LC : Efficacy of Immunological Measurement of Aspartate/ Alanine Aminotransferase with Monoclonal Antibodies in the Prediction of Liver Fibrosis

( Ki Tae Suk ),( Yun Hyeong Lee ),( Eui Yul Choi ),( Dong Joon Kim ) 대한간학회 2013 춘·추계 학술대회 (KASL) Vol.2013 No.1

Background: Enzymatic evaluation of alanine/aspartate aminotransferase (AST/ALT) has been currently used and regarded as sensitive indicator of hepatocellular damage. However, enzymatic analysis does not exactly represent the progression of diseases in the patients with liver disease. Immunoassay methods have been suggested as one of the alternatives for the replacement or supplement of the conventional enzymatic analysis. Two forms of AST (cytoplasmic [c] AST and mitochondrial [m] AST) and 3 forms of ALT (ALT1, ALT2, and ALT2_2) have been identified. We evaluated the efficacy of immunoassay method in the prediction of liver fibrosis. Methods: A total of 219 patients with CHB who underwent HVPG and liver biopsy before antiviral therapy were recruited. Blood samples were collected during HVPG. For the evaluation of immunoassay, sandwich ELISA using fluorescence labeled monoclonal antibodies was used. Liver function test including enzymatic AST/ALT and immunological (c) AST, (m) AST, ALT1, ALT2 were checked and compared according to the stage of liver disease. Results: The mean levels of AST and ALT were 121±157 IU/L and 210±279 IU/L. Mean HVPG score of all patients was 4.7±2.5 mmHg. The distribution of the METAVIR fibrosis stages was as follows: F0=6 (3%), F1=52 (24%), F2=88 (40%), F3=45 (20%), and F4=28 (13%). According to the stage of liver fibrosis, HVPG score and mean levels of AST, mAST, ALT1, ALT1+ALT2, AST+cAST+Mast, and ALT+ALT1+ALT2 were significantly different. Mean levels of cAST+mAST, ALT1+ALT2, AST+cAST+mAST, and ALT+ALT1+ALT2 were significantly increased according to the METAVIR stage. Enzymatic AST/ALT level did not show the difference. Conclusions: Sandwich ELISA method for determining serum AST/ALT level showed positive correlation with the stage of liver fibrosis. Therefore, immunoassay method may be useful as a method to predict the stage of liver fibrosis.

Fibrosis-4 index, a predictor for prognosis of hepatocellular carcinoma patients after curative hepatectomy even in hepatitis B virus dominant populations

Sang Oh Yun,김종만,Jinsoo Rhu,Gyu Seong Choi,Jae Won Joh 대한외과학회 2023 Annals of Surgical Treatment and Research(ASRT) Vol.104 No.4

Purpose: Liver fibrosis plays an important role in the development of hepatocellular carcinoma (HCC) and determining its prognosis. Although many staging systems and liver reserve models have been developed without the intention of predicting prognosis of HCC, some studies have investigated their prognostic values in HCC after curative liver resection (LR). The aim of this study is to evaluate prognostic value of non-invasive biomarkers after curative LR. Methods: Between 2006 and 2013, HCC patients underwent LR were included and total 962 patients were enrolled. All non-invasive biomarkers (fibrosis 4 index (FIB-4), aspartate aminotransferase-to-platelet ratio index (APRI), aspartate aminotransferase-to-alanine aminotransferase ratio (AAR), AAR-to-platelet ratio index (AARPRI), and albumin-bilirubin (ALBI) score) were measured at the time of HCC diagnosis. To binarize each biomarker, an optimal cut-off value for fibrosis stage was selected using the value of minimum distance from the left-upper corner of the receiver operating characteristic curve with a specificity >60%. We performed Cox regression analysis on 2-year recurrence-free survival (RFS) and overall survival (OS). Results: The area under curve values for FIB-4 and APRI were the largest for fibrosis stage compared to other biomarkers, 0.669 (95% confidential interval (CI), 0.610–0.719) and 0.748 (95% CI, 0.692–0.800), respectively. Between those two indices, FIB-4 is considered a statistically significant prognostic factor of RFS in HCC patients after LR. The HR for 2-year RFS and OS were 1.81 (95% CI, 1.18–2.77; P = 0.007) and 2.36 (95% CI, 0.99–5.65; P = 0.054), respectively. Conclusion: FIB-4 is identified as a statistically significant predictor of HCC prognosis after curative LR even in HBV dominant populations.

