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// Yuki Haga 1 , Tatsuo Kanda 1,2 , Shin Yasui 1 , Masato Nakamura 1 , Yoshihiko Ooka 1 , Koji Takahashi 1 , Shuang Wu 1 , Shingo Nakamoto 1,3 , Makoto Arai 1 , Tetsuhiro Chiba 1 , Hitoshi Maruyama 1 , Osamu Yokosuka 1 , Nobuo Takada 4 , Mitsuhiko Moriyama 2 , Fumio Imazeki 1,5 and Naoya Kato 1 1 Department of Gastroenterology, Chiba University, Graduate School of Medicine, Inohana, Chuo-ku, Chiba, Japan 2 Division of Gastroenterology and Hepatology, Department of Internal Medicine, Nihon University School of Medicine, Itabashi-ku, Tokyo, Japan 3 Department of Molecular Virology, Chiba University, Graduate School of Medicine, Inohana, Chuo-ku, Chiba, Japan 4 Department of Internal Medicine, Toho University Sakura Medical Center, Shimoshizu, Sakura, Japan 5 Safety and Health Organization, Chiba University, Yayoicho, Inage-ku, Chiba, Japan Correspondence to: Tatsuo Kanda, email: // Keywords : hepatitis C virus; direct-acting antiviral failure; non-structural 5A (NS5A) inhibitors; resistance-associated variants; case reports Received : June 20, 2017 Accepted : December 23, 2017 Published : December 29, 2017 Abstract Background: Interferon-free treatment results in higher sustained virologic response (SVR) rates, with no serious adverse events in hepatitis C virus (HCV)-infected patients. However, in some patients with treatment-failure in HCV NS5A inhibitor-including interferon-free regimens, the treatment-emergent HCV NS5A resistance-associated variants (RAVs), which are resistant to interferon-free retreatment including HCV NS5A inhibitors, are observed. In HCV-infected Japanese patients with daclatasvir and asunaprevir treatment failure, retreatment with sofosbuvir and ledipasvir could lead to only ~70% SVR rates. Case summary: Three HCV genotype (GT)-1b-infected cirrhotic patients who discontinued the combination of daclatasvir and asunaprevir due to adverse drug reactions within 4 weeks; retreatment with sofosbuvir and ledipasvir combination could result in SVR in these patients without RAVs. One HCV GT-1b-infected cirrhotic patient who discontinued the combination of daclatasvir and asunaprevir due to viral breakthrough at week 10; retreatment with sofosbuvir and ledipasvir combination for this patient with the treatment-emergent HCV NS5A RAV-Y93H resulted in viral relapse at week 4 after the end of the treatment. Conclusion: Retreatment with sofosbuvir and ledipasvir is effective for HCV GT-1b patients who discontinue the combination of daclatasvir and asunaprevir within 4 weeks. The treatment response should be related to the existence of treatment-emergent HCV NS5A RAVs, but may not be related to the short duration of treatment.

Background The utility of molecules derived from cancer cells as biomarkers of the pathological status in biliary tract and pancreatic cancers is still limited. Soluble LDL receptor relative with 11 ligand-binding repeats (sLR11), a molecule released from immature cells, has been shown to be a circulating biomarker for early stage hematological malignancies. Methods We have evaluated the pathological significance of bile sLR11 levels in 147 samples from 72 patients with biliary tract cancer (BTC), pancreatic cancer (PC), or benign diseases. Results The bile sLR11 levels in the cancer patients were significantly increased compared with those in patients without cancer, independent of cytological detection of cancer cells in bile. The average bile sLR11 levels in cancer patients were significantly higher than in those with benign diseases, while levels of bile carbohydrate antigen 19-9 (CA19-9) and carcinoembryonic antigen (CEA) were not different. LR11 protein was found to be highly expressed in the BTC and PC cells. The LR11 transcript levels in cholangiocarcinoma and pancreatic cancer cell lines were sharply induced during proliferation and significantly increased under hypoxic conditions. Conclusions Therefore, sLR11 levels in bile may be indicative of cancer cell conditions and may serve as potential novel biomarker in patients with BTC and PC.

We describe a case of a 57-year-old man who, immediately after a right parietal ischemic stroke, showed urodynamically determined bladder sensory decrement during filling and an underactive detrusor during voiding, both of which were ameliorated during the course of his treatment. The lower urinary tract symptom (LUTS) occurs in stroke in up to 60% of patients, when it involves the frontal and insular cortices. In addition, LUTS does occur in parietal stroke as seen in our patient, presumably by sensory deafferentiation within the brain that is relevant to the central regulation of the micturition reflex.

