1.Development of a Method for the Immunological Measurement of Aspartate Aminotransferase with Monoclonal Antibodies.
Sunga CHOI ; Dong Joon KIM ; Eui Yul CHOI
The Korean Journal of Hepatology 2003;9(2):135-144
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 activities 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 microgram/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 microgram/L was CVs < 7% and CVs < 6%, respectively. In the normal population, the mean AST concentration was 35.5 microgram/L. The mean AST concentration in patients with liver disease was 266.5 microgram/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.
Antibodies, Monoclonal
;
Aspartate Aminotransferase, Cytoplasmic/*blood/immunology
;
Fluorescent Antibody Technique/*methods
;
Humans
;
Liver Diseases/diagnosis
2.Aspartate and alanine aminotransferase in Fasciola hepatica.
Sun Hyo PARK ; Nyon Soo KWON ; Hi Sung LEE ; Chul Yong SONG
The Korean Journal of Parasitology 1983;21(1):41-48
The activity and distribution of aspartate aminotransferase (EC 2.6.1.1) and alanine aminotransferase (EC 2.6.1.2) in adult Fasciola hepatica have been studied. Fasciola hepatica was fractionated by differential centrifugation into nuclear, mitochondrial and cytosolic fractions. The activity of GOT and GPT was measured by the method of Reitman and Frankel. Isozyme patterns of those enzyme were also examined by DEAE-cellulose column chromatography. The results obtained were as follows: The activity of aspartate and alanine aminotransferase was about 0.55 unit and 0.92 unit per 1 g of Fasciola hepatica, respectively. The activity of those enzymes was relatively low compared with those in mammalian tissues. The distribution of aspartate aminotransferase in the subcellular organelles showed that 71 percent of the activity was in cytosolic, 24 percent in mitochondrial and 5 percent was in nuclear fraction. About 22 percent of the total alanine aminotransferase activity was found in the mitochondrial fraction, about 66 percent in the cytosolic fraction. Aspartate aminotransferase from cytosolic fraction was separated into two types of isozymes, whereas alanine aminotransferase from cytosolic fraction gave only one active peak on DEAE-cellulose column chromatography.
parasitology-helminth-trematoda
;
Fasciola hepatica
;
biochemistry
;
enzyme
;
aspartate aminotransferase
;
alanine
;
aminotransferase
;
alanine aminotransferase
3.Association of Ki67 with raised transaminases in hepatocellular carcinoma.
Phaik-Leng Cheah ; Lai-Meng Looi ; Abdul Rahman Nazarina ; Kein-Seong Mun ; Khean-Lee Goh
The Malaysian journal of pathology 2008;30(2):103-7
Transaminase enzymes, alanine (ALT) and aspartate transaminase (AST), have been reported to be raised and implicated to have prognostic value in hepatocellular carcinoma (HCC). Ki67, a marker of cellular proliferative activity, has also been noted to be increased in HCC. A study was conducted at the Department of Pathology, Faculty of Medicine, University of Malaya, Kuala Lumpur to determine the possible association of proliferative activity, as determined by Ki67, with the transaminase enzymes. 31 cases of histologically diagnosed HCC who underwent tumour resection were retrieved from departmental archives. The patients' ages ranged between 40 to 79 years with a mean of 58.3 years. There was a male preponderance with M:F = 2.9:1. Ethnic Chinese formed 83.9% of the cases. 4 microm sections, cut from the formalin-fixed, paraffin-embedded tumour tissue block of each case, were immunohistochemically stained with Ki67 (DAKO monoclonal MIB-1) using the commercial DakoCytomation EnVision+System-HRP kit. The latest ALT and AST levels, assayed within 7 days prior to tumour resection, were retrieved from the patients' case records. 24 (77.4%) HCC demonstrated elevation of either ALT and/or AST. 27 (87.1%) HCC were immunopositive for Ki67. Ki67 immunoexpression was significantly correlated with raised transaminases (p<0.05). Hypothetically, the mechanism by which this phenomenon may occur may simply be release of transaminases due to destruction of hepatocytes by the cancer. Thus rising levels of the transaminases could signal a more rapid growth of the tumour and these routinely performed tests can be of prognostic value in management of HCC patients.
