Optimization of assay conditions for, and the selected tissue distribution of, alanine aminotransferase and aspartate aminotransferase of English sole, Parophrys vetulus Girard

Optimization of assay conditions for serum alanine aminotransferase (ALAT, GPT) and aspartate aminotransferase (ASAT, GOT) are described for English sole, Parophrys vetulus . Although most conditions for measurement of these fish enzymes parallel conditions in mammals, the optimal pH is lower for both these enzymes. Liver represents a potentially large source of ASAT activity, potential sources for ALAT activity are liver and kidney. As in mammals, the use of these enzymes as indicators of fish liver and kidney dysfunction may be warranted.     

Use of enzymes for the diagnosis of alcohol-related organ damage

Elevated levels of serum enzymes are frequently associated not only with alcohol-related organ damage but also with excessive alcohol consumption and alcoholism without significant tissue injury. However, both in the early detection of alcoholism as well as also in the diagnosis of alcohol-related diseases the sensitivities and specificities of these enzyme markers vary considerably. They may be influenced by nonalcohol-related diseases, enzyme-inducing drugs, nutritional factors, metabolic disorders, age, smoking, etc. Consequently, we have neither a single laboratory test--enzyme marker--nor a test combination that is reliable enough for the exact diagnosis between alcohol- and nonalcohol-related organ damage. In most cases it is possible to determine the tissue from which the elevated enzyme is derived, but only occasionally enzyme changes reflect the quantity of the tissue injury. Gamma-glutamyltransferase (GGT) is the most widely used laboratory marker of alcoholism and heavy drinking, detecting 34-85% of problem drinkers and alcoholics. However, the unspecificity of increased serum GGT limits its use for general screening purposes. Its value in the follow-up of various treatment programs, however, is well established. An elevated level of serum aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT) in an alcoholic or a heavy consumer indicates alcohol-induced organ damage. The use of test combinations significantly improves the information received with single serum enzyme determinations. An ASAT/ALAT ratio greater than 1.5 can be considered as highly suggestive for the alcoholic etiology of the liver injury. Still better discrimination between alcoholic and nonalcoholic origin of the liver disease may be achieved by the determination of the ratio of GGT to alkaline phosphatase. If this ratio exceeds 1.4 the specificity of the finding in favor for alcoholic liver injury is 78%. The determination of the mitochondrial isoenzyme of ASAT also improves the diagnostic value of ASAT determination. The ratio of mitochondrial isoenzyme to total over 4 is highly suggestive for alcohol-related liver injury. In general, however, the determination of serum activities of other enzymes such as ornithine carbamyl transferase, lactate dehydrogenase, isocitrate dehydrogenase, sorbitol dehydrogenase, alcohol dehydrogenase, guanase, aldolase, alkaline phosphatase or glutathione S-transferase do not significantly improve the diagnostic information obtained with more conventional laboratory markers of liver injury.(ABSTRACT TRUNCATED AT 400 WORDS)     

Serum alanine aminotransferase to aspartate aminotransferase ratio and degree of fatty liver in morbidly obese patients

We evaluated the change in serum alanine aminotransferase (ALT; EC 2.6.1.2) to serum aspartate aminotransferase (AST; EC 2.6.1.; ALT/AST) ratio with the degree of fatty liver in morbidly obese patients. A total of 31 patients were included in the study. Fatty liver was graded as 0 to 4+. The mean and SD of AST and ALT were not significantly different between groups of patients with various grades of fatty liver. There was, however, a significant correlation between the ALT/AST ratio and the degree of fatty infiltration of the liver. This, we believe, implies damage mainly to the plasma membrane allowing loss of cytoplasmic enzymes rather than loss of mitochondrial enzymes.     

