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Effect of Fusion Procedure on the Development of Embryos Produced by Somatic Cell Nuclear Transfer in Hanwoo (Korean Cattle) (한우에서 융합방법이 체세포 핵이식 수정란의 발달에 미치는 영향)

  • Im, G.S.;Yang, B.S.;Park, S.J.;Chang, W.K.;Park, C.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.365-373
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    • 2000
  • The purpose of this study was to investigate the effects of the fusion pulses and fusion media on fusion rate and the development of embryos produced by somatic cell nuclear transfer in Hanwoo (Korean cattle). Nuclear donor cumulus and fetal fibroblast cells were cultured in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at 38.5$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. The in vitro matured oocytes were enucleated and then the isolated donor cells were introduced. The cumulus cell and cytoplast were fused using one pulse of 70 volts for 40$mutextrm{s}$, two pulses of 70 volts for 40$mutextrm{s}$ and one pulse of 180 volts for 15$mutextrm{s}$. The fetal fibroblast cell and cytoplast were fused using one pulse of 180 volts for 15$mutextrm{s}$ or 30$mutextrm{s}$. The cumulus cell and cytoplast were fused using mannitol and Zimmerman cell fusion medium (ZCFM) as a fusion medium. The fused embryos were activated after the fusion with 10 $\mu$M calcium ionophore for 5 min and 2 mM 6-dimethyl- aminopurine for 3 h. The nuclear transfer embryos were cultured in 500 ${mu}ell$ well of modified CR1aa supplemented with 3 mg/$m\ell$ BSA in th $\varepsilon$ four well dish cove red with mineral oil. After 3 days culture, culture medium was changed into modified CRlaa medium containing 1.5 mg/$m\ell$ BSA and 5% FBS for 4 days. The incubation environment was 5% $CO_2$, 5% $O_2$, 90% $N_2$ at 38.5$^{\circ}C$. When the cumulus cells were fused with enucleated oocytes by three different fusion pulses, one pulse of 180 volts for 15 $mutextrm{s}$ yielded the highest fusion rate and developmental rate to blastocyst among the pulses (P<0.05). When the fetal fibroblast cells were fused with enucleated oocytes, one pulse of 180 volts for 30$mutextrm{s}$ yielded significantly higher fusion rate compared with that for 15 $mutextrm{s}$(P<0.05). The present result indicates that the fusion rate between karyoplast and cytoplast was affected by the cell type and the optimal fusion condition was different according to cell type or size. When the fusion was conducted by the use of mannitol and ZCFM, the fusion rate was 71.2% and 65.8%, respectively. The developmental rates to blastocyst were 37.8% and 39.8%, respectively. There was no significant difference between two fusion media in the developmental rate of cumulus cell nuclear transfer embryos. These results indicate that optimal electric current should be selected according to cell type.

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"Jungmo2510", Forage Rye Cultivar of Early-Heading and Resistance to Lodging (조숙성이고 도복에 강한 사일리지용 호밀 품종 '중모2510' 개발)

  • Han, O.K.;Ku, J.H.;Ahn, J.W.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.21 no.1
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    • pp.61-70
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    • 2019
  • "Jungmo2510", a rye cultivar, Secale cereal L., was developed by National Institute of Crop Science, RDA in 2015. It was developed from open pollination from within 10 rye cultivars or lines including "Chochun" in 1995. The line "SR95POP-S1-523-1-5-5-4-7-3-B-16-3-19" was selected for its excellent agronomic appearance and was placed in yield trials for two years from 2011 to 2012. The line was designated "Homil55" and was placed in regional yield trials at the four locations around Korea from 2013 to 2015, during which time the name "Jungmo2510" was given. This cultivar is an erect plant type and leaves of short and broad size with a green color, a yellow colored, medium-diameter culm, and a yellowish brown-colored, medium-size grain. The heading date of "Jungmo2510" was April 16, which were 2 days earlier than that of "Gogu". "Jungmo2510" also showed similar to winter hardiness and greater resistance to lodging compared to those of the check cultivar. Over three years, the average dry matter yield of "Jungmo2510" was 802 kg 10a-1 , which was harvested in late April and was lower than that of the check cultivar "Gogu" (825 kg). The seed productivity of "Jungmo2510" was approximately 481 kg 10a-1 , which was 2.4% less than that of the check. "Jungmo2510" was higher to than "Gogu" in term of protein content (9.1% and 8.0%, respectively), total digestible nutrients(TDN)(57.5% and 55.5%, respectively), and TDN yield 10a-1(419 kg and 392 kg, respectively). This cultivar is recommended as a fall sowing crop in areas where the average daily minimum-mean temperatures are higher than -12 ℃ in January, and as a winter crop for whole-crop forage before the planting of rice or green manure around Korea.