Aim To evaluate the effectiveness of probiotic therapy for suppressing relapse in patients with inactive ulcerative colitis (UC). Methods Bio-Three tablets, each containing 2 mg of lactomin (Streptococcus faecalis T-110), 10 mg of Clostridium butyricum TO-A, and 10 mg of Bacillus mesentericus TO-A, were used as probiotic therapy. Sixty outpatients with UC in remission were randomly assigned to receive 9 Bio-Three tablets/day (Bio-Three group) or 9 placebo tablets/day (placebo group) for 12 mo in addition to their ongoing medications. Clinical symptoms were evaluated monthly or on the exacerbation of symptoms or need for additional medication. Fecal samples were collected to analyze bacterial DNA at baseline and 3-mo intervals. Terminal restriction fragment length polymorphism and cluster analyses were done to examine bacterial components of the fecal microflora. Results Forty-six patients, 23 in each group, completed the study, and 14 were excluded. The relapse rates in the Bio-Three and placebo groups were respectively 0.0% vs 17.4% at 3 mo (P = 0.036), 8.7% vs 26.1% at 6 mo (P = 0.119), and 21.7% vs 34.8% (P = 0.326) at 9 mo. At 12 mo, the remission rate was 69.5% in the Bio-Three group and 56.6% in the placebo group (P = 0.248). On cluster analysis of fecal flora, 7 patients belonged to cluster I, 32 to cluster II, and 7 to cluster III. Conclusion Probiotics may be effective for maintaining clinical remission in patients with quiescent UC, especially those who belong to cluster I on fecal bacterial analysis.

Some patients infected with hepatitis C virus (HCV) genotype 2 could be cured with treatment shorter than 24 weeks using peginterferon plus ribavirin, but there are still treatment-refractory patients. Direct-acting antivirals (DAAs) are not currently available for HCV genotype 2 patients, different from genotype 1 patients, in clinical practice. We investigated 29 HCV genotype 2-infected Japanese patients who had been previously treated and failed to clear HCV. We retreated them with peginterferon alfa-2a plus ribavirin and measured HCV RNA level to assess the efficacy and safety of this treatment in patients who had failed previous therapy. We found that retreatment of HCV genotype 2-infected Japanese patients with peginterferon alfa-2a plus ribavirin for 24-48 weeks led to 60 to 66.6% sustained virological response (SVR) in patients previously treated with (peg-)interferon monotherapy and to 69.9% SVR in relapsers previously treated with peginterferon plus ribavirin. Attention should be paid to certain patients with unique features. Selection of patients according to their previous treatment could lead to optimal therapy in HCV genotype 2 treatment-experienced patients.

Background: Infliximab (IFX) is an antitumor necrosis factor (TNF)-α antibody used to treat Crohn's disease (CD). However, antibodies to IFX (ATI) emerge, which can impair its efficacy. A fluid-phase enzyme immunoassay (FP-EIA) was established for measuring serum functional IFX (f-IFX) in CD patients receiving maintenance IFX. Methods: In 31 patients, 16 had maintained response (GI) and 15 had lost response to IFX despite good initial response (GII) were selected. Serum f-IFX was measured just before and immediately after IFX infusion and the values together with CD activity index (CDAI) and C-reactive protein (CRP) were compared. Results: IFX therapy in GI and GII were 1.8 ± 1.2 years and 2.7 ± 1.5 years, respectively, while the median dose frequency was 56 days in GI and 29 days in GII. Our FP-EIA for f-IFX showed TNF-α binding increasing with the IFX dose, which was suppressed by antibodies to IFX. On the infusion day, CRP and CDAI in GII were significantly higher than in GI, while median trough f-IFX for GI and GII were 4.7 μg/mL and 6.3 μg/mL, respectively. The median f-IFX immediately after IFX infusion for GI and GII were 149.5 μg/mL and 126.3 μg/mL, respectively (P = 0.0488), and binary logistic regression showed conditional maximum likelihood estimate to be −0.0258 (P = 0.0395), supporting association of low postinfusion f-IFX to the loss of response. Conclusions: FP-EIA could accurately measure f-IFX. High serum ATI strongly impacted f-IFX levels immediately after an infusion. The postinfusion f-IFX level was associated with clinical response. f-IFX level should be valuable in decision-making to optimize treatment efficacy. (Inflamm Bowel Dis 2010)

In this study, poly (ether ether ketone) /carbon nanotubes (CNTs) nano composites were fabricated by using multi-walled carbon nanotubes (MWNTs) as the filler. Tensile properties of nano composites at before and after glass transmission temperature were studied. At room temperature, the addition of 15 wt% MWNTs increased the tensile modulus (0.10.3% strain) by 89% and the yield stress by 19%. At after glass transmission temperature (200) , they increased by 163% and 42%, respectively. The experimental results showed that the reinforcement of CNTs are more effective for high temperature than room temperature. DMA results showed an increase in storage modulus with increasing MWNTs loading fraction at both below and above the glass transition temperature Tg (around 145oC). Scanning electron microscope and transmission electron microscope images showed that the MWNTs were well dispersed in the matrix and orientated one direction.

