• Title/Summary/Keyword: Evaluation level

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EVALUATION OF OSTEOGENIC ACTIVITY AND MINERALIZATION OF CULTURED HUMAN DENTAL PAPILLA-DERIVED CELLS (배양된 치유두 유래세포의 조골활성 및 골기질 형성의 평가)

  • Park, Bong-Wook;Byun, June-Ho;Choi, Mun-Jeoung;Hah, Young-Sool;Kim, Deok-Ryong;Cho, Yeong-Cheol;Sung, Iel-Yong;Kim, Jong-Ryoul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.29 no.4
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    • pp.279-288
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    • 2007
  • In the present study, we focused on stem cells in the dental papilla of the tooth germ. The tooth germ, sometimes called the tooth bud, is the primordial structure from which a tooth is formed. The tooth germ consists of the enamel organ, the dental papilla, and the dental follicle. The dental papilla lies below a cellular aggregation of the enamel organ. Mesenchymal cells within the dental papilla are responsible for formation of dentin and pulp of a tooth. Tooth germ disappears as a tooth is formed, but that of a third molar stays in the jawbone of a human until the age of 10 to 16, because third molars grow slowly. Impacted third molar tooth germs from young adults are sometimes extracted for orthodontic treatment. In the present study, we evaluated the osteogenic activity and mineralization of cultured human dental papilla-derived cells. Dental papillas were harvested from mandible during surgical extraction of lower impacted third molar from 3 patients aged 13-15 years. After passage 3, the dental papilla-derived cells were trypsinized and subsequently suspended in the osteogenic induction DMEM medium supplemented with 10% fetal bovine serum, 50 g/ml L-ascorbic acid 2-phosphate, 10 nM dexamethasone and 10 mM -glycerophosphate at a density of $1\;{\times}10^6\;cells/dish$ in a 100-mm culture dish. The dental papilla-derived cells were then cultured for 6 weeks and the medium was changes every 3 days during the incubation period. Dental papilla-derived cells showed positive alkaline phosphatase (ALP) staining during 42 days of culture period. The formation of ALP stain showed its maximal manifestation at day 7 of culture period, then decreased in intensity during the culture period. ALP mRNA level was largely elevated at 1 weeks and gradually decreased with culture time. Osteocalcin mRNA expression appeared at day 14 in culture, after that its expression continuously increased in a time-dependent manner up to day 28. The expression remained constant thereafter. Runx2 expression appeared at day 7 with no detection thereafter. Von Kossa-positive mineralization nodules were first present at day 14 in culture followed by an increased number of positive nodules during the entire duration of the culture period. Osteocalcin secretion was detectable in the culture medium from 1 week. The secretion of osteocalcin from dental papilla-derived cells into the medium greatly increased after 3 weeks although it showed a shallow increase by then. In conclusion, our study showed that cultured human dental papilla-derived cells differentiated into active osteoblastic cells that were involved in synthesis of bone matrix and the subsequent mineralization of the matrix.

Geographical Structure of View Point in the Landscape Experience on Mountain in Yangsan City (경관체함이 발생하는 조망점의 지리구조에 관한 연구;양산시 소재 산지를 대상으로)

