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The optimal conditions to improve retrovirus-mediated transduction efficiency to NIH 3T3 cells (레트로바이러스(retrovirus)의 NIH 3T3 세포로의 유전자 전달효율을 증가시키기 위한 적절한 조건들)

  • Lee, Jun Ah;Lee, Kang-Min;Lee, Hyun Jae;Lee, Yun Jeong;Kim, Dong Ho;Lim, Jung Sub;Park, Kyung-Duk
    • Clinical and Experimental Pediatrics
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    • v.50 no.10
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    • pp.1011-1017
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    • 2007
  • Purpose : We tried to assess the optimal conditions to improve low transduction efficiency and their effect on target cells. Methods : Cultured NIH 3T3 cells were incubated with retroviral vectors bearing an enhanced green fluorescent protein (eGFP) gene. We varied the ratio of viral vectors to target cells (1:1-1:8) and the number of transfections (${\times}1$, ${\times}2$), and compared transduction efficiencies. Also, the effects of polybrene on transduction efficiency and viability of target cells were assessed. Transduction of the eGFP gene was evaluated by observing NIH 3T3 cells under a fluorescence microscope and efficiencies were measured by the percentage of eGFP positive cells using FACscan. Results : As the ratio of retroviral vectors to target cells increased, transduction efficiency was greatly improved, from 7% (1:1) to 38% (1:4). However, transduction efficiency did not increase any more when the ratio increased from 1:4 to 1:8. Cells transfected twice showed higher transduction efficiencies than cells transfected once, at a ratio of 1:8. The eGFP gene transduced to NIH 3T3 cells sustained its expression during repeated passages. However, after the third passage (day 9), the percentage of eGFP positive cells began to decline. The degree of this decline in eGFP expression was lower in cells transfected twice than in cells transfected once (P<0.05). The addition of polybrene did not have any toxic effect on NIH 3T3 cells and greatly increased transduction efficiency (P=0.007). In addition to vector component, transduction efficiency was very sensitive to culture confluence. Cells cultured and transfected in 24-well plate showed higher transduction efficiency, although cells cultured in 6- well plate proliferated more (P=0.024). Conclusion : Our data could be used as a basis for retrovirus-based gene therapy. Further study will follow using human cells as target cells.

Stellite bearings for liquid Zn-/Al-Systems with advanced chemical and physical properties by Mechanical Alloying and Standard-PM-Route

