Cholylglycine Research Articles

Usefulness of collagen type IV in the detection of significant liver fibrosis in nonalcoholic fatty liver disease

Introduction/aimsLiver fibrosis assessment is a key issue in the evaluation of nonalcoholic fatty liver disease (NAFLD) patients. In the present study, we aimed to validate a noninvasive marker panel to assess significant and advanced fibrosis in these patients. Method126 biopsy-proven NAFLD patients were included. NAFLD diagnosis was based on histological criteria. Fibrosis stages were determined according to NASH-Clinical Research Network criteria. Clinical and laboratorial data were collected during the interval of three months before or after liver biopsy. Histological fibrosis stages were classified as significant fibrosis (F2-F4) and advanced fibrosis (F3-F4). Five serum biomarkers [hyaluronic acid (HA), collagen type IV (cIV), procollagen type III (PC III), laminin (LN) and cholylglycine (CG)] were assessed by chemiluminescence immunoassays. ResultsMost patients were female (61.61%), mean age: 55.7 ± 9.13 years old and mean BMI was 32.1 ± 5.9 kg/m2. Prevalence of diabetes, dyslipidemia, arterial hypertension, and metabolic syndrome was 68.75%, 82.29%, 63.54% and 81.05%, respectively. Patients with cIV above 30 ng/mL had a 5.57-times (IC: 1.86–16.69) the chance of having significant fibrosis and 7.61-times (IC: 2.27–25.54) the chance of having advanced fibrosis versus patients with values below 30 ng/mL. HA, PC III, LN and CG did not detect the presence of significant and advanced fibrosis. The AUROC of clV for detection of significant (0.718) and advanced fibrosis (0.791) was better than that of other serum biomarkers. ConclusionType 4 collagen could predict the presence of significant and advanced fibrosis in NAFLD patients and it would be a useful tool in routine clinical practice.

Reusable optofluidic point-of-care testing platform with lyophilized specific antibody for fluorescence detection of cholylglycine in serum.

A reusable optofluidic point-of-care testing platform (OPOCT) was successfully constructed through integrating evanescent wave fluorescence technology into an all-fiber-based optofluidic system. The compact design of the OPOCT allows it to be portable and suitable for on-site sensitive determination of biomarkers in serum without complicated and costly procedures. The sensitivity of 90.9pM for antibody determination is observed thanks to the high transmission efficiency of excitation light and fluorescence in the OPOCT. The affinity constant between cholylglycine (CG) and anti-CG antibody was quantified using this platform based on the proposed theory. Using the lyophilized fluorescence-labeled specific antibody and reusable fiber optic biosensor, the OPOCT is applied to the one-step sensitive determination of CG in serum, which eliminates the dearth associated with liquid reagent handling, disposable biosensors, and user intervention. A limit of detection of 0.025μg/mL for CG is obtained, which is far more than adequate for meeting diagnostic requirements. The matrix effect of serum samples on the evanescent wave-based optofluidic biosensor can be effectively reduced by simple dilution of serum samples. The performance of the OPOCT also compared favorably with that of a commercial turbidimetric inhibition immunoassay through analyzing multiple serum samples. This platform is ready to expand to measure any other biomarker by using its specific antibody. The simplicity, sensitivity, cost-effectiveness, and robustness of the OPOCT enable the early diagnosis of disease and making a timely clinical decision. Graphical abstract .

Cholylglycine determination by an automated chemiluminescence immunoassay: preliminary results in the intrahepatic cholestasis of pregnancy

