단계적 온도 하강법을 이용한 췌도세포 냉동보존법

정인경,오승훈,김병준,양태영,이병완,하창영,노정현,정재훈,민용기,이명식,이문규,김광원 대한당뇨병학회 2002 Diabetes and Metabolism Journal Vol.26 No.1

연구배경:최근 당뇨병의 새로운 치료법으로 시도되고 있는 췌도이식은 충분한 췌도수의 확보와 췌도생존율을 높이기 위한 면역억제제 사용이 제한점이 되고 있다. 본 연구의 목적은 이식 전 충분한 췌도 수의 확보를 위해 분리한 췌도를 냉동보존하는 방법을 확립하고 냉동보존한 백서 췌도세포의 시험관내 그리고 생체내 기능을 조사하였다. 방법:분리한 백서의 췌장소도를 48시간 배양한 후 한 시험관당 췌도세포 1000개씩 나누었다. 냉동보존은 6개의 시험관에 DMSO를 첨가한 후 초 냉각(supercooling), 핵화(nucleation)단계를 거친 후 99% isopropanol과 액체질소가 들어있는 dewer를 이용하여-0.25℃/분의 냉각속도로 -40℃까지 단계적으로 얼린후-70℃ 액체질소 탱크에 보관했다. 해동은 냉동시킨 vial들을 액체 질소 태으에서 꺼내 37℃ 항온조에 담가 급격히 해동시킨 후, 원심분리하여 상층액을 제거하고 각 vial에 0.75M sucrose 용액을 가한 후, 10% fetal calf serum이 함유된 RPMI 1640 media에서 배양하였다 각각 6개의 시험관에서 해동한 췌도들을 광학현미경 및 형광현미경하에서 췌도의 모양과 생존율에 대해 조사하고 인슐린 정적반응을 알아보았다. 또한 분리한 췌도를 냉동보존하지 않고 이식한 경우를 대조군(6마리)과 생체내 기능을 비교하였다. 결과:① 냉동보존후 획득한 췌도의 수와 생존율 해동후 획득한 췌도의 수는 해동시킨 당일날이 902±21, 24시간 배양 후에는 857±16, 72시간 후에는 817±18개로 점차 감소되었다. AO/PI 염색상 각 췌도의 생존율은 냉동 전을 100으로 하였을 때 해동당일, 24시간 후, 72시간 후가 각각 60±5, 80±5, 90±5%로 해동후 3일간 배양하였을 때 냉동전의 수준으로 회복하였다. ② 냉동보존후 췌도의 포도당에 대한 정적 인슐린 분비능:냉동직후 감소된 경향을 보였으나 해동후 3일간 배양한 췌도의 인슐린 분비는 냉동전과 통계적으로 의미있는 차이가 없이 냉동보존 전의 수준으로 회복되었다. ③ 냉동보존후 췌도의 포도당에 대한 동적 인슐린 분비능:냉동보존한 췌도를 해동후 3일째의 인슐린 동적 분비능은 냉동 전과 마찬가지로 자극 인슐린의 반응의 제1기와 2기가 잘 관찰되었다. ④ 냉동보존한 췌도세포 이식 후 혈당 변화:스트렙토조토신으로 유도된 당뇨병 쥐에 췌도이식 후 혈당은 냉동보존한 췌도이식군에서 대조군에 비해 혈당의 조절효과가 더 오래 지속되었다. 결론:소동물에서 단계적 온도 하강법을 이용한 췌도세포 냉동보존법을 확립하였으며 이는 기능, 구조 및 생존율에 큰 이상을 보이지 않았으므로 장차 사람의 췌도세포 동종이식시 부족한 췌도세포수를 극복하고 면역반응을 줄일 수 있는 매우 유용한 방법이 될 것으로 판단된다. Background : Although islet transplantation has been attempted to reverse the state of diabetes, achieving a critical number of islets and modulating the immune response limit the success of islet transplantation. Cryo-preservation of islets offers many important benefits for islet transplantation by collecting islets with a wide variety of HLA phenotypes and islet MHC expression. The aims of this study was to determine the optimal conditions for cryo-preservation by using a controlled cooling method and to evaluate in vitro and in vivo functional properties of the cryo-preserved islets. Methods : Collagenase-isolated, Ficoll-purified islets were cultured for 48 hours. They were aliquoted into freezing tubes (1000 islets per tube), equilibrated with 2M dimethyl sulfoxide (DMSO) in three steps, supercooled, nucleated, and controll-cooled at rate of 0.25℃/min to - 40℃ prior to storage at - 196℃. Rapid thawing and removal of DMSO with 0.75 M sucrose preceded 48 hour of culture and the morphology, viability, glucose-induced insulin secretion, and in vivo function of rats transplanted with cryopreserved islets was reexamined. Results : ① Recovery was 90.2±0.2%, 85.7±0.1% and 81.7±0.1% immendiately after, 24 hours and 72 hours after thawing respectively. The viability was 60±5%, 80±5%, 90±5% immediately after, 24 hours and 72 hours after thawing respectively. ② The glucose-stimulated-insulin secretion (GSIS) tended to decrease immediately after thawing, but GSIS increase to the level of pre-cryopreservation 72 hours after thawing. ③ The in dynamic GSIS, the first and the second phase of insulin secretion were well preserved in islets cultured for 72 hours after thawing. ④ The cryopreserved islets were cultured for 3 days and transplanted into renal sub-capsular space of streptozotocin (STZ) induced diabetic rats. The duration of normoglycemia in the STZ-induced diabetic rats transplanted with cryopreserved islets was significantly longer than of the fresh islets. Conclusion : The optimal condition of cryopreservation using the controlled cooling method was established in rat pancreatic islets. This cryopreservation method can be a feasible approach for human islet transplantation (J Kor Diabetes Asso 26:64~74, 2002).