  • Zoz, H.;Benz, H.U.;Huettebraeucker, K.;Furken, L.;Ren, H.;Reichardt, R.
    • Proceedings of the Korean Powder Metallurgy Institute Conference
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    • 2000.04a
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    • pp.9-10
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    • 2000
  • An important business-field of world-wide steel-industry is the coating of thin metal-sheets with zinc, zinc-aluminum and aluminum based materials. These products mostly go into automotive industry. in particular for the car-body. into building and construction industry as well as household appliances. Due to mass-production, the processing is done in large continuously operating plants where the mostly cold-rolled metal-strip as the substrate is handled in coils up to 40 tons unwind before and rolled up again after passing the processing plant which includes cleaning, annealing, hot-dip galvanizing / aluminizing and chemical treatment. In the liquid Zn, Zn-AI, AI-Zn and AI-Si bathes a combined action of corrosion and wear under high temperature and high stress onto the transfer components (rolls) accounts for major economic losses. Most critical here are the bearing systems of these rolls operating in the liquid system. Rolls in liquid system can not be avoided as they are needed to transfer the steel-strip into and out of the crucible. Since several years, ceramic roller bearings are tested here [1.2], however, in particular due to uncontrollable Slag-impurities within the hot bath [3], slide bearings are still expected to be of a higher potential [4]. The today's state of the art is the application of slide bearings based on Stellite\ulcorneragainst Stellite which is in general a 50-60 wt% Co-matrix with incorporated Cr- and W-carbides and other composites. Indeed Stellite is used as the bearing-material as of it's chemical properties (does not go into solution), the physical properties in particular with poor lubricating properties are not satisfying at all. To increase the Sliding behavior in the bearing system, about 0.15-0.2 wt% of lead has been added into the hot-bath in the past. Due to environmental regulations. this had to be reduced dramatically_ This together with the heavily increasing production rates expressed by increased velocity of the substrate-steel-band up to 200 m/min and increased tractate power up to 10 tons in modern plants. leads to life times of the bearings of a few up to several days only. To improve this situation. the Mechanical Alloying (MA) TeChnique [5.6.7.8] is used to prOduce advanced Stellite-based bearing materials. A lubricating phase is introduced into Stellite-powder-material by MA, the composite-powder-particles are coated by High Energy Milling (HEM) in order to produce bearing-bushes of approximately 12 kg by Sintering, Liquid Phase Sintering (LPS) and Hot Isostatic Pressing (HIP). The chemical and physical behavior of samples as well as the bearing systems in the hot galvanizing / aluminizing plant are discussed. DependenCies like lubricant material and composite, LPS-binder and composite, particle shape and PM-route with respect to achievable density. (temperature--) shock-reSistibility and corrosive-wear behavior will be described. The materials are characterized by particle size analysis (laser diffraction), scanning electron microscopy and X-ray diffraction. corrosive-wear behavior is determined using a special cylinder-in-bush apparatus (CIBA) as well as field-test in real production condition. Part I of this work describes the initial testing phase where different sample materials are produced, characterized, consolidated and tested in the CIBA under a common AI-Zn-system. The results are discussed and the material-system for the large components to be produced for the field test in real production condition is decided. Outlook: Part II of this work will describe the field test in a hot-dip-galvanizing/aluminizing plant of the mechanically alloyed bearing bushes under aluminum-rich liquid metal. Alter testing, the bushes will be characterized and obtained results with respect to wear. expected lifetime, surface roughness and infiltration will be discussed. Part III of this project will describe a second initial testing phase where the won results of part 1+11 will be transferred to the AI-Si system. Part IV of this project will describe the field test in a hot-dip-aluminizing plant of the mechanically alloyed bearing bushes under aluminum liquid metal. After testing. the bushes will be characterized and obtained results with respect to wear. expected lifetime, surface roughness and infiltration will be discussed.

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The Change of Christian Pre-Service Early Childhood Teachers through Development of Bible-Based Early Childhood Language Education Activities (성경에 기초한 유아 언어 교육 활동 개발을 통한 기독 예비 유아 교사의 변화)

  • Kim, Min-Jung
    • Journal of Christian Education in Korea
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    • v.61
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    • pp.165-201
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    • 2020
  • The purpose of this study is to explore the development of language education of Christian early childhood education by exploring the change of pre-Christian preschool teachers through the development of biblical language education activities. Interviews, surveys, action plans, and reflections of 19 Christian education students who participated in the development of bible-based language education activities for children based on the language of early childhood language, were conducted from September 3 to December 28, 2018. The data were collected through a portfolio. By analyzing the collected data, the key categories were derived and categorized. For the objectification of data analysis and interpretation, two thematic and early childhood education specialists were identified. As a result, the preparatory Christian teacher experience for the development of bible-based langage education activities for young children was categorized into cognitive change, personality change and practical change. First, through the development of bible-based early childhood language education activities, Christian pre-service early childhood teachers have brought cognitive changes as 'processes not outcomes', 'integration not separation', 'living non-curriculum' and 'meaning not effect'. In developing bible-based language education activities for young children, Christian pre-service early childhood teachers experienced a cognitive change in the 'process of language education activities' rather than the developmental achievements and results of early childhood language education. Christian pre-service early childhood teachers recognized the necessity of 'integration of listening-speaking-reading-writing', not the separation of early childhood language education. They recognized the importance of 'informal language education in kindergarten life', as well as teacher-centered formal language education. In addition, they have made a cognitive change that 'child-centered meaningful language education experience' is more important than the effectiveness of early childhood language education. Second, in developing bible-based language education activities for young children, Christian pre-service early childhood teachers showed personality changes as 'confident teachers', 'professional teachers', and 'teachers with reflective thoughts and attitudes'. Finally, in developing bible-based language education activities for young children, Christian pre-service early childhood teachers recognized the power of positive language and practiced it to form habits of using the right language and to link Christian education with early childhood education. Through the development of bible-based language education activities for young children, Christian pre-service early childhood teachers are equipped with the heart attitude and enthusiasm required to become true early childhood teachers for young children in unpredictable educational conditions and rapidly changing educational realities. Teacher efficacy has improved. In the future, it is expected that various teacher education programs linking Christian education and early childhood education will be continuously and systematically implemented.

