片野 義明

Mutations in carboxy-terminal part of E2 including PKR/<br /> eIF2 alpha phosphorylation homology domain and <br /> interferon sensitivity determining region of nonstructural <br /> 5A of hepatitis C virus 1b: Their correlation with response to interferon monotherapy and viral load.

Mutations in carboxy-terminal part of E2 including PKR/<br /> eIF2 alpha phosphorylation homology domain and <br /> interferon sensitivity determining region of nonstructural <br /> 5A of hepatitis C virus 1b: Their correlation with response to interferon monotherapy and viral load.

Background/Aims: Hepatitis C virus induces various clinical features in a host depending on duration of the viral infection. Methodology: We investigated peripheral lymphocyte subsets in patients with three different stages of hepatitis C virus infection: 5 patients with acute hepatitis, 10 with chronic hepatitis unassociated with cirrhosis, and 10 with cirrhosis. Peripheral lymphocytes were double-stained with multiple fluorescent antibody combinations: anti-CD3 plus anti-gamma delta T cell receptor; anti-CD19 plus anti-CD5; anti-CD4 plus anti-CD45RA; or anti-CD8 plus anti-CD11b. Triple staining was performed with fluorescent antibodies against CD4, interferon gamma, and interleukin-4. Both staining protocols were followed by flow cytometric analysis. Results: Acute hepatitis patients had a high proportion of CD3+ T cells with increased CD4+CD45-RAT helper and CD8+CD11b- cytotoxic T cells. Compared to this group, chronic hepatitis patients showed a decrease in CD4+ cells and an increase in CD19+ B cells and interleukin-4-producing Th2 cells. Cirrhotic patients showed decreased circulating lymphocytes and a low proportion of CD8+ cells accompanied by a decrease in cytotoxic T cells. Furthermore, their lymphocyte profiles showed decreases in primordial lymphocyte subpopulations (T cells with gamma delta T cell receptors and B cells with CD5). Conclusions: Although the same pathogenic agent was involved, immune dynamics differed greatly according to duration of viral infection.

Background/Aims: Serum aminotransferase, a sensitive marker of hepatocellular damage, often poorly correlates with the severity of damage. Serum nuclear matrix protein (NMP), a structural protein released from dead cell nuclei, is investigated as a candidate marker of organ damage in liver disease. Methodology: Serum NMP and aminotransferase levels of 134 patients with various liver diseases and 26 healthy individuals were examined. Results: Patients with chronic viral hepatitis showed slightly higher NMP levels (17.8 U/mL; 95% Cl 15.0-20.5 U/mL) than those of healthy individuals (6.05 U/mL; 95% Cl 4.82-7.27 U/mL). Their NMP values had no correlation with aminotransferase levels. NMP levels were similar irrespective of liver disease progression, whereas aminotransferase values decreased in parallel with progression. Patients with autoimmune hepatitis or primary biliary cirrhosis who were under an appropriate treatment as well as individuals with fatty liver showed no elevation of serum NMP levels. Patients with acute viral hepatitis showed very high NMP levels (38.8 U/mL; 95%Cl 27.6-50.0 U/mL) that correlated with serum aminotransferase levels in their sera. Conclusions: In chronic liver diseases, the serum NMP level elevates to various degrees independent from the degree of aminotransferase elevation. Serum NMP, putatively representing the number of dead cells, is a candidate as an indicator of organ damage severity in liver disease.

A 65-year-old man diagnosed with hepatitis C virus-positive hepatitis and severe valvular heart disease was scheduled to undergo cardiac valve replacement. We then found hepatocellular carcinoma in the liver. Because of his severe cardiac dysfunction, we treated him surgically with radiofrequency ablation for the hepatocellular carcinoma only. We continued medical treatment of the heart disease. He hoped to undergo with cardiac surgery one year later for the cardiac dysfunction. There was no evidence of tumor recurrence. We informed him that cardiac surgery requiring extracorporeal circulation might lead to tumor recurrence. He agreed to cardiac valve replacement, and the surgery was successful. Recurrent hepatocellular carcinoma was found in the liver 1 month after the surgery. Over the next month, the tumor progressed rapidly, showing portal vein thrombi. We believe the use of extracorporeal circulation in particular triggered the rapid growth of the recurrent hepatocellular carcinoma. This is the first report of a recurrent hepatocellular carcinoma associated with hepatitis C virus that progressed extensively after cardiac surgery.