To examine the association between the interleukin 28B (IL-28B) genotype and treatment response in hepatitis C virus (HCV)-infected patients with persistently normal alanine aminotransferase (PNALT).We compared the treatment response of HCV-infected patients with PNALT to that of patients with non-PNALT. Between February 2010 and April 2013, 278 patients infected with HCV were enrolled in this study. All of the patients were treated with peginterferon-alpha 2a or 2b plus ribavirin. In addition, 180 μg of peginterferon alpha-2a or 1.5 μg/kg peginterferon alpha-2b per week plus weight-based ribavirin (600-1000 mg/d) were typically administered for 24 wk to HCV genotype 2-infected patients or for 48-72 wk to HCV genotype 1-infected patients. In all of the patients, the IL-28B rs8099917 genotype was determined using a TaqMan single-nucleotide polymorphism assay. HCV RNA was measured using the COBAS TaqMan HCV test.Female patients were dominant in the PNALT group (P0.0001). Among 72 HCV genotype 1-infected patients with PNALT, the early virologic response (EVR) rates (P0.01) and the sustained virologic response (SVR) rates (P0.01) were higher in patients with the IL-28B TT genotype than in those with the IL-28B TG/GG genotype. In HCV genotype 1-infected patients with PNALT, multivariate logistic-regression analysis showed that SVR was independently predicted by the IL-28B rs8099917 TT type (P0.05) and having an EVR (P0.01). The IL-28B rs8099917 TT genotype strongly correlated with treatment response in HCV genotype 1-infected Asian patients with PNALT.The IL-28B genotype may be useful for selecting HCV genotype 1-infected patients with PNALT who should receive interferon-based treatment.

Penicillium abeanum lipase was purified to homogeneity as judged by non-denaturing and denaturing polyacrylamide gel electrophoresis. The enzyme is a monomeric protein with a molecular mass of approximately 28 kDa and a pI of 3.4. The optimal pH at 25°C and the optimal temperature at pH 7.5 were 7–8, and 25–30°C, respectively. The enzyme hydrolyzes the 1- and 3-position ester bonds at nearly nine times the rate of cleavage at the 2-position. The enzyme showed lower activities towards ester bonds of polyunsaturated fatty acids as compared with those of other acids, and was proved effective for increasing the concentration of docosahexaenoic acid (22:6) in tuna oil.

Experiments were conducted to obtain row-by-row heat transfer data during condensation of downward-flowing HCFC123 vapour in staggered bundles of horizontal two-dimensional fin tubes. Two conventional low-fin tubes and two finned tubes with fin shapes close to a new class of high performance fin were tested. The heat transfer characteristics were compared with previous results for in-line bundles of the same test tubes. The decrease in heat transfer due to condensate inundation was less significant for the staggered tube bundle. The highest heat transfer performance was provided by the staggered bundle of one of the newly developed finned tubes. Comparison was also made with theoretical predictions for stagnant vapour and predictions of empirical equations for bundles of smooth tubes. The theoretical predictions compared well with experimental data at a low vapour velocity and low-to-medium condensate inundation rates.

AIM: To examine the epidemiological data, hematological safety and treatment responses of peginterferon-alpha 2a plus ribavirin therapy for hepatitis C. METHODS: Between March 2008 and February 2011, 196 hepatitis C virus (HCV) genotype 1 infected Japanese (127 treatment-naive and 69 treatment-experienced patients) patients treated with peginterferon-alpha 2a plus ribavirin were enrolled. We examined the epidemiological data and treatment responses were retrospectively analyzed in terms of hematological safety. HCV RNA was measured by the COBAS TaqMan HCV test. RESULTS: Overall sustained virological response (SVR) rates of treatment-naive and treatment-experienced patients were 56% and 39%, respectively. Multivariate logistic regression analysis showed that SVR was attained independently of early virological response in both treatment-naive and treatment-experienced patients. SVR rates did not differ between the pretreatment hemoglobin < 13 g/dL and ≥ 13 g/dL groups. However, in treatment-naive patients, the SVR rate of the pretreatment platelet count < 130000/µL group was significantly lower than that of the pretreatment platelet count ≥ 130000/µL group. CONCLUSION: Attention should be paid to potential thrombocytopenia in the treatment of chronic hepatitis C patients.

The efficacy of infliximab (IFX) has validated the role of TNF-α in the immunopathogenesis of Crohn's disease (CD). However, antibodies to IFX emerge, which impair its efficacy. This study investigated factor(s) associated with the loss of response (LOR) to IFX and how IFX non-responders may be treated.Seventy-four patients, 36 IFX responders (GI) and 38 with LOR (GII) were included. Trough IFX level, CD activity index (CDAI) and immunological markers during IFX maintenance therapy were measured. Adsorptive granulocyte/monocyte apheresis (GMA) was applied to patients with LOR.The durations of CD, 9.3 ± 5.5 yr and IFX therapy, 3.4 ± 2.0 yr in GII were longer vs GI (P=0.02, P=0.01). Similarly, C-reactive protein (P0.0001) and CDAI (P0.0001) in GII were higher. The median trough IFX was 4.7 μg/mL in GI and 8.4 μg/mL in GII, while the dose frequency was 8 weeks in GI and 4 weeks in GII. Soluble interleukin-2 receptor (sIL-2R) was higher in GII vs GI (P0.001). Seropositive rates of anti-nuclear antibodies (ANA) and circulating immune complexes (CIC) in GII were 50.0% and 68.4%, significantly higher vs GI (P0.05, P0.01). Patients with LOR duration1.5 yr showed higher CDAI and sIL-2R (P0.05) vs patients with LOR duration1.5 yr. Fifteen GII patients received GMA plus IFX combination and 46.7% responded. IL-10 increased in GMA-responders (P0.05), while CIC and ANA decreased (P=0.0237, P=0.0463).Patients with LOR to IFX had dysregulated immune response despite uncompromised trough IFX level. Further, inadequate T-cell differentiation by IFX was suggested. GMA appeared to benefit LOR patients by immunoregulation.

