Expression of oncogene products HER2/Neu and Ras and fibrosis-related growth factors bFGF, TGF-beta, and PDGF in bile from biliary malignancies and inflammatory disorders.

The expression of several growth factors and K-ras gene mutation in bile were studied to better understand the pathogenesis and improve early diagnosis of bile duct cancers. Bile samples were collected from 12 cholangiocarcinomas (CLC), 10 ampullary cancers (APC), 3 gallbladder cancers (GBC), 7 pancreatic cancers (PNC), 9 biliary tract infection (BTI), 8 biliary stone disease (ST), and 5 normal controls (NC). The highest mean value of TGF-beta in bile was in patients with BTI; the mean levels of bFGF and PDGF were highest in CLC, and patients with APC and CLC had higher expression of HER2/Neu than other groups. In bile, a K-ras gene codon 12 mutation was found in 5 of 6 (83%) cases of CLC by the PCR-RFLP method. The results suggest overexpression of bFGF, PDGF, and HER2/Neu and the presence of K-ras mutation are important for carcinogenesis of bile duct cancers, and detection of the above abnormalities in bile is helpful for early diagnosis.

Protective effect of melatonin on renal injury of rats induced by bile duct ligation.

Oxygen radicals have been implicated in the pathogenesis of renal injury induced by extrahepatic cholestasis. We conduct this study to investigate whether melatonin can have a protective effect against such injury. Either normal saline or gentamicin with or without melatonin was injected into rats that received either a bile duct ligation or a sham operation. The serum levels of malondialdehyde and total antioxidative activity were measured. The kidney was fixed for histologic scoring of renal injury. The serum malondialdehyde level was highest in the rats that received both bile duct ligation and gentamicin injection. Treatment with melatonin significantly increased the serum total antioxidative activity and reduced the serum malondialdehyde concentration. The mean score of renal injury, highest in the rats receiving bile duct ligation and gentamicin injection, was significantly reduced with melatonin treatment. By reducing the systemic oxygen radicals, supplementation with antioxidants exerts a protective effect on the renal injury induced by extrahepatic cholestasis.

Human biliary beta-glucuronidase activity before and after relief of bile duct obstruction: is it the major role in the formation of pigment gallstones?

BACKGROUND AND AIMS: The bacterial beta-glucuronidase (bBG) can deconjugate conjugated bilirubin to form calcium bilirubinate gallstone. Yet, the role of the human biliary beta-glucuronidase (hBG) in the pathogenesis of pigment gallstone formation still remains unsolved. METHODS: Hepatic bile was collected from bile-duct-obstructed patients on the day of, and 3 days after, biliary drainage. Patients were divided into pigment-stone (PS) group (n = 34) and stone-free (SF) group (n = 29). All patients of the PS group had the complication of cholangitis. The concentrations of bile contents and the activities of bBG and hBG were measured. RESULTS: The activities of hBG and bBG in bile obtained on the day of biliary drainage were higher in the PS group than in the SF group (activities corrected for bile salt concentration: hBG 128.7 +/- 340.0 vs 13.1 +/- 25.0 U/mmol; bBG 12.5 +/- 22.2 vs 4.6 +/- 7.7 U/mmol, P < 0.05). This difference disappeared after biliary drainage. The changes of enzyme activity in the bile of the SF group were unremarkable before and after biliary drainage. The mean concentrations of bile pigments and free calcium in the PS group were lower than those in the SF group. CONCLUSIONS: An increase in the activity of hBG may be a secondary response, developed after bile duct inflammation because it was elevated only when the bile duct obstruction was associated with infection.

Melatonin prevents pigment gallstone formation induced by bile duct ligation in guinea pigs.

