Measurement of Total Plasma Homocysteine in Patients with Chronic Renal Failure Using HPLC/FLD

Kyung-Ok Lee(이경옥),Bo-Kyung Kang(강보경),Hyung-Jin Roh(노형진),Kwang-Suk Ryoo(류광석),Jeong-Yeon Yoo(유정연),Man-Jeong Paik(백만정),Kang-Hyeob Lee(이강협) 대한의생명과학회 1997 Biomedical Science Letters Vol.3 No.1

본 연구에서는 한국인 만성신부전증 환자에서 심혈관계 질환의 조기진단을 위한 생화학적 표지자로서 homocysteine의 임상적 유용성을 검토하기 위하여, 만성신부전증 환자와 건강인에서 HPLC/FLD (high performance liquid chromatography/fluorescence detector)를 이용하여 혈장 homocysteine 농도를 측정하고 그 결과를 비교하였다. 본 실험방법의 회수율은 98.6±5.8%를 나타내었으며, 0.2 n㏖/L보다 낮은 농도까지 측정이 가능하였고, 2~50 n㏖/㎖까지 직선성이 성립하였다 (correlation coefficient =0.9997). 한국인 중 건강한 정상인 (20명)과 만성신부전증 환자 (90명)에서 혈장 homocysteine은 각각 6.81±1.54 n㏖/㎖과 27.28±14.94n ㏖/㎖이었으며, 환자군에서는 정상인군에 비하여 약 4 배 정도 높은 수치를 나타내었다 (p<0.05). 본 실험의 결과로 볼 때 HPLC/FLD를 이용한 homocysteine 측정은 예민도와 재현성이 높아 routine 실험실 방법으로 유용성이 높을 것으로 생각되며, 또한 혈장 homocysteine의 측정은 만성신부전증 환자의 주요한 사망원인이 되는 폐쇄성 동맥질환을 조기에 진단하거나, 혹은 homocysteine 농도를 저하시키는 치료를 실시한 후 치료효과를 판단하기 위한 생화학적 marker로 활용될 수 있을 것으로 기대된다. Cardiovascular disease has been the leading cause of death among patients with chronic renal failure. Many reports have been described that homocysteine is one of the independent risk factor to the occulsive vascular disease. In this study, HPLC/FLD (high performance liquid chromatography-fluorescence detector) technique was used to measure homocysteine level in patients with chronic renal failure and normal control group. The detection limit and recovery of total plasma homocysteine using HPLC/FLD were 98.6±5.8% and 0.2 n㏖/㎖, respectively. The linearity of this method was established in concentration range of 2~50 n㏖/㎖ (correlation coefficient=0.9997). The concentrations of total plasma homocysteine were 6.81 ± 1.54 n㏖/㎖ and 27.28 ± 14.94 n㏖/㎖ in normal control (n=20) and patient group (n=90), respectively (p<0.05). In this study, the HPLC/FLD method showed high sensitivity and reproducibility for a routine clinical laboratory testing. Moreover determination of homocysteine level in plasma might be useful for a biochemical marker for predicting the cardiovascular diseases and for monitoring of therapeutic effect of lowering homocysteine in patients with chronic renal failure.

보문 : 버섯 추출물이 인체 병원성 균에 미치는 항균활성의 특성

김성태 ( Sung Tae Kim ),이강협 ( Kang Hyeob Lee ),민태진 ( Tae Jin Min ) 한국균학회 2003 韓國菌學會誌 Vol.31 No.2

Iodochitin에 합성에 관한 연구

안병제 ( Byung Jae Ahn ),김현준 ( Hyun Jun Kim ),최강혁 ( Kang Hyuk Choi ),이강협 ( Kang Hyeob Lee ),한상문 ( Sang Mun Han ),김용범 ( Yong Beom Kim ),성용길 ( Yong Kiel Sung ),민태진 ( Tae Jin Min ),유국현 ( Kook Hyun Yu ) 한국키틴키토산학회 2004 한국키틴키토산학회지 Vol.9 No.2