Advanced chronic hepatitis C patients with sustained virolological response by antivirals remain at risk for hepatocellular carcinoma (HCC). We investigated the incidence of HCC during and immediately after peginterferon-alfa-2a and ribavirin (RBV) treatment in patients with chronic hepatitis C in Japan. HCC was detected in 8 of 238 patients during and after these treatments (mean follow-up period: 572 ± 252 days). In conclusion, occurrence of HCC is not a rare event during and immediately after peginterferon-alfa-2a plus RBV treatment. In cases with cirrhosis, higher α-fetoprotein levels, old age, or a previous history of HCC treatment, clinicians should be especially alert for the possible development of HCC during and immediately after peginterferon-alfa-2a and RBV treatment. Clinicians should regularly check for the possible development of HCC even in chronic hepatitis C patients under treatment.

Effects of interferon (IFN) treatment on 91 patients with chronic HCV-related liver diseases in three hospitals were analyzed. HCV-RNA encoding the NS5 region of HCV was detected by the RT-PCR method, and HCV genotyping was performed by slot-blot hybridization using the type-specific cDNA probes on HCV-NS5. HCV titers in blood before treatment with IFN were determined by the multicyclic PCR method. HCV-NS5 was positive in 61 out of 91 patients and the HCV-K1 genotype was found in 40 patients, whereas the HCV-K2 genotype was found in 21. To summarize results from all three hospitals, 52.3% of patients recovered completely and 33.3% recovered partially in the K2 group, in each of these the percentage was significantly higher than in the K1 group. In addition, the percentage of the patients showing no response to IFN treatment was significantly higher in the K1 group than in the K2 group. In the HCV-NS5-negative group, the percentage of patients who did not respond was significantly lower than in the K1 group. In the K1 group, a complete recovery tended to be found more frequently in patients with lower titers of HCV. However, some patients with low titers of HCV did not respond to treatment. The differences of HCV titers between the groups showing different responses to IFN treatment was not statistically significant. In the K2 group, HCV titers were significantly lower than those in the K1 group; however, the effects of IFN were not related to the titers of HCV in K2 group. Two patients with liver cirrhosis in the K2 group had a complete or partial recovery, whereas four patients with liver cirrhosis in the K1 group did not respond to IFN treatment. These results indicate that the HCV genotype is the most important determinant of the effects of IFN treatment.

Two actinomycete strains, MS16-10G and MS16-10W, isolated from soil and producing an antibiotic, were examined and assigned to the genus Streptomyces, because they have a gray aerial mass, spiral spore chains, and a spiny spore surface; form a melanoid pigment on tyrosine agar, on peptone-yeast extract-iron agar, and in tryptone-yeast extract broth; and have cell wall chemotype I. The DNA base compositions and DNA-DNA reassociation of the isolates were compared with those of four strains of authentic species of the genus Streptomyces: two synonymous strains of Streptomyces cyaneus(Streptomyces afghaniensis and Streptomyces echinatus) and one strain each of Streptomyces chromofuscus and Streptomyces antibioticus showing similar morphological characteristics. The results indicated that the two isolates differed from the four authentic species and represent a single new species for which the name Streptomyces bungoensis is proposed. The type strain is MS16-10G, which has been deposited in the collection of the Fermentation Research Institute, Agency of Industrial Sciences and Technology, Tsukuba, Japan, with accession no. FERM 8432T.

Interferon is commonly used for treatment of type C hepatitis, but the effects are variable and many factors may be responsible. Hepatitis C virus (HCV) can be classified into 4 types, PT, Kl, K2a and K2b. Therefore, the responses to interferon treatment in patients with the different HCV genotypes were analyzed. Twenty-four patients with type C hepatitis were treated with 3 to 10 million units of various types of interferon for more than 8 weeks. HCV-RNA encoding the NS5 region (HCV-NS5) was positive in these 24 patients, 16 of which were classified with the Kl type and 8 with the K2 type of HCV. In all patients except for 2, HCV-NS5 became negative within 3 weeks of treatment without relation to the HCV genotypes. Serum alanine aminotransferase levels were normalized in 7 out of 8 patients in the K2 group and in 4 out of 16 patients in the Kl group at the end of 8 weeks. At the 24th week, ALT levels were normalized in 5 out of 6 patients in the K2 group, and in one out of 9 patients in the K1 group. The percentage of patients exhibiting a good response was significantly higher in the K2 group than in the K1 group at both observation periods. During the post-treatment periods, relapse following complete response was found in 3 patients in the K2 group and in one patient in the K1 group. The final effects of interferon were significantly better in the K2 group than in the K1 group. These results indicate that the effects of interferon were significantly better in patients in the K2 group than in patients in the K1 group. Normalization of serum ALT levels was found in two patients with cirrhosis in the K2 group, but in none of the cirrhosis patients in the Kl group, suggesting that effects of interferon may not be related to the progression of liver disease in patients in the K2 group. These results suggest that genotypes of HCV are one of the major determinants of the effect of interferon treatment.

