• Title/Summary/Keyword: $P_2O_5$

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Electron Microscopic Studies on Olfactory Bulbs in the Vertebrates by Phylogenetics (계통발생에 따른 척추동물의 뇌후구에 대한 전자현미경적 연구)

  • Choi, W.B.;Chung, Y.H.;Seo, J.E.
    • Applied Microscopy
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    • v.15 no.2
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    • pp.31-68
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    • 1985
  • Authors are trying to unveil the ultrastructural organization of olfactory bulb, which has been summerized under light microscopic level or communicated only in some detail in different view point until now. For the critical point of view, since the phylogenetical approach will give the ultimate value in the correlative study between structural and functional bases (Brodal, 1969), the present study was carried out light and electron microscopic analyses of the structures of the neurons and synaptic organizations in olfactory bulbs from different animals in phylogenetical scale. We selected each one species from five animal classes: the house rabbit(Oryctolagus cuniculus var. domesticus [Gmelin]) from Mammalia, the domestic fowl (Gallus gallus domesticus Brisson) from Aves, the viper (Agkistrodon hylys [G.P. Pallas]) from Reptilia, a frog (Bombiana orientalis Boulenger) from Amphibia and the crussian carp (Carassius carassius [Linne]) from Pisces. For light microscopic study, samples were fixed in 10% formalin and paraffin sections were stained with hematoxylin-eosin. For the electron microscopic study, the tissues were fixed by perfusion through the heart or immersion with 1% paraform-aldehyde-glutaraldehyde mixture (phosphate buffer, pH 7.4), and final tissue block trimmed under dissecting microscope were osmicated (1% OsO4), they were embedded in Araldite or Epon 812, and ultrathin sections were made by LKB-V ultratome following the inspection of semi-thin sections stained with toluidine blue-borax solution. Ultra-thin sections contrasted with uranyl acetate and lead citrate were observed with JEM 100CX electron microscope. We have summerized our morphological analyses as follows: 1. The olfactory bulb of rabbit, viper and frog shows the eight layers of fila olfactoria, glomerular, external granular, external plexiform, mitral cell, internal plexiform, internal granular, medullary but domestic fowl shows the five layers of glomerular, fibrillar, mitral, granular and medullary and the three layers of fibrilla, glomerular and medullary in crussian carp. The sharpness of demarcation between the layers shows deferential tendency according to phylogenetical order. 2. Mitral cells of vertebrate have large triangular or oval shape with spherical nuclei which contain not so much chromatin. The cytoplasm contains numerous cell organelles, of which Nissl's bodies or granular endoplasmic reticula arranged as parallel strands. Development of granular endoplasmic reticula were declined as the phylogentical grade is going lower. 3. Tufted cells of all animal are mostly spindle or polygonal contour and contain oval nuclei which located in periphery of cytoplasm. The nuclei of rabbit, fowl, viper and frog has relatively space chromatin, but a nucleus of crussian carp contain irregularly aggregated chromatin in karyoplasm. Their cytoplasmic volume and cell organelle contents are in between those of mitral cell and granular cell. They contain moderate amount of mitochondria, granular endoplasmic reticula, a few Golgi complex, polysomes, lysosome, etc. 4. Granule of cells of all the vertebrate amimals studied exhibit similar features; cells and their dense nuclei show spherical or oval contour, and they have the thin rim of cytoplasm which contain only a few cell organelles. 5. In rabbit, the soma of mitral cells were in contact with boutons with two types of synaptic vesicles, that is, round and flat vesicles, especially flat vesicles in boutons were showing reciprocal synapses. However, in domestic fowls, vipers, frogs and crussian carps, there were found boutons showing only spherical synaptic vesicles. 6. The boutons containing round synaptic vesicles were made contact with the some of tufted cell of olfactory bulb in the rabbits, fowls, vipers and frogs, but no synaptic boutons were observed in soma of tufted cells in crussian carps. In the frogs, there were observed dendrites were contact with the soma of tufted cells. 7. In the neuropils of plexiform, granular and glomerular layers olfactory bulbs in the vertebrate, the synapses were axo-large dendrites, axo-median and small dendrites, dendrodendritic, and axo-axonal contacts. However, in the neuropil of crussian carps, synapses were observed only in glomerular layer.