Free radical-mediated oxidative stress has been implicated in the genesis of gallstone in vitro. This study was designed to examine the oxidative stress changes during pigment gallstone formation and to investigate whether melatonin (MLT) could act as a chemopreventive agent for cholelithiasis in a guinea pig model. The common bile duct of guinea pigs was ligated with or without MLT pretreatment. Animals were studied on day 7, 9, 12, and 14 after surgery. Stone and/or sludge developed in ligated guinea pigs without MLT. Fourier transform infrared spectra of the sludge showed the presence of calcium bilirubinate, whose peak height per milligram of sludge gradually increased with time after ligation. Total antioxidant activity (TAA) in bile of guinea pigs at day 14 after ligation reduced to one third of the level in sham-operated controls (P <.001). In addition, the bile of ligated guinea pigs had increased pH (P <.001), bile salts (P <.01), and malondialdehyde (MDA) (P <.05), compared to sham controls. Pretreatment of guinea pigs with MLT at a dose of 1,000 microg/kg significantly decreased the incidence of pigment gallstone formation at day 14 after ligation, as compared to no pretreatment (0/7 vs. 8/10). MLT also reverted the ligation-induced changes in biliary bile salts, pH, MDA, and TAA to control levels. These in vivo findings support a causative role of oxidative stress in the bile duct ligation-induced pigment gallstone formation. Antioxidants may prove useful in preventing pigment gallstone formation in humans.

Selection of lower cutoff point of [13C]urea breath test is helpful to monitor H. pylori eradication after proton pump inhibitor-based triple therapy.

Our objectives were to test the efficacy of [13C] urea breath test (UBT) for H. pylori infection in patients before and after proton pump inhibitor (PPI) based triple therapy, and thus to trace the optimal cutoff value of UBT to monitor H. pylori eradication; and to analyze the histologic bacterial density and distribution of H. pylori in patients with equivocal UBT. A total of 441 dyspeptic cases patients enrolled and completed the study design, including 120 noninfected and 321 H. pylori-infected patients. All 441 cases had received the same protocol of UBT. in which the baseline and 15-min gas samples after ingestion of 100 mg 13C-labeled urea were analyzed for excess 13CO2/12CO2 ratio (ECR). In addition, a first endoscopy was performed in each patient to evaluate the initial status of H. pylori by culture and histology. Of the 321 H. pylori-infected patients, 286 received a second endoscopy and a second UBT six weeks after completing any one of four regimens of the PPI-based triple therapy to document the success of H. pylori eradication. During both sessions of endoscopy, topographic gastric biopsies for histology were sampled to evaluate the distribution and density of H. pylori. Based on the diagnostic standard by culture and histology, the sensitivity and specificity of the first UBT achieved most optimally was 97.5% and 96.7%, respectively, by setting the cutoff point of ECR at 4.0. In contrast, using the same cutoff point of 4.0, the sensitivity and specificity of the second UBT in patients after therapy achieved just 80% and 97.6%, respectively. By applying cutoff points downward of 4.0, 3.5, 3, and 2.5 for the second UBT, the sensitivity was elevated to 80%, 82.8%, 88.6%, and 94.3%, respectively, while the specificity was preserved at more than 95.2%. The overall eradication rate of H. pylori was 87.8% (251/286). Seven of 35 patients with failure of therapy had equivocal ECR at the second UBT (range 2-5), and this accounted for the false negative result. All seven patients had low bacterial densities, and three patients had bacteria distributed only in high body near the cardia. In conclusion, selection of a lower cutoff value of ECR at 2.5 is helpful to maintain the diagnostic accuracy of UBT for monitoring the H. pylori eradication. The equivocal ECR of UBT after therapy would be related to the low bacterial load and its distribution.

Serologic response to lower-molecular-weight proteins of H. pylori is related to clinical outcome of H. pylori infection in Taiwan.