Reset of Cultivation Zones of Green Manure Crops under Paddy Field in South Korea: Focused on Minimum Temperature and Drainage Class (국내 주요 녹비작물 논재배 적지 재설정: 1월 평균 최저기온 및 배수등급을 기준으로)

  • Kim, Min-Tae;Kim, Kwang Seop;Park, Ki Do;Ryu, Jin-Hee;Choi, Jong-Seo;Ku, Ja-Hwan;Kim, Suk-Jin;Lee, Choon-Woo;Lee, Kyungdo;Kang, Hang-Won
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.1
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    • pp.91-96
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    • 2015
  • Stable production of green manure crop is one of the critical factor for successful implementation of environmental-friendly agriculture in South Korea. Most green manure crops including hairy vetch cultivated in South Korea must be passed cold winter season. This study was conducted to establishment of cultivation zones on green manure crops under paddy field in South Korea using on minimum temperature and drainage class. In order to build the spatial distribution map, hairy vetch, triticale, barley, rye, and clover as green manure crop were selected and mean of daily minimum temperature and drainage class in paddy field were considered as limiting factor for stable growth of green manure crop. And cultivation zone on green manure crops was divided into two categories; 'Availability area' and 'Safety area'. Low limit temperature on green manure crops in this study was the following. Temperature on 'Availability area' was $-10^{\circ}C$, $-8^{\circ}C$, $-6^{\circ}C$, and $-20^{\circ}C$ for hairy vetch (including triticale), barley, clover, and rye respectively. And temperature on 'Safety area' was lower $-2^{\circ}C$ compared to that of 'Availability area' exception rye which was the same degree. Rye was expected to safely cultivated the almost whole paddy field (99.71%) of South Korea. 'Safety area' among the whole paddy field for hairy vetch, barley, and clover were 86.58, 53.06, and 15.76% respectively. The spatial distribution map of green manure crops reestablished in this study could be used the policy and agricultural management for environmental-friendly agriculture using green manure crops in South Korea.

Comparisons of Growth, Heading and Grain Filling Characteristics between Wet-hill-seeding and Transplanting in Rice (벼 무논점파재배와 이앙재배의 생육, 출수 및 등숙특성 비교)

  • Shon, Ji-Young;Lee, Chung-Kuen;Kim, Jun-Hwan;Yoon, Young-Hwan;Yang, Won-Ha;Choi, Kyung-Jin;Choi, Min-Gyu;Park, Hong-Kyu;Ko, Jong-Cheol;Kim, Yeon-Gyu;Kim, Chung-Kon;Yang, Woon-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.57 no.2
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    • pp.151-159
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    • 2012
  • Direct seeding of rice is a time- and laborsaving method, compared to machine transplanting. However, the area planted to direct-seeded rice is decreasing because of instability of seedling establishment, lodging, occurrence of weedy rice, and deterioration of grain quality. A newly developed wet-hill-seeding of rice, a type of direct seeding method, has shown features of growth similar to machine transplanted rice. This study was carried out to compare tillering pattern, heading characteristics and grain filling traits between transplanting and wet-hill-seeding in rice. Tillering of wet-hill-seeded rice was more vigorous than that of transplanted rice showing maximum tiller number 30~100% higher than transplanted rice. Wet-hill seeded rice showed later heading and shorter heading duration than transplanted rice. To find out the relationship between heading characteristics and grain filling traits, every panicle was tagged for flowering date, and the panicles on each heading date were inspected for grain filling traits. Wet-hill seeded rice and transplanted rice exhibited no significant difference in culm length, panicle length, the percentage of grain filling and perfect brown rice on each heading date during total heading period. Therefore, we conclude that wet-hill-seeding method is not inferior to machine transplanting in terms of seedling establishment, growth, grain filling and head rice yield.

