• Bulletin of the Korean Chemical Society
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• v.34 no.8
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• pp.2419-2424
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• 2013

Guanosine-5'-diphosphate 3'-diphosphate (ppGpp) serves as alarmone in bacterial stringent responses. In this study, an affinity column was constructed by immobilizing ppGpp to NHS-Sepharose for isolating ppGpp-binding proteins. A novel ppGpp-binding protein, YjgA, was isolated and characterized by MALDI-TOF MS (matrix-assisted laser desorption ionization-time-of-flight mass spectrometry) coupled with two-dimensional gel electrophoresis. YjgA and truncated forms of YjgA were cloned and over-expressed in BL21 (DE3). The binding affinity of YjgA to ppGpp was determined by equilibrium dialysis. The interaction of YjgA with ppGpp was very specific, considering that the dissociation constant of YjgA with ppGpp was measured as $5.2{\pm}2.0{\mu}M$, while the affinities to GTP and GDP were about 60 and 30 times weaker than ppGpp. Expression of yjgA gene in Escherichia coli K-12 MG1655 was examined by reverse transcription polymerase chain reaction (RT-PCR). RT-PCR results revealed that yjgA was expressed from early to late stationary phase. The yjgA deletion mutant exhibited decreased cell number at stationary phase compared to parent strain and the over-expression of YjgA increased the cell number. These results suggested that YjgA might stimulate cell division under stationary phase. In most prokaryotic genome, about half of the protein candidates are hypothetical, that are expected to be expressed but there is no experimental report on their functions. The approach utilized in this study may serve as an effective mean to probe the functions of hypothetical proteins.

• Clean Technology
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• v.20 no.2
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• pp.108-115
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• 2014

Various metal chlorides and acid catalysts in ionic liquid solvent were investigated to directly convert cellulose into 5-hydroxymethylfurfural (5-HMF). Metal chlorides containing Sn(II), Zn(II), Al(III), Fe(III), Cu(II), and Cr(III) were used and acidic ionic liquid immobilized on silica gel as an acid catalyst and commercial acid catalysts (sulfuric acid, chloric acid, Amberlyst-15,DOWEX50x8) were used for comparison studies. The acid strength and amount of acid catalysts were probed with Hammett indicator. The selectivity and yield of 5-HMF were determined with reaction temperature, reaction time and catalyst ratio. A catalyst containing $CrCl_3-6H_2O$ and $SiO_2-[ASBI]HSO_4$ showed the highest selectivity and it was found that this catalyst had higher activity than commercial solid acid catalysts such as Amberlyst-15 and DOWEX50x8. The selectivity of 5-HMF appeared to be mainly dependent on the acid strength and catalyst ratio, it was found that levulinic acid was produced from 5-HMF by rehydration.

• Journal of Ginseng Research
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• v.35 no.2
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• pp.209-218
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• 2011

Ginseng has been used as a general tonic agent to invigorate the human body as an adaptogenic agent. In a previous report, we have shown that ginseng contains a novel glycolipoprotein called gintonin. The main function of gintonin is to transiently enhance intracellular free $Ca^{2+}$ $[Ca^{2+}]_i$ levels in animal cells. The previous method for gintonin isolation included multiple steps using organic solvents. In the present report, we developed a simple method for the preparation of crude gintonin from ginseng root as well as stem and leaf, which produced a higher yield of gintonin than the previous one. The yield of gintonin was 0.20%, 0.29%, and 0.81% from ginseng root, stem, and leaf, respectively. The apparent molecular weight of gintonin isolated from stem and leaf through sodium dodecyl sulfate polyacrylamide gel electrophoresis was almost same as that from root but the compositions of amino acids, carbohydrates or lipids differed slightly between them. We also examined the effects of crude gintonin from ginseng root, stem, and leaf on endogenous $Ca^{2+}$-activated $Cl^-$ channel (CaCC) activity of Xenopus oocytes through mobilization of $[Ca^{2+}]_i$. We found that the order of potency for the activation of CaCC was ginseng root > stem > leaf. The $ED_{50}$ was $1.4{\pm}1.4$, $4.5{\pm}5.9$, and $3.9{\pm}1.1$ mg/mL for root, stem and leaf, respectively. In the present study, we demonstrated for the first time that in addition to ginseng root, ginseng stem and leaf also contain gintonin. Gintonin can be prepared from a simple method with higher yield of gintonin from ginseng root, stem, and leaf. Finally, these results demonstrate the possibility that ginseng stem and leaf could also be utilized for ginstonin preparation after a simple procedure, rather than being discarded.

