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Ultrasonic Degradation of Endocrine Disrupting Compounds in Seawater and Brackish Water
So Young Park,Jong Sung Park,Ha Yoon Lee,Ji Yong Heo,Yeo Min Yoon,Kyung Ho Choi,Nam Guk Her 대한환경공학회 2011 Environmental Engineering Research Vol.16 No.3
In this study, a series of experiments was conducted on the relative degradation of commonly known endocrine-disrupting compounds such as bisphenol A (BPA) and 17α-ethinyl estradiol (Ee₂) in a single-component aqueous solution using 28 and 580 kHz ultrasonic reactors. The experiments were conducted with three different types of model water: deionized water (DI), synthetic brackish water (SBW), and synthetic seawater (SSW) at pH 4, 7.5, and 11 in the presence of inert glass beads and humic acids. Significantly higher sonochemical degradation (93-97% for BPA) occurred at 580 kHz than at 28 kHz (43-61% for BPA), regardless of water type. A slightly higher degradation was observed for Ee₂ compared to that of BPA. The degradation rate of BPA and Ee₂ in DI water, SBW, and SSW after 30 min of ultrasound irradiation at 580 kHz increased slightly with the increase in pH from 4 (0.073-0.091 min-1 for BPA and 0.081-0.094 min-1 for Ee₂) to 7.5 (0.087-0.114 min-1 for BPA and 0.092?0.124 min-1 for Ee₂). In contrast, significant degradation was observed at pH 11 (0.149-0.221 min-1 for BPA and 0.147-0.228 min-1 for Ee₂). For the given frequencies of 28 and 580 kHz, the degradation rate increased in the presence of glass beads (0.1 mm and 25 g) for both BPA and Ee₂: 0.018-0.107 min-1 without beads and 0.052-0.142 min-1 with beads for BPA; 0.021-0.111 min-1 without beads and 0.054-0.136 min-1 with beads for Ee₂. A slight increase in degradation of both BPA and Ee₂ was found as the concentration of dissolved organic carbon (DOC, humic acids) increased in both SBW and SSW: 0.107-0.115 min-1 in SBW and 0.087-0.101 min-1 in SSW for BPA; 0.111-0.111 min-1 in SWB and 0.092-0.105 min-1 in SSW for Ee₂. After 30 min of sonicating the humic acid solution, DOC removal varied depending on the water type: 27% (3 mg L-1) and 7% (10 mg L-1) in SBW and 7% (3 mg L-1) and 4% (10 mg L-1) in SSW.
연구논문 : 만성 바이러스간질환에서 간섬유화의 다양한 비침습 혈청표지자검사의 비교
김선민 ( Sun Min Kim ),손주현 ( Joo Hyun Sohn ),김태엽 ( Tae Yeob Kim ),노영욱 ( Young Wook Roh ),은창수 ( Chang Soo Eun ),전용철 ( Yong Cheol Jeon ),한동수 ( Dong Soo Han ),오영하 ( Young Ha Oh ) 대한간학회 2009 Clinical and Molecular Hepatology(대한간학회지) Vol.15 No.4
목적: 만성 바이러스간질환에서 병의 경과와 예후를 평가하고 치료반응을 예측함에 있어 간섬유화의 진단은 중요하다. 최근 간섬유화 평가에 표준방법이지만 침습적 간생검을 대신하여 여러 가지 비침습적 진단법이 이용되고 있고, 또한 새로운 검사법들이 개발되고 있다. 저자들은 만성 B형 및 C형간염 환자를 대상으로 임상에서 광범위하게 흔히 시행하는 혈액검사와 임상 소견을 이용한 아래와 같은 진단법들만으로 간섬유화를 얼마나 정확하게 평가할 수 있는지 알아보고, 단일 검사로 간섬유화를 평가하는 데 유용하다고 알려져 있는 hyaluronic acid(HA)와 IV형 콜라겐 농도와 비교하여 그 임상적 유용성을 평가하였다. 대상과 방법: 2002년 3월부터 2007년 2월까지 만성 바이러스간 질환으로 간생검을 시행받은 225명의 환자(HBV 180명, HCV 43명, HBV+HCV 2명)를 대상으로 분석하였다. 간생검을 시행하는 날에 말초혈액검사, 혈액응고검사, 혈청생화학검사를 시행하고, 혈중 HA, IV형 콜라겐 농도를 측정하였다. 간섬유화의 정도는 F0(섬유화가 없음), F1(문맥역 섬유화), F2(문맥주변부 섬유화), F3(섬유성 격막) 및 F4(간경변증)의 4단계로 구분하였다. 대상 환자를 F0-1, F2-4 혹은 F0-2, F3-4의 두 집단으로 분류하여 두 집단을 구분하고자 할 때 AAR(AST/ALT ratio), API(age-platelet index), APRI(AST to platelet index), CDS(cirrhosis discriminant score), platelet count, HA, IV형 콜라겐의 예측능을 area under the receiver operating characteristic curve (AUROC)값을 이용하여 비교하였다. 결과: 대상환자의 섬유화의 단계는 F0 집단은 17명, F1 집단은 40명, F2 집단은 61명, F3 집단은 74명, F4 집단은 33명이었다. 의미 있는 간섬유화를 F2 이상으로 판단할 때, 의미 있는 섬유화의 예측에 대한 AUROC 값은 APRI=0.822, CDS=0.776, platelet count=0.773, API=0.756, HA=0.749, IV형 콜라겐=0.718, AAR=0.642 순이었고, F3 이상의 광범위한 섬유화 예측에 대한 AUROC값은 CDS=0.835, platelet count=0.795, API=0.794, HA=0.766, AAR=0.711, IV형 콜라겐=0.697, APRI=0.691 순으로 관찰되었다. 