• Journal of Microbiology and Biotechnology
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• v.22 no.3
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• pp.390-399
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• 2012

Endoglucanase gene egIV was cloned from Trichoderma viride AS 3.3711, an important cellulose-producing fungus, by using an RT-PCR protocol. The egIV cDNA is 1,297 bp in length and contains a 1,035 bp open reading frame encoding a 344 amino acid protein with an estimated molecular mass of 35.5 kDa and isoelectronic point (pI) of 5.29. The expression of gene egIV in T. viride AS 3.3711 could be induced by sucrose, corn straw, carboxymethylcellulose (CMC), or microcrystalline cellulose, but especially by CMC. The transcripts of egIV were regulated under these substrates, but the expression level of the egIV gene could be inhibited by glucose and fructose. Three recombinant vectors, pYES2-xegIV, $pYES2M{\alpha}$-egIV, and $pYES2M{\alpha}$-xegIV, were constructed to express the egIV gene in Saccharomyces cerevisiae H158. The CMCase activity of yeast transformants $IpYES2M{\alpha}$-xegIV was higher than that of transformant IpYES2-xegIV or $IpYES2M{\alpha}$-egIV, with the highest activity of 0.13 U/ml at induction for 48 h, illustrating that the modified egIV gene could enhance CMCase activity and that $MF{\alpha}$ signal peptide from S. cerevisiae could regulate exogenous gene expression more effectively in S. cerevisiae. The recombinant EGIV enzyme was stable at pH 3.5 to 7.5 and temperature of $35^{\circ}C$ to $65^{\circ}C$. The optimal reaction condition for EGIV enzyme activity was at the temperature of $55^{\circ}C$, pH of 5.0, 0.75 mM $Ba^{2+}$, and using CMC as substrate. Under these conditions, the highest activity of EGIV enzyme in transformant $IpYES2M{\alpha}$-xegIV was 0.18 U/ml. These properties would provide technical parameters for utilizing cellulose in industrial bioethanol production.

• Fisheries and Aquatic Sciences
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• v.17 no.1
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• pp.37-46
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• 2014

Fresh sea squirt meat requires a modified processing and preservation process because it has a short shelf life due to its high moisture content and strong proteolytic enzyme activity. In this study, bottled sea squirt meat prepared in vegetable oil (BSMO) to enhance the consumer acceptability was exposed to ${\gamma}$-ray (Co60, 10 kGy/h) irradiation to extend the shelf life without the use of a heating process. Response surface methodology was used to determine the optimal mixing ratio of BSMO containing 5% dehydrated fresh meat. Texture analysis and nutritional evaluation were also performed on a control and BSMO samples. The volatile basic nitrogen (VBN) content and total cell count were measured to determine the shelf life of irradiated BSMO products during chilled storage at $4^{\circ}C$ for 60 days. According to a panel of 10 trained tasters (aged 20-29 years), the optimal mixing formulation was 80 g meat in 60 mL of mixed vegetable oil (30 mL of olive oil and 30 mL of sesame oil). The highest rated formulation, according to a panel of nine trained tasters (aged ${\geq}30$ years), was 80 g meat in 60 mL of mixed vegetable oil (42 mL of olive oil and 18 mL of sesame oil). Moisture, ash, and protein contents in BSMO did not change significantly (P < 0.05) compared with the control. A higher lipid content ($0.84{\pm}0.23$ to $2.13{\pm}0.61$; P < 0.05) was observed due to the presence of vegetable oil on the surface of BSMO. The vegetable oil raised the hardness, springiness, cohesiveness, gumminess, chewiness, and resilience of BSMO. BSMO products remained edible after 50 days of storage at $4^{\circ}C$ based on the VBN content (BSMO 1: $27.92{\pm}0.96$ mg/100 g, BSMO 2: $24.84{\pm}1.95$ mg/100 g) and total cell count (BSMO 1: $4.60{\pm}0.80$ log CFU/mL, BSMO 2: $3.65{\pm}0.20$ log CFU/mL) when compared with standard levels of VBN (25.00 mg/100 g) and total cell count (5 log CFU/mL), respectively. The results showed that irradiated BSMO products could help to expand the processed seafood market and increase the popularity of seafood among the younger generations.

