Efficacy and safety of prednisolone in patients with autoimmune hepatitis.

A retrospective cohort study involving 29 Japanese patients with autoimmune hepatitis (AIH) was performed to clarify factors that predict the efficacy of prednisolone and the occurrence of various serious adverse effects. Independent predictors were identified by logistic analysis and with use of the Cox proportional hazard model. Responses to prednisolone were noted in 28 patients, who were classified into the complete remission group (52%) or the relapse group (48%). Multivariate analysis identified alanine aminotransferase, alkaline phosphatase, and immoglobulin G levels as independent predictors of relapse. The adverse effects most frequently observed were diabetes mellitus (37.9%), psychiatric/ neurologic symptoms (34.5%), and circulatory symptoms (34.5%). Predictive factors included lactate dehydrogenase, albumin, and fasting blood glucose levels for diabetes mellitus, alkaline phosphatase and C-reactive protein for psychiatric/ neurologic symptoms, and autoimmune hepatitis score and lactate dehydrogenase for circulatory symptoms. Selection of an optimal treatment method for individual patients may be possible after the risks of relapse and adverse effects have been estimated.

Molecular differences between sporadic serrated and conventional colorectal adenomas.

PURPOSE: The purpose is to compare the molecular characteristics of serrated adenomas (SAs) with those of conventional adenomas (CADs) and hyperplastic polyps (HPs). EXPERIMENTAL DESIGN: We evaluated the proliferative activity and molecular alterations in 47 SAs (25 pure-type and 22 mixed-type), 71 CADs, and 23 HPs. RESULTS: The proliferative activity of SAs, as evaluated by Ki-67 expression, was intermediate between CADs and HPs. There was no significant difference in the incidence of KRAS or p53 mutations between the three histological groups. In the microsatellite instability (MSI) analysis, 21% of SAs (9 of 43) showed MSI at two or more loci (MSI-H); corresponding values were 5% of CADs (3 of 64) and 8% of HPs (1 of 13; SAs versus CADs, P = 0.0125). MSI-H was more likely to be found in pure-type SAs (36%; 8 of 22) than in mixed-type SAs (5%; 1 of 21; P = 0.0212). Loss of hMLH-1 expression was found in 8 of 9 SAs with MSI-H. The incidence of BRAF or KRAS mutations was 36 and 15% of SAs, respectively; the combined incidence of BRAF and KRAS mutations occurred in 49% of SAs. However, there was no significant difference in the incidence of BRAF or KRAS mutations between SAs with and without MSI-H. CONCLUSIONS: Genetic instability is more frequently implicated in the tumorigenesis of SAs, especially pure-type SAs, than in that of CADs. In contrast, activation of the Ras/Raf/MEK/MAP kinase cascade by BRAF or KRAS mutation, independently of the genetic instability, may be associated with the progression of about half of SAs.

Colorectal carcinoma with special reference to growth pattern classifications: clinicopathologic characteristics and genetic changes.

BACKGROUND: Many colorectal carcinomas are known to develop from preexisting polypoid adenomas; however, they can also develop from so-called "flat adenomas". To elucidate the growth patterns of flat- or depressed-derived colorectal carcinomas, we investigated the clinicopathologic characteristics and genetic changes of invasive carcinomas. METHODS: Seventy-five colorectal carcinomas were classified into three groups: 46 upward growth (UG) type, 22 downward growth (DG) type, and 7 lateral growth (LG) type. All of them had histologically infiltrated the submucosa (SM) and muscularis propria (MP). Ki-ras mutation was examined by polymerase chain reaction-single-strand conformation polymorphism analysis, and overexpression of p53 protein was analyzed by immunohistochemistry. RESULTS: No DG or LG carcinomas histologically demonstrated an adenomatous remnant, whereas UG carcinomas did (SM, 19 of 26; 73%; MP, 3 of 20; 15%). The percentage of tumors existing in the right colon was significantly higher in LG carcinomas (71%) than in the UG type (28%; P = 0.037). The frequency of Ki-ras mutation was significantly higher in the UG carcinomas than in the DG and LG carcinomas (52% vs 0%; P < 0.0001; and vs 0%; P = 0.014). However, the frequency of this mutation in SM-UG carcinomas with an adenomatous remnant (9 of 19; 47%) did not differ significantly from that in SM-UG carcinomas without an adenomatous remnant (3 of 7; 43%). The frequency of p53 overexpression did not differ among UG (57%), LG (57%), and DG (50%) carcinomas. CONCLUSIONS: These results suggest that UG carcinomas develop on the basis of the adenoma-carcinoma sequence, while the development of DG and LG carcinomas is different from that of UG carcinomas.