The high prevalence of hepatitis C virus (HCV) markers in alcoholic liver cirrhosis (AL-LC) and hepatocellular carcinoma (HCC) suggests a close aetiopathogenic relationship between alcoholic liver disease (ALD) and HCV infection. In the present study, HCV markers in ALD were measured by the highly sensitive methods, and the changes of sequential HCV markers after abstinence in ALD patients were analysed in order to elucidate the effect of alcohol on HCV. Antibodies to HCV-related antigen were determined using the first or second generation test kit. HCV-RNA genomes encoding the NS-5 region were detected using the RT-PCR method. In the HCV-NS5 negative serum, HCV genomes of the 5'-noncoding region were detected using the two-stage PCR method. Titres of HCV-RNA were measured by multiple cyclic PCR and cDNA dot blotting. Typing of HCV genomes was carried out on the PCR product from the NS-5 region by slot blot hybridization using type-specific cDNA probes, or by restriction fragment length polymorphisms analysis. In alcoholic fibrosis and alcoholic hepatitis, the prevalence of HCV markers was low, suggesting that the main aetiological factor is alcohol but not HCV in these types of ALD. HCV markers were positive in the half of the patients with AL-LC, and in more than 80% of patients with AL-CH and AL-HCC, indicating that HCV infection closely relates to these types of ALD. The ratio of the K1 type to the K2 type of HCV genomes was 4:1 in all types of NANB liver disease.(ABSTRACT TRUNCATED AT 250 WORDS)

Progression of alcoholic liver disease is closely related to drinking habits. However, prognosis of alcoholic liver disease is not determined just by drinking habits, but also by other factors. In this study, the roles of alcohol-altered liver membrane antibody and hepatitis C virus infection were analyzed in alcoholic patients who were followed up for various lengths of time. Serial changes of liver histological appearance were analyzed in 39 patients with alcoholic liver disease who were followed for long periods (49.7 +/- 34.3 mo) and who underwent liver biopsy at least two times. Prognoses of 35 patients with alcoholic cirrhosis who were followed for more than 1 yr were also evaluated. Development of cirrhosis in alcoholic liver disease was significantly higher in the alcohol-altered liver membrane antibody-positive patients than in the patients negative for this antibody. On the other hand, hepatitis C virus markers were not related to development of cirrhosis. However, hepatocellular carcinoma developed more frequently in the hepatitis C virus marker-positive patients. In patients with cirrhosis, the cumulative rates of hepatocellular carcinoma development were significantly higher in the hepatitis C virus marker-positive patients than in the marker-negative patients. Cumulative survival rates in the hepatitis C virus marker-positive patients were significantly lower than rates in the hepatitis C virus marker-negative patients, even in noncancer patients. However, such differences were not observed in the alcohol-altered liver membrane antibody-positive and alcohol-altered liver membrane antibody-negative cirrhotic patients.(ABSTRACT TRUNCATED AT 250 WORDS)

A microorganism producing a solvent-tolerant lipase was identified as Fusarium (F.) heterosporum. The lipase was purified from the culture filtrate to homogeneity as judged by disc-PAGE and SDS-PAGE. The purification included SP-Sephadex chromatography, gel filtration and isoelectric focusing, and the recovery yield was 38%. The lipase was a monomeric protein with a molecular weight of 31 kDa estimated by SDS-PAGE, and a pI of 7.0. The optimum pH at 40°C and optimum temperature at pH 5.6 were 5.5–6.0 and 45–50°C, respectively, when olive oil was used as the substrate. The lipase was stable over a pH range of 4–10 at 30°C for 4 h, and up to 40°C at pH 5.6 for 30 min. Furthermore, the enzyme was not inactivated even after incubation at 30°C in 50% solvent such as dimethylsulfoxide (DMSO), hexane, benzene and ether for 20 h. The activity did not decrease in a reaction with stirring in a mixture containing 50% DMSO or dimethylformamide. The lipase preferably reacted on middle-chain fatty acid triglycerides (6≤C≤12), and cleaved only 1,3-ester bonds of triolein. The enzyme had an N-terminal sequence of Ala-Val-Thr-Val-Thr-Thr-Gln-Asp-Leu-Ser, which has not previously been found in any other protein. We compared the properties of lipases from F. heterosporum and another strain F. oxysporum.