Background: One of the most frequent reasons to determine total bile acids (TBA) in a routine laboratory is for the diagnosis and management of intrahepatic cholestasis of pregnancy (ICP). The analytical methods that can be carried out for TBA determination are enzymatic and immunometric methods. However, the enzymatic methods suffer of significant cross-reactivity with ursodeoxycholic acid (UDCA), the principal drug administered in the ICP management. The aim of this paper is to evaluate the clinical utility of a new fully-automated chemiluminescence immunoassay (CLIA) to measure cholylglycine (CG) in the ICP. Methods: Two cohorts of patients with a singleton pregnancy, in the third trimester, were retrospectively recruited. The first cohort (controls, n=56, 21–48 years) was represented by patient hospitalized, for a different reason respect to ICP. The second cohort (ICP women, n=32, 22–45 years) was composed of women with a final diagnosis of ICP, before the treatment with UDCA. Results: Intra-assay and intra-laboratory imprecisions were less than 6%. The 95% CI of CG in the control cohort were 0.9–9.1 µmol/L. The levels of CG in the ICP women cohort ranged from 2.3 to 57.4 µmol/L. The optimal cut-off of CG for ICP was 5.5 µmol/L (sensitivity of 84% and specificity of 92%). Conclusions: CG determination by CLIA method has demonstrated a reliability from the analytical point of view. The obtained diagnostic accuracy for ICP was similar to the traditional assays carried out for TBA determination. Moreover, the turnaround time of CG analysis allows a rapidly answer to clinical suspects of cholestasis in pregnancies ensuring the patient-foetus safety.

Effects of intrahepatic cholestasis of pregnancy on hepatic function, changes of inflammatory cytokines and fetal outcomes.

The effects of intrahepatic cholestasis of pregnancy (ICP) on hepatic function, changes of inflammatory cytokines and fetal outcomes were studied. In total, 663 pregnant women admitted to Daqing Longnan Hospital from July 2016 to December 2017 were selected. There were, 40 cases with ICP enrolled in the observation group, and 40 normal pregnant women were recruited in the normal group. They were also grouped according to hepatic function and inflammatory cytokines, with 40 cases in each group. Neonatal Apgar scores were recorded. The correlations of serum cholylglycine (CG) in pregnant women with umbilical artery systolic-to-diastolic (S/D) ratio in the third trimester of pregnancy, the alanine aminotransferase level, the high-sensitivity C-reactive protein (hs-CRP) level, neonatal Apgar score and gestational week were analyzed. The birth weight in the observation group was lighter than that in the normal group (P<0.05); the gestational week at birth was earlier than that in the normal group (P<0.05); Apgar score at birth was lower than that in the normal group (P<0.05), and the levels of inflammatory cytokines were higher than those in the control group (P<0.05). Apgar scores of newborns at birth and at 1 and 5 min after birth in the normal hepatic function and normal inflammatory cytokine groups were higher than those in the abnormal hepatic function group (P<0.05). The serum CG level in pregnant women was positively correlated with umbilical artery S/D ratio, the alanine aminotransferase level and the hs-CRP level in the third trimester of pregnancy, but negatively correlated with neonatal Apgar score and gestational week. Among patients with ICP, the higher the GG level in the body is, the higher the alanine aminotransferase, inflammatory cytokine and umbilical artery S/D ratio will be, which may cause lower neonatal Apgar score, neonatal asphyxia and premature delivery.

A cholylglycine sensor based on 1,2-naphthoquinone-4-sulphonic acid sodium (NQS)/β-cyclodextrin-graphene oxide modified electrode

In this paper, a novel, simple and sensitive cholylglycine (CG) electrochemical sensor was prepared based on the specific recognition of 1, 2-naphthoquinone-4-sulphonic acid sodium salt (NQS) to CG for the first time. It was constructed with β-cyclodextrin-graphene oxide (β-CD-GO) on glassy carbon electrode (GCE) and modified with NQS to obtain NQS/β-CD-GO/GCE. A decrease of the oxidation peak current of NQS arising from the specific binding of CG and NQS through a nucleophilic substitution reaction was used for indirect detection of CG. The performances of the proposed sensor were investigated by cyclic voltammetry (CV), differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS). The difference of the oxidation peak current was linear with the CG concentration in the range of 0.2-60.0 μM and the detection limit of CG was 0.061 μM. Moreover, the CG sensor showed good repeatability and high stability and has been applied to the determination of CG in human serum with satisfactory recoveries. So, a new method has been provided for detecting electrochemically inert material CG.