결론: 만성 바이러스간질환에서 간섬유화를 평가하는 데 임상 소견 및 혈액검사를 이용한 방법들이 비침습적 진단법 중 단독 검사로도 유용한 것으로 알려져 있는 혈중 HA와 IV형 콜라겐 농도와 비교하여 우월하거나 대등하였다. 특히 APRI는 다른 인자들에 비해 F2 이상의 의미 있는 섬유화를 예측하는 데 가장 유용하였고, CDS는 F3 이상의 광범위한 섬유화를 예측하는 데 가장 유용하였다. Background/Aims: The aim of this study was to determine the clinical performances of noninvasive serum markers for the prediction of liver fibrosis in chronic viral liver diseases. Methods: We analyzed a total of 225 patients with chronic viral liver diseases (180 with hepatitis B virus, 43 with hepatitis C virus, and 2 with hepatitis B+C virus) who underwent a liver biopsy procedure at the Hanyang University Guri Hospital between March 2002 and February 2007. Serum was also obtained at the time of liver biopsy. Liver fibrosis was staged according to the scoring system proposed by the Korean Study Group for the Pathology of Digestive Diseases. Various noninvasive serum markers were evaluated, including the aspartate aminotransferase (AST)/alanine aminotransferase (ALT) ratio (AAR), age-platelet (AP) index, AST/platelet ratio index (APRI), cirrhosis discriminant score (CDS), platelet count, hyaluronic acid (HA), and type IV collagen. Results: There were 17, 40, 61, 74, and 33 patients at stages F0, F1, F2, F3, and F4, respectively. The overall diagnostic accuracies of each marker, as determined by the area under receiver operating characteristics curves, were APRI=0.822, CDS=0.776, platelet count=0.773, AP index=0.756, HA=0.749, type IV collagen=0.718, and AAR=0.642 for predicting significant fibrosis (≥F2); and CDS=0.835, platelet count=0.795, AP index=0.794, HA=0.766, AAR=0.711, type IV collagen=0.697, and APRI=0.691 for predicting extensive fibrosis (≥F3). Conclusions: Conclusions: All noninvasive serum markers evaluated in this study were useful for predicting significant or extensive liver fibrosis in chronic viral liver diseases. In particular, APRI was most useful for the prediction of significant fibrosis, and CDS was most useful for the prediction of extensive fibrosis. (Korean J Hepatol 2009;15:454-463)
동충하초 투여가 최대하 운동 시 지방연소에 미치는 영향
김철우,이용수,이상현,이상호,하민수,윤영조,이재일 한국스포츠리서치 2003 한국 스포츠 리서치 Vol.14 No.4
The purpose of this study was to evaluate effects of Cordyceps militaris on fat oxidation during submaximal exercise. Fifteen healthy male collegiate students(ingestion group, N=7; control group, N=8) participated as subjects after signing an informed consent following overnight, subjects completed maximal graded execise test on treadmill to determine exercise intensity(50%VO2max). To evaluate the fat oxidation using indirect calorimeter, it was performed submaximal treadmill to determine exercise lasting. 40min at 50%VO2max after 5day and consumed Cordyceps militaris on separate days. This study was used 2×4 two way repeated ANOVA to analyze physiological and metabolic variables between two groups and metabolic variables between two groups and four exercise periods(l0min, 20min, 30min 40min), and used Scheffe to post-hoc test. Significance was set at α=.05. The results of this study were as followed: First, there were significantly highest in ingestion group for V02, percentage of fat oxidetion(%fat), fat per kilocarolies(Fkcal), and total per kilocarolies(Tkcal)(p<.05), but respiratory exchange ration(RER) and heart rete(HR) were significantly lower in ingestion group(p<.05). Second, RER, %fat, Fkcal were significantly difference in 10, 20, 30, 40min of exercise period(p<.05). This results suggest that there is the effect of Cordyceps militaris on fat oxidation submaximal exercise. Additionally, to evaluate effect of Cordyceps militaris as ergogenic aid, it should be required the future study that included extend subjects, various dosage, and chronic consumption.