This paper presents a comparison of measured condensation heat transfer coefficients for refrigerant CFC-11 and its proposed replacement HCFC-123 flowing down a 3×15 (columns×rows) in-line bundle of horizontal finned tubes. The vapor velocity at the tube bundle inlet (based on minimum flow cross section) ranged from 0.9 to 10 m/s, and the condensation temperature difference from 1.3 to 19K. Heat transfer measurements were made on a row-by-row basis. Generally, the heat transfer coefficient for HCFC-123 was about 10% lower than that for CFC-11. The heat transfer enhancement due to vapor shear was much smaller than that in the case of a smooth tube bundle. At a high vapor velocity, the enhancement was most significant for the top row and decreased sharply with increasing row number. The decrease in the heat transfer coefficient due to condensate inundation was also much smaller for the finned tube bundle. The measured heat transfer coefficient for the smallest vapor velocity agreed well with the theoretical prediction for a stagnant vapor.

We recently classified the hepatitis C virus (HCV) into 4 types (HCV-PT, -K1, -K2a and -K2b) according to differences in nucleotide sequences. It was found that HCV-PT, the prototype reported from the U.S.A., was rare in Japan, suggesting that distribution of HCV genotypes may be different in various countries. The prevalence of HCV genotypes was therefore compared in different countries. Genotyping of HCV was performed by slot-blot hybridization analysis using cDNA probes specific to each type of HCV or by restriction fragment length polymorphism analysis. In 121 Japanese non-cancer patients, the prevalence of HCV genotypes was 77.7% for HCV-K1, 16.5% for HCV-K2a and 5.0% for HCV-K2b. HCV-PT was detected in only 1 patient (0.8%). The prevalence in 43 Japanese hepatocellular carcinoma (HCC) patients was 74.4% for HCV-K1, 18.6% for HCV-K2a and 4.7% for HCV-K2b. HCV-PT was found in only 1 sample. In 19 European non-cancer patients, HCV-PT was found in 42.1% and HCV-K1 was found in 52.6%. HCV-K2 was not found. All 7 samples from European HCC patients were HCV-K1, indicating a significantly higher prevalence than in non-cancer patients. In 13 Brazilian non-cancer patients, the distribution pattern was similar to that of the Europeans. In 10 samples from the U.S.A., HCV-PT was found in 70% and HCV-K2 was found in 1 sample. In 18 Chinese noncancer patients, HCV-K1 was found in 44.4%, HCV-K2a in 50.0% and HCV-K2b in 5.6%. HCV-PT was not found. Two samples from Chinese HCC patients were HCV-K1. These results indicate that the distribution patterns of HCV genotypes vary widely among different countries. Prevalence of HCV-K1 was high in every country except for the U.S.A., suggesting that HCV-K1 may be the major type of HCV worldwide. HCV-PT was occasionally found in the Japanese and Chinese samples but was more common in the samples from Western countries. On the other hand, HCV-K2 was rarely found in samples from Western countries but was commonly found in the Japanese and Chinese samples. These differences suggest that HCV-PT may be the Western type and HCV-K2 may be the Oriental type.

Summary. Extremely low levels of serum hepatitis C virus (HCV) RNA can be detected by COBAS TaqMan HCV test. To investigate whether the COBAS TaqMan HCV test is useful for measuring rapid virological response (RVR) and early virological response (EVR) to predict sustained virological response (SVR), we compared the virological response to PEG-IFN-alfa 2a plus RBV in 76 patients infected with HCV genotype 1 when undetectable HCV RNA by the COBAS TaqMan HCV test was used, with those when below 1.7 log IU/mL HCV RNA by COBAS TaqMan HCV test was used, which corresponded to the use of traditional methods. Among the 76 patients, 28 (36.8%) had SVR, 13 (17.1%) relapsed, 19 (25.0%) did not respond, and 16 (21.0%) discontinued the treatment due to side effects. The positive predictive values for SVR based on undetectable HCV RNA by COBAS TaqMan HCV test at 24 weeks after the end of treatment [10/10 (100%) at week 4, 21/23 (91.3%) at week 8 and 26/33 (78.7%) at week 12] were superior to those based on

わが国でのC型肝炎ウイルス(HCV)は,その塩基配列の差から4型に分けられるが,HCV各型の意義については知られていない.そこで,HCV-RNA陽性の肝疾患患者71例にいて,その遺伝子型を2種類の方法で分析した.同時に,スペインの13例,中国の11例の非癌患者についても同様に分析した.非癌患者58例では,K1が44例(75.9%), K2aが9例(15.5%),K2bが4例(6.9%)であり,PTは輸入血液製剤で感染した1例(1.7%)のみであった.肝癌患者13例ではK1が11例(84.6%)と多く,特に大酒家では全例がK1であった.スペイン例ではK2は1例もなく,PTが5例(38.5%), K1が7例(53.8%)であった.一方,中国例ではPTは1例もなく,K1が6例(54.5%), K2aが4例(36.4%), K2bが1例(9.1%)であった.このように,HCVの型別頻度は国によって大きく異なるが,その頻度からみて,K1がHCVの原型で,PTは西洋型の,K2は東洋型の亜型であり,型別分布の差が,各国でのC型肝炎の病態の差に関係していると推定された.