The influence of feeding on gastric acid suppression in Helicobacter pylori-positive patients treated with a proton pump inhibitor or an H2-receptor antagonist after bleeding from a gastric ulcer.

BACKGROUND: This study investigated the influence of feeding on gastric acid suppression in Helicobacter pylori-positive patients treated with intravenous infusions of proton pump inhibitors (PPIs) or with H2-receptor antagonists (H2-RAs) after bleeding from a gastric ulcer. METHODS: Forty-nine H. pylori-positive patients with bleeding gastric ulcers (44 men and 5 women) were divided into four groups: one group received an H2-RA while fasting, one group received an H2-RA while eating regularly, one group received a PPI while fasting, and one group received a PPI while eating regularly. Intragastric pH was monitored during fasting and nonfasting to calculate the pH 3 and pH 4 holding times and the mean pH. RESULTS: During a 24-h fast, the pH 3 and pH 4 holding times and the mean pH were significantly higher in patients administered omeprazole (PPI; 93.2 +/- 9.2%, 90.6 +/- 11.1%, and 6.9 +/- 0.6, respectively) than in those administered ranitidine (H2-RA; 61.0 +/- 27.5%, 55.8 +/- 29.1%, and 4.8 +/- 1.3, respectively; P<0.001 for all). Results were similar during feeding (PPI meal, 98.9 +/- 2.6%, 98.3 +/- 3.7%, and 6.9 +/- 0.3; H2-RA meal, 59.8 +/- 17.6%, 49.7 +/- 18.0%, and 4.3 +/- 0.7, respectively; P<0.001 for all). In addition, the pH 3 and pH 4 holding times and the mean pH in the H2-RA meal group were not significantly lower than those in the H2-RA group (P=0.999, P=0.865, and P=0.687, respectively). The values in the PPI and PPI meal groups were similar (P=0.872, P=0.777, and P>0.999, respectively). CONCLUSIONS: Gastric acid suppression during the administration of an H2-RA or a PPI soon after the cessation of gastric bleeding was scarcely affected by feeding. It may well be that H. pylori-positive patients with bleeding gastric ulcer can resume a regular diet and return to work soon after bleeding ceases.

Risk factors for infections in cirrhotic patients with and without hepatocellular carcinoma.