Visualization of cholylglycine hydrolase activities through nickel nanoparticle-assisted liquid crystal cells

Cholylglycine (CG) is an extremely important biomarker used to diagnose liver abnormalities. Herein, we report a novel nickel nanoparticle (NiNP)-assisted liquid crystal (LC) cell system that can be used to visualize the enzymatic activities between cholylglycine hydrolase (CGH) and CG. When doped with trace amounts of NiNPs, the nematic LC, 4-cyano-4′-pentylbiphenyl (5CB), exhibited a uniform homeotropic alignment that provided dark optical outputs under a polarizing optical microscope (POM), at the designed LC sensing cells. These cells were sandwiched between a pretreated gold surface and an indium tin oxide (ITO) glass surface. In this system, the LC molecules were disordered, producing bright optical signals when CG was introduced to the binding CGH immobilized gold surface. We determined the CG detection limit in the “10 pM” level, and quantitatively defined a working range of 0.1nM to 1μM. The mechanism of the topographical changes on the reacted surface that led to these results was also verified by atomic force microscopy (AFM). This innovative technique, with its simplified path, shows great advances when compared to existing commercially available enzymatic-based bioassays, and offers an easy-to-use principal allowing the fabrication of practical LC-based sensors for CG detection.

Optical imaging of cholylglycine by using liquid crystal droplet patterns on solid surfaces

Determining cholylglycine (CG) levels is of great importance in the detection of liver abnormalities. In this study, we present a novel liquid–crystal (LC)-based method assisted with cholylglycine hydrolase (CGH) for the optical detection of CG levels. The detection method is based on the disruption of the orientations of a nematic LC, 4-cyano-4′-pentylbiphenyl (5CB), doped with dodecyl aldehyde. Aldehyde-doped 5CB droplets were placed on pre-treated glass slides and patterned with solutions of interest. When in contact with a small drop of a CG aqueous solution, bright fan-shaped LC droplet patterns were observed under polarizing optical microscopy, indicating a planar orientation of LC at the aqueous/LC interface. However, aldehyde with short-alkyl chain couples with glycine released from the enzymatic reaction between CG and CGH forming amphiphilic Schiff bases, which display dark cross patterns of LC, suggesting a homeotropic orientation of the LC. This system may offer a highly sensitive and methodologically simple approach to determine CG levels for clinical diagnostics and commercial applications.

Study on the placental expression of MDR3 in intrahepatic cholestasis of pregnancy

目的 探讨妊娠期肝内胆汁淤积症(intrahepatic cholestasis of pregnancy,ICP)胎盘中胆盐载体磷脂输出泵(multidrug resistance p-plycoprotein,MDR3)的表达,以及母、胎血中总胆汁酸(total bile acids,TBA)和甘胆酸(cholylglycine,CG)水平的变化;进一步分析胎盘胆盐载体对胆汁酸水平的调节作用及其与ICP的关系.方法ICP(ICP组)及正常妊娠(对照组)孕妇各20例.母血、胎儿脐静脉血中TBA、CG水平的测定分别采用放射免疫法和速率法;胎盘组织中胆盐载体MDR3 mRNA的表达采用荧光定量PCR法.结果(1) ICP组母血、脐血中的TBA、CG水平均高于对照组(P＜0.05).ICP组母血TBA水平高于脐血(25.8±16.6和8.6±5.5)μnol/L,CG水平高于脐血(3416.1±1986.0和821.8±673.2)μg/dL(P＜0.05);对照组母血和脐血TBA水平(3.4±2.5和4.4±3.3)μmol/L,CG水平(342.7±234.9和309.3±145.2)μg/dL比较,差异无统计学意义(P＞0.05).(2)ICP和正常妊娠各期胎盘组织中均有MDR3 mRNA的表达.ICP组MDR3 mRNA表达量低于对照组(0.80±0.67和162.63±334.33)拷贝/ng(P＜0.05).(3)ICP组、对照组胎盘中MDR3 mRNA表达量与母血、脐血中TBA和CG水平均无相关性(r=-0.60～0.165,P＞0.05).结论ICP胎盘胆盐载体MDR3 mRNA表达降低,可能引起ICP胎盘对胆汁酸的转运障碍、胎儿体内胆汁淤积,可能是引起ICP胎儿窘迫的原因。