총경동맥 폐쇄시간에 따르는 국소 뇌혈류 변화 : 실험적 연구 An Experimental Cat Model
강준기,성태경,조병일,백민우,김문찬,허춘웅,하영수,송진언 대한신경외과학회 1983 Journal of Korean neurosurgical society Vol.12 No.3
The microvasculature of the brain is also quite susceptible to ischemic insult, and substantial portions of the brain are not reperfused after restoration of the blood supply following overtime of critical ischemic periods. The purpose of this series of experiments was to determine the effects of ischemia on subsequential regional cerebral blood flow measurements and cortical electric activities following reperfusion after ischemia and also to define the proper time of vascular occlusion without irreversible neural damage. Cerebral ischemia was induced in cat by bilateral common carotid occlusions for periods of 10, 30, to 60 minutes, and the blood supply was reperfused for 3 hours after clamp-off. Regional cerebral blood flow(rCBF) was measured by hydrogen clearance technique following ischemia, restoration of blood supply and electroencephalogram recovery could be predicted according to the rCBF. Forty adult cats weighing 2.7 to 4.0㎏ were used in this study. The animals were divided into 4 groups of 10 cats each : normal control, 10 min-clamped, 30 min-clamped, and 60 min-clamped groups. The results obtained were as follows : 1) The mean rCBF was 24.6±7.0㎖/100g/min in control group. 2) Bilateral carotid occlusions resulted in a reduction of the rCBF(12.4±4.1㎖/100g/min) to 50% of control flow on both hemispheres. 3) Sequential changes of the rCBF after reperfusion : (1) There was restored the rCBF(21.3±5.1㎖/100g/min) to control flow in the 10 minutes-clamped group. (2) There was a 85% recovery of control flow in the 30 minutes-clamped group. (3) There was a only 25% recovery of control flow in the 60 minutes-clamped group. 4) A close correlation was found between cortical electrical activity and rCBF suggesting a threshold relationship. (1) The changes of cortical electric activity began to notice at rCBF less than 17.4±4.7㎖/100g/min. (2) The recovery of cortical electric activity noted at rCBF more than 10.2±2.3㎖/100g/min. 5) There was no evidence of ischemic involvement at the cortex, white matter and basal ganglia in the 10 minutes clamped group, but demonstrated a dense wedge shaped infarct at the cortex and uncus herniation in the 60 minutes clamped group. The rCBF and cortical electric activity restored to normal values in reperfusion within 10 minutes after occlusion of both common carotid arteries.