  • Kang, Young-Jo;Cha, Young-Chae;Cha, Myeong-Sook
    • Journal of the Korean Institute of Landscape Architecture
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    • v.36 no.3
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    • pp.75-84
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    • 2008
  • The purpose of this research is to find the visual characteristics and geographic structure in a landscape of mountains in Yangsan City. In order to carry out this study, 40 view points of 6 mountains in Yangsan City were selected to experience the mountainscape. The results are as follows. The mountainscape of Yangsan City was confirmed from the view point to the object of view as ranging in middle distance reverse, the angle of elevation 11${\sim}13^{\circ}$ which give the impression of confronted to the mountain, and the horizontal angle 50${\sim}90^{\circ}$ which gives the impression of vastness. This shows that mountainscapes are perceived impressively when seeing the mountain with the meaning to feel the texture of the mountain as a confrontational relationship rather than a pictorial relationship. The geographic structure of the view point which occurs in scenes of impressive experience is classified into eight types and its characteristics are investigated. The first type is corridor, and it emerges in a long narrow linear structure made from the surrounding environment with buildings or street trees. The second is tunnel in which a vision and light hidden while passing underneath a bridge, high-level road, or inside a tunnel can be seen. Third, the maze is formed by the mountain shape. The hill spur emerges when the direction of traffic changes after turning the coner of a building or mountain edge. The hill ground emerges at the top of a hill slope. Next, parallel emerges when the observer and the mountain are in the same direction of process. Confrontation occurs when confronting an isolated mountain. Finally, the view point emerges when passing through major points such as a bridge or square and the boundary of a village or city. This research arranged visual conditions that create impressive mountainscapes. Geographic characteristics in terms of Types that make possible to experience mountainscapes were described. The results of this research will be basic data collected for the management and preservation of mountainscapes and for landscape effect evaluation. Furthermore, this research suggests theoretical evidence to preserve and manage geographic structures that create view experiences as well as to preserve the landscape in terms of view points.

Understanding Contemporary Interstate Rivalries: Consensus Rivalries and Rivalry Termination (숙적관계 국가들에 대한 연구 - 숙적관계 종식에 영향을 미치는 주요 요인들을 중심으로 -)