• Korean Journal of Food Science and Technology
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• v.16 no.2
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• pp.228-234
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• 1984

A partial hydrolysis of soybean protein isolate was carried out by using pepsin and trypsin. The degree of hydrolysis was evaluated by chemical analysis, viscometric measurements and gel electrophoresis. The functional properties of the hydrolyzates such as flow behavior, emulsion properties and foaming properties were evaluated. A selective hydrolysis of 11S protein fraction by pepsin was observed from the SDS-PAG electrophoresis. The changes in the molecular weight distribution by different conditions of enzyme hydrolysis were evaluated. The changes in the intrinsic viscosity of the protein hydrolylate by reaction time were highly correlated to the contents of TCA soluble protein and 0.03 M $CaCl_2$ soluble nitrogen. The degree of hydrolysis ($DH_{TCA}$, $DH_{Ca}$) were used to evaluate the effect of enzyme treatment on the functional properties of the hydrolyzate. The apparent viscosity and emulsion capacity and stability of the protein solution decreased as DH increased, while the foaming capacity increased linearly with the increasing DH.

• Journal of Life Science
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• v.17 no.10
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• pp.1321-1329
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• 2007

Deer antler tissue contains the most rapidly growing bone in the animal kingdom. Thus, it is likely that growing antler tissue is a rich source of local paracrine bone-stimulating factors. Growth factors, at least the insulin-like growth factor (IGF), control the bone-remodelling process. In this study, we tried to isolate and purify IGF-I from fresh antler tissue by the routine isolation and purification of protein. The purification involved ammonium sulfate precipitation, DEAE-Sepharose CL-60 ion-exchange chromatography, CM-Sepharose CL-6B ion-exchange chromatography, and Sephadex G-50 chromatography. Purified fractions from each step were analyzed by high-performance liquid chromatography (HPLC), SDS polyacrylamide gel electrophoresis (SDS-PACE), Dot-blot, and Western-blot methods. Furthermore, the quantification of partially purified IGF-I was calculated by enzyme-linked immunosorbent assays (ELISA) using antibody to human recombinant IGF-1. SDS-PAGE analysis of the final fraction yielded two molecular bands and the signal band was at 12 kDa on the Western-blot film. This purified IGF-I fraction showed a peak at retention time of eight min. The quantity of IGF-I in 20 g deer antler tissue as starting weight was calculated using a standard curve to be 2910 ng/ml, and total IGF-I amount is 0.291 g. The results show that IGF-I, which can be found in deer antler, can be partially purified and quantified by classic protein isolation methods.

• Journal of Korean society of Dental Hygiene
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• v.6 no.4
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• pp.455-467
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• 2006

