• Korean Journal of Microbiology
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• v.47 no.3
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• pp.209-217
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• 2011

Archaeal communities were investigated using 454 pyrosequencing technology based on 16S rRNA gene in 11 samples collected from six different full-scale anaerobic digesters. Observed operational taxonomic units (OTUs) estimated from the archaeal 16S rRNA gene sequences were 13-55 OTUs (3% cutoff) which was corresponded to 29-89% of Chao1 richness estimates. In the anaerobic digesters there were archaeal sequences within the orders Thermoproteales, Thermoplasmatales, Desulfurococcales as well as within the orders Methanomicrobiales, Methanobacteriales, Methanococcales, Methanosarcinales, and Methanocellales, which are known to produce methane. Among these orders, Methanococcales known to produce methane using hydrogen was the predominant taxon and constituted 51.8-99.7% of total sequences. All samples showed a very similar community structure (Pearson correlation coefficient=0.99) except for one sample based on a heat map analysis. In addition, canonical correspondence analysis correlating archaeal communities to the environmental variables demonstrated that digester temperature and total solids removal rate were the two important explanatory variables. Overall results suggested that environmental and operational variables of anaerobic digester are important factors determining archaeal diversity and community structure.

• The Korean Journal of Mycology
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• v.47 no.4
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• pp.359-371
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• 2019

To optimize the expression and secretion of ferritin protein associated with ion storage in the mushroom, Pleurotus eryngii, a recombinant secretion vector, harboring the ferritin gene, was constructed using a pPEVPR1b vector under the control of the CaMV 35S promoter and signal sequence of pathogen related protein (PR1b). The ferritin gene was isolated from the T-Fer vector following digestion with EcoRI and HindIII. The gene was then introduced into the pPEVPR1b secretion vector, and it was then named pPEVPR1b-Fer. The recombinant vector was transferred into P. eryngii via Agrobacterium tumefaciens-mediated transformation. The transformants were selected on MCM medium supplemented with kanamycin and its expression was confirmed by SDS-PAGE and western blotting. Expression of ferritin protein was optimized by modifying the culture conditions such as incubation time and temperature in batch and 20 L airlift type fermenter. The optimal conditions for ferritin production were achieved at 25℃ and after incubating for 8 days on MCM medium. The amount of ferritin protein was 2.4 mg/g mycelia, as measured by a quantitative protein assay. However, the signal sequence of PR1b (32 amino acids) seems to be correctly processed by peptidase and ferritin protein may be targeted in the apoplast region of mycelia, and it might not be secreted in the culture medium. The iron binding activity was confirmed by Perls' staining in a 7.5% non-denaturing gel, indicating that the multimeric ferritin (composed of 24 subunits) was formed in P. eryngii mycelia. Mycelium powder containing ferritin was tested as a feed additive in broilers. The addition of ferritin powder stimulated the growth of young broilers and improved their feed efficiency and production index.

• Applied Biological Chemistry
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• v.36 no.2
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• pp.105-110
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• 1993

Inulin fructotransferase from Enterobacter sp. S45 was purified with DEAE-cellulose column chromatography and fast protein liquid chromatography. The purified enzyme gave a single band on polyacrylamide gel electrophoresis. The molecular weight was estimated to be 42,800 by SDS-polyacrylamide gel electrophoresis. The optimal pH and temperature for the enzyme reaction were pH 5.5 and $55^{\circ}C$, respectively. $Mg^{2+}$ activated the enzyme activity, but $Fe^{3+}$, $Cu^{2+}$, $Hg^{2+}$ significantly inhibited. After exhaustive digestion of inulin by the enzyme, DFA III, sucrose, 1-kestose and nystose were produced. Sucrose, 1-kestose, raffinose and melezitose can't be used as substrates by the enzyme, but nystose and 1-F-fructofuranosyl nystose were hydrolysed. The Km and Vmax for inulin of the enzyme were 1.4 mM and $0.196\;{\mu}mole/min$, respectively.