  • Oh, Soon-Kun
    • Strategy21
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    • s.32
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    • pp.222-270
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    • 2013
  • 서구 근대 국제관계학의 태동은 "전쟁의 원인"을 밝혀내려고 했던 유럽 지식인들의 학문적 도전에서부터 비롯되었다고 할 수 있다. 1, 2차 세계대전의 잔혹함을 겪으면서 이들은 전쟁(특히 전쟁의 원인)에 대한 좀 더 과학적인 연구를 통해 어떻게 하면 국가들 간의 전쟁을 예방할 수 있을까에 집중해 왔다. 이러한 학문적 경향을 반영한 것이 잘 알려진 미국 미시간 대학의 "전쟁 상관성 연구 프로젝트(Correlates of War Project)"이다. 이는 나폴레옹 전쟁이 끝나고 비엔나체제가 시작된 1815년 이후 국가들 간 발생한 모든 전쟁 관련 자료를 데이터베이스화하여,국제관계학자들이 전쟁(Interstate War)이나 군사분쟁(Militarized Interstate Disputes)을 정량 또는 정성적으로 연구할 수 있도록 다양한 정보들을 제공하고 있다. 최근 Paul F. Diehl, William R. Thompson과 같은 학자들은 전쟁을 분석단위로 하는 전쟁의 원인(Causes of War)을 연구하는 학풍에서 벗어나, 국가들 간의 분쟁과 전쟁을 연구하기 위해 숙적관계(Rivalry)라는 새로운 분석의 단위를 제시하였다. 숙적관계는 국제관계에 있어서 지속적으로 분쟁 또는 전쟁을 일으키는, 즉 무력분쟁의 긴 역사를 가지고 있는, 두 국가를 일컫는다. 국가들 간의 숙적관계는 학자들에 따라 Interstate Rivalry, Enduring Rivalry, Strategic Rivalry 등 다양한 정의와 성격을 갖고 있다. 이러한 국가들의 사례로는 2차대전 이전까지 프랑스-독일 관계, 2차대전 이후 중동지역 이스라엘과 아랍국가들 간의 관계, 냉전기 미소관계, 인도-파키스탄 및 남북한 관계 등을 예로 들 수 있다. 이렇게 널리 알려진 숙적관계 외에도 남미의 칠레-아르헨티나, 에콰도르-페루와 아프리카의 소말리아-에티오피아 등 학자들에 따라 1815년 이후 약 200개의 숙적관계를 제시하고 있다. 숙적관계에 대한 연구는 기존의 전쟁의 원인 중심이었던 정량적 국제분쟁 연구에 두 국가의 분쟁역사를 포함시키는 정성적 연구를 접합시키고 있다. 본 연구는 1945년 이후 숙적관계 국가들의 관계종식과 관련하여, 거시론적이고 전체론적 접근방법(Macro - level Holistic Approach)을 제시하고 있다. 먼저 1945년 이후 발생한 국가들 간의 숙적관계 종식(Rivalry Termination)을 이해하기 위해, 거시적 관점에서 숙적관계를 발생시키게 된 역사적 원인을 살펴보고 있다. 특히 1945년 이후 숙적관계와 관련된 중요한 요인들 중에서 전쟁, 영토분쟁, 그리고 근대국가 형성에 대해 살펴보고 다음과 같은 두 가지 명제(proposition)를 제시한다. 첫째, 1945년 이후의 숙적관계 형성과 지속에는 2차대전 이후 독립국가 형성, 헌팅턴이 주장한 제3의 물결(민주화), 그리고 냉전기 미소경쟁 등의 역사적 배경이 영향을 주었다는 것이다. 둘째, 이러한 역사적 배경에 더하여 1945년 이후 숙적관계는 전쟁에 의해 시작된 숙적관계와 전쟁 없이 시작된 숙적관계 등 2가지로 나눌 수 있으며, 전쟁에 의해 시작되고 영토분쟁과 근대국가 형성 문제를 내포하고 있는 숙적관계가 그렇지 않은 숙적관계에 비해 오랫동안 지속되며 관계해결이 어렵다는 것이다. 앞서 제시된 2가지 명제들과 관련하여 본문에서는 다양한 학자들이 제시한 숙적관계 정의들에 일치하는 23개의 숙적관계(Consensus Rivalries)를 선정하여 이들에 대한 비교분석(Descriptive Analysis)을 실시하였다. 이들 사례들을 1945년 이전과 이후로 나누어, 숙적관계 형성과 종식에 있어 핵심요소인 국가들 간 힘의 차이(Power Relations), 분쟁의 주요 원인(Primary Conflict Issue), 숙적관계에 있어 다른 국가들과의 연계성(Rivalry Linkage), 전쟁의 횟수와 시기 등을 통해 비교하였다. 숙적관계의 종식과 관련하여 약소국 간의 숙적관계(Minor Dyad)가 오래 지속되고, 영토분쟁(Territorial Disputes)이 숙적관계를 지속시키는 주요인이며, 다른 숙적관계와의 연계성이 적은 숙적들이 오래 지속된다는 비교분석 결과가 나왔다. 또한 전쟁의 횟수는 숙적관계 종식에 큰 영향을 미치지 못하지만 전쟁으로 인해 시작된 숙적관계가 그렇지 않은 경우보다 더욱 오래 지속되는 것으로 확인되었다. 끝으로 2차대전 이전의 숙적관계는 대부분 전쟁을 통해 종식되었지만, 1945년 이후에는 전쟁 없이도 숙적관계가 종식된 경우가 많았음을 보여 주고 있다. 본 연구의 주목적은 "1945년 이후 형성된 숙적관계를 어떻게 종식시킬 수 있을 것인가?"라는 규범적 논제를 통해, 23개의 주요 숙적관계를 발굴하여 거시적, 역사적 관점에서 비교분석함에 있다. 이는 특히 우리나라가 처한 현실과 관련하여 중요한 시사점을 갖는다. 남북관계가 갖는 특수성보다는 1945년 이후 국제정치 역사속에서 발생한 숙적관계 현상이라는 일반성의 틀에서 남북관계를 이해하고자 했다. 남북관계를 베트남, 예멘, 독일 등 분단국가의 사례들과만 비교 연구하는 제한된 시각에서 벗어나, 인도-파키스탄, 그리스-터키, 에콰도르-페루 등 유사한 숙적들 간의 관계들과 비교하는 새로운 연구의 장을 제시하고자 한다. 따라서 앞으로 숙적관계 종식에 대한 더욱 다양화된 사례연구를 통해 한반도 분쟁해결에 필요한 새로운 교훈을 얻을 수 있을 것이다. 예를 들면 2차대전 이후 발생한 국경을 접하고 있는 비강대국들의 평화적인 숙적관계 종식에 대한 사례연구는 남북한이 앞으로 지향해야 할 방향을 제시해 줄 수 있을 것이다. 끝으로 본 연구는 특정한 정책적 함의를 도출하기 보다는 숙적관계와 관련된 하나의 거시적 이론를 제시하고, 주요 숙적관계 국가들에 대한 비교설명을 통해 현존하는 숙적관계 해결을 위한 하나의 분석의 틀을 제시하는 것으로 국제분쟁 연구에 기여하고자 하였다.

