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Purification and Characterization of β-N-Acetylhexosaminidase from Rice Seeds

  • Jin, Yu-Lan;Jo, Yu-Young;Kim, Kil-Yong;Shim, Jae-Han;Kim, Yong-Woong;Park, Ro-Dong
    • BMB Reports
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    • v.35 no.3
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    • pp.313-319
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    • 2002
  • N-Acetyl-$\beta$-D-hexosaminidase ($\beta$-HexNAc'ase) (EC 3.2.1.52) was purified from rice seeds (Oryza sative L. var. Dongjin) using ammonium sulfate (80%) precipitation, Sephadex G-150, CM-Sephadex, and DEAE-Sephadex chromatography, sequentially. The activities were separated into 7 fractions($F_1-F_7$) by CM-Sephadex chromatography. Among them, F6 was further purified to homogeneity with a 13.0% yield and 123.3 purification-fold. The molecular mass was estimated to be about 52 kDa on SDS-PAGE and 37.4 kDa on Sephacryl S-300 gel filtration. The enzyme catalyzed the hydrolysis of both p-nitrophenyl-N-acetyl-$\beta$-D-hexosaminide (pNP-GlcNAc) and p-nitrophenyl-N-acetyl-$\beta$-D-hexosaminide (pNP-GalNAc) as substrates, which are typical properties of $\beta$-HexNAc'ase. The ratio of the pNP-GlcNAc'ase activity to the pNP-GalNAc'ase activity was 4.0. However, it could not hydrolyze chitin, chitosan, pNP-$\beta$-glucopyranoside, or pNP-$\beta$-glucopyranoside. The enzyme showed $K_m$, $V_{max}$ and $K_{cat}$ for pNP-GlcNAc of 1.65 mM, $79.49\;mM\;min^{-1}$, and $4.79{\times}10^6\;min^{-1}$, respectively. The comparison of kinetic values for pNP-GlcNAc and pNP-GalNAc revealed that the two enzyme activities are associated with a single binding site. The purified enzyme exhibited optimum pH and temperature for pNP-GlcNAc of 5.0 and $50^{\circ}C$, respectively. The enzyme activity for pNP-GlcNAc was stable at pH 5.0-5.5 and $20-40^{\circ}C$. The enzyme activity was completely inhibited at a concentration of 0.1 mM $HgCl_$ and $AgNO_3$, suggesting that the intact thiol group is essential for activity. Chloramine T completely inhibited the activity, indicating the possible involvement of methionines in the mechanism of the enzyme.

Characteristics of the media under a self-propelled compost turner in button mushroom cultivation (양송이버섯 재배시 자주식 배지교반기 활용 배지의 특성 및 수량성)

  • Lee, Chan-Jung;Yu, Byeong-Kee;Park, Hye-sung;Lee, Eun-Ji;Min, Gyeong-Jin
    • Journal of Mushroom
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    • v.18 no.3
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    • pp.274-279
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    • 2020
  • This study was conducted to investigate the characteristics of the medium used on the composting step, comparing the excavator agitator with the self-propelled turner. The temperature of the outdoor composting medium tended to increase rapidly after flipping in the turner. The late composting medium temperature was maintained at the excavator treatment area (farm practice), and the late composting effect progressed. During the field composting stage, various microorganisms such as Bacillus spp., Actinomycetes, fluorescent Pseudomonas spp., and filamentous fungi were distributed in the medium, and the density of aerobic bacteria involved in the decomposition of the medium was increased. Under high-temperature composting conditions, blue fungi, and mesophilic actinomycetes were inhibited or killed. Thermophilic actinomycetes, which play an important role in decomposing organic matter, showed higher densities than those observed in farm practices in the self-propelled turner process. The length of rice straw was slightly shorter when the self-propelled turner was used, and the water content did not show any significant difference between treatments. The a and b values tended to increase as the inverter was turned over. The CN ratio of the composting broth was lowered from 23.1 to 16.2 for the 5th turnover in the context of farming practices, and from 23.3 to 16.9 in the context of the self-propelled turner. The yield of each treatment was increased by 20% in 1 period, 28% in 2 periods, and 26% in 3 periods; the overall yield was 23%.