흰쥐에서 심정지후 Pentoxifylline이 뇌의 허혈-재관류 손상에 미치는 효과

정시경,김영민,오동렬,최경호,박승현,이운정,박규남,유은영,이원재,김세경 大韓應急醫學會 1999 대한응급의학회지 Vol.10 No.2

Background: Two major events occurring in the cerebral hemodynamics after successful resuscitation from cardiac arrest are reactive hyperemia and postischemic hypoperfusion. In this study, we examined the effect of Pentoxifylline(PTX) on the rat brain following cardiac arrest. Methods: Fourteen rats were anesthetized and artificially ventilated. Cardiac arrest was produced by chest compression and clamping of tracheal tube for 3 minutes in ketamine anesthetized rats. Circulation was restored by standard cardiopulmonary resuscitation methods. In 7 rats, PTX 10mg/kg was infused at 10min after cardiac arrest(PTX group). In the other 7 rats, same amount of normal saline was infused(control group). Results: In both groups, hemodynamic variables, neurologic deficit(ND) score and histopathologic findings of hippocampal CA1 neurons were observed. Hemodynamic variables and ND score were not significantly different between two groups. Delayed ischemic neurons of hippocampal CA1 were decreased in PTX group(2.2±2.4%) compared with control group(9.1±1.2%). Conclusions: We conclude that PTX prevented development of delayed ischemic neurons in hippocampal CA1 after cardiac arrest. PTX may be useful in emergency situations following cardiac arrest.

3차원 전산화단층촬영 영상을 이용한 얼굴 연조직 두께 계측

정호걸,김기덕,한승호,신동원,허경석,이제범,박혁,박창서 대한구강악안면방사선학회 2006 Imaging Science in Dentistry Vol.37 No.1

Purpose : To evaluate accuracy and reliability of program to measure facial soft tissue thickness using 3D computed tomographic images by comparing with direct measurement. Materials and Methods : One cadaver was scanned with a Helical CT with 3 mm slice thickness and 3 mm/sec table speed. The acquired data was reconstructed with 1.5 mm reconstruction interval and the images were transferred to a personal computer. The facial soft tissue thickness were measured using a program developed newly in 3D image. For direct measurement, the cadaver was cut with a bone cutter and then a ruler was placed above the cut side, The procedure was followed by taking pictures of the facial soft tissues with a high-resolution digital camera. Then the measurements were done in the photographic images and repeated for ten times. A repeated measure analysis of variance was adopted to compare and analyze the measurements resulting from the two different methods. Comparison according to the areas was analyzed by Mann-Whitney test. Results : There were no statistically significant differences between the direct measurements and those using the 3D images (p>0.05). There were statistical differences in the measurements on 17 points but all the points except 2 points showed a mean difference of 0.5 mm or less. Conclusion : The developed software program to measure the facial soft tissue thickness using 3D images was so accurate that it allows to measure facial soft tissues thickness more easily in forensic science and anthropology. (Korean J Oral Maxillofac Radiol 2006; 36 : 49-54)

IL-β의 인슐린 분비 자극효과와 그 과정에 관여하는 인자들

정인경,오승훈,강동묵,정재훈,민용기,이명식,이문규,김광원 대한당뇨병학회 2002 Diabetes and Metabolism Journal Vol.24 No.4