Effect of Grass Filter Strips on NO3-N in Runoff from Forage Cropland (사료작물 재배지에서 초지식생대를 이용한 NO3-N 저감효과에 관한 연구)

  • Jo, Nam-Chul;Kim, Won-Ho;Seo, Sung;Yoon, Sei-Hyung;Lee, Ki-Won;Choi, Ki-Choon;Jung, Min-Woong
    • Journal of Animal Science and Technology
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    • v.53 no.1
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    • pp.51-57
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    • 2011
  • The performance of grass filter strips (GFS) in abating $NO_3$-N concentrations from the forage cropland was tested in an experiment on the 10% slope in Grassland and Forages Division, National Institute of Animal Science, Rural Development Administration (RDA) from October 2007 to September 2009. Forage croplands with rye-corn double cropping system applied with chemical fertilizer and livestock manure (LM) were compared in a natural condition. The plots were hydrologically isolated in a randomized block layout of 3 treatments $\times$ 2 factors $\times$ 3 replicates. Main plots consisted of the length of GFS, such as 0 m, 5 m, 10m and 15m. Sub plots consisted of the type of LM, such as chemical fertilizer (CF), cattle manure (CM) and swine manure (SM). Dry matter yields of rye and corn increased significantly in order of CF > CM > SM (p<0.05). Concentrations of $NO_3$-N in surface runoff water were reduced as the length of GFS increased. Especially, GFS with 10 m and 15m reduced $NO_3$-N concentrations significantly compared to that with 0 m and 5 m (p<0.05). The results from this study suggest that GFS improved the removal and trapping of manure nutrients from forage croplands.

Pollution Characteristics of PM2.5 Observed during Winter and Summer in Baengryeongdo and Seoul (겨울 및 여름철 백령도와 서울에서 측정한 PM2.5 오염 특성)

  • Yu, Geun-Hye;Park, Seung-Shik;Park, Jong Sung;Park, Seung Myeong;Song, In Ho;Oh, Jun;Shin, Hye Jung;Lee, Min Do;Lim, Hyung Bae;Kim, Hyun Woong;Choi, Jin Young
    • Journal of Korean Society for Atmospheric Environment
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    • v.34 no.1
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    • pp.38-55
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    • 2018
  • Hourly measurements of $PM_{2.5}$ mass, organic and elemental carbon (OC and EC), and water-soluble ionic species were made at the air quality intensive monitoring stations in Baengryeongdo (BR) and Seoul (SL) during the winter (December 01~31, 2013) and summer (July 10~23, 2014) periods, to investigate the increase of $PM_{2.5}$ and secondary ionic species and the reasons leading to their increase during the two seasons. During winter, $PM_{2.5}$ and its major chemical species concentrations were higher at SL than at BR. Contribution of organic mass to $PM_{2.5}$ was approximately 1.7 times higher at BR than at SL, but the $NO_3{^-}$ contribution was two times higher at SL. Total concentration of secondary ionic species ($SO{_4}^{2-}$, $NO_3{^-}$, and $NH_4{^+}$) at BR and SL sites accounted for 29.1 and 40.1% of $PM_{2.5}$, respectively. However, during summer, no significant difference in chemical composition of $PM_{2.5}$ was found between the two sites with the exception of $SO{_4}^{2-}$. Total concentration of the secondary ionic species constituted on average 43.9% of $PM_{2.5}$ at BR and 53.0% at SL. A noticeable difference in chemical composition between the two sites during summer was attributed to $SO{_4}^{2-}$, with approximately twofold concentration and 10% higher contribution in SL. Low wind speed and high relative humidity were important factors in secondary formation of water-soluble ionic species during winter at SL, resulting in $PM_{2.5}$ increase. While the secondary formation during summer was attributed to strong photochemical processes in daytime and high relative humidity in nighttime hours. The increase of $PM_{2.5}$ and its secondary ionic species during the winter haze pollution period at SL was mainly caused either by long-range transport (LTP) from the eastern Chinese regions, or by local pollution. However, the increased $SO{_4}^{2-}$ and $NO_3{^-}$ during summer at SL were mainly caused by LTP, photochemical processes in daytime hours, and heterogeneous processes in nighttime hours.