BACKGROUND: Patients with liver cirrhosis (LC) frequently have complications with bacterial infections, and these infections increase the mortality rate. However, a detailed analysis of infections associated with LC patients has not yet been performed. METHODS: We analyzed 325 patients with LC with and without hepatocellular carcinoma (HCC) who were hospitalized between 1997 and 1999. RESULTS: Infections developed in 70 (21.5%) patients and 48 (68.6%) of these developed infections during hospitalization. The mortality rate of 28.6% (20/70) in patients with infectious complications was higher than that of 12.5% (32/255) in patients without infectious complications. Forty (57.1%) of the 70 patients had infections of unknown causes; 11 (15.7%) had sepsis; 6 (8.6%) had intravenous hyperalimentation (IVH) infection; 3 (4.3%) each had spontaneous bacterial peritonitis (SBP), liver abscess, and cholecystitis; and 4 (5.7%) had other infections. Bacterial cultures of blood were prepared from 73 of the 325 patients (22.5%), and were positive in 22 of the 73 patients (30.1%). Of these 22 culture-positive patients, 11 had sepsis, 6 had IVH infection, 2 had liver abscess, 1 had cholecystitis, 1 had pneumonia, and 1 had decubitus ulcer. Gram-positive bacterial strains were detected most frequently, in 16 of the 24 strains isolated. Univariate analysis revealed significant differences between the groups with and without infectious complications with regard to hepatitis B virus infection, Child-Pugh classification, ascites, esophageal varices, survival rate, total-bilirubin (T-Bil), albumin (Alb), lactate dehydrogenase (LDH), total cholesterol (T-chol), and prothrombin time (PT). On multivariate analysis, the Alb level was selected as a significant independent factor contributing to the development of infections. CONCLUSIONS: Patients with advanced cirrhosis with low Alb levels should be carefully treated, and the administration of broad spectrum antibiotics covering gram-positive bacteria needs to be considered in the treatment of infections.

Effect of IS-741 on ethionine-induced acute pancreatitis in rats: relation to pancreatic acinar cell regeneration.

BACKGROUND: Tissue destruction arising from neutrophil infiltration of the pancreas and other organs in acute pancreatitis is supposed to be suppressed by IS-741. We studied the effect of IS-741 on acute pancreatitis induced by DL-ethionine in rats. METHODS: Rats fed with a low protein diet for 11 days received daily intraperitoneal administration of DL-ethionine (20 mg/100 g) for the last 4 days of the diet. To evaluate the therapeutic effect on ethionine-induced pancreatitis, IS-741 (10 mg/kg s.c.) was administered every 8 h beginning after the second ethionine injection (IS group). An equal volume of saline was used for control rats as alternative to IS-741 (control group). The rats were killed 1, 3, 5, and 7 days after the last injection of ethionine. Blood was collected to measure concentrations of the inflammatory cytokine, interleukin-8. Histologic and biochemical examinations of the pancreas were performed. The pancreatic weight, DNA content, and protein levels were determined. The pancreas was histologically examined. RESULTS: Pancreatic tissue in the control group showed marked infiltration of inflammatory cells, and acinar cell necrosis was widespread 1 day after the last injection of ethionine (day 1). The severity of acute pancreatitis was alleviated in rats treated with IS-741 (IS group). Pancreatic wet weight and DNA content in the IS group were higher than those in the control group on day 1. Pancreatic protein level per DNA in the IS group was higher than that in the control group on day 7. The plasma interleukin-8 level in the control group was higher than that in the IS group on day 5. CONCLUSIONS: Therapeutic administration of IS-741 ameliorated ethionine-induced acute pancreatitis in rats, and IS-741 could be a useful drug to treat patients with severe acute pancreatitis.

Prevalence of antimitochondrial antibody in Japanese corporate workers in Kanagawa prefecture.

BACKGROUND: The prevalence of antimitochondrial antibody (AMA) in humans and its relationship to the development of primary biliary cirrhosis (PBC) are not well known. We have estimated the frequency of AMA in the general population, and studied its association with PBC. METHODS: We studies 1714 corporate workers (median age, 48 years; range, 30 to 59 years) who had an annual health check from 1998 to 1999 at Kawasaki Social Insurance Hospital in Japan. We used an indirect immunofluorescence method for screening serum AMA. We applied the prevalence of AMA-positive people in the study group to the general population in Japan. Then the inferred AMA-positive population was compared to the actual number of patients with PBC in statistics published by the Japanese Government. RESULTS: AMA was detected in 11 of 1714 people (0.64%; 95% confidence interval, 0.26% to 1.02%). All these 11 sera reacted with 2-oxoacid-dehydrogenase complex by immunoblotting. Of these 11 individuals, none had subjective symptoms, all had normal serum bilirubin levels, and 6 had abnormal liver function test results. Using published statistics for the Japanese population, we inferred that there were approximately 336,472 AMA-positive people in Japan from age 30 to 59 years. The number of patients with symptomatic PBC recorded by the nationwide epidemiological survey of the Japanese Government was 2459. Thus, we inferred the rate of symptomatic PBC among AMA-positive persons to be about 0.73% (2459/336,472). CONCLUSIONS: AMA is not a rare antibody in the general population, but few people develop recognizable PBC even if they have AMA.