An Analysis of Recruitment Importance and Priority of According to the introduction of NCS(National Competency Standards) in Sports Public Institution (NCS(국가직무능력표준) 도입에 따른 스포츠계열 공공기관의 채용 중요도 및 우선순위 분석)

  • Kim, Dong-Man
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.5
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    • pp.1409-1417
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    • 2020
  • The purpose is to increase the likelihood of physical education students for employment in public institutions by examining the priority for hiring sports-related public institutions. The subject of the study was purposeful sampling of a total of 11 persons including 4 sports professors, 3 NCS experts in sports field, 2 judges from public sports institutions, and 2 personnel in charge of hiring public institutions. Through this process, from January 3 to March 12, 2020, the importance of priority was analyzed using hierarchical structure analysis using the main factors of NCS vocational basic competency. All data are coded so that statistical processing can be performed. Using SPSS/PC (ver. 21.0) for Windows, the hierarchical structure analysis was used for frequency analysis and priority determination. First, communication skills (.231), organizational comprehension skills (.177), resource management skills (.128), interpersonal skills (.110), vocational ethics (.082), problems in the major areas of recruitment of sports-related public institutions Solving ability (.061), information ability (.056), mathematical ability (.054), self-development ability (.052), and description ability (.049) were analyzed in order. Second, in terms of evaluation items, communication is communication skills (.442), mathematical skills are basic computation skills (.512), problem solving skills are thinking skills (.722), self-development skills are self-management skills (.587), Resource management ability was analyzed in order of time management ability (.531), interpersonal relationship ability as teamwork ability (.382), information ability in computer use ability (.677), technical ability in technology understanding ability (.599).

Effect of Liquid Fertilizer Application using Fish-meal, Bone-meal and Sesame oil-cake on Seed Germination and Growth of Tomato (어분, 골분 및 참깨박을 이용한 발효액비 제조에 따른 무 발아 및 토마토 생육에 미치는 영향)

  • An, Nan-Hee;Lee, Sang-min;Cho, Jung-Rai;Lee, Cho-Rong;Kong, Min-jae
    • Journal of the Korea Organic Resources Recycling Association
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    • v.27 no.4
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    • pp.61-70
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    • 2019
  • This study aimed to investigate the physicochemical characteristics of fish meal, bone meal, and sesame oil cake, which are readily available by-products from agriculture and fisheries, during the process of liquid fertilizer fermentation, and to examine the effects of liquid fertilizer application on seed germination and growth of tomatoes. During processing the fermentation for liquid fertilizers by using fish meal, bone meal, and sesame oil cake liquid fertilizers, the pH of the fertilizer increased in the order of bone meal > fish meal > sesame oil cake, and the concentration increased rapidly up to 30 days in all types of liquid fertilizer. The nitrogen content of the liquid fertilizers increased as fermentation progressed in the order of fish meal > bone meal > sesame oil cake. The phosphorus content increased as fermentation progressed and the highest was 1.0 % in the liquid fertilizer of sesame oil cake. The germination rate and its index of radish seeds were compared for different dilutions of each of the liquid fertilizers. Excluding the 10-fold dilution of the fish meal and oil cake liquid fertilizer, all the treatment groups showed a germination rate ≥ 95 % and the germination index tended to increase with dilution rate of liquid fertilizers. For responses of tomato growth, there were no significant differences among the liquid fertilizer treatment groups; however, the organic content, microbial density, and microbial biomass C in the soil were higher than chemical fertilizer treatment. These results demonstrated that there were differences in the characteristics of liquid fertilizers depending on the materials used, and that liquid fertilizer can be used for nutrition management for the organic crop cultivation.

Estimation of PM10 Source Contributions on Three Cities in the Metropolitan Area by Using PMF Model (PMF 모델을 이용한 수도권 내 3개 도시에서의 PM10 오염원의 기여도 추정)

  • Lee, Tae-Jung;Huh, Jong-Bae;Yi, Seung-Muk;Kim, Shin-Do;Kim, Dong-Sool
    • Journal of Korean Society for Atmospheric Environment
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    • v.25 no.4
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    • pp.275-288
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    • 2009
  • The Korean government strengthened the environmental polices to manage and enhance Metropolitan Area air quality, and also has enforced "Special Act on Seoul Metropolitan Air Quality Improvement (SASMAQI)" issued in Dec. 2004. Recently government expanded the Seoul Metropolitan Air Quality Management District (SMAQMD) to the outskirts satellite cities of Seoul area through the "Revised Law Draft of SASMAQI". The SMAQMD has been alloted the allowable emission loads to the local governments on the basis of the carrying $PM_{10}$ capacity. However, in order to establish the effective air quality control strategy for $PM_{10}$, it is necessary to understand the corresponding sources which have a potential to directly impact ambient $PM_{10}$ concentration. To deal with the situations, many receptor methodologies have been developed to identify the origins of pollutants and to determine the contributions of sources of interests. The objective of this study was to extensively identify $PM_{10}$ sources and to estimate their contributions at the metropolitan area. $PM_{10}$ samples were simultaneously collected at the 3 semi-industrialized local cities in the Seoul metropolitan area such as Hwasung-si, Paju-si, and Icheon-si sites from April 15 to May 31, 2007. The samples collected on the teflon membrane filter by one $PM_{10}$ cyclone sampler were analyzed for trace metals and soluble ions and samples on the quartz fiber filter by another sampler were analyzed for OC and EC. Source apportionment study was then performed by using a positive matrix factorization (PMF) receptor model. A total of 6 sources were identified and their contributions were estimated in each monitoring site. Contribution results on Hwasung, Paju, and Icheon sites were as follows: 33%, 27%, and 27% from soil source, 26%, 26%, and 21% from secondary aerosol source, 11%, 11%, and 12% from biomass burning, 12%, 6%, and 5% from sea salt, 7%, 15%, and 19% from industrial related source, and finally 11%, 15%, and 16% from mobile and oil complex source, respectively. This study provides information on the major sources affecting air quality in the receptor sites and thus it will help to manage the ambient air quality in the metropolitan area by establishing reasonable control strategies, especially for the anthropogenic emission sources.