Ultrasonographic Measurement of Articular Cartilage Thickness of Medial Femoral Condyle in Knee Osteoarthritis (슬관절 골관절염에서 초음파를 이용한 대퇴 내과 관절 연골의 두께 측정)

  • Kim, Jung-Man;Kim, Tae-Hyung;Im, Dong-Sun;Kang, Min-Gu;Lee, Kyu-Jo
    • The Journal of Korean Orthopaedic Ultrasound Society
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    • v.4 no.1
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    • pp.7-14
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    • 2011
  • Purpose: The purpose of this study was to analyze the correlation between the medial joint space on weight bearing simple X-ray and the ultrasonographic articular cartilage thickness. Materials and Methods: The articular cartilage thickness of 91 osteoarthritic knees of 73 patients were measured with weight bearing simple X-ray and ultrasonography between June 2010 and September 2010. Male were 13 and female were 60. Right knees were 35, left knees were 20 and bilateral involvements were 18. Medial joint spaces on X-ray were measured in full extension view and $45^{\circ}$ PA(Rosenberg) view. Femoral cartilage thicknesses at and those at the point between the middle 1/3 and posterior 1/3 area were measured by ultrasonography, that were scanned coronally at $30^{\circ}$ flexion and $130^{\circ}$ flexion respectively. The results were analyzed statistically by Pearson correlation test and Bland-Altman plot test. Results: Medial joint spaces measured in full extension view and femoral articular cartilage thicknesses at the point between the anterior 1/3 and middle 1/3 area measured with sonography were statistically correlated and medial joint spaces measured in $45^{\circ}$ PA view and those at the point between the middle 1/3 and posterior 1/3 area measured with sonography were also correlated (P<0.05, $SD{\pm}2$). Conclusion: The measurement of articular cartilage thicknesses by the ultrasonography was easy even in the patient who are unable to take weight bearing view and was thought to be a useful diagnostic and follow up method to examine the degenerative change in addition to simple radiographic examination in the knee osteoarthritis.

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The immunogenicity and reactogenicity of Td booster vaccination in Korean preadolescents, aged with 11-12 years old (국내 11-12세 소아에서 Td 백신 추가접종의 면역원성과 안전성 평가)