• Journal of Microbiology and Biotechnology
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• v.21 no.8
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• pp.808-817
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• 2011

Because of its elevated cellulolytic activity, the filamentous fungus Trichoderma harzianum has a considerable potential in biomass hydrolysis applications. Trichoderma harzianum cellobiohydrolase I (ThCBHI), an exoglucanase, is an important enzyme in the process of cellulose degradation. Here, we report an easy single-step ion-exchange chromatographic method for purification of ThCBHI and its initial biophysical and biochemical characterization. The ThCBHI produced by induction with microcrystalline cellulose under submerged fermentation was purified on DEAE-Sephadex A-50 media and its identity was confirmed by mass spectrometry. The ThCBHI biochemical characterization showed that the protein has a molecular mass of 66 kDa and pI of 5.23. As confirmed by smallangle X-ray scattering (SAXS), both full-length ThCBHI and its catalytic core domain (CCD) obtained by digestion with papain are monomeric in solution. Secondary structure analysis of ThCBHI by circular dichroism revealed ${\alpha}$- helices and ${\beta}$-strands contents in the 28% and 38% range, respectively. The intrinsic fluorescence emission maximum of 337 nm was accounted for as different degrees of exposure of ThCBHI tryptophan residues to water. Moreover, ThCBHI displayed maximum activity at pH 5.0 and temperature of $50^{\circ}C$ with specific activities against Avicel and p-nitrophenyl-${\beta}$-D-cellobioside of 1.25 U/mg and 1.53 U/mg, respectively.

• Journal of Korean Academy of Nursing
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• v.14 no.2
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• pp.93-111
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• 1984

The main purpose of this study is to develop a ratio scale measuring level of pain using Korean pain terms. The specific purposes of this study are to identify the degree of pain of each pain term in each subclass: to classify each subclass in terms of dimensions of pain; and to analyze factors of the Korean pain ratio scale clustering together. One hundred an4 fifty eight pain terms which were originally identified as representative terms and their synonyms were used for data collection. Fifty eight nursing professors ana sixty one medical doctors who have contacted with patients having pain were asked to rate the weight of each pain term on a visual analogue scale. Subclasses in which ranks of pain terms were same f s findings in two previous studies were 1) thermal 3 am 2) cavity pressure, 3) single stimulating pain, 4) radiation pain. and 5) chemical pain. Subclasses in which ranks of pain terms were confused were 1) incisive pressure, and 2) cold pain. Subclasses in which one new pain term was added were 1) inflammatory-repeated pain, 2) punctuate pressure, 3) constrictive pressure, 4) fatigue-related pressure, and 5) suffering-relate4 pain. Subclasses in which two new pain terms were added were 1) traction pressure, 2) peripheral nerve pain, 3) dull pain, 4) pulsation-related pain, 5) digestion-related pain, 6) tract pain, and 7) punishment-related pain. Subclass in which 3 new pain terms were included was fear-related pain. Rating scores of 5 words in 4 subclasses were significantly different between the normal group and the extreme group of subjects in terms of subjective rating. Only one word among 6 words was that newly added to the scale. Rating scores of 12 words in 9 subclasses were significantly different between doctor group and nursing professor group. Among these 12 words, only 3 were those newly added to the scale. In comparison of these 12 words, mean scores of the nursing professors were always 7 to 16 points higher than those of the medical doctors. In the analysis of judgement of subjects in terms of dimensions of pain terms, subclasses of dull pain, cavity pressure, tract pain and cold pain were suggested to be included in the miscellaneous dimension. As a result of factor analysis of the ratings given to 96 pain words using principal components analysis without iteration and with varimax rotation limiting the number of factors to 4, factors of severe pain (factor I) mild-moderate pain (factor II) , causative pain (factor III) and temperature-related pain(factor IV) were extracted with the factor loading above 0.388. When the pain words were re-arranged on the bases of factor loading above 0.368, number of factors decreased to only first two factors. Maximum score of pain word in factor II was 46.17 and the minimum score of the factor I was 45.36. Further studies are needed to identify the validity, reliability, sensitivity and practicability of this ratio scale using patients having various sources of pain.