The study aimed to examine the serum serological response among H. pylori-infected patients with various upper gastrointestinal diagnoses; to ascertain whether it could be predictive to the diagnostic outcome of dyspepsia. One hundred seventy H. pylori-infected patients with dyspeptic symptoms but without previous treatment were enrolled, including those with duodenal ulcer disease (N = 47), gastric ulcer (N = 23), nonulcer dyspepsia (N = 60), gastric cancer (N = 34), and MALToma (N = 6). Sera from dyspeptic patients without H. pylori infection (N = 33) were used as controls. During endoscopy, gastric biopsies were taken for CLO-test, histology, and culture for the detection of H. pylori infection, defined by a positive culture or positive results of both CLO-test and histology. Total H. pylori IgG antibody was tested by an ELISA method. Antibody responses to specific H. pylori proteins were tested by a western blotting system. Of patients with H. pylori-infected gastroduodenal diseases, 76.5%, 42.9%, 23.6%, 46.7%, 84.1%, 76.5%, 82.9%, and 32.4% on average, showed responses to the 116-kDa (CagA), 89-kDa (VacA), 60-kDa, 45-kDa, 35-kDa, 30-kDa, 26.5-kDa, and 19.5-kDa H. pylori-specific proteins, respectively. A significant association was found between the serological response to 19.5-kDa and 26.5-kDa proteins and malignant outcome of H. pylori infection (P<0.02). Among patients without malignancy, the absence of a band at 19.5 kDa was statistically associated with the presence of an ulcer (P<0.05). The presence of serum antibody against CagA is not different between patients with ulcer and with malignancy in clinical diagnosis. The serum test for detecting antibodies against lower-molecular-weight proteins of H. pylori, such as those of 19.5 and 26.5 kDa, could be useful to identify H. pylori-infected patients at risk of peptic ulcer or malignancy.

The effects of bile duct obstruction on the biliary secretion of ciprofloxacin in piglets.

OBJECTIVE: The aim of this study was to evaluate the effect of bile duct obstruction on biliary secretion of ciprofloxacin and to look for useful parameters to guide clinical use of antibiotics in patients with acute cholangitis. METHODS: Twenty-five Landrace piglets were used in this study. Their common bile ducts were ligated, with a duration ranging from 1 to 7 days. The changes of liver biochemical tests, intrabiliary pressure, diameter of common bile duct, and concentration of ciprofloxacin in bile were compared among the piglets with different severity of bile duct obstruction. RESULTS: The bile-to-serum ratio of the ciprofloxacin concentration was 586.4% +/- 140.3% before the ligation of bile duct and was reduced significantly to 94.5% +/- 118.0% after 1-day obstruction. The biliary secretion of ciprofloxacin was greatly affected by the intrabiliary pressure in that the bile-to-serum ratio of ciprofloxacin concentration was reduced to 12.2% +/- 14.8% when the intrabiliary pressure reached to 32 cm H2O. The change in the intrabiliary pressure was correlated with the diameter of common bile duct, with a correlation coefficient of 0.90 (p < 0.001). CONCLUSIONS: Biliary secretion of ciprofloxacin is decreased in proportion to the increase of intrabiliary pressure. In obstruction, a markedly dilated common bile duct usually indicates high intrabiliary pressure, and thus biliary concentrations of antibiotics will be lower than expected. Hence, our observations suggest that choosing an antibiotic with high antimicrobial activity, or establishing a biliary drainage to lower the intrabiliary pressure, would be beneficial for patients suffering from cholangitis with a dilated biliary tree.

Enhanced dissolution of gallstone by combining ethanol with two commonly used cholelitholytic solvents.