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Management Guidelines and the Structure of Vegetation in Natural Monuments Koelreuteria Paniculata Community (천연기념물 모감주나무군락의 식생구조와 관리제언)

  • Shin, Byung Chul;Lee, Won Ho;Kim, Hyo Jeong;Hong, Jeum Kyu
    • Korean Journal of Heritage: History & Science
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    • v.43 no.1
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    • pp.100-117
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    • 2010
  • This study analyzed vegetation structure of natural monuments Koelreuteria paniculata community in search of a conservation and management plan. Plant sociological analysis of Koelreuteria paniculata community indicates that it can be classified into Achyranthes japonica subcommunity and Rhodotypos scandens subcommunity and Trachelospermum asiaticum var. intermedium subcommunity. While Koelreuteria paniculata community of Ahnmyeondo is composed of sub tree layer and herb layer, those of Pohang and Wando are composed of tree layer, Sub tree layer, shrub layer, herb layer. The results of tree vitality analysis showed that those in Ahnmyeondo appeared to be relatively low when compared to those in Pohang and Wando-gun. This can be understood in two different aspects: disease and insects vulnerability due to a relatively simple structure and lack of competitive species, and decreased vitality / natural branch losses due to crown competition arising from high density. The result of soil characteristics analysis showed that soil texture, soil pH, organic matter, $p_2O_5$, exchange positive ion were sufficient for tree growth while total nitrogen was not, so that discretion would be needed for fertilizer application. As there were damages of disease and inscet, but only for 10~15% of the entire area; it still requires consistent preconsideration. The study suggests the management methods for preservation of Koelreuteria paniculata community. First, securing designated areas is necessary in order to minimize environment deterioration due to surrounding development. Especially, for sections with decreased areas, expansion of designated areas through land purchase should also be considered. Second, artificial interference may affect the livestock. Therefore, monitoring of artificial interference is necessary, based on which protection projects must be conducted. Third, from analysis of young plants which influence the maintenance mechanisms of Koelreuteria paniculata community, a decrease compared to the prior year was observed; investigation is needed. Therefore, an active management policy through status examination of livestock such as germination and young plants is necessary.

Effects of Salt Stress on Dry Matter, Glucose, Minerals Content and Composition in Potato (Solanum tuberosum L.) (염스트레스가 감자(Solanum tuberosum L.)의 건물, 환원당, 무기성분의 함량 및 조성에 미치는 영향)

  • Im, Ju Sung;Kim, Mi Ok;Hong, Me Soon;Kim, Mi Suk;Cheun, Chung ki;Park, Yeong Eun;Cho, Ji Hong;Cho, Kwang Soo;Chang, Dong Chil;Choi, Jang Gyu;Lee, Jong Nam
    • Korean Journal of Environmental Agriculture
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    • v.38 no.1
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    • pp.38-46
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    • 2019
  • BACKGROUND: Salinity is one of the main environmental stresses deteriorating qualities as well as yields of food crops. This study was conducted to identify the effects of salt stress on dry matter ratio, glucose content, and mineral content and composition in potatoes (Solanum tuberosum L.). METHODS AND RESULTS: Four potato varieties were grown in plastic pots (diameter 20 cm and height 25 cm) with three salinity levels (EC: 1.0, 4.0, and 8.0 dS/m) in a glasshouse. Dry matter ratio, specific gravity, starch, and glucose content in tubers harvested at 90 days after sowing were analyzed. Also, mineral contents (T-N, T-C, $P_2O_5$, $K^+$, $Ca^{2+}$, $Mg^{2+}$, $Na^+$) in stem, leaf, and tuber were investigated and statistically analyzed for analysis of variance (ANOVA). Dry matter ratio, specific gravity, and starch content in tubers were reduced in all varieties as the salt concentration increased. Glucose content tended to decrease according to the salt concentration. In ANOVA analysis of mineral contents, there were significant differences in $K^+$ and $Mg^{2+}$ of leaf and stem, in $Na^+$ of leaf and tuber, and also in $Ca^{2+}$ of leaf by the interactions of variety and salinity. In the case of $K^+/Na^+$ and $Ca^{2+}/Na^+$, the stem was more sensitively influenced by the salt treatment than the leaf or the tuber. The $K^+/Na^+$ and $Ca^{2+}/Na^+$ decreased in leaf, stem, and tuber of four varieties, as the salt concentration became higher. The decreasing level varied according to the varieties. 'Kroda' and 'Duback' maintained relatively higher $K^+/Na^+$ and $Ca^{2+}/Na^+$ than 'Atlantic' or 'Goun' under the salt stress conditions. CONCLUSION: The composition and accumulation of minerals in potato plant as well as dry matter ratio, starch, and glucose contents were significantly influenced by salt stress. The respond patterns were different depending on the varieties and it was related to the salt tolerance among varieties.