Adacolumn selective granulocyte and monocyte apheresis (GMA) depletes activated leukocytes in patients with ulcerative colitis (UC). However, this per se cannot fully explain the efficacy of GMA. We have investigated the effects of GMA on the expression of toll-like receptors (TLRs) and plasma interleukin-8 (IL-8). Twenty-two patients with clinical activity index (CAI) of 5–17, 15 with total colitis and 7 with left-sided colitis, were included. Each patient could receive up to 10 GMA sessions, at 1 or 2 sessions per week. GMA was added to the patients’ ongoing medication following a relapse or worsening UC, but no additional medication was given. Further, at entry and pre-GMA, blood samples were taken for full blood cell count, expression of TLRs on leukocytes, and plasma IL-8. Seventy-five percent of patients achieved remission after the 10th session (CAI, ≤4; P < 0.005) and there was a marked fall in C-reactive protein (P < 0.01), plasma IL-8 (P < 0.001), and granulocytes (P < 0.05) but an increase in lymphocytes (P < 0.05). The expression of TLR2 on granulocytes was down-modulated (P < 0.05) together with suppression of inflammatory cytokines produced by peripheral blood leukocytes. In conclusion, GMA appears to be an effective adjunct therapy to induce remission in the majority of patients, who are then spared from excess drug therapy. The procedure is associated with sustained immunomodulation. Control studies should strengthen these findings.

We have reported that hepatitis C virus (HCV) RNA encoding the NS5 region (HCV-NS5) became negative following abstinence in some patients with alcoholic liver disease (ALD). In order to clarify the relationship between alcohol misuse (abuse) and the replication of HCV, serial changes of HCV-NS5 and the titer of HCV (HCV-titer) in blood after abstinence were studied in 11 male patients with ALD. Disappearance of HCV-NS5 after abstinence was found in five out of 11 patients (45%) and HCV-titer in blood decreased in all of these cases. In one patient, HCV-NS5 became positive and HCV-titers increased again following resumption of alcohol misuse. The etiology of liver disease in heavy drinkers was subdivided into three categories: alcohol alone, alcohol and HCV, and HCV alone, according to the diagnostic criteria of the Japanese research group for ALD. The etiology in four patients was a combination of alcohol and HCV, and was HCV alone in the remaining seven patients. In all four patients of the alcohol and HCV group, and one out of seven patients in the HCV alone group, HCV-NS5 became negative and HCV-titer decreased after abstinence. The decrease in HCV-titer was not related to the HCV genotypes. These results suggest that alcohol misuse may enhance the replication of HCV.

Hepatitis C virus (HCV) genomes were recently detected in biological materials, and variations of nucleotide sequences were reported. In the present study, typing of the HCV genomes was performed in 91 HCV-RNA-positive patients and the clinical features of patients with different types of HCV were compared. From the nucleotide sequences of the cDNA fragments, HCV can be divided into at least two types: HCV-K1-PT and HCV-K2. All cDNAs amplified from 91 patients were hybridized with cDNA probes of either HCV-K1-PT or HCV-K2. HCV-K1-PT was found in about 80% of the patients, and HCV-K2 was found in about 20% of the patients. These results indicate that types of HCV are limited to two types, i.e., K1-PT and K2, and the major type is HCV-K1-PT, at least in Japan. Detection rate of antibodies to C-100-3 protein were not different between the patients having HCV-K1-PT and HCV-K2, indicating that the antibodies may develop in HCV-related patients without relation to the types of the HCV genomes. Prevalence of the two types of HCV were nearly the same in various forms of NANB-related liver disease. However, the prevalence was somewhat different in alcoholic liver disease. HCV-K2 was found in patients younger than the patients with HCV-K1-PT. Frequency of a history of blood transfusion tended to be lower and the initial response to interferon treatment was clearly better in patients having HCV-K2 versus patients having HCV-K1-PT. These results suggest the possibility that clinical features due to HCV-K1 may be somewhat different from those due to HCV-K1-PT. However, the number of patients examined was too small to allow a definite conclusion, indicating a necessity for further study with a larger number of patients.

Recently, we reported that hepatitis C virus (HCV) can be classified genetically into two types, HCV-K1 and HCV-K2, which show 67% and 71% identity at the nucleotide and amino acid sequence levels in a 340 bp region which encodes the NS5 gene Gly-Asp-Asp motif. To develop a rapid method to classify the genomes of HCV isolates, we identified restriction fragment length polymorphisms (RFLPs) in reverse transcriptase-polymerase chain reaction products encoding a portion of the NS5 gene. AluI and AccII enabled HCV to be classified into the K1 and K2 types, and Sau96I enabled classification into the K1 type, and the K2a and K2b subtypes. These RFLPs also generally allow Japanese isolates to be distinguished from the prototype (PT, an isolate from the U.S.A.), which is a K1 type. Sequence analysis of the 5′-untranslated regions of Japanese isolates revealed near identity between the K1 type and PT, and 93 to 94% identity between the K1 and K2 types, indicating that there are type K1- and K2-specific RFLPs in this region. Our results suggest that the nucleotide sequences of the K1 and K2 types are different throughout the HCV genome. The incidence of HCV types K1, K2a and K2b, and PT in 50 samples was 74%, 16%, 8% and 2%, respectively.