BACKGROUND/AIMS: Contact dissolution therapy is one of the non-surgical treatments for patients with gallstone. Among the various solvents, methyl tert-butyl ether (MTBE) is used for cholesterol gallstone, while tetrasodium ethyl-dimethyl tetraacetate (EDTA-4Na) solution is used to dissolve calcium bilirubinate stones. However, the contents of gallstone cannot be precisely predicted while they are still present in the human body. This study was designed to test if the MTBE and EDTA can be mixed together and to test the solubility of different kinds of gallstone in each original solution and mixture. METHODOLOGY: Each 0.1 gm of mixed cholesterol stone, brown stone and pigment stone from 18 patients was used. Pure ethanol was chosen to enhance the miscibility between the organic phase of MTBE and the aqueous phase of EDTA. The contents of gallstone after dissolution were examined with scanning electromicroscopy. RESULTS: We found the mixture of ethanol, MTBE and EDTA to be the most efficient solvent in gallstone dissolution in comparison with the other two original solvents. The mixture reached a dissolution percentage of 97.96 +/- 1.00, 88.96 +/- 6.51 and 67.75 +/- 14.26 for cholesterol, brown and black pigment gallstone, respectively. CONCLUSIONS: We concluded that ethanol can be used to mix the MTBE and EDTA with good preservation in their litholytic effects on gallstone. The ethanol-MTBE-EDTA solvent is, therefore, a promising universal cholelitholytic agent which deserves further tests for its safety and efficacy in the in vivo study.

Biliary sludge and pigment stone formation in bile duct-ligated guinea pigs.

We studied the effects of bile stasis in a guinea pig model of pigment gallstone. The common bile ducts of guinea pigs were partially ligated, and the guinea pigs killed one or two weeks later. Biliary sludge or stones were examined with the Fourier transform infrared spectroscopy and the scanning electromicroscopy. The bile was analyzed for pH, free calcium, bile acids and bilirubin fractions, and the activities of both bacterial and endogenous beta-glucuronidase. After bile duct ligation, calcium bilirubinate precipitates or stones formed in all except one of the animals studied. The bile pH and the proportion of unconjugated bilirubin rose after bile duct ligation, with a concomitant fall of bilirubin monoglucuronide. The activity of bacterial beta-glucuronidase decreased after ligation, while the activity of endogenous beta-glucuronidase rose at week 2. Our results imply that precipitation of calcium bilirubinate in this animal model was induced by an increased bile pH and the nonenzymatic hydrolysis of conjugated bilirubin.

Computer morphometry for quantitative measurement of liver fibrosis: comparison with Knodell's score, colorimetry and conventional description reports.

Liver fibrosis is currently described quite subjectively or, at best, semiquantitatively by scoring systems. In order to measure the severity of liver fibrosis quantitatively and to compare this with established methods, such as Knodell's scoring system, the colorimetric method and conventional description reports, we undertook the present study. A personal computer with an image grabber card and a microscope equipped with a computer-controlled slide-driver was used for computer morphometry. The principle behind morphometry is based on the different colours of hepatocytes and fibres following staining with Masson's trichrome stain. There were 31 patients (25 male, six female) recruited into the present study with a mean +/- SD age of 41.6 +/- 15 years (range 24-66 years). Of these patients, 16 had chronic hepatitis B, 12 had chronic hepatitis C and three were alcoholics. Colorimetric methods and Knodell's fibrosis score were performed according to established protocols. Conventional description reports were obtained from reviews of patient charts. The results from computer morphometry were highly correlated with results from the colorimetric method, with a correlation coefficiency gamma = 0.85 (P<0.0001). The results from computer morphometry also correlated with both Knodell's scoring system (gamma = 0.69; P<0.001) and conventional description reports (gamma = 0.46; P<0.01). Results from Knodell's scoring system were significantly correlated with computer morphometry, as follows: score 0, 2.7 +/- 1.4; score 1, 5.7 +/- 1.2; score 2, 7.7 +/- 2.3; score 3, 10.7 +/- 3.2; score 4, 21.8 +/- 14.1. The trend was statistically significant by the Wilcoxon rank sum test. In conclusion, our computerized morphometry system is a reliable tool for the evaluation of the severity of liver fibrosis and can be used as a tool for the objective quantification of liver fibrosis.

Indicators of liver excretory function in patients undergoing biliary decompression for obstructive jaundice.