A Comparative Study on Family Support, Self-esteem, and Health Status between the Institutionalized Elderly People and the Home-staying Ones (시설노인과 재가노인의 가족지지, 자존감 및 건강상태 비교연구)

  • Kim, Kwuy-Bun;Lee, Kyung-Ho
    • Journal of East-West Nursing Research
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    • v.5 no.1
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    • pp.36-49
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    • 2000
  • This study aims to provide the fundamental data for substantial nursing intervention in the elderly through a comparative appreciation on family support, self-esteem, and health status between the institutionalized elderly people and the home-staying ones. The subjects of this study are the institutionalized 108 elderly people of E and C Public Homes and the home-staying 109 elderly ones of O-Nho In Jeong(a kind of public recreational facilities for the aged) over the age of 65. The instruments for this research are based upon the tool(11 items, 5 points for each) for measuring family support developed by Choi, Young Hee(1984), a self-esteem scale done by Rosenberg (1965), the tools(20 items) for checking the health status of the elderly done by Lee, Young-Ja(1989). The sampling for this study has been carried on from July, 2000 until November, 2000. Questionnaire data were drawn up by personal interviews. The analyses of collected data are based on general characteristics calculated at the rate of 100 percentage to the average, t-test, ANOVA(some difference on a level with p<.05 being subsequently confirmed by DMR) for family support, self-esteem and health status, and Pearson Correlation to verify the hypothetical correlation among the subjects' family support, self-esteem and health status. The results of this study are as follows: 1. The difference between two groups in the light of family support, self-esteem and health status. (1) Family support - The rate of the family support that the institutionalized elderly people perceive turned out to be 22.13, that of the home-staying ones 30.99. (2) Self-esteem - The rate of the self-esteem that the former perceives proved to be 25.59, that of the latter 32.28. (3) Health Status - The rate of the health status that the former perceives turned out to be 39.67, that of the latter 51.60. 2. Family support, self-esteem, health status in terms of demographic characteristic (1) Family support - The group of institutionalized elderly people shows a tendency to be chiefly influenced by the death or life of the spouse and the number of the children; the group of the home staying ones to be chiefly influenced by the educational level (2) Self-esteem - The group of institutionalized elderly people shows a tendency to be chiefly influenced by educational level; the group of the home staying ones to be chiefly influenced by the amount of pocket money, the pocket money provider and the family main supporter. (3) Health Status - The group of institutionalized elderly people shows a tendency to be chiefly influenced by educational level; the group of the home staying ones to be chiefly influenced by age, the death or life of spouse, religion, and the educational level. 3. Correlation among family support, self-esteem, and health status The rate of correlation between family support and health status proved to be the highest (r=.549). After came the rate of correlation between health status and self-esteem, which turned out to be(r=.506). The last came the rate of correlation between family support and self-esteem, which proved to be(r=.406). According to this study, there is a conspicuously close correlation among family support, self-esteem, and health status for the elderly. Thus, it would be indispensable to seek out a variety of nursing intervention ways how the elderly could promote family support, self-esteem, and health status.

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Effect of Various Nitrogen Fertilizers on Rice Yield in Direct Seeding on Dry Paddy (건답 직파재배 논에서 질소비료 형태에 따른 시용효과)

  • Park, Chol-Soo;Jung, Yeong-Sang;Lee, Ho-Jin;Joo, Jin-Ho;Seo, Jun-Han
    • Korean Journal of Soil Science and Fertilizer
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    • v.33 no.5
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    • pp.318-324
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    • 2000
  • Field experiment was conducted on a sandy clay loam paddy field in the Experimental Farm of the Kangwon National University for four years from 1995 to 1998 to evaluate the effect of different nitrogen fertilizers on soil property and rice yield. The size of each experimental plot was 75 m2. Basal application of fertilizer to the paddy field was on late April, and rice (cv. Odae) was sown with drill seeding on early May. Irrigation was done at 3 leaf stages after rice seeding. Electrical conductivity in soils treated with phosphate- and resin coated slow release fertilizers were relatively greater than those of the other fertilizer application. Available $P_2O_5$ concentration in soils after rice harvest increased significantly compared that in soil before field trial. Rice yield in transplanting culture were $6.516kg\;ha^{-1}$. The rice yield in the direct seeded paddy was 81.9 percents of the yield in the transplanted paddy. The rice yield in phosphate coated slow release fertilizer was the highest among various fertilizer trials in directly seeded dry paddy, while rice yield was the lowest in the organic fertilizer plus urea application. Rice yield between organic fertilizer plus urea and organic fertilizer application were not significantly different. The green manuring effect of rye in the direct seeded rice paddy in Chuncheon was low due to low temperature and short growth period.