Pancreatic regeneration after ethionine-induced acute pancreatitis in rats lacking pancreatic CCK-A receptor gene expression.

BACKGROUND: We examined the effects of cholecystokinin (CCK) on the development of ethionine-induced pancreatitis and pancreatic recovery. We used Otsuka Long-Evans Tokushima Fatty (OLETF) rats, a model lacking pancreatic CCK-A receptor gene expression. METHODS: Ethionine-induced pancreatitis was induced in the 7-week-old male OLETF rats and in a control group that does not lack the pancreatic CCK-A receptor, Long-Evans Tokushima Otsuka (LETO) rats. The two groups were maintained on a low-protein diet for 11 days. During the last 4 days of the low-protein diet, dl-ethionine 20 mg/100 g body weight was administered intraperitoneally once daily. Histologic and biochemical examinations of the pancreas were performed, and plasma CCK concentrations were measured on days 1, 4, and 7 after the last ethionine administration. RESULTS: Pancreatic histologic scores for inflammation, hemorrhage, and necrosis in the LETO and OLETF rats were highest on days 1 and 4, respectively. Pancreatic weight, DNA content, and protein level per DNA content in both groups decreased during the low-protein diet, and recovery signs were delayed in the OLETF rats. The highest plasma CCK concentrations in the LETO and OLETF rats were reached on days 1 and 4, respectively. CONCLUSIONS: Ethionine-induced pancreatitis developed in the OLETF rats, and their pancreatic regeneration was delayed in comparison to that in the LETO rats. Our results suggested that CCK plays an important role in the development of pancreatitis as well as in the pancreatic repair process.

Significance of measurement of high-sensitivity C-reactive protein in acute pancreatitis.

BACKGROUND: Determination of the severity of acute pancreatitis is difficult in the early phase after onset, and we often encounter difficulties in making decisions to initiate intensive care during the early phase. Therefore, there is real need for a simple and inexpensive method that can precisely evaluate the severity of acute pancreatitis. METHODS: In the present study, we measured serum C-reactive protein (CRP) levels in 20 patients with acute pancreatitis, using a high-sensitivity CRP (hs-CRP) assay method. RESULTS: CRP levels were as low as 1.0, 0.4, and 0.3 mg/dl in cases 2, 3, and 9, respectively, with severe acute pancreatitis. These three patients were hospitalized within 24 h after the onset of pancreatitis. Cases 2, 3, and 9 showed high hs-CRP levels, of 209 000, 68 600, and 154 000 ng/ml, respectively, and their interleukin (IL)-6 levels were above 500 pg/ml. The mean hs-CRP level was 222 760 +/- 32 197 ng/ml in patients with severe acute pancreatitis and 22 798 +/- 8216 ng/ml in patients with mild to moderate pancreatitis, with a significantly higher level in the severe cases. Cases 14, 16, and 20, with mild to moderate pancreatitis, had hs-CRP levels of 83 400, 1800, and 55 400 ng/ml, respectively. CONCLUSIONS: Measurement of hs-CRP levels is a simple and inexpensive method. The hs-CRP levels were found to significantly increase in the early phase of severe acute pancreatitis, suggesting that hs-CRP could possibly serve as an early indicator of the progression of acute pancreatitis into a serious state.

A patient with hepatitis C-related cirrhosis and hepatocellular carcinoma who was cured with an orthotopic liver transplantation and interferon therapy.