Aspartate aminotransferase assay
;
Alanine aminotransferase measurement
;
Primary carcinoma of the liver cells
;
prognostic
;
Excision
4.Changes of SGOT and SGPT enzym activities in blood of experimental mice after long-time exposing to Bassa
Journal of Medical and Pharmaceutical Information 2000;(4):29-31
100 white mice were divided in two groups, 50 mice for each one. The experimental group was exposed to Bassa 50EC with dose of 30 mg/kg body weight orally during 60 days. The other one was taken distilled water with a similar dose and time. The results showed that SGOT and SGPT enzyme activities in blood of experimental mice increased after exposing to Bassa.
Aspartate Aminotransferases
;
Alanine Transaminase
;
blood
5.A Clinical Study of Hypotensive Effect of Tripamide(Normonal(R)) in Essential Hypertension.
Chang Soo LEE ; Young Gun YOON ; Jeong Gwan CHO ; Jung Chaee KANG ; Ock Kyu PARK
Korean Circulation Journal 1984;14(1):157-163
The hypotensive effect of tripamide(Normonal(R)) were evaluated in 31 cases of essential hypertension. Fifteen to thirty mg of tripamide per day were administered continuously for 8 weeks. The results were as follows; 1) The systolic blood pressure was significantly lowered from 175+/-15 mmHg(Mean+/-SD) before treatment to 144+/-17 mmHg after treatment(p<0.01), and the diastolic blood pressure was significantly lowered from 106+/-11 mmHg before treatment to 90+/-12 mmHg after treatment(p<0.01). 2) The systolic blood pressure was lowered 20 mmHg or more in 25 out of 31 cases(80.7%), and the diastolic blood pressure was decreased 10 mmHg or more in 25 out of 31 cases(80.7%) at the end of 8 weeks treatement. 3) There was no significantl differences in the level of SGOT, serum cholesterol, serum electrolytes(Na+, K+, Cl-), BUN, creatinine and urinc acid before and after treatment with tripamide. 4) In most cases the subjective symptoms disappeared or became easier after tripamide treatment. 5) Profound weakness was developed in 2 cases during tripamide administration.
Aspartate Aminotransferases
;
Blood Pressure
;
Cholesterol
;
Creatinine
;
Hypertension*
6.Precise application of Beichaihu and Nanchaihu in classical formulas.
Yu-Ping SHE ; Pei-Pei ZHANG ; Jing-Yan HU ; Yong-Hua GUO ; Qian CAO ; Xiang YIN ; Xiu-Hua HU ; Ting ZHAO ; Feng-Bo LIU ; Yuan ZHANG ; Sheng-Li WEI
China Journal of Chinese Materia Medica 2023;48(3):681-688
To maintain the precision and stability of the efficacy of classical formulas, this study compared the origins and specifications of Bupleuri Radix and revealed the precise application regularity of Bupleurum chinense(Beichaihu) and Bupleurum scorzonerifolium(Nanchaihu) in classical formulas. The efficacy and indications of formulas with Bupleuri Radix as the sovereign drug in the Treatise on Cold Damage and Miscellaneous Diseases(Shang Han Za Bing Lun) were investigated. The difference in the efficacy of Bupleuri Radix as well as the differences in the chemical composition, and liver-protecting and lipid-lowering effects of the decoctions of Beichaihu and Nanchaihu were analyzed with LC-MS technology based on the CCl_4-induced liver injury model in mice and sodium oleate-induced HepG2 hyperlipidemia cell model. The results showed that seven classical formulas with Bupleuri Radix as the sovereign drug in the Treatise on Cold Damage and Miscellaneous Diseases were mainly used in the treatment of digestive, metabolic, immune, circulatory, and other diseases. Bupleuri Radix mainly played the functions of protecting the liver, benefiting the gallbladder, and lowering the lipid, and had different focuses in different formulas. There were 14 differential components in the decoctions of Beichaihu and Nanchaihu, and the chemical structures of 11 components were identified, including 10 saponins and one flavonoid. The results of the liver-protecting efficacy experiment showed that compared with the Nanchaihu decoction, Beichaihu decoction could reduce the serum aspartate aminotransferase(AST) activity in liver injury model mice(P<0.01). The results of the lipid-lowering efficacy experiment proved that Beichaihu and Nanchaihu decoctions both showed highly significant differences in lowering the total cholesterol(TC) and triglyceride(TG) content in HepG2 cells(P<0.01), and Nanchaihu decoction was superior to Beichaihu decoction in lowering the lipid. The results of this study preliminarily proved that there were differences in chemical composition, and liver-protecting and lipid-lowering effects of Beichaihu and Nanchaihu decoctions, indicating that it was necessary to determine the precise origin of Bupleuri Radix in the clinical formulation of traditional Chinese medicine. The study provides a scientific basis for both precise clinical medication and purpose-based accurate quality evaluation of traditional Chinese medicine in clinical application.