단일 환아에서만 미숙아망막병증이 발생한 쌍둥이 간의 관련인자 비교 분석

임재완(Jae Wan Lim),유재호(Jae Ho Yoo),이승욱(Seung Uk Lee),이상준(Sang Joon Lee),남기엽(Ki Yup Nam) 대한안과학회 2016 대한안과학회지 Vol.57 No.10

목적: 쌍둥이 미숙아 중에서 한 환아에게만 미숙아망막병증이 발생한 쌍을 비교하여 미숙아망막병증과 관련된 인자를 알아내고자 하였다. 대상과 방법: 고신대학교 복음병원에서 미숙아로 태어난 쌍둥이들 중 한 환아만 미숙아망막병증(1단계 이상)으로 진단 받은 13쌍 26명을 대상으로 의무기록을 후향적으로 분석하였다. 미숙아망막병증 유무에 따라 쌍둥이를 ‘미숙아망막병증군’과 ‘미숙아망막병증이 없는 군’으로 나누었다. 태어날 당시의 임신나이, 성별, 몸무게, 아프가점수, 시행된 치료, 혈액검사 결과, 신생아 황달, 패혈증, 호흡곤란 증후군, 기관지폐 형성이상, 동맥관개존증, 신생아괴사성장염 등 신생아 합병증 여부 등을 비교 분석하였다. 결과: 출생 시 시행된 다양한 처치들과 미숙아망막병증 발생과 연관이 있는 것으로 알려져 있는 출생 후 합병증들에 대해서 분석하였으나 두 군 간 유의한 차이는 없었다. 혈액 검사상에서는 혈소판이 미숙아망막병증군에서 191 (±46) ×103/μL, 미숙아망막병증이 없는 군에서 240 (±77) ×103/μL로 나타나 미숙아망막병증군에서 유의하게 낮았고(p=0.046) 아스파라진산 아미노전이효소는 미숙아 망막병증군에서 36 (±26.6) IU/L, 미숙아망막병증이 없는 군에서 22 (±5.9) IU/L로 미숙아망막병증 군에서 유의하게 높은 것으로 확인되었다(p=0.019). 결론: 한 환아에게만 미숙아 망막병증이 발생한 쌍둥이들을 대상으로 한 이번 연구에서 혈소판 수치와 아스파라진산 아미노전이효소가 미숙아망막병증의 발생과 관련이 있는 인자로 확인되었다. 선별 검사 시에 고려해야 할 인자로 생각되나 임상적으로 적용하기 위해서는 추후 더 많은 환아를 대상으로 한 전향적인 연구가 필요할 것으로 사료된다. <대한안과학회지 2016;57(10):1592-1597> Purpose: To analyze related factors of retinopathy of prematurity by comparing between premature twins in which retinopathy developed on one twin. Methods: A retrospective survey consisting of 13 premature twins in which retinopathy of prematurity (stage 1 or more) developed on one twin was performed. All twins were born in Kosin University Gospel Hospital. The twins were separated into two groups according to whether they had retinopathy of prematurity: the retinopathy of prematurity (ROP) group and non-ROP group. The twins’ gestational age, weight, sex, Apgar score, treatments, blood tests, and neonatal complications were investigated. Results: There were no significant differences between the twins except platelet count and aspartate aminotransferase. Platelet count was 191 (±46) ⅹ103/μL in the ROP group and 240 (±77) ⅹ103/μL in the non-ROP group, a significant difference (p = 0.046). Aspartate aminotransferase was 36 (±26.6) IU/L in the ROP group and 22 (±5.9) IU/L in the non-ROP group, a significant difference (p = 0.019). Conclusions: In conclusion, we found platelet count and aspartate aminotransferase to be significant factors related to development of retinopathy of prematurity. It is thought that these factors should be considered when screening for ROP, although a larger prospective study is be needed before the results can be applied in clinical practice. J Korean Ophthalmol Soc 2016;57(10):1592-1597

Immunological measurement of aspartate/alanine aminotransferase in predicting liver fibrosis and inflammation

김현정,Sang Yeol Kim,Suk Pyo Shin,Young Joo Yang,Chang Seok Bang,백광호,김동준,함영림,Eui Yul Choi,석기태 대한내과학회 2020 The Korean Journal of Internal Medicine Vol.35 No.2