BACKGROUND/AIMS: The recovery of liver function after biliary drainage in patients with obstructive jaundice may be different depending on the severity and duration of the obstruction. We conducted this study to determine whether there are any clinical factors that can be used to monitor the course of recovery. METHODOLOGY: Serum and bile from 12 patients were collected for biochemical testing on the day of drainage and every 3 days for 6 days. Liver function was evaluated by the indocyanine green retention test (ICG R15) before and 6 days after decompression. Patients with an ICG R15 reduction ratio of less than 50% were considered to have a poor recovery (group 1, n = 6), while a good recovery was indicated by a reduction ratio higher than 50% (group 2, n = 6). Sequential data were compared between the groups and correlated with the results of the ICG test. RESULTS: After drainage, the patients in group 1 had less bile acid excretion on day 3 (1.0 +/- 0.8 vs. 3.4 +/- 1.1 mmol/day, p < 0.05), a slower reduction ratio of serum bilirubin on day 3 (0.38 +/- 0.14 vs. 0.60 +/- 0.12, p < 0.05) and more biliary output on day 6 (1.11 +/- 0.25 vs. 0.60 +/- 0.25 L/day, p < 0.05). The ICG R15 reduction ratio was well correlated with the bilirubin reduction ratio, the bile volume and the amount of excreted bile acids checked on day 3 (gamma = 0.73, -0.71 and 0.74, respectively, p < 0.01). CONCLUSIONS: The presence of choleresis implies ductular cell hyperplasia, while decreased excretion of bile acids and a slow reduction of hyperbilirubinemia represents severe liver damage. Both conditions are sequelae of prolonged obstruction; therefore, they might indicate a long and poor recovery.

Implications of Helicobacter pylori serological titer for the histological severity of antral gastritis.

BACKGROUND AND STUDY AIMS: We attempted to determine whether the serological titer of anti-Helicobacter pylori (HP) immunoglobulin (IgG) would be capable of predicting the presence of ulcer, or would correlate with the histological grading of gastritis in patients with dyspepsia. PATIENTS AND METHODS: One hundred eighty-three dyspeptic patients were prospectively included in the study after panendoscopy. Each patient underwent blood sampling for anti-HP IgG titer, and antral biopsy for both a rapid urease test (CLO) and histology. The severity of antral gastritis was semi-quantitated for acute and chronic inflammation scores (range 0-3). RESULTS: The HP findings were positive in 157 patients (85.5%), and their histological inflammation scores and serological titer were higher than those of HP-negative patients (P < 0.05). Based on the endoscopic findings, these 157 patients were classified into ulcer (n = 109) and non-ulcer dyspepsia (n = 48) subgroups. The mean chronic inflammation score in the ulcer subgroup was higher than that in the non-ulcer dyspepsia subgroup (1.77 vs. 1.28, P < 0.001). However, on the basis of only the titer itself, there was no cut-off value for serological titer to predict the presence or absence of ulcer in HP-infected patients. As the scores for either acute or chronic inflammation increased, the mean serological titer rose (acute inflammation score 0-3: 0.63, 0.78, 0.93, 1.39; chronic inflammation score 0-3: 0.18, 0.56, 0.88, 0.91). CONCLUSIONS: The titer of HP serology does not provide a method for predicting the presence of ulcer in patients with HP infection, but may indirectly offer evidence of the severity of histological changes.

Bacterial density of Helicobacter pylori predicts the success of triple therapy in bleeding duodenal ulcer.