• YAKHAK HOEJI
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• v.37 no.5
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• pp.511-519
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• 1993

The short and variabke transit of drug throught GI tracj and the inter-and intra-subject variations of the transit restrict the sustained drug absorption after oral adminstration. These restrictions may be solved by retaining the dosage forms in the stomach. Then the dosage form will act as a platform which releases the drug slowly and makes the GI absorption occur for a long time. In this study, as the platforms, PVP hydrogels were synthesized by chemical and y-irradiation method in the cylindrical test tube. The chemical method means the synthesis of the hydrogel by heating the mixed solution of N-vinyl-2-pyrrolidone [monomer], acrylated albumin [crosslinking agent], 2, 2'-agobis(2-methylpropionitrile) [initiator] and proxyphylline [drug] at $65^{\circ}C$ for 5 hr. The $\gamma$-irradiation method means the synthesis of the hydrogel by irradiation with $^{60}$ Co $\gamma$-ray of the mixed solution of the monomer, acrylated albumin, and flurbiprofen [drug] at room temperature with total 0.2 Mrad for 3 hr. Our intention is to design the hydrogel tablet (diameter : 1.20 cm, thickness : 0.60 cm) which swells in the gastric fluid after oral administration to such a size that passing through the pylorus could be inhibited during the period of drug release. After releasing drug, the hydrogel should be degraded by the enzymeatic digestion in the stomach, or by hydrolysis and eventually solubilized. Thus, in votro tests were performed to examine the factors that affect swelling and drug release from the PVP hydrogels. Experimental results show that the hydrogels swell to a size larger than the diameter of the pylorus(l.3$\pm$0.7 cm) and the hydrogel prepared by the chemical method is digested by pepsin. But the hydrogel prepared by the $\gamma$-irradiation method was not digested by the pepsin and just collapsed with time. Thus, the swelling of the hydrogel synthesized by $\gamma$-irradiation was independent albumin acrylation time and pepsin concentration. But drug content and radiation dose affected the swelling and drug release kinetics of the hydrogel. Drug release from the hydrigels was prolonged up to about 24 hr. Therefore, it was concluded that by adjusting these factors, the albumin-crosslinked PVP hydrogel synthesized by $\gamma$-irradiation method is expected to be retained in the stomach for up to 60hr and be a potential platform of drugs for long-term GI absorption.

• Journal of the Korea Organic Resources Recycling Association
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• v.20 no.1
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• pp.71-77
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• 2012

In order to find a feasibility of applying an agricultural biomass to the anaerobic digestion system, the effects of pre-treatment of rice straw was investigated by different sizes,temperatures, ultrasonic frequencies, and with/without NaOH treatment on the change of soluble organic matter, which is generated most dominantly in Korea. It was observed that SCOD(soluble chemical oxygen demand) as the index for the decomposition of rice straw increased with the smaller particle size, higher reaction temperature and alkali treatment. With treatment of 5% NaOH, it was shown that the highest concentrations of SCOD were observed at 9,000 mg $L^{-1}$ and 6,000mg $L^{-1}$ at $35^{\circ}C$ and $55^{\circ}C$, respectively. Agitating with ultrasonic irradiation could be enhanced SCOD of rice straw less than 3 cm with 40 kHz of ultrasound. Optimal RPM in this study was at 150 rpm regardless of reaction temperatures.

• Korean Journal of Microbiology
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• v.37 no.4
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• pp.245-252
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• 2001

Erm proteins, MLS (macrolide-lincosamide-streptogramin B) resistance factor proteins, show high degree of amino acid sequence homology and comprise of a group of structurally homologous N-methyltransferases. On the basis of the recently determined structures of ErmC` and ErmAM, ErmSF was divided into two domains, N-terminal end catalytic domain and C-terminal end substrate binding domain and attempted to overexpress catalytic domain in E. coli using various pET expression systems. Three DNA fragments were used to express the catalytic domain: DNA fragment 1 encoding Met 1 through Glu 186, DNA fragment 2 encoding Arg 60 to Glu 186 and DNA fragment 3 encoding Arg 60 through Arg 240. Among the pET expression vectors used, pET 19b successfully expressed the DNA fragment 3 and pET23b succeeded in expression of DNA fragment 1 and 2. But the overexpressed catalytic domains existed as inclusion body, a insoluble aggregate. To assist the soluble expression of ErmSF catalytic domains, Coexpression of chaperone GroESL or Thioredoxin and lowering the incubation temperature to $22^{\circ}C$ were attempted, as did in the soluble expression of the whole ErmSF protein. Both strategies did not seem to be helpful. Solubilization with guanidine-HCl and renaturation with gradual removal of denaturant and partial digestion of overexpressed whole ErmSF protein (expressed to the level of 126 mg/ι culture as a soluble protein) with proteinase K, nonspecific proteinase are under way.