• Korean Journal of Community Nutrition
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• v.7 no.2
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• pp.167-176
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• 2002

To assess the dietary fiber intake of preschool children in Busan and to evaluate the relationship between of the intake of dietary fiber and nutrient intake. Nutrient intake using 24 hour recall, and total dietary fiber (TDF) intake based on tables of TDF of common Korean floods developed by the modified Prosky Method, were estimated for 176 preschool children. The mean daily intakes of TDF, and TDF after adjusting energy intake, were 10.20 g and 7.69 g/1,000 kcal, respectively. The mean daily intakes of TDF for children aged 1-3 and 4-6 years were 9.20 g and 11.08 g, respectively. The range of TDF intake was 1.86 to 22.16 g. The major sources of TDF were cereals (31.0%), vegetables (18.9%) and fruits (11.9%). The TDF intake showed positive correlations with nutrient adequacy ratios (NAR) of iron and Vitamin $B_1$, (p < 0.05, p < 0.05). The TDF intake per 1,000 kcal showed negative correlations with the NARs of protein, calcium, phosphate, iron, Vitamin A, vitamin $B_1$, Vitamin $B_2$, and niacin (p < 0.001, p < 0.001, p < 0.001, p < 0.05, p < 0.001, p < 0.001, p < 0.001, p < 0.001) and with the mean adequacy ratio (MAR, p < 0.001). When children were stratified into quartiles ($Q_1-Q_4$) on the basis of their fiber intake per 1,000 kcal, their NARs for calcium, phosphate, iron, Vitamin A, Vitamin $B_1, Vitamin $B_2$ and niacin (p < 0.05, p < 0.05, p < 0.001, p < 0.05, p < 0.05, p < 0.001, p < 0.001), and their MAR (p < 0.001) were significantly lower in the children with higher fiber intake per 1,000 kcal (the upper quartile). The NARs fur calcium (0.63), iron (0.60), Vitamin A (0.66), Vitamin $B_2$(0.74), niacin (0.64), Vitamin C (0.65) and the MAR (0.74) were lower than 0.75 in the children with fiber intakes of more than 9.25 g per 1,000 local (0,), the highest fiber intake per 1,000 kcal. Based on these results, the mean TDF intake of children was higher than the age (yr)+5g , the minimum recommended level for American children. Meals with a fiber intake of more than 9.25 g per 1,000 local ($Q_4$) could cause a decreased nutritional status for minerals and vitamins. The result of this study could contribute to the establishment of Recommended Dietary Allowances (RDA) for dietary fiber for Korean Preschool children.

• Journal of Periodontal and Implant Science
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• v.30 no.4
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• pp.757-779
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• 2000

The aim of our study is to achieve complete periodontal tissue regeneration by the application of BMP and resorbable membrane. Three beagle dogs aged over one and half years and weighed 14 to 16 kg were used in this study. Mandibular 1st, 2nd premolars were extracted bilaterally. Horizontal furcation defects were induced around 3rd, 4th premolars bilaterally. BMP-4 were applied in the right side with resorbable membranes and only resorbable membranes were applied in the left side respectively. Each animal was sacrificed at 2, 4, and 8weeks, after regenerative surgery. Specimens were prepared with Hematoxylin-Eosin stain and Goldner's modified Masson Trichrome stain for light microscopic evaluation. The results were as follows: 1. At 2 weeks after regenerative surgery, downgrowth of junctional epithelium was observed both in the membraneapplied site and BMP-4-and-membrane-applied site. 2. At 4 weeks after regenerative surgery, resorbable membranes were completely resolved, therefore would not prevent downgrowth of junctional epithelium. New bone formation, new cementum formation and Sharpey's fiber were observed in BMP-4-andmembrane-applied site. 3. At 8 weeks after regenerative surgery, downgrowth of junctional epithelium was observed in the membrane-applied site. But, new cementum formation was observed in the same site. The extensive regeneration of new bone, new cementum and remarkable formation of Shapey's fiber were showed in BMP-4-and-membrane-applied site. 4. Resorbable membranes were resolved via the cell-mediated processes. 5. Periodontal tissue regeneration were better achieved in the BMP-4-andmembrane-applied site than in the membrane-applied site. Within the above results, BMP-4 may have the strong capability to form the new bone and resorbable membrane may be able to prevent the bony ankylosis. However, resolution rate of resorbable membrane may not be enough to protect rapid epithelial downgrowth for ideal periodontal regeneration. In conclusion, I suggest BMP-4 may have the strong possibility to be utilized in the clinical periodontal treat-

• Journal of the Korean Chemical Society
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• v.54 no.5
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• pp.567-572
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• 2010