Biochemical alterations induced by a new phosphorothionate (RPR-II) in tissues of male and female rats

This study was conducted to investigate the effect of a new phosphorothionate, the methyl ester of 2-butenoic acid-3-diethoxy phosphinothioyl (RPR-II) on membrane bound target enzymes aspartate amino transferase (ASAT), alanine amino transferase (ALAT) and RBC acetylcholinesterase (AChE) in different tissues of male and female albino wistar rats when treated orally with 0.014 (low), 0.028 (medium) and 0.042 (high) mg/kg daily for a period of 90 days. Repeated administration of RPR-II caused significant increase of ASAT and ALAT enzymes in serum, liver and kidney and significant decrease was recorded in lung in both male and female rats when measured after 45 and 90 days of treatment. This compound also caused significant inhibition of RBC AChE indicating its effect on nerve synapsis. Females were more susceptible than males with regard to ASAT and ALAT levels in serum and liver and also in kidney ASAT, whereas reverse trend was recorded in lung ALAT, suggesting sexual dimorphism in the treated rats. These studies also indicated that the levels of these affected enzymes were recovered to normal conditions after 28 days of post treatment (withdrawal study). Positive correlation was observed with regard to these enzymes between serum, liver and kidney, whereas in case of serum and lung a negative correlation was recorded. These enzymes profile elucidates lung necrosis whereas in other tissues the level of enzymes increased showing an adaptive mechanism due to the chemical stress.     

Blood thiamine and thiamine phosphate ester concentrations in alcoholic and non-alcoholic liver diseases.

Thiamine state was investigated in patients with alcoholic liver disease, patients with various non-alcoholic liver diseases, and controls using a direct technique (thiochrome assay) to measure thiamine, thiamine monophospate, and the active coenzyme thiamine pyrophosphate in whole blood after isolating the fractions by ion exchange chromatography. Overall nutrition was similar in all groups as assessed by anthropometry, and no patient had clinical evidence of thiamine deficiency. There was no significant difference among the groups in mean concentration of any form of thiamine. The scatter was much greater in patients with alcoholic liver disease but only 8.7% had biochemical thiamine deficiency (defined as a blood concentration of the active coenzyme greater than 2 SD below the mean control value). An unexpected finding was of abnormally high total thiamine concentrations (greater than 2 SD above the mean control value) in 17.4% of patients with alcoholic liver disease, the highest concentrations being found in two patients with severe alcoholic hepatitis and cirrhosis. The ratio of phosphorylated to unphosphorylated thiamine was calculated as an index of phosphorylation and, although the mean did not differ significantly among the groups, the range was greatest in alcoholic liver disease. The lowest ratios occurred in the two patients with severe alcoholic hepatitis, but neither had evidence of thiamine pyrophosphate deficiency. Contrary to studies using indirect assay techniques, these results suggest that thiamine deficiency is unusual in well nourished patients with alcoholic liver disease. The new finding of unexpectedly high thiamine concentrations in some patients may be due to abnormalities of hepatic storage or release in liver disease, particularly in severe alcoholic hepatitis. There was no convincing evidence of impaired thiamine phosphorylation in any patients with liver disease. Conclusions from studies using indirect assays on the prevalence and mechanisms of thiamine deficiency in liver diseases may not be valid.     

Lymphocytotoxicity in alcoholic hepatitis: evalutation of effector cells

Alcohol induced liver damage has been recently suggested to occur in some cases as a consequence of a true autoimmune reaction. However the nature of the effector cells is still debated. Microlymphocytotoxicity "in vitro" test against rabbit hepatocytes has been performed in 29 patients with alcoholic cirrhosis and 8 with acute alcoholic hepatitis. The diagnosis has been done on clinical and histological grounds. The mean cytotoxicity index (CI) of subjects with alcoholic cirrhosis was 15.12 +/- 14.18 and with acute alcoholic hepatitis 62.49 +/- 16.33 (normal range: 0 -37.90). To further characterize the effector cells in our system, the test has been performed again either using T-enriched fractions of peripheral lymphocytes or after pronase-induced digestion of SmIg of total lymphocytes in 5 patients with acute alcoholic hepatitis and high CI. In both instances a disappearance of citotoxic activity has been observed. These data suggest: a) autoimmune reaction can occur only in patients with acute alcoholic liver damage; b) non T-cells are responsible for cytotoxicity; c) the immune reaction appears to be of cell-mediated antibody-dependent type.     