To clarify the pathogenetic role of acetaldehyde in the development of alcoholic liver disease, genotyping of aldehyde dehydrogenase-2 genes was performed and the clinical features of the alcoholic liver disease patients with different genotypes were compared. Genotyping of aldehyde dehydrogenase-2 was performed in 47 patients with alcoholic liver disease using the polymerase chain reaction and slot-blot hybridization. Of the 47 patients with alcoholic liver disease, 40 were homozygous for the normal aldehyde dehydrogenase-2 gene and the remaining seven cases were heterozygous for the normal and mutant aldehyde dehydrogenase-2 genes. No homozygote was found for the mutant aldehyde dehydrogenase-2 genes. Daily alcohol intake was less than 100 gm in all heterozygotes without relation to the type of alcoholic liver disease. On the other hand, all but four patients homozygotic for the normal aldehyde dehydrogenase-2 gene drank more than 100 gm alcohol/day. The mean daily alcohol intake in the heterozygotes was significantly lower than that in the normal homozygotes. The incidence of alcoholic fibrosis tended to be lower in the heterozygotes than in the normal homozygotes (14.2% vs. 52.5%). On the other hand, the incidence of alcoholic hepatitis and/or cirrhosis tended to be higher in the heterozygotes than in the normal homozygotes. These results indicate that alcoholic liver disease develops even with moderate amounts of alcohol intake in heterozygotes of the aldehyde dehydrogenase-2 genes, in which acetaldehyde metabolism in the liver is impaired and liver damage in the heterozygotes is more severe than that in the normal homozygotes, suggesting that habitual drinkers who are heterozygotes of the aldehyde dehydrogenase-2 genes may be at high risk for alcoholic liver disease.

To verify the existence of chronic hepatitis induced by alcohol, the clinicopathological features of chronic hepatitis in heavy drinkers were studied using various viral markers. Histological features of chronic active hepatitis were seen in 27 heavy drinkers. These patients were divided into four groups. The AL group (seven cases) consisted of alcoholics who were negative for both hepatitis C antibody and HBsAg; the HB group (four cases) was positive for HBsAg; the HC1 group (seven cases) was positive for hepatitis C antibody but negative for hepatitis C virus–RNA genome; and the HC2 group (nine cases) was positive both for hepatitis C antibody and hepatitis C virus–RNA genome. Serum AST and ALT activity declined during 4 wk of abstinence in most patients in the AL group and in the HC1 group. The response of serum AST and ALT to abstinence was poor in most patients in the HB group and the HC2 group. Serum desialo-transferrin and alcohol liver membrane antibodies were detected more frequently in the sera of patients in the AL group and HC1 group. A trend toward increased frequency of centrilobular ballooning existed in the AL group, but this did not reach statistical significance. These results suggest that chronic active hepatitis in patients in the AL group, in whom markers of HBV and hepatitis C virus were absent, may be caused by alcohol. Patients in the HC1 group who had hepatitis C antibody but not hepatitis C virus–RNA may represent cases where both alcohol and hepatitis C virus are involved. (HEPATOLOGY 1991;13:876–881.)

Recently, the assay system of anti-hepatitis C virus antibody (HCV-Ab) was developed. However, there is no clinically useful method to detect hepatitis C virus (HCV) itself. The authors recently developed a method to detect the HCV-RNA genome in plasma using polymerase chain reaction (PCR). In the present study, the specificity of this assay in detecting HCV infection was investigated. Freshly obtained 1 ml plasma specimens from 100 patients with various liver diseases and from 11 control subjects were studied. In patients with non-A, non-B (NANB) hepatitis-related liver diseases, HCV-RNA was detected in 2 out of 7 cases of acute hepatitis, in 29 out of 31 cases of chronic hepatitis, in 17 out of 21 cases of cirrhosis and in 2 out of 6 cases of hepatocellular carcinoma. On the other hand, no HCV-RNA was detected in 15 cases of various types of alcoholic liver diseases, in 12 cases of hepatitis B related liver diseases, and in 11 controls. HCV-RNA was detected in 2 of 6 drinkers with chronic hepatitis. The prevalence of HCV-RNA was not closely related to a history of blood transfusions. These results suggest that our method for HCV-RNA is specific for HCV infection and HCV infection is the likely etiology of most chronic NANB hepatitis cases. The clinical usefulness of our method is illustrated by the fact that we were able to study 100 patients and needed only 1 ml plasma per HCV-RNA assay.

These results indicate that there are at least two subtypes of HCV genomes and that the major type is HCV-K1, HCV-K2 is the minor type and the prototype HCV is scarcely found in Japan.