5,10-Methenyltetrahydrofolate synthetase from chicken liver was purified through 30-70% ammonium sulfate fractionation, Q Sepharose Fast Flow anion exchange and Source 15Phe hydrophobic interaction chromatography. Specific activities of cell extract, ammonium sulfate, Q Sepharose Fast Flow and Source 15Phe were 0.0085, 0.031, 0.80 and 1.27 U/mg, respectively. Purification fold activities of cell extract, ammonium sulfate, Q Sepharose Fast Flow and Source 15Phe were 1, 3.7, 94.1 and 149.4, respectively. HPLC gel permeation chromatography and SDS-polyacrylamide electrophoresis experiments indicated that the enzyme is a monomeric protein with a molecular weight of 22.8 kDa. Km for 5-methyl THF and Mg-ATP were $7.1\;{\mu}M$ and $63\;{\mu}M$, respectively. Optimum temperature and pH were $30^{\circ}C$ and 6.0, respectively. The data for metal ion specificity and stoichiometry showed that the maximum activity was obtained with a 1:l. ratio of $Mg^{2+}$. The ATP and Km values increased in the order of MgATP, MgCTP, MgUTP and MgGTP, and the maximum activities also decreased in the same order, indicating MgATP as the most efficient substrate. The enzyme was chemically modified only by tetranitrometane and 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide, indicating that tyrosine and carboxylate are present in the active site.

• Korean Journal of Community Nutrition
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• v.16 no.1
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• pp.51-61
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• 2011

Elevated serum concentration of inflammation markers is known as an independent risk factor of metabolic syndrome (MS) and dietary intake is an important factor to control MS. The purpose of this study was to investigated the hypothesis that inflammatory indices are associated with dietary intake and diet quality index-international (DQI-I) in subjects with MS. A cross-sectional study was conducted on 156 men and 73 postmenopausal women with MS, defined by three or more risk factors of the modified Adult Treatment Panel III criteria. Serum levels of high sensitive C-reactive protein (hs-CRP), adiponectin were examined and nutrients intake and DQI-I were assessed using a semiquantitative food frequency questionnaire. The total DQI-I score was significantly higher in female subjects ($65.87{\pm}9.86$) than in male subjects ($62.60{\pm}8.95$). There was a positive association between hs-CRP and polyunsaturated fatty acid intake (p < 0.05) and a negative association between adiponectin and lipid (p < 0.05), total sugar (p < 0.01), and total fatty acids (p < 0.05). When the subjects were divided into 5 groups by quintile according to serum adiponectin and hs-CRP level, there was no association between DQI-I score and hs-CRP levels. Moderation score of DQI-I was significantly higher in highest quintile group than the lower quintile groups. Therefore, our results provide some evidence that dietary intake and diet quality are associated with inflammation markers and dietary modification might be a predictor to decrease risk for metabolic syndrome complications. However further research is needed to develop the dietary quality index reflecting the inflammatory change by considering the dietary habit and pattern of Koreans.

• Polymer(Korea)
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• v.33 no.4
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• pp.389-395
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• 2009

To develop the carrier of hydrophobic antifungal agents based on low molecular weight water-soluble chitosan (LMWSC), LMWSC was chemically modified with deoxycholic acid (DA) which is one of the bile acid as a hydrophobic group. The nanoparticles (WSCDA) using DA conjugated LMWSC were characterized using dynamic light scattering (DLS) and transmittance electron microscope (TEM). The particle size of WSCDA ranged from 250 to 350 nm and increased with the number of DA substitution. The loaded itraconazole as an antifungal agent WSCDA nanoparticles (WSCDA-ITCN) were prepared by solvent evaporation method. The drug content and the loading efficiency were investigated approximately $9{\sim}10%$ and $61{\sim}68%$ by UV spectrophotometer, respectively. The release of drug from nanoparticles was slow and showed sustained release characteristics. Based on the results of release study that the higher DA contents in WSCDA, the slower the releasing rate, the WSCDA-ITCN could be used as an excellent antifungal agent.

• Korean Journal of Food Science and Technology
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• v.29 no.6
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• pp.1151-1157
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• 1997

The chemical identities of purified polysaccharide, methylan, were analyzed by various chemical methods. The polysaccharide contained 79%(w/w) sugar, 6% protein, and 16% organic acids such as uronic acid, pyruvic acid, and acetic acid. With proceeding fermentation, the acids content in methylan increased from 10% at 34 hr to 17% at 72 hr, and the viscosity of methylan in the same concentration also increased. The correlation between viscosity and acid content in methylan was studied using chemically or biologically modified methylan. Methylan with a high content of pyruvic acid exhibited a high apparent and an intrinsic ·viscosity. When the pyruvic acid content of methylan with the same content of uronic acid was increased 1%, apparent viscosity and intrinsic viscosity increased 290 cp and 6 dL/g, respectively. Methylan with a high content of uronic acid exhibited a high apparent and an intrinsic viscosity. When the uronic acid content of methylan with the same content of pyruvic acid was increased 1%, apparent viscosity and intrinsic viscosity increased 85 cp and 1.5 dL/g, respectively. It was found that the increased viscosity of methylan resulted from the increased content of organic acids in methylan, and pyruvic acid was more an important factor contributed to the increase of methylan viscosity than uronic acid.