Th17及相关细胞因子与酒精性肝硬化患者肝脏硬度值、肝功能及脂联素水平的相关性分析

摘要 目的：探讨Th17及相关细胞因子与酒精性肝硬化患者肝脏硬度值（LSM）、肝功能及脂联素（APN）水平的相关性分析。方法：以2018年1月~2020年1月本院收治的酒精性肝硬化患者72例作为酒精性肝硬化组,同期在本院进行体检的健康志愿者100例作为健康对照组。对比两组研究对象Th17、白介素-17（IL-17）、LSM、肝功能指标、APN水平的差异,采用Pearson检验评估酒精性肝硬化患者Th17及IL-17与病情相关指标的相关性。结果：酒精性肝硬化组患者的Th17细胞分布比例、IL-17水平、LSM均高于健康对照组（P<0.05）。外周血中谷丙转氨酶（ALT）、谷草转氨酶（AST）、总胆红素（STB）的水平高于健康对照组,白蛋白（ALB）的水平低于健康对照组,血清中APN的水平低于健康对照组（P<0.05）。相关性分析发现,酒精性肝硬化患者Th17细胞分布比例及IL-17水平与LSM、ALT、AST、STB的水平呈正相关,与ALB、APN的水平呈负相关（P<0.05）。结论：酒精性肝硬化患者Th17及其细胞因子IL-17表达水平异常增高,可能在反映病情相关指标异常变化程度方面具有积极作用。     

Assessment of toxicological effects of mancozeb in male rats after chronic exposure

Mancozeb, an ethylenebisdithiocarbamate fungicide was administered orally to male rats at doses 0, 500, 1000 and 1500 mg/kg/day for 90, 180 and 360 days produced dose dependent signs of poisoning, loss in body weight gain and mortality. However the signs of toxicity and mortality were more pronounced initially at 0-90 days as compared to 90-360 days of treatment period. A significant increase in the relative weight of liver and slight decrease in the kidney weight were observed in animals exposed to mancozeb (1000 and 1500 mg/kg/day) for 180 and 360 days associated with pathomorphological changes in liver, brain and kidney. Mancozeb has produced significant enzymatic changes in the activities of aspartate aminotransferase (ASAT), alanine aminotransferase (ALAT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and acetylcholinesterase (AChE) throughout the period of study in a dose dependent manner. The alterations in the activity of enzymes associated with pathomorphological changes suggest that the chronic exposure of mancozeb produced significant toxicological effects in rats.     

Nitrite toxicity in Indian major carps: sublethal effect on selected enzymes in fingerlings of Catla catla, Labeo rohita and Cirrhinus mrigala

The effects of 96-h sublethal exposure of nitrite (1, 2, 4, 8 and 10.4 mg l(-1)) on selected enzymatic activities in serum and tissues of fingerlings of catla (Catla catla), rohu (Labeo rohita) and mrigal (Cirrhinus mrigala) were studied for the first time in these species. All three species responded almost identically to nitrite exposure. With increasing nitrite concentration, reduction in activities was observed in acetylcholinesterase (AChE) in brain and liver; alkaline phosphatase (ALP) in serum, brain and gill; and acid phosphatase (ACP) in gill, while progressive increase in alanine aminotransferase (ALAT) and aspartate aminotransferase (ASAT) activities in brain, gill and serum, and ACP activity in serum and brain was observed. Lactate dehydrogenase (LDH) activity increased in gill, liver, kidney, brain and serum of all three species with increasing nitrite concentration up to 8 mg l(-1) followed by reduction at 10.4 mg l(-1). The study revealed nitrite stress causing alteration in activities of all measured tissue and serum enzymes in the fingerlings, and so stresses the need for proper management of this particular nutrient in water during carp culture.     

Epidemiology of serum aminotransferase activities in the elderly.