Background/Aims: Enzymatic analysis of aspartate/alanine aminotransferase (AST/ALT) does not exactly represent the progression of liver fibrosis or inflammation. Immunoassay for AST (cytoplasmic [c] AST/mitochondrial [m] AST) and ALT (ALT1/ALT2) has been suggested as one alternatives for enzymatic analysis. The objective of this study was to evaluate the efficacy of immunoassay in predicting liver fibrosis and inflammation. Methods: A total of 219 patients with chronic hepatitis B (CHB) who underwent hepatic venous pressure gradient (HVPG) and liver biopsy before antiviral therapy were recruited. Serum samples were prepared from blood during HVPG. Results of biochemical parameters including enzymatic AST/ALT and immunological assays of cAST, mAST, ALT1, and ALT2 through sandwich enzyme-linked immunosorbent assay (ELISA) immunoassay with fluorescence labeled monoclonal antibodies were compared with the results of METAVIR stage of live fibrosis and the Knodell grade of inflammation. Results: METAVIR fibrosis stages were as follows: F0, six (3%); F1, 52 (24%); F2, 88 (40%); F3, 45 (20%); and F4, 28 patients (13%). Mean levels of AST and ALT were 121 ± 157 and 210 ± 279 IU/L, respectively. Mean HVPG score of all patients was 4.7 ± 2.5 mmHg. According to the stage of liver fibrosis, HVPG score (p < 0.001, r = 0.439) and ALT1 level (p < 0.001, r = 0.283) were significantly increased in all samples from patients with CHB. ALT (p < 0.001, r = 0.310), ALT1 (p < 0.001, r = 0.369), and AST (p < 0.001, r = 0.374) levels were positively correlated with Knodell grade of inflammation. Conclusions: ALT1 measurement by utilizing sandwich ELISA immunoassay can be useful method for predicting inf lammation grade and fibrosis stage in patients with CHB.