Background/Aims: Hepatitis C virus induces various clinical features in a host depending on duration of the viral infection. Methodology: We investigated peripheral lymphocyte subsets in patients with three different stages of hepatitis C virus infection: 5 patients with acute hepatitis, 10 with chronic hepatitis unassociated with cirrhosis, and 10 with cirrhosis. Peripheral lymphocytes were double-stained with multiple fluorescent antibody combinations: anti-CD3 plus anti-gamma delta T cell receptor; anti-CD19 plus anti-CD5; anti-CD4 plus anti-CD45RA; or anti-CD8 plus anti-CD11b. Triple staining was performed with fluorescent antibodies against CD4, interferon gamma, and interleukin-4. Both staining protocols were followed by flow cytometric analysis. Results: Acute hepatitis patients had a high proportion of CD3+ T cells with increased CD4+CD45-RAT helper and CD8+CD11b- cytotoxic T cells. Compared to this group, chronic hepatitis patients showed a decrease in CD4+ cells and an increase in CD19+ B cells and interleukin-4-producing Th2 cells. Cirrhotic patients showed decreased circulating lymphocytes and a low proportion of CD8+ cells accompanied by a decrease in cytotoxic T cells. Furthermore, their lymphocyte profiles showed decreases in primordial lymphocyte subpopulations (T cells with gamma delta T cell receptors and B cells with CD5). Conclusions: Although the same pathogenic agent was involved, immune dynamics differed greatly according to duration of viral infection.

Background/Aims: Serum aminotransferase, a sensitive marker of hepatocellular damage, often poorly correlates with the severity of damage. Serum nuclear matrix protein (NMP), a structural protein released from dead cell nuclei, is investigated as a candidate marker of organ damage in liver disease. Methodology: Serum NMP and aminotransferase levels of 134 patients with various liver diseases and 26 healthy individuals were examined. Results: Patients with chronic viral hepatitis showed slightly higher NMP levels (17.8 U/mL; 95% Cl 15.0-20.5 U/mL) than those of healthy individuals (6.05 U/mL; 95% Cl 4.82-7.27 U/mL). Their NMP values had no correlation with aminotransferase levels. NMP levels were similar irrespective of liver disease progression, whereas aminotransferase values decreased in parallel with progression. Patients with autoimmune hepatitis or primary biliary cirrhosis who were under an appropriate treatment as well as individuals with fatty liver showed no elevation of serum NMP levels. Patients with acute viral hepatitis showed very high NMP levels (38.8 U/mL; 95%Cl 27.6-50.0 U/mL) that correlated with serum aminotransferase levels in their sera. Conclusions: In chronic liver diseases, the serum NMP level elevates to various degrees independent from the degree of aminotransferase elevation. Serum NMP, putatively representing the number of dead cells, is a candidate as an indicator of organ damage severity in liver disease.

A 65-year-old man diagnosed with hepatitis C virus-positive hepatitis and severe valvular heart disease was scheduled to undergo cardiac valve replacement. We then found hepatocellular carcinoma in the liver. Because of his severe cardiac dysfunction, we treated him surgically with radiofrequency ablation for the hepatocellular carcinoma only. We continued medical treatment of the heart disease. He hoped to undergo with cardiac surgery one year later for the cardiac dysfunction. There was no evidence of tumor recurrence. We informed him that cardiac surgery requiring extracorporeal circulation might lead to tumor recurrence. He agreed to cardiac valve replacement, and the surgery was successful. Recurrent hepatocellular carcinoma was found in the liver 1 month after the surgery. Over the next month, the tumor progressed rapidly, showing portal vein thrombi. We believe the use of extracorporeal circulation in particular triggered the rapid growth of the recurrent hepatocellular carcinoma. This is the first report of a recurrent hepatocellular carcinoma associated with hepatitis C virus that progressed extensively after cardiac surgery.

Hemochromatosis is a genetically heterogeneous condition. Mutations in the recently described hemojuvelin gene were found in patients with juvenile hemochromatosis, who usually manifest clinical signs of iron overload, including cardiomyopathy and hypogonadism, in their teens and early 20s. In this report, we describe three Japanese patients who showed typical clinical and hepatic histological damage compatible with hemochromatosis at around 50 years of age. Genetic analyses showed that all three patients carried mutations in the hemojuvelin gene. The first patient was homozygous for a novel mutation (745G &gt; C [D249H]), and the second and third patients from the same family were homozygous for another novel mutation (934C &gt; T [Q312X]). No mutations in their HFE, hepcidin, transferrin receptor 2, or ferroportin genes were found. One patient had chronic infection with Helicobacter pylori. The age at initial presentation of hemojuvelin-hemochromatosis occurs over a wider range than previously described. (c) 2005 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

