, the intensity of staining was not equal in the secretory and ciliated cells. After primary cell culture of fallopian tube epithelial cells, ring cloning was used to isolate colonies of ciliated epithelial cells, distinct from non-ciliated epithelial cells. The expression of TLRs 1-10 was examined by quantitative real-time polymerase chain reaction, and protein localization was confirmed by immunostaining. The function of the TLRs was determined by interleukin (IL)-6 and IL-8 production in response to TLR2, TLR3, TLR5, TLR7, and TLR9 ligands. Results: Fallopian tube epithelial cells expressed TLRs 1-10 in a cell-type-specific manner. Exposing fallopian tube epithelial cells to TLR2, TLR3, TLR5, TLR7, and TLR9 agonists induced the secretion of proinflammatory cytokines such as IL-6 and IL-8. Conclusion: Our findings suggest that TLR expression in the fallopian tubes is cell-type-specific. According to our results, ciliated cells may play more effective role than non-ciliated cells in the innate immune defense of the fallopian tubes, and in interactions with gametes and embryos.