Effect of Long-Term Pyridoxine Depletion on Asparte Aminotransferase and Pyridoxal 5'-Phosphate of Rat Liver Mitochondrial and Cytosolic Fractions (장기간의 Pyridoxine 부족이 쥐 간의 Mitochondria 및 Cytosolic Fraction 에 있는 Aspartate Aminotransferase 및 Pyridoxal 5'- Phoshate에 미치는 영향)

  • 임경숙
    • Journal of Nutrition and Health
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    • v.19 no.4
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    • pp.246-254
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    • 1986
  • Weanling female Sprague Dawley rats were fed d diets containing 22mg pyridoxine. BCI/kg diet (control diet) and l.2mg pyridoxine. BCI/kg diet (deficient diet). One control group and one defi­c dent group were fed their diet throughout growth, g gestation and lactation. After the pups were born and weaned, the deficient group was divided into two groups. One switched to control diet(supple­I mented group) and the other continued the same d deficient diet( deficient group) until 10 week -old. The liver mitochondrial and cytosolic asparate a aminotransferase activity and pyridoxal phosphate content were determined in offspring rats. The aspartate aminotransferase activities in both liver mito$\phi$ondrial and cytosolic fractions of den­d cient group were significantly lower than those of controls, but there were no significant differences between two groups after addition of 1O^{-4}M pyri­d do뼈I phosphate to the medium. By pyridoxine s supplementation after weaning, the reduced aspar­a tate aminotnmsferase activities were only partialy I restored to control levels. The pyridoxal phospha­t te content of deficient group in Iiver mitochondr­ial and cytosoIic fractions were alo significantly different from those of controls, but readily restored by dietary supplementation. These results suggest that there is a quantitative and a qualitative changes of aspartate amino trans­f ferase and pyridoxal phosphate in liver mitochon­d drial and cytosolic fraction by long-term pyrido­x xine deficiency and these reductions can partially recovered by dietary pyridoxine supplementation after weaning.

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Comparative Analysis of Hunting System in OECD Countries (OECD 국가(國家)들의 수렵제도(狩獵制度) 비교(比較)·분석(分析))

  • Byun, Woo-Hyuk;Yoon, Seong-Il
    • Journal of Korean Society of Forest Science
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    • v.87 no.2
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    • pp.286-299
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    • 1998
  • This study was conducted to do a comparative analysis of hunting system in 16 OECD(Organization for Economic Co-operation and Development) countries. Hunting is one of the most advanced types for developing natural resources, esp. wildlife, which is one of major products from forest. There are two types of hunting system ; Revier and/or License System, around most countries which have traditional advanced hunting and higher hunter population. Licence system, which is chosen in Korea, is less efficient ways of hunting for maximizing economical value and wildlife management than Revier system does. Adopting better system for much more efficient economical condition from hunting in Korea in a future is a ultimate goal in this study. The first step for the goal is an analysis of these two system among 16 OECD countries ; United State, Japan, Germany, France, etc. Comparative researches in hunting regulations such as number of hunters, hunter ratio to population, hunting bags, game species, seasons, ammunition and hunting permit are a next step for adopting developed hunting system in Korea. Korea has a smallest hunter ratio to population (1/2,463) among 16 OECD centuries and economical value lead by hunting is estimated still very low. Revier system is a popular one in most OECD countries except United State, Japan, etc, which is useful for wildlife management and maximizing economical value of hunting. And usually rifles are permitted for big game hunting. Hunting examination and/or insurance are compulsory for achieving hunting permit. Depend on game species, yearly hunting bag and season are constructed in most OECD countries.