Animals
;
Mice
;
Liver
;
Aspartate Aminotransferases
;
Bupleurum
7.Changes of ALP , SGOT and SGPT Levels Following Halothane Anesthesia .
Korean Journal of Anesthesiology 1983;16(4):315-319
In the present study, an effort was directed to assess the changes of hepatic ensymes according to the exposed in short and long surgical cases under halothane anesthesia. 60 surgical cases were studied in two different group. Halothane anesthesia was given for less than 2 hours in the first group of 30 cases and for more than 2 hours in the second group of 30 cases. The results were compared with the normal values and summarized as follows: In the first group, the changes after halothane exposure for less than 2 hours were negligible. In the second group, SGOT and SGPT were elevated significantly and the number of enzyme-elevated cases were 5~6 greater in group 1.
Alanine Transaminase*
;
Anesthesia*
;
Aspartate Aminotransferases*
;
Halothane*
;
Reference Values
8.An Experimental Study on the Effect of Thiopental on Glutamic Oxaloacetic Transaminase Isozyme in Adult Rabbit Brain.
Ok Yong SHIN ; Kwang II SHIN ; Yong Ho CHO
Korean Journal of Anesthesiology 1983;16(4):275-283
In order to determine the enzyme activity, as expressed in Reitman-Frankel unit, of GOT isozyme present in whole homogenate, mitochondrial fraction and supernatant fraction were prepared from brain tissues of normal adult rabbit, by a differential centrifugal method. The effect of thiopental on the GOT isozyme activity in each fraction was determined and the following results were obtained. 1) The activity of GOT isozyme in whole homogenate of normal rabbit brain tissues was found to be 545+/-2.608 units/mg of wet weight whereas the corresponding figure for the supernatant GOT isozyme was 512+/-3.081 and the value for the mitochondrial GOT isozyme was found to be 34.9+/-1.224. 2) The supernatant GOT isozyme existing in a floating status within the cytoplasm accounted for 94 percent followed by 6.35 percent of mitochondrial GOT isozyme. 3) The activated-peak of mitochondrial GOT isozyme contained in the whole homogenate of adult rabbit brain tissues was found to be at #15 on the tube of elution in comparison to that of #73 for supernatant GOT isozyme, as analyzed by the DEAE-Cellulose column chromasography. 4) The supernant GOT isozyme from the thiopentaltreated brain was proportionaly distorted while mitochondrial GOT isozyme was not influenced. Fro example, treated with thiopental, the supernatant GOT isozyme was divided to be #63 & #73 on the tube in comparison to #15 for the mitochondrial GOT isozyme. 5) The activity of supernatant isozyme was proportionaly reduced as the concentration of thiopental. 6) Fifty percent inhibition dose(1se) of thiopental on the supernatant GOT isozyme was found to be 0.63mM. 7) The inhibitory effect of thiopental on the supernatant GOT isozyme was very high significantly by the statistics. 8) The mchanism by which thiopental inhibits the supernatant GOT isozyme in the adult rabbit brain was found to bh uncompetitive inhibition as its Michaelis-Menten constant Km=58.07mM demonstrated. In view of the above finding it is suggested that the thiopental inhibited selectively the activity of supernatant GOT isozyme of the adult rabbit brain tissues while it did not inhibitnificantly by the statistics. 8) The mechanism by which thiopental inhibits the supernatant GOT isozyme in the adult rabbit brain was found to bh uncompetitive inhibition as it Michaelis-Menten constant of Km=58.07 mM demonstrated. In view of the above findings it is suggested that the thiopental inhibited selectively the activity of supernatant GOT isozyme of the adult rabbit brain tissues while it did not inhibit that of mitochondrial GOT isozyme. The GOT isozyme of adult rabbit brain tissues was divided into thiopental-sensitive GOT isozyme(supernatant GOT isozyme) and thiopental insensitive GOT isozyme(mitochondrial GOT isozyme) Furthermore, it is suggested that the cellular function of the brain can be somewhat hindered, when thiopental is injected into the brain cell, while mitosis of the brain cell is not influenced.