BACKGROUND: We studied whether different initial bacterial densities of Helicobacter pylori would alter the eradication rate of H. pylori by triple therapy (amoxicillin 500 mg t.i.d. and metronidazole 500 mg t.i.d. for 14 days; bismuth subcitrate 120 mg t.i.d. for 28 days) in patients with duodenal ulcer bleeding. METHOD: One hundred thirty-six cases with duodenal ulcer bleeding and H. pylori infection (proved by rapid urease test and histology during emergency endoscopy) were studied. One hundred twenty-seven of these patients completed a course of triple therapy. In each case, anti-H. pylori IgG titer, gastric biopsies for H. pylori density (score 1 to 5), and evaluation of severity of gastritis were collected at the first endoscopy and 1 month after completion of the triple therapy. RESULTS: The ulcer healing rate was 84.3% (107 of 127) at the time of the second evaluation. The eradication rate of H. pylori was 76.4% (97 or 127). Eradication for H. pylori failed in 30 cases. In these eradication failure cases, initial serologic titer and density of H. pylori were higher than those of eradication success cases. The eradication rate of H. pylori decreased as the initial density of H. pylori increased (density of H. pylori: 1, 88.3%; 2, 83.8%; 3, 74.2%; 4, 68%; 5, 50%). At the second evaluation, the serologic titer was lower and continued to decline in eradication success cases whose mean residual titer ratio (100% x follow-up titer/initial titer) was lower than that of eradication failure cases (57.1% +/- 14.6% vs 107.1% +/- 24.1%, p < 0.001). The mean residual titer ratio also disclosed an upward trend as the density of H. pylori increased (density of H. pylori 1 to 5: 57.5%, 66.6%, 73.5%, 75.3%, 81.8%, respectively). CONCLUSIONS: We suggest routine gastric biopsy to detect both the presence of H. pylori and its density inasmuch as quantitative results may predict the usefulness of triple therapy. The higher the H. pylori density, the less effective triple therapy will be at successful eradication of H. pylori.

Sequential changes of bile contents in patients with obstructive jaundice from different etiologies.

BACKGROUND/AIMS: The decreasing of serum concentration of bilirubin and the ability of hepatocytes to excrete various biliary contents after release of obstructive jaundice are good indicators of recovery of liver function. We conducted this study to clarify whether different causes of obstructive jaundice have different effects on the biliary excretion and how they are different when obstruction is released. PATIENTS AND METHODS: Fifteen patients with obstructive jaundice undergoing percutaneous transhepatic catheter drainage or endoscopic nasobiliary drainage were classified into two groups, depending on the cause of obstruction: common bile duct stones (n = 7) and biliary tract tumors (n = 8). All patients in the gallstone group presented with acute cholangitis while only three patients in the tumor group had a positive bacteria culture in bile. Fasting biles were collected on the day of catheter placement and the 2nd, 4th, 6th, 8th, 10th day thereafter. The sequential changes of biliary concentration of bilirubin, phospholipid, cholesterol, bile salts and serum bilirubin were checked and compared between the two groups. RESULTS: The difference in the improvement of jaundice between the stone and tumor group (p > 0.05) were not significant, nor were the excretion of biliary contents after relief of obstruction. The reduction of serum bilirubin paralleled with the increased excretion of biliary bile salts and bilirubin (gamma = -0.51, p < 0.01 and gamma = -0.4, p < 0.05) in tumor group, but not in the stone group. CONCLUSIONS: The decrease of serum bilirubin and the sequential changes of bile contents after relief of obstruction are quite similar in stone and tumor induced obstructive jaundice.

C-reactive protein for the evaluation of acute abdominal pain.

The diagnostic value of C-reactive protein (CRP) was established in 143 patients with acute abdominal pain, 67 men and 76 women with mean age of 48 +/- 20 years old. Clinical and laboratory variables were collected after the patients' arrival at the emergency department. The attending clinicians did not consider the CRP value during the study period and did not use it for their management. When patients were grouped by final disposition, which was according to severity, only CRP and leukocyte count were identified as significant quantitative variables by multivariate analysis. CRP can detect the serious conditions, ie, in 79% of the hospitalized group, although specificity was 64%, the total accuracy was 73%. When elevated CRP was combined with leukocytosis, the diagnostic value was much improved, with specificity of 89% and positive predictive value of 88%. The sensitivity was improved to 90% when elevated CRP or leukocytosis was used. It is thus concluded that CRP is a helpful quantitative variable for disposition decision-making in patients with acute abdominal pain.