연구배경: IL­1β는 용량과 노출시간에 따라 인슐린분비에 대해 상반되는 효과를 가지고 있다. IL­1β의 인슐린 분비 억제효과는 제1혁명 당뇨병의 자가면역 기전과 관련되어 잘 알려져 있으나 인슐린 자극효과에 대해서는 아직 명백히 밝혀지지 않았다. 이에 저자 등은 IL­1β의 다양한 농도에 따라 백서의 췌도세포에서 인슐린 분비에 미치는 영향을 살펴보고, 그 기전으로 인슐린 생합성, iNOS의 발현, 칼슘통로의 활성도 변화여부를 알아보고자 하였다. 방법:200∼300g인 수컷 Sprague­Dawley 백서의 췌도를 변형된 Lacy&Kostianovsky's 방법으로 분리한 후 IL­1β의 다양한 농도(0, 0.5, 5, 50, 500pmol/L)에 2, 6, 24시간 노출시켜 췌도세포의 형태, 생존능을 관찰하고 인슐린 분비능 및 췌도세포내 인슐린 함량을 측정하였으며, 전전구 인슐린 mRNA발현, iNOS mRNA발현을 RT­PCR을 통해 확인하였고, 세포의 칼슘 통로 활성도 변화 여부를 측정하였다. 결과:1) IL­1β에 노출되 췌도의 생존능:2시간 노출시는 대조군과 차이가 없었으나, 6시간 고농도군과 24시간 모든 군에서 생존능이 감소되었다. 2)인슐린 분비능은 IL­1β를 2시간 5poml/L이상의 고농도와 6시간 0.5pmol/L 저농도 처리시 대조군에 비해 의미있게 증가하였으나, 6시간과 24시간 5poml/L이상의 고농도에서는 의미있게 인슐린의 분비가 억제 되었다. 3)췌도내 인슐린양의 변화는 IL­1β의 시간과 농도에 따라 배지내의 인슐린 변화와 비슷한 경향을 보였으나 통계학적으로 의미있는 차이는 없었다. 4)전전구인슐린의 mRNA발현은 2시간 50pmol/L이상 고농도의 IL­1β에서 의미있게 증가하였고, 6시간 처리군에서는 의미있는 차이는 없었으나, 24시간에서 IL­1β의 용량의존적으로 억제되었다. 5)iNOS mRNA는 IL­1β 처리 후 2시간부터 발현되기 시작하여, 6시간에 최고에 달한 후 24시간에는 점차 감소하였다. IL­1β의 처리시간과 무관하게 5poml/L이상의 고농도에서 용량에 따라 발현이 증가하였다. 6)칼슘통로 활성도는 IL­1β의 농도나 시간에 따라 유의한 차이가 없었다. 결론: IL­1β는 단시간 고용량이나 장기간 저용량에서 인슐린 분비와 생합성을 증가시키며, 이런 효과는 iNOS나 칼슘 통로 활성도 변화와는 무관한 것으로 생각된다. Background : The inhibitory effort of IL-1β on the insulin secretion has been validated in pathogenesis of type 1 diabetes, but complex results about the stimulatory effect of IL-1β have been reported. The aims of this study are to clarify the effects of IL-1βon insulin secretion of pancreatic islets and to investigate the mechanisms in terms of preproinsulin synthesis, inducible NOS expression, and calcium channel activity. Method : Islets were isolated from male Sprague-Dawley (SD) rat by modified Lacy-Kostianovsky's method. After islets were treated with different concentrations (0, 0.5, 5, 50, 500 pmol/L) and exposure time (2, 6, 24 hours) of IL-1β, morphology, viability, static stimulation of insulin to glucose, insulin content, preproinsulin mRNA expression, iNOS mRNA expression and calcium channel activity were measured. Results : 1) Viability o islets was reduced in high concentrations of long term exposure of IL-1β. 2) Insulin secretion was stimulated in islets treated with 5, 50, and 500 pmol/L of IL-1β for 2 hours. 3) Insulin content was not significantly different regardless of concentration and exposure time of IL-1β. 4) Preproinsulin mRNA expression increased in islets treated with 50, 500 pmol/L of IL-1β for 2 hours. After 24 hours, it decreased in dose dependent manner. 5) iNOS mRNA expression was detectable after 2 hours in the presence of IL-1β, peaks at 6 hour and decreased after 24 hours. It was increased above 5 pmol/L of IL-1β in dose dependent manner. 6) Activities of the voltage-dependent Ca^2+ channels were not different among groups. Conclusion : IL-1β plays a positive role in terms of insulin secretion and insulin synthesis in high concentration of short term or low concentration of long term. These effects of IL-1β might be neither dependent of iNOS pathway nor Ca^2+ channel activity (J Kor Diabetes Asso 431~443, 2000).