BACKGROUND: Our study used data collected in Chung-Hsing Village in May 1998 to explore the distribution of serum aminotransferase activities and the relationship between aminotransferase and its related factors in the elderly. METHODS: All individuals aged 65 and over were recruited as study subjects. A total of 1093 persons, out of 1774 registered residents, were contacted by face-to-face interview. The response rate was 61.6 percent. However, only 586 subjects had blood tests and completed questionnaires. Analysis in this study was based on these 586 subjects. In order to study the significant related factors of abnormal aminotransferase activities, the t-test, ANOVA, chi-square analysis, and multivariate logistic regression were used. RESULTS: There were 66 percent men and 34 percent women. The mean age was 73.1 +/- 5.3 years. The mean values of aspartate aminotransferase (AST) were 29.3 +/- 14.5 u/l in men and 27.8 +/- 10.7 u/l in women (p > .05). The mean values of alanine aminotransferase (ALT) were 30.9 +/- 25.2 u/l in men and 26.3 +/- 12.6 u/l in women (p < .01). The abnormality rates of AST (> or = 40 u/l) were 10.5 percent in men and 12.2 percent in women (p > .05). The abnormality rates of ALT (> or = 40 u/l) were 16.7 percent in men and 12.6 percent in women (p > .05). After controlling for the other covariates, the multivariate logistic regression analysis showed that the significant related factor of abnormal AST was retirement status (odds ratio 4.4; 95 percent confidence interval = 1.5-13.3; p < .01). The significant related factors of abnormal ALT were obesity (odds ratio = 2.2; 95 percent confidence interval = 1.1-4.2; p < .05) and hypertriglyceridemia (odds ratio = 2.7; 95 percent confidence interval = 1.5-4.9; p < .01). CONCLUSIONS: We raise the hypothesis that evidence of liver disease with abnormal ALT may co-vary with other indicators of chronic diseases. A large-scale investigation will be suggested in the future to demonstrate the causal-effect issue between abnormal ALT and obesity or hypertriglyceridemia.     

Biochemical evaluation of hepatic damage in subchronic exposure to malathion in rats: effect on superoxide dismutase and catalase activities using native PAGE

The aim of this study was the evaluation of the hepatic damages following a subchronic exposure to malathion, an organophosphorus (OP) insecticide. Malathion was administered intragastrically in 1 ml corn oil containing 100 mg/kg Body Weight daily for 32 days. Malondialdehyde (MDA) concentration superoxide dismutase (SOD) and catalase (CAT) activities were analysed using a non-denaturing electrophoresis. The serum activities of Pseudocholinesterase (PchE), aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT) were determined. Malathion exposure leads to a significant decrease in AchE activity, an increase in hepatic MDA, and in serum ASAT and ALAT activities. A positive correlation between serum transaminases levels and hepatic MDA was demonstrated. These results indicate that malathion exposure induced lipid peroxidation LPO, a process of degradation of membrane lipids, involving the deterioration of the cellular integrity. We have recorded a slight increase in antioxidant enzymes activities. This leads us to suggest an insufficient elimination of free radicals, causing cytotoxic effects. To cite this article: R. Rezg et al., C. R. Biologies 331 (2008).     

Plasma Constituents in the Sow

The concentrations in plasma of total protein, urea-N, Ca, Mg, Na and K and the activity of aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT) in plasma were studied in 20 sows from one week before weaning until four weeks after weaning. Ten of the sows resumed ovarian activity within 10 days after weaning, while in the other 10 sows the resumption of ovarian activity was delayed. Plasma glucose was studied in 10 sows from one week before weaning until two weeks after weaning, five of these sows had delayed resumption of ovarian function. After weaning the concentration of total protein and the ALAT activity increased, while plasma glucose and Mg decreased. Concerning ASAT, Ca, Na and K no changes were observed. A transient increase in urea-N level took place after weaning in sows with prolonged weaning-to-ovulation period. In sows with normal weaning-to-ovulation period no change in urea-N was observed. This was the only difference in blood chemistry observed between sows with normal weaning-to-ovulation period and sows with prolonged weaning-to-ovulation period.     