2010 해남, 평창, 원주지역 갈대 등 야초류의 이용실태와 사료가치 평가
서성(Sung Seo),한대덕(Dae Duk Han),장선식(Sun Sik Jang),김원호(Won Ho Kim),정민웅(Min Woong Jung),최진혁(Jin Hyuck Choi),김진숙(Jin Sook Kim),김하영(Ha Young Kim),이종경(Joung Kyong Lee) 한국초지조사료학회 2012 한국초지조사료학회지 Vol.32 No.1
본 연구는 부존 조사료자원 이용으로 수입사료를 줄이고 사료비를 절감하고자 2010년도에 전남 해남(간척지), 강원 평창(강변), 원주(섬강변), 전남 영광(와탄강변) 등 4개 지역에서 갈대와 억새 등 야초 자생지를 탐색하고, 수확·이용실태 조사와 함께 사료가치를 분석하였다. 전남 해남 해남목장 내 갈대(Phragmites communis) 위주의 야초 자생지는 300 ha로 예년에는 2회 곤포작업하여 연간 사일리지로 2,300~2,500톤(ha당 8톤)을 생산하였으나 2010년도는 1회 곤포조제로 사일리지 2,000톤(ha당 7톤)을 생산하였다. 유통가격은 롤 당 53천원으로 ㎏당 110원이었다. 곤포는 전량 자가소비하며, 외부 작업단에 롤 당 26천원의 작업비를 지불하였다. 연간 조수입은 2억 2,260만원이며, 소득은 1억원 수준(345천원/ha)이었다. 초장 150 ㎝의 갈대는 조단백질 8.4%, 상대사료가치 71.9 (건초 5등급), 건물 소화율 60.1%로 사료가치는 볏짚에 비해 약간 나은 것으로 평가되었다. 갈대 사일리지의 사료가치와 유기산 함량은 동일 지역에서 동일한 재료로 사일리지를 조제하더라도 수확시기에 따라 차이가 컸으며(CP 4.7~6.4%, RFV 62.2~78.9, 건물 소화율 41.9~53.9%), 갈대 사일리지의 품질은 수단그라스에 비해 불량하였다. 강원 평창 강변 자생갈대의 사료가치는 초장이 79, 117, 121, 142㎝로 생육이 진행됨에 따라 낮아져, 조단백질은 각각 13.9%에서 7.6%로, 상대사료가치는 90.7 (건초 3등급)에서 76.1 (건초 4등급)로, 건물 소화율은 72.9%에서 54.7%로 감소하였다. 원주 섬강 주변 야초류는 곤포수확 작업에 어려움이 있었으며, 초장 130 cm의 갈대는 조단백질 8.5%, 상대사료가치 82.3 (건초 4등급), 건물 소화율 70.2%로 사료가치는 높았으며, 220㎝의 성숙한 갈대는 조단백질 10.2%, 상대사료가치 65.1 (건초 5등급), 건물 소화율 48.9%로 볏짚 대비 조금 나은 것으로 평가되었다. 전남영광지역에서는 와탄천 주변에 자생하는 갈대를 수거 이용하고자 시도하였으나 경제성이 없어 중단하였다. 이상의 결과를 종합하여볼 때, 갈대, 억새 등 야초류는 현 상태에서 최대한 수거·이용하되, 사료가치를 유지하고 가축 기호성이 양호한 시기에 수확하는 것이 중요하며, 최소한 볏짚과 비슷하거나 그 이상의 사료가치를 기대하기 위해서는 7월 중순 이전(늦어도 7월까지)에 수확·이용하는 것이 권장된다. This study was carried out in 2010 to investigate the utilization and forage quality of native grasses, such as Phragmites communis which might reduce the cost of feeding domestic cattle. The regions surveyed were the Haenam ranch in Haenam, Pyeongchang, Wonju, and Yeonggwang. In Haenam, yearly silage production harvested from 300 ha was 2,000 MT (7 MT/ha). All of those round bale silages were self-consumed in that region, and marketing price was 50,000 55,000 won per roll (110 won/㎏). Phragmites communis of 150 ㎝ in length contained 8.4% crude protein (CP) with relative feed value (RFV) 71.9 and 60.1% in vitro dry matter digestibility (IVDMD), which was somewhat more favorable than forage quality of rice straw. The silage quality of Phragmites communis varied greatly by the time of harvest (CP 4.7 6.4%, RFV 62.2 78.9, and IVDMD 41.9 53.9%), even with the same district and of the same original forage materials. Generally, the quality of Phragmites communis silage of was poorer than that of sorghum * sudangrass hybrid. In Pyeongchang, forage quality of Phragmites communis was decreased from 13.9% to 7.6% in CP, 90.7 to 76.1 in RFV, and 72.9% to 54.7% in IVDMD, as plant was getting mature, from 79 ㎝ to 117 ㎝, 121 ㎝ or to 142 ㎝ in length. In Wonju, the quality values of Phragmites communis of 130 ㎝ in length were 8.5% CP, 82.3 RFV and 70.2% IVDMD, while those of matured grasses of 220 ㎝ in length were lower (10.2% CP, 65.1 RFV and 48.9% IVDMD), but this was a little more favorable than quality of rice straw. In Yeonggwang, feeding Phragmites communis was tried in a Hanwoo feed, but stopped due to low profitability. In conclusion, the overall quality of most native grasses including Phragmites communis in this survey was poor. Therefore, we recommend that Phragmites communis and native grasses should be harvested on June or July to obtain richer forage quality in forage values than rice straw.