Effects of Dietary Germanium Biotite in Weaned, Growing and Finishing Pigs (이유자돈, 육성돈 및 비육돈에 있어 게르마늄흑운모의 급여 효과)

  • Kwon, O.S.;Kim, I.H.;Hong, J.W.;Lee, S.H.;Jung, Y.K.;Min, B.J.;Lee, W.B.;Shon, K.S.
    • Journal of Animal Science and Technology
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    • v.45 no.3
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    • pp.355-368
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    • 2003
  • In Exp. 1, this study was conducted to determine the effect of dietary germanium biotite on growth performance and nutrient digestibility in nursery pigs. A total of sixty crossbred pigs (initial body weight 15.09$\pm$0.18kg) were used in this experiment. This study was carried out for 28 days. The five treatments were control (CON; basal diet), GB0.1 (basal diet + germanium biotite 0.1%), GB0.3 (basal diet + germanium biotite 0.3%), GB0.6 (basal diet + germanium biotite 0.6%) and GB1.0 (basal diet + germanium biotite 1.0%). For overall period, ADG and Gain/feed were not significantly different among the treatments. In Exp. 2, a study was conducted to evaluate the effect of germanium biotite as a substitute for antibiotics in growing pigs. A total of fifty five crossbred pigs (initial body weight 32.47$\pm$0.9kg) were used in this experiment. The three treatments were negative control (NC: basal diet without antibiotic), positive control (PC: basal diet + 200ppm CTC) and GB0.3 (basal diet + germanium biotite 0.3%). Pigs fed PC (17%, 385 vs 451 g/d) and GB0.3 (14%, 385 vs 438 g/d) diets grew faster(P<0.05) than pigs fed NC diet. Pigs fed PC and GB0.3 diets resulted higher(P<0.05) ADFI than pigs fed CON diet. However, pigs fed GB0.3 diet had improved gain/feed compared to pigs fed NC diet(P<0.05). Apparent digestibility of DM and N by pigs fed PC and GB0.3 diets were greater(P<0.05) than those by pigs fed NC diet. In Exp. 3, a study was conducted to determine the effect of dietary germanium biotite on growth performance, plasma characteristics, backfat thickness and fecal ammonia gas concentration in finishing pigs. A total of seventy-two finishing pigs (initial body weight 78.56$\pm$1.32kg) were used in this experiment. The treatments included 1) Control (CON; basal diet) 2) GB1.0 (basal diet + germanium biotite 1.0%), 3) GB3.0 (basal diet + germanium biotite 3.0%). Pigs fed GB1.0 diet grew faster than pigs fed CON diet and GB0.3 diet (P<0.05). Also, pigs fed CON diet showed higher(p<0.05) ADFI than pigs fed GB3.0 diet. Pigs fed GB diets had improved gain/feed compared to pigs fed CON diet(P<0.05). Total?and VLDL concentrations in plasma of pigs fed GB diets treatments were significantly decreased compared to those in pig fed CON diet(P<0.05). However, HDL-cholesterol concentration in plasma of the pig was significantly increased compared to those in pigs fed CON diet (P<0.05). Pigs fed CON diet exerted higher(P<0.05) backfat thickness than pigs fed GB1.0 (5.4%, 27.19 vs 25.71mm) and GB3.0 (16.1%, 27.19 vs 22.81mm) diets. Feces from CON treatment were higher in fecal ammonia gas concentration than faces from pigs fed GB1.0 (64.1%, 17.00 vs 6.10mg/kg)and GB3.0 (61.8%, 17.00 vs 6.50mg/kg) treatments(P<0.05). In conclusion, the results suggest that the dietary addition of germanium biotite into diets for nursery pigs did not affect growth performance. The results also suggest the possibility of germanium biotite to replace antibiotic in diets for growing pigs. In finishing pigs, dietary supplementation of germanium biotite was an effective means for improving growth performance and for decreasing Total-and LDL+VLDL-plasma cholesterols, backfat and fecal ammonia gas concentration.