• Journal of Microbiology and Biotechnology
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• v.27 no.9
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• pp.1701-1710
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• 2017

Classical swine fever virus (CSFV) is the etiologic agent of classical swine fever, a highly contagious disease that causes significant economic losses to the swine industry. The lapinized C-strain, a widely used vaccine strain against CSFV, has low growth efficiency in cell culture, which limits the productivity in the vaccine industry. In this study, a recombinant virus derived from C-strain was constructed and subjected to continuous passaging in PK-15 cells with the goal of acquiring a high progeny virus yield. A cell-adapted virus variant, RecCpp80, had nearly 1,000-fold higher titer than its parent C-strain but lost the ability to induce fever in rabbits. Sequence analysis of cell-adapted RecC variants indicated that at least six nucleotide changes were fixed in RecCpp80. Further adaption of RecCpp80 variant in swine testicle cells led to a higher virus yield without additional mutations. Introduction of each of these residues into the wild-type RecC backbone showed that one mutation, M979R (T3310G), located in the C-terminal region of E2 might be closely related to the cell-adapted phenotype. Rabbit inoculation revealed that $RecCpp40_{+10}$ failed to induce fever in rabbits, whereas $RecCpp80_{+10}$ caused a fever response similar to the commercial C-strain vaccine. In conclusion, the C-strain can be adapted to cell culture by introducing specific mutations in its E2 protein. The mutations in RecCpp80 that led to the loss of fever response in rabbits require further investigation. Continuous passaging of the C-strain-based recombinant viruses in PK-15 cells could enhance its in vitro adaption. The non-synonymous mutations at 3310 and 3531 might play major roles in the enhanced capacity of general virus reproduction. Such findings may help design a modified C-strain for improved productivity of commercial vaccines at reduced production cost.

• Korean Journal of Head & Neck Oncology
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• v.19 no.1
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• pp.25-33
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• 2003

Background and Objectives: The Herpes Simplex type 2 Defective Infectious Single Cycle virus (DISC virus) is attenuated virus originally produced as viral vaccines but are also efficient gene transfer vehicle. The main goals of this study were to examine the efficiencies of the gene transfer using DISC vectors for various head and neck squamous cell carcinoma cell lines and to evaluate the efficacy of vaccination with DISC virus carrying a immunomodulatory genes (GM-CSF) as cancer therapy in a organotopic oral cavity squamous cell cancer model. Materials and Methods : We determinated the gene transfer efficiency of DISC virus by x-gal stain method and proved gene and protein expression of DISC-GMCSF transfected SCCVII cells by RT-PCR and ELISA method. Also we evaluated the ex vivo vaccination effects of SCCVII/GMCSF (DISC-GMCSF transfected SCCVII vaccine) vaccine on preventing the recurrence of micro-residual tumor. After the vaccination of SCCVII/GMCSF, specific cytotoxic T-cell responses was evaluated by CTL assay. Results: At an MOI of 10 DISC virus showed 64-88% of transfection rates in various head and neck squamous cancer cell lines. SCCVII cells transduced by DISC virus vector (MOI=10) carrying the GM-CSF gene, produced 4.5 nanogram quantities of GM-CSF per $10^6$ cells. In vivo vaccination using tumor cells transduced ex vivo with DISC-GMCSF resulted in better protection rate against subsequent tumor recurrence in organotopic oral cavity cancer model. Although tumor free survival rate was not statistically significantly increased in vaccination group (p=0.078), tumor specific cytotocic T-cell responses were significantly increased in SCCVII/GMCSF vaccination group. Conclusion: These data demonstrate that; 1) The DISC virus vector is capable of efficient gene transfer to various head and neck squamous cancer cell lines, 2) GM-CSF secreting genetically modified tumor vaccine (SCCVII/GMCSF) efficiently protected against tumor recurrence in organotopic micro-residual oral cavity cancer model and produced tumor specific cytotoxic T-cell response. DISC virus-mediated, cytokine gene transfer may prove to be useful as a clinical therapy for head and neck cancers.