Long-term outcome of triple therapy in Helicobacter pylori-related nonulcer dyspepsia: a prospective controlled assessment.

OBJECTIVE: To ascertain whether triple therapy alters the history of Helicobacter pylori (HP)-related nonulcer dyspepsia (NUD). METHODS: Forty-one young (<45 yr) dyspeptic patients were confirmed to be HP-related NUD by serology, rapid urease test, and antral biopsy. Endoscopy excluded the presence of ulcer. These cases were randomly plotted into control (n = 21) and triple therapy (n = 20) groups. In the former group, H2 blocker was given for 2 months and then intermittent antisecretory agents for up to 1 yr. In the latter group, 20 patients received a course of triple therapy and then were treated like the control group. The symptom scores (range: 0-10) were collected on enrollment, and at the end of 2nd, 6th, and 12th months. Each case had serial tests of HP IgG ELISA titer on start, at weeks 2, 4, and 8, at the 6th month, and at the end of the 1st yr. The second endoscopy was done in the 9th wk for eradication survey, and the third endoscopy, at the end of the 1st yr to resurvey the HP status. The histological gradings of biopsy specimens, sampled on each endoscopy, were compared. RESULTS: In the triple therapy group, the rate of eradication of HP was 75%. At he end of the 2nd month, the HP-eradicated cases of the triple therapy group improved the symptom score more significantly then the noneradicated cases (2.42+/-1.37 vs.4.76+/-1.58, p <0.001). At the ends of the 6th month and 1st yr, the symptom scores of the eradicated cases improved more significantly than those of the control group (6th month, 0.61+/-1.18 vs.] 2.66+/-2.06; 1st yr, 0.82+/-1.17 vs.] 3.56+/-2.89, p <0.001). The decline trend of ELISA titers occurred only in eradicated cases and became significantly evident from the 4th wk (0.30+/-0.15 vs.] 0.49+/-0.07, p <0.05) and thereafter. Both acute and chronic pathological grading was improved in the triple group at the end of the 1st yr (acute, 1.95-0.46; chronic, 1.9-0.92; p <0,01) CONCLUSION: Compared with control therapy at 1 yr, triple therapy showed greater symptomatic, serological, and histological improvements. Therefore, triple therapy is beneficial to symptomatic HP-related NUD.

Measurement of creatine kinase isoforms by agarose gel electrophoresis in the diagnosis of myocardial infarction.

To evaluate a method to quantitate the isoforms of serum creatine kinase isoenzyme MM (CK-MM) by agarose gel electrophoresis, sera of normal subjects (n = 74) and patients with acute myocardial infarction (n = 21) and other diseases (n = 67) were studied. The within-assay imprecision (CV) for CK-MM1, -MM2, and -MM3 was 1.9%, 0.8%, and 2.2% at the activity of 79, 105, and 64 U/L (30 degrees C, CK-NAC), respectively; while the assay-to-assay imprecision was 4.8%, 3.2% and 3.9%, respectively. The method could detect 5 U/L or more of any CK-MM isoform and was linear with respect to CK activity at values less than 1100 U/L. Sera from healthy subjects (n = 74) contained mainly CK-MM1 (mean = 48.5%), with lesser amounts of CK-MM2 (mean = 30.6%) and CK-MM3 (mean = 20.8%). The central 95-percentile reference range for the ratio of MM3/MM1 was 0.12-1.34 with mean = 0.49. The sensitivity of CK-MM3/MM1 ratio greater than 1.3 in the diagnosis of acute myocardial infarction employing the first available sample was 90% at a specificity of 91%, compared with a sensitivity of 81% and specificity of 87% for the conventional CK-MB assay. At CK-MM3/MM1 ratio of 1.6 or more, the specificity increased to 96% while sensitivity remained unchanged at 90%. This procedure for the quantitation of serum CK-MM isoforms is convenient, practical and suitable for inclusion in the routine panel of cardiac tests.