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Anti-Inflammatory Effects of Volatile Flavor Extracts from Cnidium officinale and Angelica gigas (천궁(Cnidium officinale) 및 당귀(Angelica gigas) 휘발성 향기추출물의 항염증효과)

  • Choi, Sang-Won;Kim, Eun-Ok;Leem, Hyun-Hee;Kim, Jin-Kyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.8
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    • pp.1057-1065
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    • 2012
  • The effects of volatile flavor extracts of eight different herbal medicines, Juniperus rigida (JR), Saussurea lappa SL), Cnidium officinale (CO), Angelica gigas (AG), Eugenia caryophyllata (EC), Angelica tenuissima (AT), Mentha arvense (MA), and Artemisiae argyi (AA), were investigated on LPS-stimulated inflammation using Raw 264.7 cells. The volatile flavor extracts of CO and AG considerably inhibited LPS-stimulated NO, $PGE_2$, IL-6, and TNF-${\alpha}$ (except AG) production, as well as iNOS expression. Major volatile components of CO were identified as ligustilide and of ${\beta}$-eudesmol as AG by GC-MS analysis. Thus, these results suggest that the volatile extracts of CO and AG may be useful as potential therapeutic agents for inflammation-associated disorders.

Osteonectin Interacts with Human Nebulin C-terminus in Skeletal Muscle

  • Park, Eun-Ran;Kim, Hyun-Suk;Choi, Jun-Hyuk;Lee, Yeong-Mi;Choi, Jae-Kyoung;Joo, Young-Mi;Ahn, Seung-Ju;Min, Byung-In;Kim, Chong-Rak
    • Biomedical Science Letters
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    • v.13 no.4
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    • pp.263-272
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    • 2007
  • Nebulin is a giant actin binding protein (600-900 kDa) which is specific to skeletal muscle. This protein is known to regulate thin filaments length in sarcomere as a molecular template. The C-terminus of nebulin is located in the Z-disc of muscle sarcomere and is bound to other proteins such like myopalladin, titin, archvillin, and desmin. The N-terminus of nebulin binds to tropomodulin at the pointed ends of the thin filaments. In recent research, nebulin not only found in brain but also expressed in heart, stomach, and liver. So, the roles of nebulin in non-muscle tissue have been studied. However, lack of information or studies on nebulin binding proteins and nebulin function in brain are available so far. Therefore, the current study have investigated a novel binding partner of Nebulin C-terminus by using yeast two-hybrid screening with human brain cDNA library. Nebulin C-terminus, containing simple repeats, serine rich and SH3 domain, interacts with osteonectin C-terminal region. The specific interaction of nebulin and osteonectin were confirmed in vitro by using GST pull-down assay and reconfirmed in vivo by using transfected COS-7 cells with EGFP-tagged nebulin and DsRed-tagged osteonectin. Consequently, this study identified SH3 domain in nebulin C-terminus specifically binds to extracellular Ca-binding (EeC domain in osteonectin. Also, nebulin C-terminus fusion protein colocalized with osteonectin EC domain fusion protein in transfected COS-7 cells. The current study found the interaction between nebulin and osteonectin in human brain for the first time and suggested the nebulin in brain may be associated with osteonectin, as a regulator of cell cycle progression and mitosis.

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The electrical properties of PLZT thin films on ITO coated glass with various post-annealing temperature (ITO 기판에 제작된 PLZT 박막의 후열처리 온도에 따른 전기적 특성평가)

  • Cha, Won-Hyo;Youn, Ji-Eon;Hwang, Dong-Hyun;Lee, Chul-Su;Lee, In-Seok;Sona, Young-Guk
    • Journal of the Korean Vacuum Society
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    • v.17 no.1
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    • pp.28-33
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    • 2008
  • Lanthanum modified lead zirconate titanate ($Pb_{1.1}La_{0.08}Zr_{0.65}Ti_{0.35}O_3$) thin films were fabricated on indium doped tin oxide (ITO)-coated glass substrate by R.F magnetron sputtering method. The thin films were deposited at $500^{\circ}C$ and post-annealed with various temperature ($550-750^{\circ}C$) by rapid thermal annealing technique. The structure and morphology of the films were characterized with X-ray diffraction (XRD) and atomic force microscopy (AFM) respectively. The hysteresis loops and fatigue properties of thin films were measured by precision material analyzer. As the annealing temperature was increased, the remnant polarization value was increased from $10.6{\mu}C/cm^2$ to $31.4{\mu}C/cm^2$, and coercive field was reduced from 79.9 kV/cm to 60.9 kV/cm. As a result of polarization endurance analysis, the remnant polarization of PLZT thin films annealed at $700^{\circ}C$ was decreased 15% after $10^9$ switching cycles using 1MHz square wave form at ${\pm}5V$.