단일클론항체를 이용한 면역학적 Aspartate Aminotransferase 함량 측정법의 개발

최성아,김동준,최의열 대한간학회 2003 Clinical and Molecular Hepatology(대한간학회지) Vol.9 No.2

목적: 간질환 진단을 위한 많은 종류의 혈액 내표지자들 중 아미노전이효소(aspartate/alanine aminotransferase, AST/ALT)가 간세포 손상의 민감한 표지자로서 현재 임상검사에 널리 사용되고 있다. 그러나 현재 AST/ALT 검사법은 효소의 활성도를 측정하는 것으로서 효소가 그 활성을 잃어 버린다거나 효소 활성도에 영향을 미치는 여러 인자 등에 의해 효소 활성도 측정이 영향을 받게 된다. 이러한 문제점들을 극복하기 위해서 효소 활성도를 측정하는 기존의 측정법을 보완 또는 대체하여 호소의 농도를 정량할 수 있는 면역학적 진단법의 필요성이 제기되어 왔다. 대상과 방법: AST 효소 활성 대신 혈액 내에서의 효소 농도를 측정할 수 있는 새로운 분석법을 개발하기 위하여 형광으로 표지된 인간 AST 특이 단일클론항체를 이용하여 면역 크로마토그래피 방법을 적용하였다. 면역 진단 카트리지는 형광 검출 용액, 일회용 카트리지 안의 AST 검사 스트립, 그리고 형광 판독기로 구성하였고, 면역 진단 카트리지의 측정은 형광분석기를 제작하여 사용하였다. 전혈 시료 20 μL를 검사용액과 섞어서 검사 카트리지에 적하하고 12분 경과 후 형광분석기를 사용하여 측정하였다. 농도계산을 위하여 내부 계산 프로그램을 설계하엮고 시료 내 AST 농도를 제시하도록 고안하여 경도곡선의 직선성과 검출한계 그리고 검사내 정밀도 등을 계산하였다. 결과: 정상인 243명과 43명의 간질환에서 혈액을 채취하고 형광 면역 크로마토그래피법으로 시료 내의 AST를 검출하고 그 농도를 정량 하였다. 시료 내의 AST는 0-1 mg/L 범위에서 농도 의존적으로 형광세기 비율(A_T/A_c)이 비례하여 증가함을 알 수 있었고(R=0.995), AST의 최소 검출한계는 12 μg/L이며 측정가능 범위에서의 평균 회복률은 102.4%이었다. 검사 내 정밀도는 intra-, inter-assay로 50-800 μg/L 농도에서 검사되었는데 그 결과 변이계수가 각각 CV_s<7%, CV_s<6%로 신뢰할 만한 측정결과를 산출하였다. 정상군에서 평균 AST농도는 35.5 μg/L이고 간질 환자들에서는 평균 266.5 μg/L이었다. 또한 형광 면역 크로마트그래피 측정법은 그 결과가 ELISA나 생화학적인 측정결과와도 잘 일치하였다. (각각 R=0.92, 0.88; N=43). 결론: 본 연구에서는 인간 AST 특히 항체와 면역학적인 방법을 응용하여 혈액 내에서 AST 농도를 측정할 수 있는 형광면역 크로마트그래피 측정법을 개발하였다. 이 AST 농도 측정법은 소량의 전혈을 이용하여 높은 재현성을 보여 주어 AST 효소 활성도를 측정하는 현재의 일반적인 검사방법을 보완 또는 대체할 수 있는 간편하고 빠른 현장검사법으로 판단되며 다른 혈액화학검사 항목에 대해서도 새로운 접근법으로 활용될 수 있을 것으로 기대된다. Background/Aims: For laboratory diagnostics in liver diseases, many enzymes have been used for the assessment of hepatocellular function. Among them, two transaminases, alanine and aspartate aminotransferase, have been regarded as the most sensitive indicators of hepatocellular damage. However, the enhanced enzyme activites of the enzymes do not exactly indicate or represent the cause and progression of diseases in the patients with liver disease. To overcome such limitations, immunological methods have been suggested as one of the alternatives for the replacement or supplement of the conventional enzymatic analysis. Methods: In the hope of developing a new assay system for measuring the AST concentration rather than its activity, we have developed a new assay using fluorescence labeled anti-AST monoclonal antibodies. Blood was obtained from a normal population of 234 patients and 43 liver disease patients. The linearity, limit of detection, and performance of the new assay system were tested and evaluated. The comparability of assay was examined with an ELISA and biochemical assays. Results: The linearity fell in the range of 0-1 mg/L of AST (R=0.995), and the analytical detection limit was 12 μg/L of AST. The mean recovery of the control was 102.4% in a working range. The precision of the intra- and inter-assay in a range of 50-800 μg/L was CVs<7% and CVs<6%, respectively. In the normal population, the mean AST concentration was 35.5 μg/L. The mean AST concentration in patients with liver disease was 266.5 μg/L. The new assay system correlated well with an ELISA and biochemical assay for quantification of AST concentration (R=0.92 and 0.88, respectively; N=43). Conclusions: We have developed a new immunological assay using generated monoclonal antibodies to human cytosolic AST and used them for the development of a fluorescent assay measuring the enzyme mass. Cytosolic AST mass in sera could be measured reproducibly by the immunological method. In conclusion, this study has provided us with a new type of tool for an accurate measurement of the enzyme amount in circulation.

질환별 혈청 Lactate dehydrogenase와 Aspartate aminotransferase의 활성비교

김수봉 ( Soo Bong Kim ),윤수홍 ( Soo Hong Yoon ) 한국위생과학회 2004 한국위생과학회지 Vol.10 No.1

사염화탄소로 유발된 간손상에서의 효소 활성도의 변화로 본 홍화자 분획물의 간손상 보호 작용

정춘식,정기화,정정숙 德成女子大學校 藥學硏究所 1999 藥學論文誌 Vol.10 No.1

본 연구소에서는 사염화탄소로 유발된 간손상에 대한 홍화자 메탄올 추출물과 이를 계통 분획한 분획물의 보호 효과를 이미 보고하였다. 본 실험에서는 혈장과 조직 생화학적 분석으로 홍화자 분획물의 간보호 효과를 확인하며, 간보호 작용에 대한 기전을 살펴보기 위하여 약물 대사 효소에 미치는 영향을 측정하였다. 홍화자 butanol subfractions 중 BS-5 분획물은 CCl_4로 간손상을 유발시킨 흰쥐에서의 혈장 ALT와 AST 활성과 간조직 중 cholesterol and triglyceride를 감소시켰으며, 이러한 BS-5 분획물의 CCl_4로 유발된 간손상에 대해 보호 작용은 cytochrome P450 감소와 glutathione S-transferase의 활성 증가에 기인된 것으로 생각된다. Previous studies have shown that methanol extract and its butanol fraction of Carthamus tinctorius L. Semen have the hepatoprotective effect on the CCl_4-induced hepatotoxicity. The hepatoprotective effect of subfractions has been evaluated by analyzing blood and hepatocyte biochemical analyses and biotransformation enzyme analyses. Treatment of BS-5 subfraction has significantly decreased the activities of alanine aminotransferase and aspartate aminotransferase. In addition, the levels of cholesterol and triglyceride in liver have been decreased as compared with that of CCl_4 treated rats. The hepatoprotective effect of BS-5 subfraction on the CCl_4-induced hepatotoxicity would be mediated of the attenuation of the level of cytochrome P450 and the enhancement of the activity of glutathion S-transferase.