Ascitic fluid analysis in peritoneal carcinomatosis: comparison of various biochemical tests with ascitic cirrhotics.

Owing to the high false-negative rate of cytological examinations, various biochemical tests have been investigated to search for an alternative method to differentiate between the most frequent causes of ascites formation, i.e. cirrhosis of the liver and peritoneal carcinomatosis. For this purpose, serum and ascitic fluid samples of 71 ascitic patients (54 due to cirrhosis of the liver and 17 due to peritoneal carcinomatosis) were prospectively and simultaneously collected. Eleven major parameters were analyzed, and, of them, ascitic fluid protein, albumin, cholesterol, fibronectin and serum-ascitic gradient of protein and albumin were found to be useful. The serum-ascitic fluid albumin gradient was not as useful as described in the literature and, if clinically applied, the best cut-off value should be raised up to 1.5 gm/dl. Neither as useful was the serum-ascitic fluid protein gradient. Ascitic cholesterol and ascitic fibronectin were more accurate than the ascitic total protein, ascitic albumin and serum-ascitic gradient of protein and albumin in the diagnosis of peritoneal carcinomatosis. Of these tests, an ascitic fluid fibronectin concentration greater than 40 micrograms/ml was the most accurate parameter (diagnostic accuracy 96.2%) that could solely help to establish the diagnosis of peritoneal carcinomatosis.

Serum amylase, isoamylase, and lipase in the acute abdomen. Their diagnostic value for acute pancreatitis.

We evaluated the diagnostic value of serum amylase, isoamylase, and lipase for the diagnosis of acute pancreatitis from sera of patients with acute abdominal pain. Comparison was first made in condition A between 32 patients with image-proven pancreatitis and 414 patients with nonpancreatic acute abdomen (the control group), then in condition B, between 62 pancreatitis patients with or without image proof and the control group. We found (a) that patients with image-proven pancreatitis suffer a more severe clinical course than those without; (b) that the sensitivity, positive predictive value, and accuracy in condition B are higher than in condition A at any cutoff level; (c) that none of the enzyme assays is specific at the upper reference limit, but their diagnostic yields are much improved by raising cutoff levels to about three or four times the upper limit; and (d) that at these selected cutoff levels, amylase had a diagnostic value similar to p-isoamylase or lipase in both conditions (sensitivity 84% and 92% for amylase in conditions A and B, respectively; specificity 98% and 98%; positive predictive value 75% and 90%; negative predictive value 99% and 99%; accuracy 91% and 97%). In conclusion, at an appropriately selected cutoff level, amylase can be effectively used as the first-line test and isoamylase or lipase as adjunct tests for acute abdominal conditions.

Pancreatic enzymes in uremic patients with or without dialysis.

One hundred thirty blood samples from 87 patients with renal failure, but without abdominal pain, were analyzed for blood urea nitrogen (BUN), creatinine, amylase, p-isoamylase, and lipase simultaneously. We found that 74, 78, and 80% of the patients had hyperamylasemia, hyperisoamylasemia, and hyperlipasemia. None had amylase higher than five times the upper limit. A few patients (2.3%) had lipase elevated to more than 10 times the upper limit. No significant change of pancreatic enzyme level was noted as a result of hemodialysis, but a significant amount of amylase was removed from the circulation in patients receiving intermittent peritoneal dialysis. Significantly lower pancreatic enzyme levels were observed in patients with less impairment of renal function. We conclude that elevation of pancreatic enzymes in uremic patients is more frequent and more extensive than most articles indicate, and that the extent of increase is related more to renal function than to the modalities of dialysis the patients received.