Radiosynthesis of $[^{11}C]6-OH-BTA-1$ in Different Media and Confirmation of Reaction By-products. ($[^{11}C]6-OH-BTA-1$ 조제 시 생성되는 부산물 규명과 반응용매에 따른 표지 효율 비교)

  • Lee, Hak-Jeong;Jeong, Jae-Min;Lee, Yun-Sang;Kim, Hyung-Woo;Lee, Eun-Kyoung;Lee, Dong-Soo;Chung, June-Key;Lee, Myung-Chul
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.3
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    • pp.241-246
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    • 2007
  • Purpose: $[^{11}C]6-OH-BTA-1$ ([N-methyl-$^{11}C$]2-(4'-methylaminophenyl)-6-hydroxybenzothiazole, 1), a -amyloid imaging agent for the diagnosis of Alzheimer's disease in PET, can be labeled with higher yield by a simple loop method. During the synthesis of $[^{11}C]1$, we found the formation of by-products in various solvents, e.g., methylethylketone (MEK), cyclohexanone (CHO), diethylketone (DEK), and dimethylformamide (DMF). Materials and Methods: In Automated radiosynthesis module, 1 mg of 4-aminophenyl-6-hydroxybenzothiazole (4) in 100 l of each solvent was reacted with $[^{11}C]methyl$ triflate in HPLC loop at room temperature (RT). The reaction mixture was separated by semi-preparative HPLC. Aliquots eluted at 14.4, 16.3 and 17.6 min were collected and analyzed by analytical HPLC and LC/MS spectrometer. Results: The labeling efficiencies of $[^{11}C]1$ were $86.0{\pm}5.5%$, $59.7{\pm}2.4%$, $29.9{\pm}1.8%$, and $7.6{\pm}0.5%$ in MEK, CHO, DEK and DMF, respectively. The LC/MS spectra of three products eluted at 14.4, 16.3 and 17.6 mins showed m/z peaks at 257.3 (M+1), 257.3 (M+1) and 271.3 (M+1), respectively, indicating their structures as 1, 2-(4'-aminophenyl)-6-methoxybenzothiazole (2) and by-product (3), respectively. Ratios of labeling efficiencies for the three products $([^{11}C]1:[^{11}C]2:[^{11}C]3)$ were $86.0{\pm}5.5%:5.0{\pm}3.4%:1.5{\pm}1.3%$ in MEK, $59.7{\pm}2.4%:4.7{\pm}3.2%:1.3{\pm}0.5%$ in CHO, $9.9{\pm}1.8%:2.0{\pm}0.7%:0.3{\pm}0.1%$ in DEK and $7.6{\pm}0.5%:0.0%:0.0%$ in DMF, respectively. Conclusion: The labeling efficiency of $[^{11}C]1$ was the highest when MEK was used as a reaction solvent. As results of mass spectrometry, 1 and 2 were conformed. 3 was presumed.