A patient with hepatitis C virus (HCV)-related liver cirrhosis and hepatocellular carcinoma (HCC) was treated successfully with an orthotopic liver transplantation (OLT) followed by interferon therapy. The 36-year-old Japanese man was diagnosed as having liver cirrhosis in 1983. HCC was detected in 1991, and by 1994, jaundice and ascites had developed. The patient underwent OLT in June 1995, after which hepatitis C recurred, with elevated aminotransferases. His liver biopsy specimen showed chronic active hepatitis. He was given interferon-alpha three times weekly for 24 weeks in 1999. Six months after the end of the interferon treatment, the patient's serum HCV RNA became negative, with normalization of aminotransferases, and his liver histology exhibited amelioration of fibrosis and inflammation. At the present time, he remains free of HCC (more than 6.5 years after the OLT) and free of HCV RNA (more than 2.5 years since interferon therapy was completed). This is the first Japanese patient whose HCC was cured by OLT and HCV was eradicated by interferon therapy.

Clinical significance in molecular detection of p53 mutation in serum of patients with colorectal carcinoma.

Circulating DNA can be isolated from serum of patients with various carcinomas and p53 mutation can be observed in colorectal carcinoma. The aim of this study was to investigate the correlation between p53 mutation in DNA extracted from colorectal carcinoma and that in DNA extracted from serum of patients with colorectal carcinoma. The clinical significance in molecular detection of p53 mutation in serum of patients with colorectal carcinomas was also investigated. DNA was extracted from tumors and non-tumorous colorectal tissues of 46 patients with single sporadic colorectal carcinomas of stage I (n=6), stage II (n=18), stage III (n=15), and stage IV (n=7) according to the TNM classification. Circulating DNA was also extracted from the serum of the 46 patients with colorectal carcinoma and from 7 healthy volunteers for normal control. Mutations of the p53 gene were analyzed using a fluorescence-based polymerase chain reaction (PCR)-single-strand conformation polymorphism (SSCP) method. DNA sequences were determined in DNA fragments with shifted peaks by SSCP methods. Mutations in tumors were found in 22 (48%) of 46 patients, and mutations in serum were found in 3 (14%) of these 22 patients. Of 4 patients with stage IV disease, 3 (75%) had serum p53 mutation and the mutation pattern of these 3 patients was the same in both tumor and serum. No correlation was seen between p53 mutation in serum and the level of serum DNA. There was no significant difference between the presence of p53 mutation in serum and tumor size, depth of invasion, vascular invasion, or lymph node metastasis. However, liver metastasis showed significant difference (p=0.0026). The presence of p53 mutation in serum was associated with a clinically advanced stage accompanied by liver metastasis.

Dendritic cell maturation induced by delivery of ultraviolet-mediated apoptotic colorectal cancer cell lines.

BACKGROUND: Dendritic cells (DCs) play a critical role in the immune response. The aim of this study was to investigate apoptotic tumor cells as an antigen source for DC maturation. MATERIALS AND METHODS: We compared the efficacy of ultraviolet (UV)-irradiation with that of gamma-irradiation in the induction of apoptosis of tumor cells. Phenotypic and functional changes of DCs were analyzed after co-incubation with UV-irradiated tumor cells. RESULTS: UV-irradiation (1.8 J/cm2) was a more reliable method of inducing apoptosis than gamma-irradiation (20,000 rad). The expression of costimulatory molecules on DCs was upregulated after co-incubation with UV-irradiated tumor cells. When we performed allogeneic mixed lymphocyte reaction assay, UV-irradiated tumor cells-pulsed DCs stimulated allogeneic T lymphocytes more efficiently than DCs pulsed with gamma-irradiated cells (HT29, p = 0.0419 and WiDr, p = 0.0076). CONCLUSION: UV-irradiation reliably induces apoptosis in cultured colorectal cancer cells. DCs mature in function and expression of costimulatory molecules after co-incubation with apoptotic tumor cells. The clinical implications warrant further study.