이민수,김용구,김영훈,연병길,오병훈,윤도준,윤진상,이철,정희연,강병조,김광수,김동언,김명정,김상훈,김희철,나철,노승호,민경준,박기창,박두병,백기청,백인호,손봉기,손진욱,양병환,양창국,우행원,이정호,이종범,이홍식,임기영,전태연,정영조,정영철,정인과,정인원,지익성,채정호,한상익,한선호,한진희,서광윤 大韓神經精神醫學會 1998 신경정신의학 Vol.37 No.1
연구목적 : 본 시험의 목적은 임상시험 시작전에 연구자들을 대상으로 PANSS Workshop을 통하여 PANSS, ESRS에 대한 국내에서의 표준화 작업을 구축하고 새로운 정신병 치료제인 리스페리돈의 효과와 안정성을 재확인하여 리스페리돈 사용에 대한 적정화를 이루는데 있다. 연구방법 : 1996년 4월부터 1996년 9월까지 국내 39개 대학병원 정신과에 입원중인 혹은 증상이 악화되어 입원하는 정신분열병 환자 377명을 대상으로 다시설 개방 연구를 시행하였다. 1주일간의 약물 배설기간을 가진후, 리스페리돈을 8주간 투여하였고, 기준점, 1주, 2주, 4주, 그리고 8주후에 평가되었다. 용량은 제1일에는 리스페리돈 1mg씩 1일 2회, 제2일에는 2mg씩 1일 2회, 제3∼7일에는 3mg씩 1일 2회 투여하였다. 이후 환자의 임상상태에 따라 임의로 증량할 수 있으며, 최대 일일 16mg을 초과하지 않도록 하였다. 추체외로 증상을 조절하기 위한 투약을 허용하였다. 임상증상 및 부작용의 평가는 PANSS(Positive and Negative Syndrome Scale), CGI(Clinical Global Impression) 그리고 ESRS(Extrapyramidal Symptom Rating Scale)을 사용하였다. 연구결과 : 377명중 343명(91%)이 8주간의 연구를 완결하였다. 치료 종결시점인 8주후 PANSS 총점수가 20% 이상 호전된 경우를 약물 반응군으로 정의할때, 약물반응군은 81.3%였다. 리스페리돈에 반응하는 예측인자로는 발병연령, 이전의 입원 횟수, 유병기간이 관련 있었다. 리스페리돈은 1주후부터 PANSS양성, 음성, 및 일반정신병리 점수상에 유의한 호전을 보여 효과가 빨랐다. CGI의 경우도 기준점에 비해 1주후부터 유의한 감소를 나타내었다. ESRS의 경우, 파킨슨 평가점수는 기준점과 비교해 투여 1주, 2주, 4주후 유의하게 증가되었다가 8주후 기준점과 차이가 없었다. Dystonia 평가점수는 1주후만 유의한 증가를 보였으며, dyskinesia 평가점수는 유의한 차이가 없었다. 혈압, 맥박수의 생명징후 및 일반 혈액학 검사, 생화학적 검사, 심전도 검사에서 유의한 변화는 없었다. 결 론 : 이상의 다시설 개방 임상 연구를 통해 리스페리돈은 정신분열병 환자에서 양성증상뿐만 아니라 음성증상 및 전반적인 증상에도 효과적인 것으로 사료된다. 보다 명확한 평가를 위해서는 다른 항정신병약물과의 이중맹검 연구가 필요할 것으로 생각되며, 또한 장기적 치료에 대한 평가도 함께 이루어져야 하겠다. Objective : The purpose of this study was to investigate the efficacy and safety of risperidone in the treatment of Korean schizophrenic patients. Method : This multicenter open study included 377 schizophrenic patients drawn from 39 university hospitals. After a wash-out period of 1 week, the schizophrenic patients were treated with risperidone for 8 weeks and evaluated at 5 points ; at baseline, and 1, 2, 4 and 8 weeks of treatment. The dose was increased from 2mg/day(1mg twice daily) to 6mg/day(3mg twice daily) during the first week and adjusted to a maximum of 16mg/day over the next 7 weeks according to the patient's clinical response. Medication to control extrapyramidal symptoms was permitted. The psychiatric and neurological status of the patients was assessed by PANSS, CGI, and ESRS scales. Results : 343(91%) of 377 patients completed the 8-week trial period. Clinical improvement, as defined by a 20% or more reduction in total PANSS score at end point, was shown by 81.3% of patients. The predictors of response to risperidone were associated older age, shorter duration of illness, fewer previous hospitalization. Risperidone had rapid onset of action ; a significant decrease of the total PANSS and three PANSS factor(positive, negative, general), and CGI was already noticed at the end of first week. For the ESRS, parkinsonism rating scores were significantly increased until week 4 comparing with baseline. Dystonia rating scores were significantly increased until week 1, and dyskinesia rating scores were not significantly changed during the study. Laboratory parameters including vital sign, EKG, hematological, and biochemical values showed no significant changes during the trial. Conclusions : This study suggests that risperidone is generally safe and effective against both the positive and negative symptoms in our group of patients.