Adult*
;
Aspartate Aminotransferases*
;
Brain*
;
Cytoplasm
;
DEAE-Cellulose
;
Humans
;
Mitosis
;
Thiopental*
9.Aspartate aminotransferase activity in the pulp of teeth treated for 6 months with fixed orthodontic appliances.
Rita VEBERIENE ; Dalia LATKAUSKIENE ; Vilma RACINSKAITE ; Neringa SKUCAITE ; Vita MACHIULSKIENE
The Korean Journal of Orthodontics 2015;45(5):261-267
OBJECTIVE: To measure aspartate aminotransferase (AST) activity in the pulp of teeth treated with fixed appliances for 6 months, and compare it with AST activity measured in untreated teeth. METHODS: The study sample consisted of 16 healthy subjects (mean age 25.7 +/- 4.3 years) who required the extraction of maxillary premolars for orthodontic reasons. Of these, 6 individuals had a total of 11 sound teeth extracted without any orthodontic treatment (the control group), and 10 individuals had a total of 20 sound teeth extracted after 6 months of orthodontic alignment (the experimental group). Dental pulp samples were extracted from all control and experimental teeth, and the AST activity exhibited by these samples was determined spectrophotometrically at 20degrees C. RESULTS: Mean AST values were 25.29 x 10(-5) U/mg (standard deviation [SD] 9.95) in the control group and 27.54 x 10(-5) U/mg (SD 31.81) in the experimental group. The difference between these means was not statistically significantly (p = 0.778), and the distribution of the AST values was also similar in both groups. CONCLUSIONS: No statistically significant increase in AST activity in the pulp of mechanically loaded teeth was detected after 6 months of orthodontic alignment, as compared to that of teeth extracted from individuals who had not undergone orthodontic treatment. This suggests that time-related regenerative processes occur in the dental pulp.
Aspartate Aminotransferases*
;
Aspartic Acid*
;
Bicuspid
;
Dental Pulp
;
Orthodontic Appliances*
;
Tooth*
10.Voriconazole Therapeutic Drug Monitoring is Necessary for Children with Invasive Fungal Infection.
Hyun Mi KANG ; Soo Young KANG ; Eun Young CHO ; Kyung Sang YU ; Ji Won LEE ; Hyoung Jin KANG ; Kyung Duk PARK ; Hee Young SHIN ; Hyo Seop AHN ; Hyunju LEE ; Eun Hwa CHOI ; Hoan Jong LEE
Korean Journal of Pediatric Infectious Diseases 2014;21(1):9-21
PURPOSE: To determine the clinical significance of voriconazole therapeutic drug monitoring (TDM) in the pediatric population. METHODS: Twenty-eight patients with invasive fungal infections administered with voriconazole from July 2010 to June 2012 were investigated retrospectively. Fourteen received TDM, and 143 trough concentrations were analyzed. All 28 patients were assessed for adverse events and treatment response six weeks into treatment, and at the end. RESULTS: Out of 143 samples, 53.1% were within therapeutic range (1.0-5.5 mg/L). Patients administered with the same loading (6 mg/kg/dose) and maintenance (4 mg/kg/dose) dosages prior to initial TDM showed highly variable drug levels. Adverse events occurred in 9 of 14 patients (64.3%) in both the TDM and non-TDM group. In the TDM group, voriconazole-related encephalopathy (n=2, 14.3%) and aspartate aminotransferase (AST) or alanine aminotransferase (ALT) elevation (n=8, 57.1%) occurred with serum levels in the toxic range (>5.5 mg/L), whereas blurred-vision (n=2, 14.3%) occurred within the therapeutic range (1.18 mg/L and 3.9 mg/L). The frequency of voriconazole discontinuation due to adverse events was lower in the TDM group (0.0% vs. 18.2%, P=0.481). Overall, 57.2% of the patients in the TDM group versus 14.3% in the non-TDM group showed clinical response after 6 weeks (P=0.055), whereas 21.4% in the TDM group versus 14.3% in the non-TDM group showed response at final outcome (P=0.664). In the TDM group, >67.0% of the serum levels were within therapeutic range for the first 6 weeks; however 45.5% were within therapeutic range for the entire duration. CONCLUSION: Routine TDM is recommended for optimizing the therapeutic effects of voriconazole.
Alanine Transaminase
;
Aspartate Aminotransferases
;
Child*
;
Drug Monitoring*
;
Humans
;
Retrospective Studies