Hemochromatosis is a genetically heterogeneous condition. Mutations in the recently described hemojuvelin gene were found in patients with juvenile hemochromatosis, who usually manifest clinical signs of iron overload, including cardiomyopathy and hypogonadism, in their teens and early 20s. In this report, we describe three Japanese patients who showed typical clinical and hepatic histological damage compatible with hemochromatosis at around 50 years of age. Genetic analyses showed that all three patients carried mutations in the hemojuvelin gene. The first patient was homozygous for a novel mutation (745G &gt; C [D249H]), and the second and third patients from the same family were homozygous for another novel mutation (934C &gt; T [Q312X]). No mutations in their HFE, hepcidin, transferrin receptor 2, or ferroportin genes were found. One patient had chronic infection with Helicobacter pylori. The age at initial presentation of hemojuvelin-hemochromatosis occurs over a wider range than previously described. (c) 2005 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

We treated 665 patients with chronic hepatitis C with interferon (IFN) monotherapy and 288 with combined IFN and ribavirin. At the baseline, age (53.8 +/- 11.1 vs. 49.7 +/- 10.5 years, p&lt;0.0001) and activity (p=0.0207) as well as fibrosis (p=0.0270) were higher in patients who received combination therapy than in those receiving monotherapy. Compliance to treatment (64.2 vs. 62.1%, p&lt;0.0001) and discontinuation were more frequent (18.1 vs. 14.5%, p&lt;0.0001) in patients with combination therapy than in those with monotherapy. Patients with combination therapy with genotype 2 infection achieved sustained viral response (SVR) at a rate of 77.0%, regardless of viral loads, in contrast to those with genotype 1 infection, of whom only 24.4% gained SVR. Of patients with combination therapy, reduction (42.6 vs. 29.0%, p=0.0453) and discontinuation (34.0 vs. 21.6%, p=0.0414) of ribavirin were more frequently required in the 47 patients &gt;= 65 years than in the 241 patients &lt;65 years. Although a trend for higher SVR to combination therapy was observed in patients aged &lt;65 than in those aged &gt;= 65 years (39.4 vs. 25.2%), the difference was not significant (p=0.0819). In patients with genotype 1 infection, IFN monotherapy in addition to the 24-week combination therapy increased the SVR rate (18.3 vs. 42.6%, p=0.0003). A decrease in SVR was observed with an increased body mass index in patients who received combination therapy. Copyright (C) 2006 S. Karger AG, Basel.

We treated 665 patients with chronic hepatitis C with interferon (IFN) monotherapy and 288 with combined IFN and ribavirin. At the baseline, age (53.8 +/- 11.1 vs. 49.7 +/- 10.5 years, p&lt;0.0001) and activity (p=0.0207) as well as fibrosis (p=0.0270) were higher in patients who received combination therapy than in those receiving monotherapy. Compliance to treatment (64.2 vs. 62.1%, p&lt;0.0001) and discontinuation were more frequent (18.1 vs. 14.5%, p&lt;0.0001) in patients with combination therapy than in those with monotherapy. Patients with combination therapy with genotype 2 infection achieved sustained viral response (SVR) at a rate of 77.0%, regardless of viral loads, in contrast to those with genotype 1 infection, of whom only 24.4% gained SVR. Of patients with combination therapy, reduction (42.6 vs. 29.0%, p=0.0453) and discontinuation (34.0 vs. 21.6%, p=0.0414) of ribavirin were more frequently required in the 47 patients &gt;= 65 years than in the 241 patients &lt;65 years. Although a trend for higher SVR to combination therapy was observed in patients aged &lt;65 than in those aged &gt;= 65 years (39.4 vs. 25.2%), the difference was not significant (p=0.0819). In patients with genotype 1 infection, IFN monotherapy in addition to the 24-week combination therapy increased the SVR rate (18.3 vs. 42.6%, p=0.0003). A decrease in SVR was observed with an increased body mass index in patients who received combination therapy. Copyright (C) 2006 S. Karger AG, Basel.