• Journal of Periodontal and Implant Science
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• v.32 no.4
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• pp.865-873
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• 2002

The primary cause of tooth loss after 30 years of age is periodontal disease. Destruction of alveolar bone by periodontal disease is done by bone resorbing activity of osteoclasts. Understanding differentiation and activation mechanism of osteoclasts is essential for controling periodontal disease. The purpose of this study is to identify the possible effects of Vitamin D and cytokines affecting osteoclasts and its precursor cells. Four to six week-old mice were killed and humerus, radius, tibia and femur were removed aseptically and washed two times with Hank's solution containing penicillin-streptomycin and then soft tissue were removed. Bone marrow cells were collected by 22 gauge needle. Cells were cultured in Hank's solution containing 1 mg/ml type II collagenase, 0.05% trypsin, 41mM EDTA. Supernatant solution was removed 5 times after 15 minutes of digestion with above mentioned enzyme solution, and remained bone particles were maintained in alpha-MEM for 15 minutes and $4^{\circ}C$ temperature. Bone particles were agitated for 1 minute and supernatant solution containing osteoclast precursor cells were filtrated with cell stainer. These separated osteoclast precursor cells were dispensed with 100-mm culture dish by $1{\times}10^7$ cells unit and cultured in ${\alpha}$- MEM containing 20 ng/ml recombinant human M-CSF, 30 ng/ml recombinant human soluble osteoclast differentiation factor and 10% fetal calf serum for 2 and 7 days. Total RNA of osteoclast precursor cells were extracted using RNeasy kit. One ${\mu}g$ of total RNA was reverse transcribed in $42^{\circ}C$ for 30 minutes using SuperScriptII reverse transcriptase. Expression of transcribed receptors of each hormone and cytokine were traced with 1 ${\mu}l$ of cDNA solution by PCR amplification. Vitamin D receptor WAS found in cells cultured for 7 days. TNF-${\alpha}$ receptor was found in cells cultured for 2 days and amount of receptors were increased by 7 days. IL-1 type I receptor was not found in cells cultured 2 and 7 days. But, IL-1 receptor type II was found in cells cultured for 2 days. TGF-${\alpha},{\beta}$type I receptor was found in cells cultured 2 and 7 days, and amount of receptors were increased by 7 days of culture. These results implies Vitamin D and cytokines can affect osteoclasts directly, and affecting period in differentiation cycle of osteoclasts is different by Vitamin D and cytokines.

• Hwankyungkyoyuk
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• v.11 no.2
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• pp.130-143
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• 1998

This study was conducted as a preliminary step to establish a model for the practical application of environmental education related to food wastes collected from elementary school foodservices. Methods of treatment and present recycling status of food wastes in elementary schools in Kyonggido were surveyed. The consciousness of dieticians who have been serving for elementary school foodservices was also analyzed through the survey questions concerning environmental education for the reduction and recycling of food wastes. The results derived from this survey were as follows: The major portion of food wastes from elementary school foodservices was constituted with vegetables and soup, and an average amount of food wastes per day was highest in June and July. Therefore environmental education related to reduction of wastes was needed in the first semester of school terms, and the possible reduction of food wastes achieved through the proper planning of school foodservice menu was also needed. In most cases food wastes were collected and treated by animal growing farmers or composted by machines in school. In operating composting machines microorganism inoculant was mostly used but dieticians pointed out the problem of a nasty odor, insects, and high energy consumption. This situation means it has not been operated efficiently and suggests an efficiency problem of machine composting in elementary schools because composting itself is based on the aerobic digestion and high temperature fermentation which kills insects and harmful microorganisms. Elementary school dieticians in Kyonggido were aware that food wastes cause main pollution problem, and that food wastes are valuable resources which can be recycled, and recycling of food wastes is inevitable. But more than half of the schools surveyed have not been reused food wastes in school, so a proper model for recycling and reuse of food wastes in school grounds was thought to be needed. Environmental education programs related to food wastes have not been peformed in more than half of the schools surveyed. It was concluded that the following three plans will be helpful to reduce school food wastes. First, environmental education should be enforced, second, teachers' and dieticians' intensive teaching concerning food wastes should be needed, and finally establishment of a model for recycling and reuse of food wastes in school grounds and its application to environmental education would offer a valuable field experience to school students.