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Image and Exposure Dose in Accordance with Radiation Quality on Plain Chest Radiography (흉부촬영(胸部撮影)에서 증감지(增感紙)-필름계의 선질변화(線質變化)에 따른 감도(感度)와 화질에 관(關)한 연구(硏究))

  • Kim, Jung-Min;Kim, Dong-Huan;Hayashi, Taro;Ishida, Yuji;Maeda, Mika;Sakura, Tatsuya
    • Journal of radiological science and technology
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    • v.15 no.1
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    • pp.65-78
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    • 1992
  • Routine chest radiography is generally imaged by high voltage technique but some radiological technologists use low voltage for imaging. High voltage is usually said between $120\;kV{\sim}140\;kV$. Some RTs like using heavy filtration but others seldom like using it. However which is better for use calcium tungustate film screen system or ortho system and high contrast film or wide latitude c-type film for the exculusive use of chest radiography. We could not make a decision which is ideal method for use. In my opinion any method is not always exellent for chest radiography. In my experiments that I had at Kaken hospital in Japan last year I expect to keep the balance between image quality and diagnostic range and to reduce radiation dose for patients. My experiments are as follows. 1. We have looked into system characteristics(speed and contrast) in accordance with kVp($80{\sim}140$) and added filter($no{\sim}1/16\;VL$) in three screen film systems(BX3+CRONEX4, SRO750+MGH, SRO750+MGL). 2. We have looked into skin dose and film dose with same D=1.8 lung field density in accordance with kVp($80{\sim}140$) and added filter($no{\sim}1/16\;VL$) in three screen film systems. 3. We have compared with the evaluation between correlation of physical image quality(MTF) and optical diagnostic capability. Result are follows. 1. Speed of BX3+CRONEX4 became higher in accodance with kVp and thickness of filter but speed of ortho system was not as like regular system. Thicker filter diminished the speed over 100 kV range in SRO750+MGL. In case of SRO750+MGH speed of 1/16VL filter was looked into lower than speed of 1/4VL filter. Sensitivity of ortho system depends on tube voltage and added filter. 2. Skin dose has been detected $225\;{\mu}Gy{\sim}66\;{\mu}Gy$ in BX3+CRONEX4 from 80 kV, no filter to 140 kV, 1/16VL filter. SRO750+MGH could reduce the patient dose $1/2{\sim}1/3$ level in comparison to that of BX3+CRONEX4. 3. The higher kV was the worse MTF became the thicker filter was the worse MTF became too. MTF of BX3+CRONEX4 was detected better than MTF of SRO750+MGH but SRO750+MGH's optical detectability of small lesion in lung field came out better than that of BX3+CRONEX4. Conclusion Recently routine chest radiography is generally imaged by high voltage but it seems to be there are some questions in using of film screen combination. In high voltage chest radiography the subject contrast will come down that means latitude become wider. In this case if we select the low contrast film screen system(C or L type) the film contrast will fall down extremly and detectability of small lesion will be deteriorated. Wide latitude C, L type film has a merit of high detectability on mediastinum. Furthermore high contrast film screen system has the advantage to keep the high contrast in low density region as like mediastinum and heart shadow. Therefore in low subject contrast high voltage chest radiography we would rather choose the high contrast film screen system(H type) I think. From a view point of patient dose detectability of mediastinum and lung field. The optimum technical facter was found out 120 kV, 1/16VL filter : BX3+CRONEX4, 140 kV, 1/4VL filter : SRO750+MGH, 100 kV, 1/4VL filter : SRO750+MGL.