Increased tyrosine kinase activites may lead to the phenotypic differences of gastric cancer cells.

BACKGROUND: The aim of this study was to elucidate possible mechanisms responsible for the phenotypic difference in gastric cancer cells between the intestinal type and the diffuse type. MATERIALS AND METHODS: The gastric cancer cell lines MKN7 and KATO-III were used. Expression of adhesion molecules and protein tyrosine phosphorylation were examined by immunoblotting. Protein-protein interactions were determined by immunoblotting following immunoprecipitation. ERK activity was assessed by immunoblotting using anti-phospho-ERK antibody. RESULTS: E-cadherin expression in KATO-III was decreased compared to that in MKN7. The expression of alpha-, beta- and gamma-catenin was not significantly different; beta- and gamma-catenin were highly tyrosine phosphorylated in KATO-III but not in MKN7. In KATO-III, a number of tyrosine phosphorylated proteins, including extracellular signal-regulated protein kinases (ERKs) and epidermal growth factor receptor (EGFR), were observed irrespective of EGF stimulation and induction of ERK activity and EGFR tyrosine phosphorylation by EGF were less than that in MKN7. CONCLUSION: Increased tyrosine kinase activities may cause diminished expression of E-cadherin and tyrosine phosphorylation of catenins, resulting in the abrogated cell-cell adhesion in gastric cancer cells.

Growth-inhibitory effect of phosphatidylinositol 3-kinase inhibitor on human pancreatic cancer cells and expression of Bcl-2 family.

Pancreatic cancer is one of the most devastating malignant tumors in humans and novel modalities for treatment need to be developed. We studied the mechanism of the growth-inhibitory effect of phosphatidylinositol 3-kinase (PI 3-K) inhibitor on human pancreatic cancer cells from the point of view of expression of the Bcl-2 family proteins. Growth of AsPC-1 and COLO-357 human pancreatic cancer cells was inhibited by a phosphatidylinositol 3-kinase (PI 3-K) inhibitor, wortmannin, and this growth-inhibitory effect was more marked in medium containing 10% fetal bovine serum (FBS) than in serum-free medium. In these cells, DNA fragmentation increased with the concentration of wortmannin in a dose-dependent manner. In Panc-1 human pancreatic cancer cells, cell growth and induction of DNA fragmentation were not influenced by treatment with wortmannin at concentrations up to 25 microM. Western blot analysis showed a decrease in expression of BclXL protein in AsPC-1 and COLO-357 cells by treatment with 25 microM wortmannin and this decrease was especially prominent in AsPC-1 cells. On the other hand, the expression of BclXL protein in Panc-1 cells was not influenced by treatment with wortmannin. The expression of BclXS protein was not detected by conventional Western blotting and the expression of Bcl-2 and Bax protein was not altered by wortmannin in all three cell lines. Decrease in expression of BclXL protein could be partly involved in the growth-inhibitory effect of the PI 3-K inhibitor, wortmannin, on pancreatic cancer cells. Although the growth of Panc-1 cells was not inhibited by wortmannin, PI 3-K inhibitor could still be one of the candidates for treatment of pancreatic cancer and BclXL could be a target for gene therapy.

Expression of DNA (5-cytosin)-methyltransferases (DNMTs) in hepatocellular carcinomas.