Study on Spring Cocoon Crops with the Leaf Produced in the Mulberry Field close to the Totacco Field (개량 Mulching 담배밭 부근뽕잎이 춘잠작에 미치는 영향에 관한 연구)

  • 이상풍;김정배;김계명;박광준
    • Journal of Sericultural and Entomological Science
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    • v.16 no.1
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    • pp.67-75
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    • 1974
  • The studies are to know how much cocoon crops is damaged by the stained leaf with nicotine produced from the tobacco field cultivated in mulching system in spring season and by residual nicotine in autumn season. Furthermore, the new knowledges are to make both industries keep up with their development. In spring season mulberry Held is located higher on the West-North of tobacco held below 20 degrees of slope and with 36 per cent of East-South wind and 18 per cent of South wind blowing from tobacco fold to the mulberry fold. In addition, silkworm larvae are fed with the mulberry leaf produced in the different plots placing by the different distances, l0m, 25m, 50m, 80m, and loom far from the tobacco Held as a control and it is also considered that narcotic larvae including the dead larvae are not observed. On the other hand, it is noted that better leaf quality and abundant growth of mulberry tree is produced from the mulberry fold closer to the tobacco field and with a low slope. 1) Maximum weight of larval body at the 5th stage is damaged by the stained leaf with the nicotine up to 25m far from the tobacco held. 2) The larvae fed with the mulberry leaf in mulberry Held up to 25m far from the tobacco fold produce small number of the fresh cocoons per 1 liter. 3) Low single cocoon weight and low cocoon shell weight are produced by the poison damaged larvae fed with the mulberry. leaf up to 25m far from the tobacco field and weight of cocoon shell is damaged higher than the single cocoon weight. It is resulted in low percentage of cocoon shell. 4) Cocoon yield including the double cocoon from 10,000 larvae is decreased by the larvae fed with the stained leaf in the mulberry fold up to 25m far from the tobacco fold and 19 per cent of cocoon yield is decreased with 2.4kg of cocoon yield in l0m plot and with 2.5kg of cocoon yield in 25m plot at the first season and at the 2nd season with 1.8kg o( cocoon yield in l0m plot and with 11.5kg of cocoon yield in 25m plot, 11 per cent and 9 per cent of cocoon yield including double cocoon from 10,000 larvae is decreased, as compared with the control, respectively. With these results, it is observed that nicotine damage is occurred to the silkworm larvae if the larvae are fed with the leaf in the mulberry Held within 25m-50m far from the tobacco field.

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A Study on pluralistic Reformation for Education of Telecommunication -for Establishment of Individual System for Comm. Education- (통신교육의 계열화와 계층화 -고유한 교역의 형성을 위하여-)

  • 조정현
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.3 no.1
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    • pp.28-30
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    • 1978
  • Communication actions as a social band or Human community stick (fast) to human being ceaselessly w without stopping everywhere. All of comm. actions can be kept up and developed by the education of its own. Comm. actions have to include a character of social process, and so for it the social science should t to be some essential part of it. Therefore, Comm. education have to be schemed for achieving with a point of view of synthetical s science including technical and social factor. However, recentry Comm. education be suffered to lose of itowns essential attribute and individual i independence becausing to reduce social weight recklessly in their education It is a prindiple that Comm. science is an integrate science being composed of human, social and t technical subdepartments and so comm. education have to obey for Comm. constuctional theory, i international and social claim. Originally in Korea a educational idea and genealogy forming by the comm. scientific theory has I inherited on orthodoxy. But in 1961, communication college that is only the orthodox model of Comm. education, was f forced to close by some reckless policy and then the national administration for the Comm. education h have been weakened, and so recently it’s education became to degenerate as out of genealogy or n nonsystem alike some scattering Family. On the other side, today comm. science make to it’s modern scientific factor and to keep its l integrate level, therefore, all of educational provisions and administration for the telecomm. should t to be supplement to be fit for their plural chatacters. Comm. education have to occupy an individual educational system through the comm. theory, and t then it can be coexisted with neighbour scientific field equally and can include, connect coordinate o or effect its inference in each subfactor organically. Finally, educational system for telecommunication should to be requested as preeedence that i independent field including pluralism must be formed and sufficient autonomy be guarenteed, and s so Comm. education must be to restored its orthodox genealogy and be recovered individual system a and seIfrestraint field, and then it can be accomplished its own duty for nation and society.