오이의 品質向上과 運作障害 경감을 위한 有機物 施用 및 輪換式 栽培의 효과에 關한 硏究

丁蓮圭,梁承烈,梁元模 順天大學校 1984 論文集 Vol.3 No.1

處女地 沓田輪換 式栽培地, 3年 連作地, 5年 連作地에 園藝作物을 供試하여 各 土壤別로 標集施肥, 有機物, 및 이들의 조합處理를 함으로써 連作程度와 施肥方法에 따른 作物의 收量과 品質, 土壤의 鹽類濃度 및 土壤理化學性을 比較檢査 함으로써 現象을 하고 나아가서 連作障害를 輕減 또는 防止하기 위한 방안을 모색하기 위하여 몇가지 實驗을 수행하였던 바 그 결과를 모색하면 다음과 같다. 1. 供試土壤의 實驗前 pH는 連作年數가 길어질수록 낮아지는 경향이 있었으며 반대로 유기물, 인산, 마그네슘, K, Ca 등은 連作年數가 길어질수록 높았고 C.E.C.도 비슷한 結果를 나타냈다. 2. 土壤內의 E.C.는 試驗前의 경우, 處女地, 沓田輪換式栽培地가 名名 0.35, 0.56이었고 3年, 5年 連作地가 名名 1.20, 1.30으로 連作土壤이 월등하게 높았으며 施肥後의 경우에는 3.00에서 4.35사이로서 全체的으로 높은 수준이었으나 土壤間에 큰 差異가 없었고 降雨에 노출시켜 일정한 기간이 지난후에는 0.39에서 0.43사이로 土壤에 관계없이 원래의 상태로 쉽게 회복되었다. 3. 土壤의 連作程度나 施肥方法에 따른 오이의 蔓長, 莖의 굵기, 節間數는 處理問에差異가 없었으며 유기물이나 미량요소의 施用에 의한 뚜렷한 증가는 나타나지 않았다. 4. 오이의 收量은 5年 連作地가 가장 많았고 다음으로 處女地, 畓田輪換式栽培地, 3年 連作地 順이었는데 이것은 1果의 果重, 果長, 果俓의 증대보다 수확과수의 差異에 의한 것이었다. 5. 連作程度와 施肥方法에 따른 오이의 品質反應중 外的인 品質 즉, 1果의 果重, 果俓, 果長의 差異는 없었으나 內的인 品質중 Vitamin C 含量은 連作年數가 길어질수록 낮아졌다. This experiment was conducted to investigate the effect of application methods of organic material and minor elements, and difference of replanting year on the elevation of vegetable qualities and the reduction of replant-failure in cucumber plants which were cultivated under covering conditions. Used soils were Virgin soil, Paddy-Upland rotation soil, 3 year replanting soil, 5 year replanting soil. Application methods were Standard fertilizer, Standard fertilizer+Organic material, Standard fertilizer+Minor elements, Standard fertilizer+Organic material+Minor elements. Cucumber seeds were sown at August, 15 and transplanted at September, 15 in 1984. The results were summarized as follows; The pH value of the used soil before experiment was showed a decreasing tendency with extending the replanting year. On the other hand, the contents of the Organic material, ??, Mg, K, Ca and C.E.C. were increased with extending the replanting year. Before experiment, Electric Conductivity of 3 year replanting soil and 5 year replanting soil were notably higher than that of the non-replanting soil, Virgin soil and Paddy-Upland rotation soil. Their values were 1.20, 1.30, 0.35 and 0.56 mㆍmho respectively. But E.C. of the soil after fertilization was increased highly, from 3.00 to 4.35 mㆍmho with no relation of the replanting year. And after the soil being opened to the rainfall for about 4 weeks, the soil salinity was easily removed. Replanting year and fertilizer application methods didn't have an influence on the plant height, stem diameter and No. of node of cucumber plant. The yield was, however, affectected by it, not because of the fruit weight, diameter and length but because of the different quantity of the harvest fruits. Vitamin C content of cucumber fruit was decreased with extending the replanting year.