느타리버섯(Pleurotus ostreatus) 중의 Light-induced Mitochondrial ATP Synthase 의 효소적 특성에 관한 연구

민태진,이강협 동국대학교 자연과학연구소 1990 자연과학연구 논문집 Vol.10 No.-

Mitochondrial ATP synthase의 효소 활성에 빛이 어떤 영향을 미치는지 알아보기 위하여 느타리버섯 중의 mitochondria를 설탕 밀도 단계 기울기법에 위하여 분리 정제 하였다. 400nm에서 700nm 범위의 빛을 각각 조사하였을 때 이 효소는 490nmm이 빛을 15초 동안 쪼여 주었을 때 가장 활성이 증가하였다. 최적 조건에서 mitochondrial ATP synthase의 organic compounds 효과를 보았을 때 2,6-dichlorophenol indophenol(DCPIP)에 의해 168%, phenazine methosulfate(PMS)에 의해 224% 효소 활성이 증가하였다. 그러나 oligomycin에 의해서는 91% 효소 활성억제를 나타냈고 2,4-dinitrophenol(DNP)에 의해서는 75%, 2-heptyl-4-hydroxyquinoline-N-oxide(HQNO)에 의해서 14%의 활성억제를 나타냈다. 금속 이온 효과에서 이 효소는 Ca?에 의해 효소 활성이 35% 감소하였고 Co?에 의해서 14%, Mn?에 의해 73% 활성 감소를 나타냈다. 음이온 효과에서 이 효소는 CN?에 의해 80% 활성 감소를 보였고, SO?에 의해 52%, CO?에 의해 28% 그리고 NO?에 의해 28% 효소 활성 감소를 나타냈다. 느타리버섯의 mitochondira를 분리 정제하여 mitochondrial ATP synthaserk 가장 활성화되는 빛의 파장을 밝히고, 유기물, 양이온 및 음이온의 효과를 관찰한 결과 표고버섯 중의 mitochondrial ATP synthase의 경우와 유사함을 알 수 있었다. 따라서 mitochondrial ATP synthase는 종에 무관하게 청색광에 의해 활성화되고 그 효소적 특성도 같음을 알았다. Mitochondria of Pleurotus ostreatus was isolated and purified by stepped surcrose density gradient centrifugation in order to investigate the effects of the light on the enzymatic activity of the mitochondrial ATP synthase. This enzyme, which was illuminated by the light ranging from 400nm to 700 nm, showed that the specific activity was most increased by 490 nm for 15 sec. Effects of organic compounds of the mitochondrial ATP synthase were also investigated at the optimum conditions. The activity of this enzyme were increased to 168% by 2,6-dichlorophenol indophenol(DCPIP), 224% by phenazine methosulfate(PMS), but inhibited 91% by oligomycin, 14% by 2-heptyl-4-hydroxyquinoline-N-oxide(HQNO) and 75% by 2,4-dinitrophenol(DNP). Effects of metal ions of the mitochondrial ATP synthase were investigated at the optimum conditions. The activities of the enzyme were inhibited 35% by Ca? 14% by Co? and 73% by Mn?. For effects of anions, the activities of this enzyme were inhibited 80% by CN, 52% by SO?, 28% by each of CO? and NO?.

Measurement of Total Plasma Homocysteine in Patients with chronic Renal Failure Using HPLC/FLD

Lee,Kyung-Ok,Yoo,Jeong-Yeon,Roh,Hyung-Jin,Paik,Man-Jeong,Lee,Kang-Hyeob,Kang,Bo-Kyung,Ryoo,Kwang-Suk THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1997 Journal of biomedical laboratory sciences Vol.3 No.1