  • Lee, Soo Young;Kwak, Ga Young;Mok, Hye Rin;Kim, Jong Hyun;Hur, Jae Kyun;Lee, Kyung Il;Park, Joon Su;Ma, Sang Hyuk;Kim, Hwang Min;Kang, Jin Han
    • Clinical and Experimental Pediatrics
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    • v.51 no.11
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    • pp.1185-1190
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    • 2008
  • Purpose : This study was undertaken to evaluate the immunogenicity and reactogenicity of Td booster immunization in early preadolescents of Korea. Methods : Healthy preadolescents, who had been vaccinated with 4 or 5 doses of DTaP vaccines until 6 years old age, were enrolled in this study from August 2006 to April 2007. Diphtheria and tetanus anti-toxoid antibodies in sera were measured by ELISA just before vaccination and 4 weeks after vaccination to evaluate immunogenicity. Local and systemic adverse reactions observed for 4 weeks after vaccination to access reactogenicity. Results : 183 preadolescents were enrolled and mean age was $11.40{\pm}0.51$ years old. All subjects achieved seroprotective diphtheria and tetanus anti-toxoid antibodies (titers ${\geq}0.1IU/mL$) after Td booster vaccination. Among 183 vaccinees, 73.8% showed local adverse reactions and 37.2% systemic adverse reactions. Pain at injection site (66.1%) was the most common local reaction, and the most commonly shown systemic reaction was myalgia (17.5%). The adverse reactions were spontaneously relieved within three days after vaccination. Conclusion : Td vaccine in this study was high immunogenic and showed an acceptable tolerance in Korean preadolescents. Td booster vaccination at 11-12 years old is the most effective method to increase compliance of the vaccination and to decrease the incidence of diphtheria and tetanus.

Na, K-ATPase Activity in the Aged Erythrocytes of Hypertensive Rats (고혈압쥐 노화 적혈구에서의 Na, K-ATPase에 관한 연구)

  • Park, Chang-Kil;Hur, Gang-Min;Seok, Jung-Ho;Lee, Jae-Heun
    • The Korean Journal of Pharmacology
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    • v.27 no.1
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    • pp.33-43
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    • 1991
  • To study the age dependent change of Na, K-ATPase in the erythrocyte of hypertensive rat, 1-kidey 1-clip hypertensive rat was made by the removal of right kidney and partial ligation of left renal artery. After 4 weeks, aged erythrocyte fraction was separated by density gradient centrifugation, and Na, K-ATPase activity and $^3H-ouabain$ binding with ghost cell membrane and ouabain sensitive Rb-uptake with whole cell were measured. 1) In the hypertensive rats, blood pressure was significantly increased to 165.5/119.0 mmHg (systolic/diastolic). Mean corpuscular volume and membrane protein(mg) per $10^9RBC$ were decreased and hemoglobin content was increased in the aged erythrocyte. 2) Na, K-ATPase activity in the solution containing 110 mM NaCl and 10 mM KCI, was decreased in hypertensive rat, and decreased in aged erythrocyte of both group. 3) Ouabain sensitive Rb-uptake by low RbCl concentration(4 mM) was slightly decreased in aged erythrocyte compared to that in young erythrocyte of each group, but slightly increased in young erythrocyte in hypertensive rat compared to that in normotensive rat. 4) Ouabain sensitive Rb-uptake by high RbCl concentration(16 mM) was decreased about 30% to 50 % in aged erythrocyte in both group. And in hypertensive rat, especially in young erythrocyte it was significantly decreased compared to that in normotensive rats. 5) $^3H-ouabain$ binding at 0.13 or $1{\times}10^-6M$ ouabain concentration was slightly decreased in aged erythrocyte of normotensive rat, and significantly decreased in aged erythrocyte of hypertensive rats. 6) $^3H-ouabain$ binding at 6 or $64{\times}10^-6M$ ouabain concentration is slightly decreased in aged erythrocyte of both group, but significantly decreased in young and aged erythrocyte of hypertensive rats compared to that of normotensive rats. The present results suggest that (1) in the young erythrocyte of hypertensive rat, the alterations of Na-pump activity that slightly increased in weak stimulation and inhibited in strong stimulation, may be related to increased molecular activity and the decrease in the number of low affinity site without change in high affinity site, (2) in the aged erythrocyte of normotensive rat, inhibited Na-pump may be related to the change in molecular activity of pump. (3) And in the aged erythrocyte of hypertensive rat, it may be related to the decrease in the number of high and low affinity site as well as the change in molecular activity

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