Comparison on efficacy between Astragalus-Polygonum Anti-Fibrosis decoction and Jinshuibao ("Equation missing" ) capsule in Treating liver fibrosis of chronic hepatitis B) capsule in Treating liver fibrosis of chronic hepatitis B

Objective: To observe the efficacy of Astragalus-Polygonum Anti-Fibrosis decoction (APAFD) and Jinshuibao capsule (JSBC) in treating liver fibrosis of chronic hepatitis B.Methods: Ninety-two cases of liver fibrosis of chronic hepatitis B were randomly divided into group A and group B. Patients in group A received APAFD for 48 weeks, and in group B, they received JSBC for 48 weeks. The effects on the level change of hyaluronic acid (HA), laminin (LN), pro-collagen III (PCIII) and collagen IV (CIV) as well as liver functional tests and liver biochemical parameters before and after treatment were observed.Results: Level of serum HA, LN, PCIII and CIV in group A declined more obviously than that of group B, the difference was significant (P < 0.01). The liver functional tests such as total bilirubin (TB), alanine aminotransferase (ALT), albumin/ globulin (A/G) ratio, hepatitis related serum biochemical parameters such as cholylglycine (CG), serum ferritin (SF), prealbumin (PC) of group A were all improved more significantly than that of group B (P < 0.01).Conclusion: APAFD is more effective than JSBC in treating liver fibrosis of chronic hepatitis B and in the inhibition of hepatic inflammation, hence it is a good composite Chinese herbal preparation against liver fibrosis.

Postprandial serum cholylglycine in the detection of hepatotoxicity induced by antituberculous agents

Background: Measurement of bile acid levels has been proven helpful in the detection of hepatic abnormalities when routine liver function test results are normal or only slightly elevated. We compared the effectiveness of postprandial serum cholylglycine (CG), as determined by RIA, with commonly used liver function tests (SGOT, SGPT, alkaline phosphatase, gamma-glutamyl transpeptidase) in order to detect liver injury in patients receiving antituberculous agents. Method: We studied 100 patients receiving antituberculous agents. Laboratory tests were carried out before the initiation of therapy and after 1, 2, 3 and 8 weeks of treatment. The risk of hepatotoxicity in relation to age (>35 years) and/or the addition of pyrazinamide to the 6-month isoniazid–rifampin regimen was also assessed. Results: The percentage of patients with hepatic dysfunction detected by abnormal serum CG levels was significantly greater than that detected with conventional liver function tests. Neither age over 35 years nor the use of pyrazinamide was associated with a greater number of abnormalities in liver function tests. Conclusion: Postprandial serum CG by RIA proved to be a sensitive parameter for detecting hepatotoxicity by antituberculous agents.

Correlation of serum cholylglycine level with hepatic dysfunction in children with sickle cell anemia.

Hepatic dysfunction occurs commonly in children with sickle cell disease (SCD). Although the etiology is multifactorial, cholestasis is a prominent feature. Serum cholylglycine (CG) has been found to be a very sensitive indicator of cholestasis. Our objective was to determine whether CG levels are elevated in children with SCD and whether they are predictive of hepatic dysfunction. Blood samples were obtained from 97 children with SCD. Liver function tests were done and serum CG concentrations were measured. Patients were followed up for 2 years. Thirty-eight percent of the patients had an elevated CG level. During the 2 years of follow-up, 16% of the children with a previously elevated CG level developed abnormal liver function test results or required a cholecystectomy as compared with 13% with a previously normal CG level (p = 0.92). We conclude that although CG level was elevated in 38% of the patients with SCD, it did not appear to predict liver dysfunction during the ensuring 2 years.