정신분열병, 기분장애와 보르나 바이러스

정성훈,원승희,김상헌 대한생물치료정신의학회 1998 생물치료정신의학 Vol.4 No.2

Borna disease virus is a unique neurotropic agent that appears to have a predilection for the limbic area of the brain. In some animal species, it can produce a behavioral syndrome characterized by aggressive and passive phases. These two characteristics has suggested the relationship between schizophrenia, mood disorder and Borna disease virus. Borna disease virus can infect human brain tissue possibly contributing to the pathophysiology of specific human neuropsychiatric disorders. The authors reviewed several articles suggesting an association of Borna disease virus with schizophrenia and mood disorder.

불량 단감을 이용한 속성 감식초의 제조와 품질 평가

정용진,신승렬,강미정,서지형,원충연,김광수 동아시아식생활학회 1996 동아시아식생활학회지 Vol.6 No.2

This study was performed to analyze general components such as, organic acids, alcohols and minerals in persimmon vinegar made with quick fermented using deteriorated sweet persimmon and four commercial vinegars(rice vinegar, apple vinegar, brewed vinegar, traditional persimmon vinegar) purchased in local markets for quality evaluation. The pH and total acidity of all vinegars were in the range of 2.02~3.02 and 4.62~9.78%, respectively. The pH of quick fermented persimmon vinegar(A) was relatively higher than that of others. Acidity was the highest in brewed vinegar. Total sugar content was in the range of 0.45~6.43%. These contents were high in were high in traditional persimmon vinegar wherase low in brewed vinegar. Total nitrogen and amino-nitrogen were in the range of 0.025~0.046% and 0.015~0.029%. Organic acids were identified as acetic acid, citric acid, tartaric acid, malic acid, succinic acid, formic acid and oxalic acid. While major minerals of all vinegars were Mg, Ca, and Na, minor minerals were Zn, Cu, Mn and Fe.

SOA내에서의 XPM,FWM을 이용한 WDM 시스템 파장변환기 구현

정광원,이정렬,이승우,엄진섭 강원대학교 정보통신연구소 2001 정보통신논문지 Vol.5 No.-

In this paper, a new type of wavelength converter using XPM within a loop-mirror is proposed and demonstrated. The proposed structure decreases hardware components for implementation of XPM wavelength converter and makes it possible to get non-inverted and inverted output signals simultaneously. Also, we generate wideband pumping mode series for FWM wavelength conversion scheme and propose the effective filtering method for selecting converted wavelength from FWM outputs. The results show that the pumping mode range is over 30nm within 5dB power fluctuation, and that a pumping and a prove wave can be removed successfully by using both of Michelson interferometer and polarization controller.

5-FU로 백서 가슴샘에서 유도된 세포고사의 Comet Assay에 의한 분석

김수원,한승로,정옥,조근자,김선,김원식 충남대학교 의과대학 지역사회의학연구소 2001 충남의대잡지 Vol.28 No.1

Thymus is a lymphoid organ forming T-cells from hematogenous stem cells. Apoptosis of thymocytes in the cortex of thymus is so rapid that cannot find easily with routine histological techniques. In this experiment, Comet assay and SDS-PAGE were used to detect apoptotic changes and protein changes after 5-fluorouracil(5-FU) treatment in the thymus of the rat. The results were as follows. 1. The number of thymocytes in the cortex was markedly decreased, and many apoptotic cells with various size and shapes were found on DAT 5 in 5-FU treated group. 2. On DAT 5, new nodules with germinal centers began to appear in the peripheral cortex, and in the germinal center, there was many apoptotic bodies of various size and shapes. 3. In SDS-PAGE, a protein with molecular weight of 123kDa was increased on DAT 5 which was considered as integrins subfamily.

특수가공용 공작기계의 쇼트피닝기 모델설계에 관한 연구

신용승,이승호,서유원,정성교 서울産業大學校 1999 논문집 Vol.49 No.2

As developing the technology in industry, the materials with superior grade of stiffiness and lightness are required, and hence the higher fatigue strength characteristics are necessary. Thus, the non-traditional machining to increase fatigue strength is becoming influential as very important technology. Considering environmental problem, the shot peening process is selected as a good process to increase fatigue strength. But the data related to shot peening process are not enough in this country. Shot peening process is essential to ensure repeatability and uniformity on a part to part and lot to lot basis, while conforming to the applicable specifications. The shot material, size, shape and hardness as well as velocity and impact angle must be controlled to provide consistency in peening results. In order to effectively control the shot peening process, parameters must be addressed in the following order, such as media control, coverage control, intensity control, equipment control and so on.