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Short-Term Efficacy of Steroid and Immunosuppressive Drugs in Patients with Idiopathic Pulmonary Fibrosis and Pre-treatment Factors Associated with Favorable Response (특발성폐섬유화증에서 스테로이드와 면역억제제의 단기 치료효과 및 치료반응 예측인자)

  • Kang, Kyeong-Woo;Park, Sang-Joon;Koh, Young-Min;Lee, Sang-Pyo;Suh, Gee-Young;Chung, Man-Pyo;Han, Jung-Ho;Kim, Ho-Joong;Kwon, O-Jung;Lee, Kyung-Soo;Rhee, Chong-H.
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.5
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    • pp.685-696
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    • 1999
  • Background : Idiopathic pulmonary fibrosis (IPF) is a diffuse inflammatory and fibrosing process that occurs within the interstitium and alveolus of the lung with invariably poor prognosis. The major problem in management of IPF results from the variable rate of disease progression and the difficulties in predicting the response to therapy. The purpose of this retrospective study was to evaluate the short-term efficacy of steroid and immunosuppressive therapy for IPF and to identify the pre-treatment determinants of favorable response. Method : Twenty patients of IPF were included. Diagnosis of IPF was proven by thoracoscopic lung biopsy and they were presumed to have active progressive disease. The baseline evaluation in these patients included clinical history, pulmonary function test, bronchoalveolar lavage (BAL), and chest high resolution computed tomography (HRCT). Fourteen patients received oral prednisolone treatment with initial dose of 1mg/kg/day for 8 to 12 weeks and then tapering to low-dose prednisolone (0.25mg/kg/day). Six patients who previously had experienced significant side effects to steroid received 2mg/kg/day of oral cyclophosphamide with or without low-dose prednisolone. Follow-up evaluation was performed after 6 months of therapy. If patients met more than one of followings, they were considered to be responders : (1) improvement of more than one grade in dyspnea index, (2) improvement in FVC or TLC more than 10% or improvement in DLco more than 20% (3) decreased extent of disease in chest HRCT findings. Result : One patient died of extrapulmonary cause after 3 month of therapy, and another patient gave up any further medical therapy due to side effect of steroid. Eventually medical records of 18 patients were analyzed. Nine of 18 patients were classified into responders and the other nine patients into nonresponders. The histopathologic diagnosis of the responders were all nonspecific interstitial pneumonia (NSIP) and that of nonresponders were all usual interstitial pneumonia (UIP) (p<0.001). The other significant differences between the two groups were female predominance (p<0.01), smoking history (p<0.001), severe grade of dyspnea (p<0.05), lymphocytosis in BAL fluid ($23.8{\pm}16.3%$ vs $7.8{\pm}3.6%$, p<0.05), and less honeycombing in chest HRCT findings (0% vs $9.2{\pm}2.3%$, p<0.001). Conclusion : Our results suggest that patients with histopathologic diagnosis of NSIP or lymphocytosis in BAL fluid are more likely to respond to steroid or immunosuppressive therapy. Clinical results in large numbers of IPF patients will be required to identify the independent variables.

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A study of $Smartpeg^{TM}'s$ lifetime according to sterilization for implant stability (임플랜트 안정성을 위한 자기공명막대의 소독방법에 따른 수명에 관한 연구)