본 연구에서는 한국인 만성신부전증 환자에서 심혈관계 질환의 조기진단을 위한 생화학적 표지자로서 homocysteine의 임상적 유용성을 검토하기 위하여, 만성신부전증 환자와 건강인에서 HPLC/FLD (high performance liquid chromatography/fluorescence detector)을 이용하여 혈장 homocysteine 농도를 측정하고 그 결과를 비교하였다. 본 실험방법의 회수율은 98.6±5.8%를 나타내었으며, 0.2 nmol/L보다 낮은 농도까지 측정이 가능하였고, 2∼50 nmol/ml까지 직선성이 성립하였다 (correlation coefficient=0.9997). 한국인 중 건강한 정상인 (20명)과 만성신부전증 환자 (90명)에서 혈장 homocysteine은 각각 6.81±1.54 nmol/ml과 27.28±14.94 nmol/ml이었으며, 환자군에서는 정상인군에 비하여 약 4배 정도 높은 수치를 나타내었다 (p<0.05). 본 실험의 결과로 볼 때 HPLC/FLD를 이용한 homocysteine 측정은 예민도와 재현성이 높아 routine 실험실 방법으로 유용성이 높을 것으로 생각되며, 또한 혈장 homocysteine의 측정은 만성신부전증 환자의 주요한 사망원인이 되는 폐쇄성 동맥질환을 조기에 진단하거나, 혹은 homocysteine농도를 저하시키는 치료를 실시한 후 치료효과를 판단하기 위한 생화학적 marker로 활용될 수 있을 것으로 기대된다. Cardiovascular disease has been the leading cause of death among patients with chronic renal failure. Many reports have been described that homocysteine is one of the independent risk factor to the occulsive vascular disease. In this study, HPLC/FLD (high performance liquid chromatography-fluorescence detector) technique was used to measure homocysteine level in patients with chronic renal failure and normal control group. The detection limit and recovery of total plasma homocysteine using HPLC/FLD were 98.6±5.8% and 0.2 nmol/ml, respectively. The linearity of this method was established in concentration range of 2∼50 nmol/ml, (correlation coefficient=0.9997). The concentrations of total plasma homocysteine were 6.81±1.54 nmol/ml and 27.28±14.94 nmol/ml in normal control (n=20) and patient group (n=90), respectively (p<0.05). In this study, the HPLC/FLD method showed high sensitivity and reproducibility for a routine clinical laboratory testing. Moreover determination of homocysteine level in plasma might be useful for a biochemical marker for predicting the cardiovascular diseases and for monitoring of therapeutic effect of lowering homocysteine in patients with chronic renal failure.

느타리버섯 (Pleurotus ostreatus) 중의 Light - induced mitochondrial ATP Synthase 의 효소적 특성에 관한 연구

민태진,이강협 한국균학회 1991 韓國菌學會誌 Vol.19 No.1

Mitochondria in Pleurotus ostreatus were isolated and purified by the stepped sucrose density gradient centrifugation, to investigate the effects of the light on the enzymatic activity of the mitochondrial ATP synthase. This enzyme, which was illuminated by the light ranging from 400 ㎚ to 700 ㎚, showed that the specific activity was stimulated at 490 ㎚ for 15 sec. Effects of organic compounds on the mitochondrial ATP synthase were also investigated at the optimum conditions; The activities of this enzyme were increased to 168 percent by the addition of 2,6-dichlorophenol indophenol(DCPIP), 224 percent by phenazine methosulfate(PMS), but inhibited 91 percent by oligomycin, 14 percent by 2-heptyl-4-hydroxyquinoline-N-oxide(HQNO) and 75 percent by 2,4-dinitrophenol (DNP), respectively. Effects of metal ions of the mitochondrial ATP synthase were investigated at the optimum conditions. The activities of the enzyme were inhibited 35 percent by Ca^(2+), 14 percent by Co^(2+) and 73 percent by Mn^(2+). For effects of anions, the activities of this enzyme were inhibited 80 percent by CN^-, 52 percent by SO₄^(2+), 28 percent by each of CO₃^(2+) and NO₃^-, respectively.