Down-regulation of tumor suppressor genes by hypermethylation of 5'-CpGs is one of the important mechanisms involved in tumor development. DNA (5-cytosine)-methyltransferases (DNMTs) are enzymes that methylate the cytosine residue of CpGs, and four types have been identified (DNMT1, 2, 3a and 3b). To examine the involvement of DNMTs in hepatocellular carcinogenesis, we measured DNMT mRNAs in hepatocellular carcinomas (HCCs). mRNAs of DNMT1, 2, 3a and 3b were detected by reverse transcription-PCR analysis and quantified by a real-time PCR method in surgically resected HCCs and adjacent non-tumorous liver tissue. DNMT1 was expressed in all tissues and at a significantly higher level in HCCs than in non-tumorous liver tissue (P=0.01). DNMT2 was expressed at a low level in all tissues. DNMT3a and DNMT3b mRNA were undetectable in normal liver. DNMT3a was expressed in all HCCs and was expressed at similar levels in 60% of the non-tumorous liver tissues. DNMT3b mRNA was detected at a significantly higher level (P=0.002) in HCCs than in non-tumorous liver tissues. The amount of DNMT1, 3a and 3b mRNA was not different between HCCs with or without hypermethylation of the CDH1 promoter. These data suggest that overexpression of DNMT1 and DNMT3b contributes to hepatocellular carcinogenesis.

Endoscopic color Doppler ultrasonographic evaluation of recurrent esophagogastric varices following endoscopic injection sclerotherapy.

BACKGROUND: Endoscopic injection sclerotherapy (EIS) for treatment of esophagogastric varices is well established in Japan. However, varices may still recur unpredictably following EIS. We studied this problem using endoscopic color Doppler ultrasonography (ECDUS) and specifically examined esophagogastric blood flows. METHODS: Prophylactic EIS was performed by intravariceal injection of 5% ethanolamine oleate (EO) in 49 patients with esophageal varices secondary to liver cirrhosis. No patient had documented hepatocellular carcinoma (HCC) before EIS, and patients who developed HCC during follow-up were excluded. We performed ECDUS before EIS, and 2 weeks and 2 years later. The esophagogastric intra- and extramural venous blood flows, including flow in the azygos vein, were compared between these observations. Gastric intramural blood flow and changes in extramural gastric blood flow, including the azygos vein flow, were scored. Dynamic computer tomography (CT), ultrasonography (US), and color Doppler-ultrasonography (CDUS) were also performed before EIS and 1 month following the procedure. Thereafter, patients underwent CT and US examinations every 6 months for 2 years to detect any development of porto-systemic shunts or HCC. RESULTS: The average number of EIS procedure per patient was 3.1+/-0.8 (mean+/-SD), and the total amount of sclerosant injected was approximately 33.5+/-6.5 ml. The overall recurrence rate over the 2-year follow-up was 36.7%. The gastric intra- and extramural blood flows did not differ between those patients with or without major shunts before EIS. In patients with recurrent variceal formation, the gastric intramural blood flow score following EIS (2.1+/-0.5) was significantly higher than that in patients without recurrence (0.8+/-0.6) (P<0.01). In addition, gastric extramural blood flow score following EIS (0.8+/-0.6) was significantly lower in patients with recurrence than that in those without recurrence (1.7+/-0.5) (P<0.01). The same differences held after exclusion of patients with major shunts. The gastric intramural blood flow score in patients with recurrent variceal formation (2.1+/-0.4) was significantly higher than that in patients without recurrence score (P<0.01). Moreover, gastric extramural blood flow score in patients with recurrent variceal formation (1.0+/-0.7) was significantly lower than in patients without recurrence (1.6+/-0.5) (P<0.01). CONCLUSIONS: Two characteristics were observed in patients with recurrent cases of esophageal varices 2 weeks following EIS. The first was the maintenance of gastric intramural blood flow. The second was the absence of dilation of the gastric extramural blood vessels. These observations may enable us to predict the recurrence of esophagogastric varices using ECDUS within 2 weeks following EIS.

Immunoreactivity to pyruvate dehydrogenase complex-E2 in well-defined patients with autoimmune hepatitis: Western blot analysis.