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Analysis of Basic Factors of Self-Directed Learning for the Creative Leaning Management (창의적 학습 경영을 위한 자기주도학습 기초요인 분석)

  • Ko, Jae Lyang;Kim, Kyung Soon;Byun, Sang Hea
    • Asia-Pacific Journal of Business Venturing and Entrepreneurship
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    • v.8 no.4
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    • pp.145-159
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    • 2013
  • The purpose of this study is to analyze the structural relationship as to how learning flow and self-directed learning are linked to learning motives and academic self-efficacy in the learning setting of high school students. To accomplish such purpose, based on theoretical backgrounds and preceding research findings evaluation models were put to verification for a valid research model for this study. The initial hypothetical model was that self-directed learning ability would have a direct influence on learning motive, academic efficacy and learning flow, while having an indirect influence on learning flow with learning motive and self-efficacy acting as a mediating variable. But the hypothetical model showed low significance level between self-directed learning and learning motive, and learning motive and learning flow. Therefore, links were adjusted to create the final model within the scope that the adequacy of the model might not be compromised. To verify the model, 900 high school students in Seoul were surveyed and the collected data were statistically analyzed using AMOS v21.0 and SPSS v21.0 But 815 surveys were excluded because they were not sufficiently answered. From the analysis, it was found that self-directed learning and academic efficacy have a direct influence on learning flow while self-directed learning and academic efficacy have an indirect leaning motive and learning flow. This finding means that, in the relationship of self-directed learning and learning flow, learning motive and learning efficacy are positive factors that help high school students experience learning flow. Thus, in order to enhance the experience of self-directed learning ability of high school students, various educational endeavors are needed to draw the experience of learning flow during the regular course of study. In addition, customized educational methods and environments are required to increase academic efficacy of the students.

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Evaluation of Soil and Fertilizer Management Techniques Applied by Farmers in Forcing and Semi-forcing Cucumber Cultivation Facilities (오이 촉성재배와 반촉성재배 농가들의 토양 및 시비관리기술 평가)

  • Lee, Ju-Young;Jang, Byoung-Choon;Sung, Jwa-Kyung;Lee, Su-Yeon;Kim, Rog-Young;Lee, Ye-Jin;Park, Yang-Ho;Kang, Sung-Soo;Hyun, Byung-Keun
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.6
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    • pp.983-991
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    • 2012
  • These days, agricultural products cultivated in facilities occupy the highest percentage of agricultural output price. Specifically cucumbers have been one of the crops that farmers prefer to growing, because their prices were high. However, cucumber crop is sensitive to the soil and environments and it requires the exact crop management. In order to establish cultivation techniques for cucumbers, the current situation of cucumber cultivation was surveyed from ten cucumber farmlands; five farmlands of cucumber cultivation in forcing and five of semi-forcing practicing systems, respectably. The soil conditions were alluvial or valley in soil topology, moderately or poorly drainage in soil drainage classes, coarse loamy in soil texture family. Soil was managed with deep plowing combined with application of basal fertilizers such as compost, rice straw, oil cake, wood chip and chemical fertilizer. The whole soil was prepared in uniformly with rotary. Three major nutrients ($N-P_2O_5-K_2O$)of basal application were 815-464-529 kg $ha^{-1}$ in forcing and 197-135-151 kg $ha^{-1}$ in semi-forcing cultivation. Top dressing of fertilizer was supplied in fertigation system of macro and micro elements in 2~3 day interval with water irrigation. The average yields of cucumbers were $168t\;ha^{-1}$ with 381,000 thousand won $ha^{-1}$ in average gross profit (AGP) in forcing cultivation and $115t\;ha^{-1}$ with 177,000 thousand won $ha^{-1}$ in AGP in semi-forcing cultivation. Cucumber production during the winter season was considered to increase the gross profit because cucumber price tends to stay in high level during this time. The accumulation of soil chemicals like EC, available $P_2O_5$ and exchangeable cations could be controlled by rice straw application. The rice straw application increased soil temperature during the winter season, in exchange of soil air, and in extension of plant roots. In addition, the rice straw application somewhat affected decrease of salts accumulation.