Liver aspartate aminotransferase activity as a power function of body weight

Total liver enzyme activity (E) for aspartate aminotransferase (ASAT; EC 2.6.1.1) per gram body weight (W) decreases in six species as body weight increases from mice to cattle according to the equation E/W 0.85 = c, where c is a constant. The 0.85 power of body weight thereby provides a reference standard for the direct arithmetic comparison of liver ASAT activities in six species with different body weights. The use of W 0.85 should provide a tool in clinical studies in which data from experimental animals must be related to humans, e.g., in establishing the relationship between drug dosage (or toxicity) and body weight.     

SD大鼠酒精性脂肪肝模型的监测分析

目的：通过血清生化指标和病理学的监测分析来建立标准的SD大鼠酒精性脂肪肝动物模型。方法：选取40只SD大鼠,随机分为两组,模型组采用直接饮酒法,于第8、12和20周时检测大鼠血清生化指标：丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）、甘油三酯（TG）,并于第8、12周时随机采集5只大鼠肝组织,20周时采集剩余所有大鼠肝组织并进行病理学分析。结果：模型组于第8、12和20周时体重增长量均低于对照组（P〈0．01）,血清ALT、AST均高于对照组（P〈0．01）,第8周和12周时TG高于对照组（P〈0．01）。病理学结果显示肝组织从8周至20周呈现出酒精性脂肪肝、重度酒精性脂肪肝伴肝炎和酒精性肝纤维化等演变过程。结论：直接饮酒法可成功地复制出酒精性脂肪肝动物模型,通过监测分析可了解酒精性脂肪肝病变的整个过程,为今后建立酒精性脂肪肝和肝纤维化动物模型提供了理论参考。     

SF方案治疗肝硬化的疗效观察

目的：观察SF方案对代偿性乙型肝炎肝硬化患者的治疗作用。方法：代偿性乙肝肝硬化患者63例,分为治疗组33例,对照组30例。治疗组采用SF方案进行治疗,对照组采用常规护肝疗法,疗程为9～12个月,定期监测肝功能指标变化。结果：SF方案对肝硬化有较好的疗效,一是肝脏炎症反应减轻,肝功能恢复,血清ALT、AST、TBiL经治疗后治疗组均显著低于治疗前和同期对照组;二是阻止肝纤维化进展,肝纤维化指标明显好转;三是免疫功能得以改善,外周血CD4／CD8淋巴细胞比值回升,补体C3水平上升;四是抗病毒效果显著,HBVDNA阴转率达54．5％。结论：SF方案有显著改善患者肝功能,恢复患者免疫功能,抑制病毒增殖的作用,可扩大样本量进一步研究。     

Subcellular localization of glutamate dehydrogenases and alanine aminotransferase in epimastigotes of Trypanosoma cruzi

Abstract: The subcellular localization of NAD- and NADP-linked glutamate dehydrogenases (GDH-NAD and GDH-NADP), alanine aminotransferase (ALAT) and aspartate aminotransferase (ASAT) in epimastigotes of Trypanosoma cruzi was studied by digitonin extraction from whole cells, subcellular fractionation by differential centrifugation. A and isopycnic ultracentrifugation. All enzymes presented both a cytosolic and a mitochondrial form; in addition, GDH-NADP seems to have a third, still undefined, localization. The results are compatible with the existence of two pathways for the production of l -alanine linked to the reoxidation of glycolytic NADH, one operative in the mitochondrion and the other in the cytosol, and perhaps responsible for the existence of the two alanine pools detected by ¹³C-nuclear magnetic resonance (B. Frydman et al., Eur. J. Biocbem. 192 (1990) 363–368).     