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The Effect of Application Levels of Slurry Composting and Bio-filtration Liquid Fertilizer on Soil Chemical Properties and Growth of Radish and Corn (총각무와 옥수수 재배시 SCB액비 시용수준이 토양화학성과 생육에 미치는 영향)

  • Kang, Seong-Soo;Kim, Min-Kyeong;Kwon, Soon-Ik;Kim, Myong-Suk;Yoon, Sung-Won;Ha, Sang-Gun;Kim, Yoo-Hak
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.6
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    • pp.1306-1313
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    • 2011
  • A liquid fertilizer treated with slurry composting and biofiltration (SCB) process has been applied increasingly on agricultural field but the effects on the soil properties and crop production has not been throughly evaluated. This study was conducted to investigate the effect of the SCB application on soil chemical properties and the growth of radish and corn. SCB liquid fertilizer as a basal fertilization was treated with five levels based on $6kg\;10a^{-1}$ for radish and $10kg\;10a^{-1}$ for corn. The experimental design was the completely randomized block design with five levels and three replicates. Electrical conductivity (EC), $NO_3$-N, Exch. K and Exch. Na increased depending on the treatment levels of SCB. There were no changes in soil organic matter, Avail. $P_2O_5$, Exch. Ca and Exch. Mg. EC, $NO_3$-N and Exch. Na content decreased as precipitation increased. Especially, they decreased up to the initial condition before the treatment after the heavy rainy season in 2008. Although Exch. K decreased at the rainy season, they remained relatively higher content after the experiment on August, 2008. Fresh weight and the amount of N uptake of radish increased due to the levels of SCB, but corn did not present any significant increase. It is recommended that we need to decide the proper amount of SCB as well as the application method on the field to increase the productivity and decrease environmental stress. Additional experiments also need to clarify the effect of the trace element and heavy metal accumulations due to long term application of SCB.

Project of Improving Good Agriculture Practice and Income by Intergrated Agricultural Farming (미얀마 우수농산물 재배기술 전수사업)

  • Lee, Young-Cheul;Choi, Dong-Yong
    • Journal of Practical Agriculture & Fisheries Research
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    • v.16 no.1
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    • pp.193-206
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    • 2014
  • The objectives of the project are to increase farmers' income through GAP and to reduce the loss of agricultural produce, for which the Korean partner takes a role of transferring needed technologies to the project site. To accomplish the project plan, it is set to implement the project with six components: construction of buildings, installation of agricultural facilities, establishment of demonstration farms, dispatching experts, conducting training program in Korea and provision of equipments. The Project Management Committee and the Project Implementation Team are consisted of Korean experts and senior officials from Department of Agriculture, Myanmar that managed the project systematically to ensure the success of the project. The process of the project are; the ceremony of laying the foundation and commencing the construction of training center in April, 2012. The Ribbon Cutting Ceremony for the completion of GAP Training Center was successfully held under PMC (MOAI, GAPI/ARDC) arrangement in SAl, Naypyitaw on June 17, 2012. The Chairman of GAPI, Dr. Sang Mu Lee, Director General U Kyaw Win of DOA, officials and staff members from Korea and Myanmar, teachers and students from SAl attended the ceremony. The team carried out an inspection and fixing donors' plates on donated project machineries, agro-equipments, vehicles, computers and printer, furniture, tools and so forth. Demonstration farm for paddy rice, fruits and vegetables was laid out in April, 2012. Twenty nine Korean rice varieties and many Korean vegetable varieties were introduced into GAP Project farm to check the suitability of the varieties under Myanmar growing conditions. Paddy was cultivated three times in DAR and twice in SAl. In June 2012, vinyl houses were started to be constructed for raising seedlings and finished in December 2012. Fruit orchard for mango, longan and dragon fruit was established in June, 2012. Vegetables were grown until successful harvest and the harvested produce was used for panel testing and distribution in January 2013. Machineries for postharvest handling systems were imported in November 2012. Setting the washing line for vegetables were finished and the system as run for testing in June 2013. New water tanks, pine lines, pump house and electricity were set up in October 2013.