A Comparative Study On Accident Prediction Model Using Nonlinear Regression And Artificial Neural Network, Structural Equation for Rural 4-Legged Intersection (비선형 회귀분석, 인공신경망, 구조방정식을 이용한 지방부 4지 신호교차로 교통사고 예측모형 성능 비교 연구)

  • Oh, Ju Taek;Yun, Ilsoo;Hwang, Jeong Won;Han, Eum
    • Journal of Korean Society of Transportation
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    • v.32 no.3
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    • pp.266-279
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    • 2014
  • For the evaluation of roadway safety, diverse methods, including before-after studies, simple comparison using historic traffic accident data, methods based on experts' opinion or literature, have been applied. Especially, many research efforts have developed traffic accident prediction models in order to identify critical elements causing accidents and evaluate the level of safety. A traffic accident prediction model must secure predictability and transferability. By acquiring the predictability, the model can increase the accuracy in predicting the frequency of accidents qualitatively and quantitatively. By guaranteeing the transferability, the model can be used for other locations with acceptable accuracy. To this end, traffic accident prediction models using non-linear regression, artificial neural network, and structural equation were developed in this study. The predictability and transferability of three models were compared using a model development data set collected from 90 signalized intersections and a model validation data set from other 33 signalized intersections based on mean absolute deviation and mean squared prediction error. As a result of the comparison using the model development data set, the artificial neural network showed the highest predictability. However, the non-linear regression model was found out to be most appropriate in the comparison using the model validation data set. Conclusively, the artificial neural network has a strong ability in representing the relationship between the frequency of traffic accidents and traffic and road design elements. However, the predictability of the artificial neural network significantly decreased when the artificial neural network was applied to a new data which was not used in the model developing.

Effects of Active MA Mini-packaging on Shelf-life Maintenance during Marketing of Perilla Leaves (깻잎의 active MA 소포장이 유통기간 중 신선도 유지에 미치는 영향)

  • Jeong, Cheon-Soon;Um, Gi-Jeung;Park, Jong-Nam
    • Horticultural Science & Technology
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    • v.28 no.6
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    • pp.980-984
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    • 2010
  • Research regarding respiration rate, off-flavor related material and freshness maintenance effect of active MA mini-packaging on perilla leaves has been carried in this study. Respiration rate was highly maintained at high oxygen treatments ($CO_2:O_2:N_2$=0:30:70 and 0:50:50), higher than non-treatment and low oxygen treatments (6:2:92, 0:10:90) during the storage period, and ethylene production was relatively higher. However, off-flavor related materials, acetaldehyde and ethanol production was noticeably low. Weight losses of non-treatment and low-oxygen treatment were about 1.8 and 1.4%, respectively. At the fifth day of storage there was no weight loss. Ascorbic acid content was 13.3 mg/100 g F.W. at the first day of storage. At the third day of storage non-treatment showed the lowest value of ascorbic acid, 8.8 mg/100 g F.W. Ascorbic acid content of active MA treatment gradually decreased without a striking difference as the storage day extended. Chlorophyll content was the lowest at the fifth day of storage with non-treatment, 47.5 (SPAD-502 unit) while the active MA treatment maintained high level of chlorophyll content, 53.0. Sensory evaluation (vision) showed that marketability gradually decreased but was maintained in high oxygen treatments (0:30:70 and 0:50:50), even at the fifth day of storage. Occurrence of off-flavor in non-treatment at the fifth day of storage was extremely low, 2.6; whereas high in high oxygen treatment, 3.4. Active MA packaging maintained freshness two more days at room temperature (about $27^{\circ}C$).