Plasma clearance of cholyl-lysyl-fluorescein: a pilot study in humans

Background/Aims: Cholyl-lysyl-fluorescein is a fluorescent analogue of the natural bile acid, cholyl glycine. In vivo and in vitro studies showed that this analogue has many biological characteristics similar to cholyl glycine. In this study we analysed cholyl-lysyl-fluorescein plasma clearance in six healthy volunteers as a potential quantitative liver function test. Methods: The compound in water for injection was administered as an i.v. bolus in the dose of 0.02 mg/kg b.w. Results: The plasma elimination curve showed rapid, intermediate and slow phases of clearance. Half-life (T 1 2 time) for the first (t 1 2 1st phase ), second (t 1 2 2nd phase ) and third (t 1 2 3rd phase ) phases of elimination was 1.7±0.9 min, 6.7±1.6 min and 68±17 min, respectively. Ninety-minute plasma retention (% dose/l plasma) was 2.2%. Cholyl-lysyl-fluorescein volume of distribution and residual fluorescence after 60 min were similar to the data obtained by others for natural or radiolabelled bile acids. In five out of six healthy volunteers a 25-fold higher dose of cholyl-lysyl-fluorescein (0.5 mg/kg b.w.) was injected to estimate the safety margins of the compound. This dose was eliminated at a disappearance rate similar to that of the dose of 0.02 mg/kg b.w. and did not cause any adverse reactions. Serum liver tests measured before and after injection did not change significantly. Conclusions: This study showed that cholyl-lysyl-fluorescein clearance is similar to the clearance of endogenous natural bile acids and may potentially offer a new, dynamic test of liver function.

Synthesis and characterization of a new bile acid and platinum(II) complex with cytostatic activity

With a view to using bile acids as shuttles for delivering platinum-related cytostatic drugs to the liver, a cholylglycine(CG)-derivative of platinum(II) has been synthesized. The complex, named Bamet-H2, was characterized by elemental analysis, FT-IR, NMR, FAB-MS, and UV spectroscopy. The results indicate the following composition: C52H84N2O12ClNa Pt(II). Conductivity data suggest that the complex behaves as a sodium salt (1:1) of a complex of Pt(II) bound to one cl-, one bidentate CG moiety, and another monodentate CG moiety, i.e., Na[Pt(CG-O,N)(CG-O)Cl]. The compound is highly soluble (up to 10 mM) in water, ethanol, methanol, DMF, and DMSO. Bamet-H2 was stable in solution (either water or 150 mM NaCl solution), as measured by HPLC, up to 24 h. At this time, more than 90% of the platinum present in water or saline solutions was found to be Bamet-H2. Cytostatic activity against L1210 murine leukemia cells was found. This characteristic was stronger against rat hepatocytes in primary culture. Isolated in situ rat livers were perfused for 40 min with a recirculating medium containing 1 microM Bamet-H2, CG, or cisplatin. Uptake and excretion into bile were much greater for Bamet-H2 than for cisplatin, but less than for CG. Liver content was higher for Bamet-H2 than for cisplatin or CG. The results point to the potential usefulness of Bamet-H2 in the antitumoral therapy of neoplasias derived from liver parenchymal cells.

A correlative study on serum cholylglycine levels in hepatobiliary disease.

The serum cholylglycine (CG), alanine aminotransferase (ALT) and total bilirubin levels were studied in 210 patients with hepatobiliary disease and in 70 healthy subjects. Serum CG concentrations in all the hepatobiliary diseases were found to be significantly higher than those of their controls. Patients with abnormal increases in ALT and bilirubin levels also showed raised CG concentrations; however, some patients with normal ALT and bilirubin levels, still had markedly elevated CG values. Patients with hepatic cirrhosis had high serum CG levels, followed, in descending order, by chronic active hepatitis and chronic persistent hepatitis. In the cholecystitis and cholelithiasis cases, their CG levels were significantly higher than those of the controls but lower than the values in hepatic disease patients; however, more cholecystitis cases had abnormally high serum bilirubin levels than CG. The results also show that serum CG concentrations vary in the different hepatobiliary diseases, and that serial CG measurements are more sensitive than measuring ALT and bilirubin levels in the diagnosis of hepatic diseases. Serum CG can be used as an index for evaluating the activity of chronic hepatitis; it can also be employed as a diagnostic tool in cholecystitis and cholelithiasis.