The Study on the Factors of film's Processing Fluency Inducing Film's Preference Fluency (영화의 선호도 유창성에 영향을 미치는 영화의 처리 유창성 요인에 관한 연구)

  • Choi, Nak Hwan;Lim, Ahyoung
    • Asia Marketing Journal
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    • v.13 no.4
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    • pp.29-54
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    • 2012
  • Recently, as film has been admitted as the artistic merit and higher value-added business, the past studies on film have been lively carried on various fields. Especially, the film business is thought to be value-added business and has explored the causes of influencing spectators. However, there are no researches enough to explain what induces the choice and diffusion of the film. The film is not only hedonic products but also typical example of experiential products. So how to process film formation plays important roles in explaining the procedures of forming preference on the film and the movies spread more widely. But the great part of study of films has been concentrated on exploring hedonic factors of influencing spectator's choice. Until now there is not enough study for the relationship between experiential information processing and film preference. To explain film's preference, our study focuses on preference fluency and processing fluency that can provide an insight for our question about the relationship. In this article, to explain the procedures of processing experiential information and forming preference on the film, our study focuses on finding the relationship between film's processing fluency and film's preference fluency and explores the factors that affect film's processing fluency. To achieve the goal of this study, we distinguish factors of film's conceptual fluency from factors of film's perceptual fluency and explore the paths from the factors to film's preference fluency. The factors which have effects on perceptual fluency are hypothesized to be distinction of image expression, distinction of sound expression, correspondence between actors' image and their role. The factors which have effect on conceptual fluency are supposed to be well-organized story, suitability of lines expression. The experiments in which students were sampled at 'C' university were conducted in 2010 (december). Data collection was proceeded through questionnaires. We test the hypothesized model by using structural equation modeling(Amos 17.0). The fit indices for the model are as follows : x2=416.266(df=213, p=0.00), GFI=0.855, AGFI=0.812, RMSEA =0.069, IFI=O.925, CFI=0.920, TLI=0.905. According to the guidelines, there is evidence that our measurement model fits data. The results of empirical study are as follows. The path from film's perceptual fluency to film's preference fluency is supported(estimate: 0.223, C.R: 2.641). The path from film's conceptual fluency to film's preference fluency is supported(estimate: 0.397, C.R: 4.863). The path from distinction of image expression to film's perceptual fluency is not supported(estimate: 0.113, C.R: 1.665). The path from distinction of sound expression to film's perceptual fluency is supported (estimate: 0.190, C.R: 2.042). The path from correspondence between actors' image and their role to film's perceptual fluency is supported(estimate: 0.686, C.R: 5.566). The path from well-organized story to film's conceptual fluency is supported(estimate: 0.396, C.R: 4.023). The path from suitability of lines expression to film's conceptual fluency is supported(estimate: 0.536, C.R: 5.441). Concludingly, our study explored the influencing factors of film's processing fluency inducing film's Preference fluency. First, film's perceptual fluency and film's conceptual fluency have positive effects on film's preference fluency. Second, distinction of image expression is not significant on film's perceptual fluency, but distinction of sound expression and image's correspondence of actors' image and their role have positive effects on film's perceptual fluency. Lastly, well-organized story and suitability of lines expression have positive effects on film's conceptual fluency.