Characterization of Aspartate Aminotransferase Purified from Streptomyces fradiae

Lee, Sang-Hee,Lee, Kye-Joon The Microbiological Society of Korea 1993 미생물학회지 Vol.31 No.3

Aspartate aminotransferase (ASAT) (L-aspartate : 2-oxyoglutarate, EC 2.6. 1. 1.) from Streptomyces fradiae NRRL 2702 has been purified by acetone precipitation, DEAE-cellulose, hydroxyapatite, and preparative electrophoresis (Prep cell), of which the last was the most effective step in the purification of ASAT. The molecular mass was estimated to be 54,000 dalton by SDS-PAGE and 120,000 dalton by gel filtration chromatography. Preparative isoelectric focusing of purified ASAT resulted in one polypeptide band with a pI of 4.2, showing homogeneity and indicating that the enzyme is composed of two identical subunits. The enzyme was specific for L-aspartate as an amino donor ; the $K_{m}$ values were determined to be 2.7 mM for L-aspartate, 0.7 mM for 2-oxoglutarate, 12.8 mM for L-glutamate, and 0.15 mM for oxaloacetate. The enzyme was relatively heat-stable, having maximum activity at 55.deg.C, and it had a broad pH optimum ranging from 5.5 to 8.0. The activity of the purified enzyme was not inhibited by ammonium ions. This paper reports the first purification and characterization of the aspartate aminotransferase from a species of Streptomyces.s.

알쯔하이머병과 혈관성치매의 뇌척수액 Aspartate Aminotransferase

김돈수,곽용태 대한노인병학회 2001 Annals of geriatric medicine and research Vol.5 No.1

Declined Preoperative Aspartate Aminotransferase to Neutrophil Ratio Index Predicts Poor Prognosis in Patients with Intrahepatic Cholangiocarcinoma after Hepatectomy

Lingyun Liu,Wei Wang,Yi Zhang,Jianting Long,Zhaohui Zhang,Qiao Li,Bin Chen,Shaoqiang Li,Yunpeng Hua,Shunli Shen,Baogang Peng 대한암학회 2018 Cancer Research and Treatment Vol.50 No.2

Purpose Various inflammation-based prognostic biomarkers such as the platelet to lymphocyte ratio and neutrophil to lymphocyte ratio, are related to poor survival in patients with intrahepatic cholangiocarcinoma (ICC). This study aims to investigate the prognostic value of the aspartate aminotransferase to neutrophil ratio index (ANRI) in ICC after hepatic resection. Materials and Methods Data of 184 patients with ICC after hepatectomy were retrospectively reviewed. The cut-off value of ANRI was determined by a receiver operating characteristic curve. Preoperative ANRI and clinicopathological variables were analyzed. The predictive value of preoperative ANRI for prognosis of ICC was identified by univariate and multivariate analyses. Results The optimal cut-off value of ANRI was 6.7. ANRI was associated with tumor size, tumor recurrence, white blood cell, neutrophil count, aspartate aminotransferase, and alanine transaminase. Univariate analysis showed that ANRI, sex, tumor number, tumor size, tumor differentiation, lymph node metastasis, resection margin, clinical TNM stage, neutrophil count, and carcinoembryonic antigen were markedly correlated with overall survival (OS) and disease-free survival (DFS) in patients with ICC. Multivariable analyses revealed that ANRI, a tumor size > 6 cm, poor tumor differentiation, and an R1 resection margin were independent prognostic factors for both OS and DFS. Additionally, preoperative ANRI also had a significant value to predict prognosis in various subgroups of ICC, including serum hepatitis B surface antigennegative and preoperative elevated carbohydrate antigen 19-9 patients. Conclusion Preoperative declined ANRI is a noninvasive, simple, and effective predictor of poor prognosis in patients with ICC after hepatectomy.