  • Won, Ho-Yeon;Cho, In-Ho;Lee, Joon-Seok
    • The Journal of Korean Academy of Prosthodontics
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    • v.46 no.1
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    • pp.42-52
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    • 2008
  • Purpose: Resonance Frequency Analysis(RFA) technique can be used as an effective method in measuring the implant stability and documenting the clinical results. This technique also determines how stable the implant is before performing a prosthetic practice. Having become one the guidelines of the implant therapy whose final objective is the immediate loading, the $Osstell^{TM}$ mentor is giving a lot of information to the clinicians recently. In this communication, experiments were performed to investigate how reliable the measured ISQ values by $Osstell^{TM}$ mentor are, and to see if those are also stable even after sterilization. As five objectives: 1) How stable measured ISQ values after fixation $Smartpeg^{TM}s$ for 400 times. 2) How stable measured ISQ values after 'attach-detach'$Smartpeg^{TM}'s$ for 400 times. 3) How stable measured ISQ values after clinical sterilization methods. 4) How stable measured ISQ values after repeatedly sterilization in autoclave for 10 times. 5) What is the critical temperature which is lost the magnetism of $Smartpeg^{TM}$. Materials and Methods: Clinical sterilization methods(Autoclave sterilization, Dentistar sterilization, Ultra violet sterilization, Vacuum dry unit sterilization, Boiling water sterilization, combined $H_{2}O_{2}$ and Alcohol sterilization).$Smartpeg^{TM}s$. D3 Block bone($3{\times}9{\times}2cm$). Osstem implant(${\emptyset}4.1$-10mm).$Osstell^{TM}$ mentor. Individual experiment was used 8 number of $Smartpeg^{TM}s$ and they had measured to ISQ values of before experiment and after experiment. Results: 1. The measured ISQ values did not change after fixation $Smartpeg^{TM}s$ for 400 times. 2. There was no significant changes in the measured ISQ values of 'attach-detach $Smartpeg^{TM}s'$ for 400 times. 3. The measured ISQ values did not change after the usual clinical sterilization methods. 4. The measured ISQ values did not change after sterilization in autoclave for 10 times. 5. It was impossible to exactly measure the critical temperature which is lost the magnetism of $Smartpeg^{TM}s$. But, the results was resulted to lost its magnetism in higher temperature than $150^{\circ}C$/10 minute. Conclusion: The measured ISQ values showed insignificant differences in case of no changes in the magnetism of the $Smartpeg^{TM}s$. It seems that the $Smartpeg^{TM}s$ can be used repeatedly in every measurement if the original magnetisms of the $Smartpeg^{TM}s$ can be recognized. There seems to be no significant changes in the measured ISQ values of 'attach-detach $Smartpeg^{TM}s'$ only if the screw pitches were unimpaired. The clinical sterilization methods seems acceptable because the result was resulted to lost its magnetism in higher temperature than $150^{\circ}C$/10minute.

Clinical Features and Treatment Response in 18 Cases with Idiopathic Nonspecific Interstitial Pneumonia (특발성 비특이성 간질성 폐렴 18례의 임상상 및 치료반응)

  • Kang, Eun-Hae;Chung, Man-Pyo;Kang, Soo-Jung;An, Chang-Hyeok;Ahn, Jong-Woon;Han, Joung-Ho;Lee, Kyung-Soo;Lim, Si-Young;Suh, Gee-Young;Kim, Ho-Joong;Kwon, O-Jung;Rhee, Chong-H.
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.4
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    • pp.530-542
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    • 2000
  • Background : Nonspecific interstitial pneumonia (NSIP) has been reported recently to have shown much better response to medical treatment and better prognosis compared with idiopathic UIP. However, clinical characteristics of idiopathic NSIP discriminating it from UIP have not been clearly defined. Method : Among 120 patients with biopsy-proven diffuse interstitial lung diseases admitted to the Samsung Medical Center between July 1996 and March 2000, 18 patients with idiopathic NSIP were included in this study. Retrospective chart review and radiographic analysis were performed. Results : 1) At diagnosis, 17 patients were female and the average age was $55.2{\pm}8.4$ years (44~73 years). The average duration from development of respiratory symptom to surgical lung biopsy was $9.9{\pm}17.1$ months. Increase in bronchoalveolar lavage fluid lymphocytes ($23.0{\pm}13.1%$) was noted. On HRCT, ground glass and irregular linear opacity were observed, but honeycombing was absent in all patients. 2) Corticosteroids were initially given to 13 patients, but the medication was stopped in 3 patients due to severe side effects. Further medical therapy was not possible in 1 patient who experienced streroid-induced psychosis. Herpes zoster (n=3), tuberculosis (n=1), avascular necrosis of the hip (n=1), cataract (n=2) and diabetes mellitus (n=1) developed during prolonged corticosteroid administration. Of the 7 patients receiving oral cyclophosphamide therapy, hemorrhagic cystitis hindered one patient from continuing with the medication. 3) After medical treatment, 14 of 17 patients improved, and 3 patients remained stable (mean follow-up ; $24.1{\pm}11.2$ months). FVC increased by $20.2{\pm}11.2%$ of predicted value and the extent of ground glass opacity on HRCT decreased significantly ($15.7{\pm}14.7%$). 4) Of the 14 patients who had stopped medication, 5 showed recurrence of NSIP and 2 became aggravated during steroid tapering. All patients with recurrence showed deterioration within one year after completion of initial treatment. Conclusion : Since idiopathic NSIP has unique clinical profiles and shows good prognosis, diagnosis different from UIP, and aggressive medical treatment are needed.