An Animal Study on Electrohydraulic Type Ventricular Assist Device (전기 유압식 심실보조장치의 동물실험 연구)

  • 백완기;심상석
    • Journal of Chest Surgery
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    • v.29 no.7
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    • pp.689-699
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    • 1996
  • An animal experiment was designed for the evaluation of in vivo performance of the newly developed electrohydraulic type ventricular assist device and its influence on the left ventricular function during pal- satile left ventricular assist. Eight adult sheep were incorporated into the study and data were collected from seven sheep. Total as- sist time ranged from 69 minutes to 7 days. The performance of the device was satisfactory both in asyn- chr nous and synchronous mode within the range of given native heart rate. More than 4 liters of device output could be reached within the range of normal left atral pressure without development of negative pressure in the left atrium. Moderate to severe degree of hemolysis was noted as evidenced by significant increase of plasma free hemoglobin level after 3 days of left ventricular support along with the presence of the small amount of thrombi around the floating disc type polymer valve apparatus reflecting that further study and refinement of the device need to be done in regard of biocompatibility and thromboresistance. The hemodynamics showed increase in heart rate (p < 0.05), cardiac output and left ventricular minute work (p < 0.05) after placement of the device at the flow rate of 2.0∼2.5 Llmin. The left atrial pressure, left ventricular pressure and LV dpldt were decreased after the device placement(p < 0.05). The endocardial viability ratio and oxygen contents of the mixed ven us blood and coronary venous blood were all increased (p < 0.05) after the device placement suggesting effective unloading of the left ventricle was accomplished. The myocardial perfusion was thought improved in synchronous counterpulsation as suggested by sig- nificant increase in endocardial viability ratio and coronary venous blood oxygen content in synchronous assist mode comparing with asynchronous mode.

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Ttrosine Hydroxylase in Japanese Medaka (Oryzias latipes): cDNA Cloning and Molecular Monitoring of TH Gene Expression As a Biomarker (송사리 Tyrosine Hydroxylase: cDNA 클로닝 및 생물지표로서의 TH 유전자 발현의 분자생물학적 추적)

  • Shin, Sung-Woo;Kim, Jung-Sang;Chon, Tae-Soo;Lee, Sung-Kyu;Koh, Sung-Cheol
    • Environmental Analysis Health and Toxicology
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    • v.15 no.4
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    • pp.131-137
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    • 2000
  • The release of hazardous waste materials into the environment poses serious risks in humans and ecosystems. The risk assessment of environmental pollutants including hazardous chemicals requires a comprehensive measurement of hazard and exposure of the chemicals that can be achieved by toxicity evaluation using a biological system such as biomarkers. In this report we have tried to develop a biomarker used to elucidate a molecular basis of, and to monitor abnormal behaviors caused by diazinon in Japanese medaka (Oryzias latipes) as a model organism. First, an attempt was made to clone tyrosine hydroxylase gene from Japanese medaka that would be a candidate for a biomarker for neuronal modulations and behaviors. For monitoring experiments at behavioral and molecular biological levels, the fish were treated under different sublethal conditions of diazinon and their behavioral responses were observed . In this study we have successfully cloned a partial TH gene from the medaka fish through PCR screening of an ovary cDNA library. DNA sequencing analysis revealed that the amplified fragment was 327 bp encoding 109 amino acids. Comparing the DNA sequence of medaka TH with other species, TH gene revealed the DNA sequence was completely identical to that of rat TH. In the RT-PCR, 330 Up of mRNA was consistently amplified in all the treated samples including control There were no significant differences in the TH expression level regardless of treating concentrations (1∼5,000 ppb) and time (0∼48 hr) The reason appeared to be that RT-PCR was not performed using through a quantitative analysis normalized against an actin gene expression. Organ or tissue - specific detection of TH activity and mRNA as biomarkers will be a useful monitoring tool for neurobehavioral changes in fish influenced by toxic chemicals. Furthermore, quantitative analysis of locomotive patterns and its correlation with the neurochemical and molecular data would be highly useful in measuring toxicity and hazard ofvarious environmental pollutants.

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