Demand for appropriate health care has gradually increased in Korea. In addition, developments of community- and school-based oral health programs have also focused oral health care for the oral health promotion. Especially, school-based oral health programs are the underpinnings of promoting oral health and preventing oral diseases among schoolchildren. School-based oral health programs have had three major components: oral health education, oral health services, and a healthful environments. These included oral health education(one-to-one communication, group communication, and use of mass communication), oral examination, fluoride mouthrinsing, pit-and-fissure sealants, fluoride gel application, mechanical plaque control, and chewing xylitol candy. In this study, we evaluate the effects of oral health programs among primary schoolchildren by comparing the oral health knowledge, oral health behaviors, and perception of caries prevention procedures. Data for this study were obtained from 699 primary schoolchildren at the two primary school in Daegu, Korea. One is experimental group, N primary school, that was established school-based oral health center under supervision of Nam-gu Public Health Center, the other is control group, N' primary school, that was yet to establish school-based oral health center. We surveyed children's oral health knowledge and behaviors, and perception of caries prevention procedures using self-administrated questionnaire and then analyzed differences of each item among two groups. The brief findings of this study were summarized as follows. There are several advantage to a comprehensive school-based oral health program. (1) School-based oral health programs facilitate and increase the effectiveness of teaching oral health subjects. (2) Schoolchildren are available for prevention or treatment procedure. (3) School-based oral health center may be less threating than private dental clinic. (4) With comprehensive school-based oral health programs the decayed, missing, and filled teeth(DMFT) of schoolchildren should demonstrate a substantial and steady decrease over time(Choi et al, 2004). In conclusion, treatment is not the answer to solving children's oral health programs; rather primary prevention is the key. Many countries and communities are focusing on hoe millions of underprivileged children can be provided with health care. Schoolchildren gain the knowledge and behaviors to attain and maintain good oral health in schools. For these reasons, the role of school-based oral health center is not only important but also a necessity.

• Restorative Dentistry and Endodontics
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• v.18 no.1
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• pp.1-26
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• 1993

The purpose of this study was to investigate the effect of acid etching on the surface appearance and fracture toughness of five glass ionomer cements. Five kinds of commercially available glass ionomer cements including chemical curing filling type, chemical curing lining type, chemical curing metal reinforced type, light curing tilling type and light curing lining type were used for this study. The specimens for SEM study were fabricated by treating each glass ionomer cement with either visible light curing or self curing after being inserted into a rubber mold (diameter 4mm, depth 1mm). Some of the specimens were etched with 37% phosphoric acid for 0, 15, 30, 60, go seconds, at 5 minutes, 1 hour and 1 day after mixing of powder and liquid. Unetched ones comprised the control group and the others were the experimental groups. The surface texture was examined by using scanning electron microscope at 20 kV. (S-2300, Hitachi Co., Japan). The specimens for fracture toughness were fabricated by curing of each glass ionomer cement previously inserted into a metal mold for the single edge notch specimen according to the ASTME399. They were subjected to a three-point bend test after etching for 0, 30, 60, and 90 seconds at 5 minutes-, 1 hour-and 1 day-lapse after the fabrication of the specimens. The plane strain fracture toughness ($K_{IC}$) was determined by three-point bend test which was conducted with cross-head speed of 0.5 mm/min using Instron universal testing machine (Model No. 1122) following seven days storage of the etched specimens under $37^{\circ}C$, 100% humidity condition. Following conclusions were drawn. 1. In unetched control group, crack was present, but the surface was generally smooth. 2. Deterioration of the surface appearance such as serious dissolving of gel matrix and loss of glass particles occured as the etching time was increased beyond 15 s following Immediate etching of chemical curing type of glass ionomer cements. 3. Etching after 1 h, and 1 d reduced surface damage, 15 s, and 30s etch gave rough surface appearance without loss of glass particle of chemical curing type of glass ionomer cements. 4. Light curing type glass ionomer cement was etched by acid, but there was no difference in surface appearances according to various waiting periods. 5. It was found that the value of plane stram fracture toughness of glass ionomer cements was highest in the light curing filling type as $1.79\;MNm^{-1.5}$ followed by the light curing lining type, chemical curing metal reinforced type, chemical curing filling type and chemical curing lining type. 6. The value of plane stram fracture toughness of the chemical curing lining type glass ionomer cement etched after 5 minutes was lower than those of the cement etched after 1 hour or day or unetched (P < 0.05). 7. Light curing glass ionomer cement showed Irregular fractured surface and chemical curing cement showed smooth fractured surface.