Serum catalase enzyme activity in liver diseases

Serum catalase activity was moderately increased in fatty liver, acute alcoholic hepatitis and in the decompensated form of cardiac circulatory failure. It showed significant increase in acute yellow atrophy and in toxic hepatitis while no changes were detected in liver cirrhosis and viral hepatitis. Serum catalase activity showed a good correlation (r = 0.820) with the serum glutamate dehydrogenase activity. In accordance with our results, the inexpensive assay of serum catalase activity is suggested for the detection of severe liver cell damage.     

beta-Hexosaminidase activity in the acute phase of CCl4 poisoning in the rat

beta-Hexosaminidase (Hex) activity has been shown to be increased in the sera of patients with chronic liver diseases as well as in rats with CCl4-induced liver cirrhosis. In this study, serum and liver Hex activity was determined in rats during the acute phase of CCl4 poisoning, a widely used animal model of acute necrotic liver damage. The results showed a statistically significant decrease of Hex activity in the sera of rats 36 h after CCl4 poisoning (5.84 +/- 2.90 U/l), as compared to controls (11.58 +/- 1.35 U/l; p less than 0.001). No significant change was observed in liver tissue of CCl4-treated animals and controls. A significant correlation between the decrease in Hex and the increase in serum aspartate aminotransferase in serum was found. The results are consistent with the hypothesis that this lysosomal enzyme could be released by non-parenchymal liver cells, such as activated macrophages; its increased activity could be the expression of macrophage activation, as demonstrated in patients with chronic liver diseases.     

Influence of olive and rosemary leaves extracts on chemically induced liver cirrhosis in male rats

The current study was undertaken to evaluate the protective activity of olive and rosemary leaves extracts on experimental liver cirrhosis induced by thioacetamide (TAA) in Wistar male rats. Highly significant decline in the values of body weight gain and highly statistically increase of liver/body weight ratio were noted in rats treated with TAA. Furthermore, the levels of serum alanine aminotransferase, aspartate aminotransferase, gamma glutamyl transferase, alkaline phosphatase and total bilirubin were statistically increased. Additionally, light microscopic examination of liver sections from rats treated with TAA showed a marked increase in the extracellular matrix collagen content and bridging fibrosis was prominent. There were bundles of collagen surrounding the lobules that resulted in large fibrous septa and distorted tissue architecture. Interestingly, the findings of this experimental study indicated that the extracts of olive and rosemary leaves and their combination possess hepatoprotective properties against TAA-induced hepatic cirrhosis by inhibiting the physiological and histopathological alterations. Moreover, these results suggest that the hepatoprotective effects of these extracts may be attributed to their antioxidant activities.     

Radioimmunological determination of serum 3 beta-hydroxy-5-cholenoic acid in normal subjects and patients with liver disease.

A radioimmunoassay for the determination of 3 beta-hydroxy-5-cholenoic acid in human serum has been developed, using 3 beta-hydroxy-5-cholenoyl-thyroglobulin as immunogen and 3 beta-hydroxy-5-cholenoylglycyl-125I histamine as radioactive ligand. The association constant was 6.3 X 10(8) l/mol. Cross reactivity with other bile acids of human serum was not detectable, but was 5.6% with cholesterol. Serum sample preparation included extraction of 3 beta-hydroxy-5-cholenoic acid from serum, solvolysis of sulfates, hydrolysis of conjugates, and separation from cholesterol by thin-layer chromatography. Serum concentrations of 3 beta-hydroxy-5-cholenoic acid were 0.23 +/- SD 0.12 mumol/l and 0.21 +/- SD 0.09 mumol/l in healthy males and females, respectively. In patients with primary biliary cirrhosis the serum concentration of 3 beta-hydroxy-5-cholenoic acid and the quotient 3 beta-hydroxy-5-cholenoic acid over total 3 alpha-hydroxy-bile acids (measured enzymatically) were significantly higher (P less than 0.02) than in patients with chronic active hepatitis or alcoholic cirrhosis. Analysis of 17 sera with elevated concentration of 3 beta-hydroxy-5-cholenoic acid by radioimmunoassay and capillary gas-liquid chromatography showed a close correlation (r = 0.91, slope = 0.97) between the results of the two methods.