연구보문 : 자연과학 ; 황산처리에 의한 불투수성 황근 종자의 발아촉진에 관한 연구
서상흠 ( Sang Heum Seo ),박민우 ( Min Woo Park ),장미하 ( Mi Ha Chang ),장일웅 ( Il Ung Jang ),심상인 ( Sang In Shim ),나영왕 ( Young Wang Na ),김수영 ( Soo Young Kim ),김석현 ( Seok Hyeon Kim ) 한국국제농업개발학회 2012 韓國國際農業開發學會誌 Vol.24 No.3
The reason of poor germination rate of hamabo mallow seed was speculated using TZ test to determine the viability of seed. Seeds showed high viability via TZ test were not germinated well. The quiescent seeds did not imbibed properly due to the hard seed coat and did not show further processes required for germination. Several well-known treatments for seed coat loosening and water uptake into seed were applied and compared for their efficiency. Stratification, hot water treatment, seed coat scarification, wet-cold treatment were revealed as less effective for improving germination although those treatments were common method for breaking dormant seed. Chemical scarification with sulfuric acid, however, effectively increased germinability and promptness index of hamabo mallow seeds. Optimal time for sulfuric acid treatment was 20 min. Seeds treated for 20 min showed improved germination percentage, germination speed, and promptness index of 80%, 13, and 1969 at 30 days after imbibition, respectively, in petri dish experiment. The values were greater than control seeds showed low values (12%, 1, and 183, respectively). The similar tendency was observed in the pot experiment that sulfuric acid treated seed were sown in artificially mixed soil (loamy soil:peatmoss:sand:compost = 5:2:2:1) and placed in greenhouse. Ultrastructure analysis of chemically scarified seed with sulfuric acid for 20 min by scanning electron microscopy did not show any noticeable changes in seed coat hardness and water uptake via hilum. But the only change after sulfuric acid treatment was removal of hilum cap that close the hilum and hider imbibition. Therefore, subsequent experiment was conducted to know the relationship between removal of hilum cap and facilitated imbibition by sulfuric acid treatment and whether the hilum cap removal directly influenced to the opening of micropyle. Water uptake velocity of seed treated for 20 min in concentrated sulfuric acid for was greater than control seed. This result implies that the crucial effect of sulfuric acid treatment is the removal of hilum cap that increase water uptake via micropyle in hamabo mallow.
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).
민선홍,최봉종,김한수,김영하,박광하 강릉대학교동해안지역연구소 1993 東海岸硏究 Vol.4 No.3
1993년 2월부터 9월까지 4회에 걸쳐 명주군 군선강의 8개 지점을 대상으로 하여 수질 조사를 하였다. 수질 평가를 위하여서는 수온, pH, 용존 산소량, 화학적 산소 요구량, 생물학적 산소 요구량, 부유 물질, 암모니아성 질소, 아질산성 질소, 질산성 질소, 인산염 인, 계면 활성제, 대장균등을 측정 하였다. 본 하천의 상류쪽은 산성을 나타냈으며 하류쪽으로 갈수록 알카리성으로 나타났다. 또한 대장균수는 상대적으로 작았다. 그 원인은 광산 폐수때문인 것으로 나타났으며 하류쪽으로 갈수록 해수와 혼합되기 때문인 것으로 나타났다. The water quality was monitored from February through September, 1993 for four times on the eight sites of Gunseon Stream, Myungju. To monitor the water quality, water temperature, pH, Dissolved oxygen, Chemical oxygen demand, Biochemical oxygen demand, Suspended solid, Ammonia-nitrogen. Nitrite-nitrogen, Nitrate-nitrogen, Inorganic phosphate, Sulfactant, and E. coli MPN were chosen as parameters. The upstream was more acidic; downstream was more alkalic. The E. coli MPN value was relatively low due to the mine pollutant and seawater mixture.