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도체 현수방법에 따른 쇠고기 품질 비교

  • Park, Beom-Yeong;O, Mi-Ra;Kim, Jin-Hyeong;Jo, Su-Hyeon;Ha, Gyeong-Hui;Seong, Pil-Nam;Hwang, In-Ho;Lee, Jong-Mun;Kim, Dong-Hun;An, Jong-Nam
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2006.05a
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    • pp.146-150
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    • 2006
  • 도축 후 2일째 가열 감량은 시험구(골반골 현수)가 대조구(아킬레스건 현수)에 비하여 유의적으로 감량이 높았으나, 드립량에 있어서는 반대의 결과를 보였으며, 전체 감량은 두 처리구간 차이가 없었다. 연도의 지표인 전단력가는 대조구 $6.45kg/cm^2$로 시험구 $5.52kg/cm^2$에 비하여 유의적으로 높았으며, 골반골 현수를 통하여 도축 후 8일차의 대조구의 전단력 수준을 보여 숙성기간을 약 1주일 정도 단축이 가능하였다. 근절길이는 대조구 $1.93{\mu}m$, 시험구 $2.48{\mu}m$로 시험구가 유의적으로 길어 골반골 현수에 의한 단축 억제 효과로 볼 수 있다. 관능 평가 결과 도축후 2일차에는 대조구에 비하여 시험구가 다즙성, 연도, 향미, 기호성에서 유의적으로 높은 값을 보였으나, 도축 후 8일째에는 대조구와 시험구간에 차이를 보이지 않았다. 출하 연령과 현수방법에 따른 대퇴이두근의 물리적 특성을 비교한 결과로서 근절길이는 모든 월령에서 시험구가 대조구에 비하여 길었다. 전단력은 시험구가 대조구에 비하여 유의적으로 낮은 결과를 보였다. 골반골 현수에 의한 전단력가 감소 효과는 출하 월령이 많을수록 효과가 큰 것으로 나타났다. 드립 발생율은 대조구가 시험구에 비하여 유의적으로 높은 경향을 보였다. 가열 감량과 보수성은 처리구에 관계없이 유의적인 차이를 보이지 않았다. 관능 특성을 비교한 결과 30개월 미만의 경우에는 유의적인 차이를 보이지 않았으나, 30개월 이상에서는 처리구가 대조구에 비하여 다즙성 연도, 향미, 기호성 모두 우수한 것으로 나타났다. 특히 골반골 현수는 출하 월령이 증가됨에 따라 관능 특성의 개선율이 높은 것으로 나타났다. 수 있다.분별을 성공적으로 수행하였다.(p<0.05), 맛, 연도, 다즙성 및 전체적인 기호성은 유의한 차이가 없었다.자체를 악하다고 볼 수 없고 더구나 구원을 이 세상에서의 이탈로 볼 수 없다. 진정한 구원이란 원래 하나님이 보시기에 아름다웠던 그 세상으로의 회복을 포함한다. 이런 면에서 하나님 주권 신앙 하에서 구원이란 전 인격적인 구원, 전 우주적인 구원이 된다. 그렇기 때문에 성도는 세상의 삶과 학문, 예술, 정치, 경제, 사회를 포함한 모든 분야를 하나님의 뜻 가운데서 그 원래의 목적에 부합할 수 있도록 회복시키는 일에 적극 참여해야 한다.자체가 이를 주도하기는 사실 어려움이 있다. 그리고 대형유통점이 영업행위를 영업시간제한에서부터 출점제한에 이르기까지 규제하는 건은 심사숙고하여야 한다. 대형유통점이 국가경제 및 지역사회에 미치는 영향이 부정적인가 긍정적인가에 대해 국내외 학계와 업계에서 여전히 많은 논란이 있기 때문이다. 정부와 지자체에 의한 시장개입은 반드시 필요한 경우에 한해 합당한 방법에 의해 이루어져야 한다. 대형유통점에 대한 규제는 지역사회에 미치는 영향을 다면적으로 평가한 결과에 근거하여 이루어져야 할 것이다. 대부분의 지자체는 체계적인 평가시스템과 객관적인 통계 자료를 갖고 있지 못한 실정이다. 향후 가장 시급한 과제는 시장개방 이후 지난 10년간 대형유통점이 지역사회에 미친 영향에 관한 광범위한 통계자료를 수집하고 이를 체계적으로 분석하여 정책방향을 올바르게 설정하는 것이라 할 수 있다.i와 K. pneumoniae가 존재하며 확산 중임을 시사한다. 앞으로 CTX-M형 ESBL의 만연과 변종 CTX-M형 ESBL의 출연을 감시하기 위한 정기적인 연구와 조사가 필요한 것으로 생각한다., A2-1, B1-1, B2-1의 경우,

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Development of Assay Methods for Enterotoxin of Escherichia coli Employing the Hybridoma Technology (잡종세포종기법을 이용한 대장균의 장독소 측정법 개발)

  • Kim, Moon-Kyo;Cho, Myung-Je;Park, Kyung-Hee;Lee, Woo-Kon;Kim, Yoon-Won;Choi, Myung-Sik;Park, Joong-Soo;Cha, Chang-Yong;Chang, Woo-Hyun;Chung, Hong-Keun
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.1
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    • pp.151-161
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    • 1986
  • In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.

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