• Radiation Oncology Journal
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• v.22 no.4
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• pp.298-306
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• 2004

Purpose: The purpose of this study was to identify Radiosensitivity of proteins in tissues with different radiosensitivity. Materials and Methods: C3H/HeJ mice were exposed to 10 Gy. The mice were sacrifiud 8 hrs after radiation. Their spleen and liver tissues were collected and analyzed histologicaly for apoptosis. The expressions of radiosusceptibillty protein were analyzed by 2-dimensional electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Resilts: The Peak of apoptosis levels were $35.3{\pm}1.7{\%}$ in spleen and $0.6{\pm}0.2{\%}$ in liver at 8 hrs after radiation. Liver, radioresistant tissues, showed that the levels of ROS metabolism related to proteins such as cytochromm c, glutathione S transferase, NADH dehydrogenase, riken cDNA and peroxiredoxin Vl increased after radiation. The expression of cytochrome c increased significantly in spleen and liver tissues after radiation. In spleen, radiosensitivity tissue, the identified proteins showed a significantly quantitative alteration following radiation. It was categorized as signal transduction, apoptosis, cytokine, Ca signal related protein, stress-related protein, cytoskeletal regulation, ROS metabolism, and others. Conclusion: Differences of radiation-induced apoptosis by tissues specifted were coupled with the induction of related radiosensitivity and radioresistant proteins. The result suggests that apoptosis relate protein and redox proteins play important roles in this radiosusceptibility.

• Journal of Life Science
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• v.12 no.1
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• pp.77-86
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• 2002

Chitin, a $\beta$-1,4 polymer of N-acetyl-D-glucosamine, is one of the most abundant organic compounds in nature. Chitinase (EC 3.2.1.14) is an enzyme that degrades chitin to chito-oligosaccharides, diacetyl rhitobiose and N-acetyl-D-glucosamine. An extracellular chitinase-producing bacterial strain was isolated from soil and named to as Bacillus subtilis JK-56. Optimum culture condition of B. subtilis JK-56 for the production of chitinase was 1% chitin, 0.5% polypepton, 0.1% KCl, 0.05% MnS $O_4$.4$H_2O$, 37$^{\circ}C$, initial pH 7.0 and 40 hour culture time. When B. subtilis JK-56 was grown in the optimum medium, one major active band and two minor active bands were detected by native-PAGE and active staining of the gel. Among them, the major band was purified from the culture supernatant by 70% ammonium sulfate precipitation and native-PAGE with BIO-RAD Model 491 Prep-Cell and named as Chi-56A. Its molecular weight was estimated to be 53kDa monomer and the isoelectric point (pI) was pH 4.3. The pH and temperature for the optimum activity of Chi-56A were pH 6.0 and $65^{\circ}C$, respectively. Chi-56A was stable up to $65^{\circ}C$ and in alkaline region. Its $K_{m}$ value for colloidal chitin was 17.33g/L. HPLC analysis of the reaction products confirmed that Chi-56A was an exo type chitinase.e.

• Korean Journal of Food Science and Technology
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• v.7 no.3
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• pp.109-114
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• 1975

Domestic seaweeds Gracilaria verrucosa collected from coastal areas of Namhae, Wando and Yeosoo, Korea were subjected to the examination of yield and quality of agar prepared according to various pretreatment conditions. In alkali treatment at high temperature, higher alkali concentration for one-hour period at $90^{\circ}C$ gave rise to higher yield of agar. In acid treatment, higher yield was obtained by higher acid concentration and longer treating period. Alkali treatment at room temperature gave rise to a slightly decreased yield at higher alkali concentration and longer treating time. Total nitrogen and crude ash of agar samples were greatly decreased by pretreatments. Jelly strength, gelation ability, gelation point and viscosity of agar samples tended to increase as the alkali-treating condition of seaweed became stronger. It was shown that sulfur content of agar had a high negative correlation with jelly strength of its gel. Various alkali treatments of seaweed at room temperature showed no marked difference in agar quality and did not exhibit any good effect comparable to alkali treatment at high temperature.