Hepatitis C virus (HCV) genotype 1b comprises mainly two subtypes in Japan, each named for its geographic prevalence (Japan-specific, J type; worldwide, W type). Because the newly identified subtypes have not been fully characterized, the present study directed this issue from virological viewpoints such as hypervariable region (HVR)-1 as well as interferon (IFN) sensitivity-determining region (ISDR). Fifty chronic hepatitis patients with HCV 1b (31 men and 19 women; mean age 50.5 years) were enrolled, and J/W type was determined according to envelope 1 (E1) sequence as described previously (23 J type and 27 W type). Correlations between age, number of HVR-1 clones, HVR-1 diversity, and ISDR mutations were analyzed in J and W type patients independently. In addition, the sequences of the three HCV regions obtained for the determination of the above genetic factors were studied phylogenetically. The number of HVR-1 clones was significantly higher for J type in comparison with W type (P = 0.044). In the J type-infected patients, the ISDR mutation number was correlated inversely with HVR-1 clone number (P = 0.0001, r = -0.734) and HVR-1 diversity (P = 0.0001, r = -0.722). However, this correlation was not observed in the W type patients. W type patients showed a significant correlation between age and HVR-1 clone number (P = 0.015, r = 0.462). Phylogenetic study revealed that the nonstructural (NS) 5A sequence, which is obtained for ISDR type determination, can distinguish between J and W types. The inverse correlation in J type patients between ISDR mutations and HVR-1 complexity may explain the usefulness of the ISDR for prediction of IFN response only in Japanese patients. This suggests that the ISDR is not directly related to IFN responsiveness, but the degree of HVR-1 complexity may be more important. (C) 2004 Wiley-Liss, Inc.

Although viral load of hepatitis C virus (HCV) is a predictor of response to interferon therapy, little is known about its fluctuations. We assessed its fluctuations and their correlation with serum alanine aminotransferase (ALT) levels. Viral load was prospectively measured bimonthly for 22 months in 109 patients. In 40 patients, viral load changed more than five fold. Changes were transient and always returned to the baseline levels. ALT levels changed more than three fold in 30 patients. Changes of viral load accompanied simultaneous changes of ALT levels in only 7 of 40 patients with changes of viral load. Mean viral load in 22 months was significantly correlated with mean ALT levels inversely (r = 0.278, P = 0.0036). Mean viral load was significantly higher in 27 patients with persistently normal ALT levels (452.0 +/- 342.5 pg/ml) than in 30 patients with changes of ALT levels (202.4 +/- 215.0 pg/ml) (P = 0.0016) and than in 52 patients without changes of ALT levels (301.1 +/- 295.4 pg/ml) (P = 0.0458). Inverse correlation of viral load with ALT levels suggests that viral load is in suppression by inflammatory activity. However, changes of ALT levels infrequently accompanied simultaneous changes of viral load and vice versa, as often seen in chronic hepatitis B virus infection. (C) 2004 Elsevier B.V. All rights reserved.

Hepatitis C virus (HCV) genotype 1b comprises mainly two subtypes in Japan, each named for its geographic prevalence (Japan-specific, J type; worldwide, W type). Because the newly identified subtypes have not been fully characterized, the present study directed this issue from virological viewpoints such as hypervariable region (HVR)-1 as well as interferon (IFN) sensitivity-determining region (ISDR). Fifty chronic hepatitis patients with HCV 1b (31 men and 19 women; mean age 50.5 years) were enrolled, and J/W type was determined according to envelope 1 (E1) sequence as described previously (23 J type and 27 W type). Correlations between age, number of HVR-1 clones, HVR-1 diversity, and ISDR mutations were analyzed in J and W type patients independently. In addition, the sequences of the three HCV regions obtained for the determination of the above genetic factors were studied phylogenetically. The number of HVR-1 clones was significantly higher for J type in comparison with W type (P = 0.044). In the J type-infected patients, the ISDR mutation number was correlated inversely with HVR-1 clone number (P = 0.0001, r = -0.734) and HVR-1 diversity (P = 0.0001, r = -0.722). However, this correlation was not observed in the W type patients. W type patients showed a significant correlation between age and HVR-1 clone number (P = 0.015, r = 0.462). Phylogenetic study revealed that the nonstructural (NS) 5A sequence, which is obtained for ISDR type determination, can distinguish between J and W types. The inverse correlation in J type patients between ISDR mutations and HVR-1 complexity may explain the usefulness of the ISDR for prediction of IFN response only in Japanese patients. This suggests that the ISDR is not directly related to IFN responsiveness, but the degree of HVR-1 complexity may be more important. (C) 2004 Wiley-Liss, Inc.