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The Increased Expression of Gelatinolytic Proteases Due to Cigarette Smoking Exposure in the Lung of Guinea Pig (기니픽에서 흡연 노출에 의한 젤라틴 분해 단백 효소의 발현 양상에 관한 연구)

  • Kang, Min-Jong;Lee, Jae-Ho;Yoo, Chul-Gyu;Lee, Choon-Taek;Chung, Hee-Soon;Seo, Jeong-Wook;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.4
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    • pp.426-436
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    • 2001
  • Background : Chronic obstructive pulmonary disease(COPD) is one of the major contributors to morbidity and mortality among the adult population. Cigarette smoking(CS) is undoubtedly the single most important factor in the pathogenesis of COPD. However, its mechanism is unclear. The current hypothesis regarding the pathogenesis of COPD postulates that an imbalance between proteases and antiproteases leads to the destructive changes in the lung parenchyma. This study had two aims. First, to evaluate the effect of CS exposure on histologic changes of the lung parenchyme, and second, to evaluate the effect of CS exposure on the expression of the gelatinolytic enzymes in BAL fluid cells in guinea pigs. Methods : Two groups of five guinea pigs were exposed to the whole smoke of 20 commercial cigarettes per day, 5 hours/day, 5 days/week, for 6weeks, and 12 weeks, respectively, using a smoking apparatus. Five age-matched guinea pigs exposed to room air were used as controls. Five or more sections were microscopically extamined(${\times}400$) and the number of cellular infiltration of the alveolar wall was measured in order to evaluate the effect of CS exposure on the histologic changes of lung parenchyme. The statistical significance was analyzed by a linear regression method. To evaluate the expression of the gelatinolytic enzymes in intraalveolar cells, BAL fluid was obtained and the intraalveolar cells were separated by centrifugation (500 g for 10 min at $4^{\circ}C$). Two sets of culture plates were loaded with $1{\times}10^6$ intraalveolar cells. One plate, contained O.1mM EDTA, a inhibitor of matrix metalloproteases(MMPs), and the other plate had no EDTA. Both plates were incubated for 48 hours at $37^{\circ}C$. After incubation, gelatinolytic protease expression in the supernatants was analyzed by gelatin zymography. Results : At the end of CS exposure, the level of blood carboxy Hb had increased significantly(4.1g/dl in control group, 24g/dl immediately after CS exposure, 18g/dl 30 min after CS exposure, 15g/dl 1 hour after CS exposure). Alveolar inflammatory cells were identified in the CS exposed guinea pigs. The number of alveolar cellular cells observed in a microscopic field ($400{\times}$) was $121.4{\pm}7.2$, $158.0{\pm}20.2$, $196.8{\pm}32.8$, in the control, the 6 weeks, and the 12 weeks group, respectively. The increased extent of inflammatory cellular infiltration of the lung parenchema showed a statistically significant linear relationship with the duration of CS exposure(p=0.001, $r^2=0.675$). Several types of gelatinolytic enzymes in the intraalveolar cells of CS exposed guinea pigs were expressed, of which some were inhibited by EDT A. However, the gelatinolytic enzymes were not expressed in the control groups. Conclusion : CS exposure increases inflammatory cellular infiltration of the alveolar wall and the expression of gelatinolytic proteases in guinea pigs. EDTA inhibits some of the gelatinolytic proteases. These findings suggest a possibility that CS exposure may increase MMP expression in the lungs of guinea pigs.

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