Anti-mitochondrial antibodies (AMA) are frequently detected in sera from patients with primary biliary cirrhosis (PBC). Major autoantigens for AMA have been identified as members of the 2-oxoacid dehydrogenase enzyme complex family, with pyruvate dehydrogenase complex (PDC)-E2 showing strongest reactivity to AMA in PBC patients. Recently, anti-PDC-E2 has been found in patients with other diseases. Since frequency and significance of anti-PDC-E2 in patients with autoimmune hepatitis (AIH) remain obscure, we measured anti-PDC-E2 in sera from well-defined AIH cases by Western blotting using bovine heart mitochondrial protein and recombinant PDC-E2 protein as antigen sources. All 55 enrolled patients fulfilled the international diagnostic criteria for definite or probable AIH. Anti-PDC-E2 positivity showed concordance between native and recombinant antigens. Anti-PDC-E2 was detected in nine of 55 sera from AIH patients (16%). Variables including alkaline phosphatase (ALP) and IgM concentrations, effects of prednisolone, and pathologic findings concerning bile ducts showed no significant differences between anti-PDC-E2-positive and anti-PDC-E2-negative AIH patients. These data indicate that detection of anti-PDC-E2 is not rare in defined AIH, but anti-PDC-E2-positive AIH does not represent an intermediate entity in a clinical spectrum between AIH and PBC.

Differential effect of cytotoxic T lymphocyte variant epitopes on generation and cytotoxicity in chronic hepatitis C virus infection.

Human leukocyte antigen (HLA) B44-restricted, hepatitis C virus (HCV)-specific cytotoxic T lymphocytes (CTLs) recognize HCV nucleoprotein amino acid residues 88-96 as an epitope. We previously reported the existence of variant peptide sequences at the epitope locus in three of 27 patients with HCV infection and HLA B44. Here we studied the effects of the variant peptide sequences on generation and cytotoxicity of CTLs. Two of the three variant peptides generated CTLs poorly although they activated well the cytotoxicity of CTLs. Such a differential activation of proliferation and cytotoxicity may contribute to the emergence of HCV with variant epitopes for CTLs.

Expression of hepatitis C virus core protein associated with malignant lymphoma in transgenic mice.

Hepatitis C virus (HCV) is a major causative agent for chronic liver diseases leading to hepatocellular carcinoma (HCC) and has also been suggested to be a possible etiologic factor for different lymphoproliferative diseases, including mixed cryoglobulinemia (MC) and B-cell non-Hodgkin's lymphoma (NHL). To understand the roles of HCV core protein in the pathogenesis of HCV related diseases, we produced two lines of the transgenic mice (HC82310 and HC9053) that express the HCV core transgene. One of the lines, HC9053, developed malignant lymphoma (ML, follicular center cell type) with a high frequency (80%) at the ages over 20 months. Hepatocellular adenoma was also observed in this line of transgenic mouse. We demonstrated expression of HCV core protein and mRNA in the liver of transgenic mice, and also detected the core mRNA in the enlarged lymph nodes of the transgenic mice which developed ML. These results suggest that the core protein may play an important role in the development of ML, and that the HC9053 transgenic mice provide suitable models for understanding the mechanism of HCV-related lymphoproliferative diseases.

Characterization of DNA polymorphisms in the E-cadherin gene (CDH1) promoter region.

E-cadherin is a tumor suppressor involved in epithelial cell-cell interactions. Some of the nucleotide variation in the 5'-promoter region of the gene influences transcriptional efficiency. We investigated single nucleotide polymorphisms (SNPs) in the promoter-exon 1 region of the E-cadherin gene (CDH1) using fluorescence-based PCR-single-strand conformation polymorphism (PCR-SSCP) analysis. We detected four kinds of polymorphisms between nucleotides -516 and +12, numbering from the translation initiation site. SNPs were localized at -472G-->GA, -288T-->deltaT, -285C-->A, and -54G-->C. Variants -472GA and -285A were frequently found in controls, but the -288deltaT and -54C are rare variants. We examined the effects of these variants on transcription. The activity of promoters containing the variants -288deltaT, -285A, or -54C was lower than the activity of promoters with the major variants, as assayed by a luciferase reporter gene. Variants -472G and -472GA displayed the same promoter activity. The decreased transcriptional activity from variant promoters affects the expression of E-cadherin.