Although viral load of hepatitis C virus (HCV) is a predictor of response to interferon therapy, little is known about its fluctuations. We assessed its fluctuations and their correlation with serum alanine aminotransferase (ALT) levels. Viral load was prospectively measured bimonthly for 22 months in 109 patients. In 40 patients, viral load changed more than five fold. Changes were transient and always returned to the baseline levels. ALT levels changed more than three fold in 30 patients. Changes of viral load accompanied simultaneous changes of ALT levels in only 7 of 40 patients with changes of viral load. Mean viral load in 22 months was significantly correlated with mean ALT levels inversely (r = 0.278, P = 0.0036). Mean viral load was significantly higher in 27 patients with persistently normal ALT levels (452.0 +/- 342.5 pg/ml) than in 30 patients with changes of ALT levels (202.4 +/- 215.0 pg/ml) (P = 0.0016) and than in 52 patients without changes of ALT levels (301.1 +/- 295.4 pg/ml) (P = 0.0458). Inverse correlation of viral load with ALT levels suggests that viral load is in suppression by inflammatory activity. However, changes of ALT levels infrequently accompanied simultaneous changes of viral load and vice versa, as often seen in chronic hepatitis B virus infection. (C) 2004 Elsevier B.V. All rights reserved.

The prevalence and clinical implications of occult hepatitis B virus (HBV) infection were investigated in the Japanese patients with hemophilia in whom a high prevalence of infection with transfusion-transmissible viruses has been reported. HBV DNA was detected in the sera of 22 of 43 (51.2%) patients with hemophilia who were negative for HBV surface antigen (HBs), indicating that these patients had occult HBV infection. No factor, including age, type or severity of hemophilia, presence of HBs or HBV core (HBc) antibody, or coinfection with hepatitis C virus (HCV) or human immunodeficiency virus (HIV) was associated with occult HBV infection, except for high anti-HBc titer and/or coinfection with HCV genotype 1 (1a or 1b). In general, occult HBV infection did not appear to have significant clinical implications. However, in patients in whom HBV was detected by PCR specific for the surface (S)-region, higher alanine aminotransferase levels were observed. The genotype of the occult HBV in the present study was exclusively the domestic type indigenous to Japan (genotype C), suggesting a different route of transmission for HBV in comparison to HCV and HIV in this population. (C) 2004 Wiley-Liss, Inc.

The prevalence and clinical implications of occult hepatitis B virus (HBV) infection were investigated in the Japanese patients with hemophilia in whom a high prevalence of infection with transfusion-transmissible viruses has been reported. HBV DNA was detected in the sera of 22 of 43 (51.2%) patients with hemophilia who were negative for HBV surface antigen (HBs), indicating that these patients had occult HBV infection. No factor, including age, type or severity of hemophilia, presence of HBs or HBV core (HBc) antibody, or coinfection with hepatitis C virus (HCV) or human immunodeficiency virus (HIV) was associated with occult HBV infection, except for high anti-HBc titer and/or coinfection with HCV genotype 1 (1a or 1b). In general, occult HBV infection did not appear to have significant clinical implications. However, in patients in whom HBV was detected by PCR specific for the surface (S)-region, higher alanine aminotransferase levels were observed. The genotype of the occult HBV in the present study was exclusively the domestic type indigenous to Japan (genotype C), suggesting a different route of transmission for HBV in comparison to HCV and HIV in this population. (C) 2004 Wiley-Liss, Inc.

Background/Aims: Branched-chain alpha-keto acid dehydrogenase (BCKDH) complex catalyses the committed step in the branched-chain amino acid (BCAA) catabolic pathway. In many cases of liver failure, the serum BCAAs/aromatic amino acids ratio (Fisher's ratio) decreases, and BCAAs have been administered to patients with liver failure to correct this ratio. We conducted an animal study to examine whether the effects on hepatic BCKDH complex differ between acute liver failure (ALF) and chronic liver failure (CLF). Methods: ALF and CLF was induced in rats by a single high-dose injection and 21 weeks of repeated low-dose injections of carbon tetrachloride, respectively. Plasma BCAA and branched-chain alpha-keto acid (BCKA) levels, and activities and protein amounts of hepatic BCKDH complex and kinase were measured. Results: ALF was characterized by elevated plasma BCAA and BCKA levels and decreased hepatic BCKDH activity. CLF was characterized by decreased plasma BCAA and BCKA levels and increased hepatic BCKDH activity. This increase in BCKDH activity in CLF was associated with the decreased BCKDH kinase, which is responsible for the BCKDH inactivation. Conclusions: The results obtained in the present study suggest that BCAA catabolism is suppressed in ALF and increased in CLF. (C) 2003 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.