Bacterial deconjugation and enterohepatic circulation of norursocholic acid conjugates in rats

Experiments were performed to define the metabolism of norusocholic acid (nUC) conjugates and to quantify to what extent the bile acid pool can be enriched in these bile acids. In vitro incubations of norusocholylglycine (nUCG) and -taurine (nUCT) with small intestinal or cecal content showed deconjugation with only cecal content. Cholylglycine (CG) was deconjugated by small intestinal and cecal content. Infusion of nUCG and CG showed that only a small proportion of nUCG was deconjugated after 24 h of enterohepatic circulation, whereas all CG was deconjugated. When nUCT was administered orally, deconjugation was shown to take place mainly in the cecum. Chronic feeding of nUCT enriched the bile acid pool with only 20% nUCT. We conclude that nUC conjugates are deconjugated primarily by bacteria in the cecum and colon, in contrast to CG, which, in addition to cecum and colon, is deconjugated in the distal small intestine. nUCT and its metabolites do not enrich in the circulating bile acid pool mainly for the following reasons: 1) nUC conjugates have a low affinity for the ileal transport system; 2) nUC, even if formed by deconjugation, is not passively absorbed at a sufficient rate; 3) the small amount of norursodeoxycholic acid formed from nUC is glucuronidated in the liver and glucuronide conjugates do not undergo enterohepatic circulation; and 4) nUC conjugates do not suppress bile acid biosynthesis.

The effect of calcium and sodium ion concentration on the properties of dilute aqueous solutions of glycine conjugated bile salts: Phase behavior and solubility products of the calcium salts of the common glycine conjugated bile acids

A systematic examination was made of the effect of calcium and sodium ion concentrations on the phases present at 25°C and neutral pH of dilute aqueous solutions of four synthetically prepared, naturally occurring glycine conjugated bile salts: chenodeoxycholylglycine (CDCG), ursodeoxycholylglycine (UDCG), cholylglycine (CG), and deoxycholylglycine (DCG). Approximate solubility products [ a Ca 2+ · (B −) 2] were measured, using the assumption that the precipitates had the simplest possible stoichiometric composition for calcium bile salts. Values obtained were: CDCG, 0.2 × 10 −8; DCG, 1 × 10 −8; and UDCG, 4 × 10 −8. The value for CG was considerably higher. Precipitation was remarkably slow in all cases and often occurred after the intermediate formation of metastable, nonmicellar viscoelastic gels. Equilibrium was often not reached after 20 months. Precipitation was prevented or reduced by calcium ion binding to micelles (whose formation was favored by increased sodium ion concentration) and also by binding to monomers, i.e., by the formation of a soluble complex in the absence of micelles. Addition of phosphatidylcholine to a CDCG solution favored micelle formation and reduced precipitation. A mixture of bile salts simulating that present in human bile (CDCG:CG:DCG, 2:2:1) showed little change upon addition of calcium ions and only solutions low in Na + and close to the critical micellar concentration gave a glassy precipitate depleted in CG. Thus, the rarity of precipitation in bile of calcium glycine conjugated bile salts in vivo despite stoichiometric concentrations of both components much in excess of solubility products is due to: (1) lowered bile salt activity by micelle formation, (2) lowered calcium activity by binding to bile salt micelles and monomers (in addition to other biliary constituents), (3) lowered activity of both ions because of the high ionic strength of bile, and (4) slowness of precipitation and formation of metastable gels rather than crystalline precipitates.

Serum bile acid concentrations during pregnancy and their relationship to obstetric cholestasis

We have prospectively studied changes in serum postprandial cholylglycine (CG) concentration during 297 pregnancies. We found an increase in CG concentration from 0.3 μmol/L at 15 weeks' pregnancy to 0.6 μmol/L at 40 weeks' pregnancy. Although this increase was statistically significant (p < 0.005), median concentrations of CG remained well within the normal range (0–1.5 μmol/L). However, 10% of the group showed markedly elevated serum CG concentrations at 30 weeks' pregnancy, and the CG level in this group continued to rise during the third trimester. Pruritus was significantly more common in the group with elevated CG concentrations (48%) than in the group with normal CG levels (20%) (p < 0.005). Serum CG was a much more sensitive predictor of pruritus during pregnancy than other biochemical liver tests. Elevated CG levels were found more commonly in Mediterranean and Asian patients than patients of other ethnic origins (p < 0.025). No statistically significant associations were found between elevated CG concentrations and maternal age, number of previous pregnancies, pruritus during previous pregnancies, contraceptive-induced cholestasis, and fetal maturity. We conclude (a) that obstetric cholestasis is probably much more common than previously suspected and (b) that consideration should be given to the measurement of serum bile acids in all pregnant individuals with unexplained pruritus.

Evaluation of postprandial serum bile acid response as a test of hepatic function.

Commercial assays for serum bile acids (SBA) have made this measurement practical. The purpose of this study was to examine the utility of SBA measured every 30 min after a standardized meal in controls and in patients with acute viral hepatitis, cholestasis, and anicteric cirrhosis. In five controls, repeated examination of the area under the bile acid curve (AUC) was not statistically different, whereas the fasting and 2-hr postprandial levels were significantly different. In the group of patients with anicteric cirrhosis, AUC identified disease in 18/20 using total serum bile acids (TSBAs) and in 15/20 using cholylglycine (CG). AUC can be calculated from three samples obtained at 0, 60, and 120 min without losing the sensitivity achieved with seven serial samples. SGOT, alkaline phosphatase, and serum albumin were compared for sensitivity to the total SBA response curve in 20 patients with anicteric cirrhosis. SGOT and alkaline phosphatase identified only 50% and 55% as abnormal and serum albumin was less sensitive. Using total SBA, combining the fasting level and AUC identified 100% as abnormal; using CG, 85% of these patients were detected. As a stepwise cost-effective approach, the fasting level of SBAs can identify most patients with anicteric liver disease. In cases with normal fasting levels where liver disease is suspected, the three-point AUC determination may identify additional patients.

Profiles of Serum Bile Acids in Live Diseases

Cholylglycine (CG) and sulfolithocholylglycine (SLCG) in fasting and postprandial serum were determined in patients with liver diseases by radioimmunoassay. In normal controls, fasting CG and SLCG were 54.67±68.66ng/100ml and 16.61 ±12.84ng/100ml respectively, while postprandial CG and SLCG were 61.21 ±37.29ng/100ml and 21.95± 15.9ng/100ml respectively.In liver disease, serum bile acids were elevated. The greatest increase was found in acute viral hepatitis but moderate or slight increase was also found in chronic active hepatitis, liver cirrhosis, and hepatoma. Fasting bile acids seem to be a sensitive index for hepatocellular dysfunction but not for differential diagnosis of liver diseases.In liver diseases except hepatoma, fasting CG and SLCG correlated significantly with total bilirubin, albumin, GOT, GPT, and alkaline phosphatase but not with cholesterol.Insignificant elevation of bile acids was found postprandially in patients with liver diseases as well as normal controls and postprandial bile acids were not more sensitive than fasting ones.

Rapid assays based on immobilized bioluminescent enzymes and photographic detection of light-emission

Sensitive assays have been developed for adenosine 5'-triphosphate, the reduced form of nicotinamide adenine dinucleotide, cholyl glycine and alcohol, with immobilized and co-immobilized preparations of bacterial and firefly luciferase as reagents. With high-speed (ASA 20000) instant photographic film as detector, picomole amounts of the various analytes can be detected rapidly. The simplicity and convenience of the analytical combination of co-immobilized bioluminescent enzymes and photographic film for the detection of light make this an ideal technique for rapid screening tests.