Seiji Nakai

Combination therapy of bezafibrate and ursodeoxycholic acid in primary biliary cirrhosis: a preliminary study

Combination therapy of bezafibrate and ursodeoxycholic acid in primary biliary cirrhosis: a preliminary study

Variable phase-contrast fluorescence spectrometry for fluorescently stained cells

This letter describes the spectroscopic measurements of fluorescently stained cells. Variable phase-contrast fluorescence spectrometry was used on fluorescently stained cells to achieve high two-dimensional spatial resolution. Phase shift interferometry by autocorrelation interference made it possible to measure fluorescence spectra in the field of view without the separation of wavelengths, as in the case of a conventional dispersive spectrometer. In this letter, the authors describe the experimental characteristics of fluorescence spectra generated from fluorescently stained cells and verify that the fluorescence spectra of the stained area in the cells can be measured by our method.

Efficacy of zinc administration in patients with hepatitis C virus-related chronic liver disease.

Objective. Zinc supplementation has been shown to contribute to inhibition of liver fibrosis and improvement in hepatic encephalopathy. However, little is known about the anti-inflammatory effect of zinc on hepatitis C virus (HCV)-related chronic liver disease (CLD). We therefore examined the effects of zinc administration on inflammatory activity and fibrosis in the liver of patients with HCV-related CLD. Material and methods. Polaprezinc, a complex of zinc and l-carnosine, was administrated at 225 mg/day for 6 months to 14 patients with HCV-related CLD, in addition to their ongoing prescriptions. Peripheral blood cell counts, liver-related biochemical parameters, serological markers for liver fibrosis, HCV-RNA loads, and serum levels of zinc and ferritin were evaluated before and after zinc administration. Results. Serum zinc concentrations were positively correlated with hepatic reserve before zinc supplementation. A significant increase in serum zinc level was observed after zinc supplementation (64±15 versus 78±26 mg/dl, p=0.0156). Treatment with polaprezinc significantly decreased serum aminotransferase levels (aspartate aminotransferase (AST): 92±33 versus 63±23 IU/l, p=0.0004; alanine aminotransferase (ALT): 106±43 versus 65±32 IU/l, p=0.0002), whereas alkaline phosphatase levels were significantly increased (305±117 versus 337±118 U/l, p=0.0020). Serum ferritin levels were significantly decreased by treatment with polaprezinc (158±141 versus 101±80 ng/ml, p=0.0117). The reduction rate of ALT levels by polaprezinc was positively correlated with that of ferritin (r2=0.536, p=0.0389). There was a tendency toward a decrease in serum type IV collagen 7S levels after treatment with polaprezinc. However, administration of polaprezinc did not affect peripheral blood cell counts, other liver function tests, or HCV-RNA loads. Conclusions. These findings suggest that polaprezinc exerts an anti-inflammatory effect on the liver in patients with HCV-related CLD by reducing iron overload.

Reduced expression of cell cycle regulator p18INK4C in human hepatocellular carcinoma

Cyclins, cyclin‐dependent kinases (Cdks), and Cdk inhibitors (CdkIs) are frequently altered in human cancer. p18INK4C, a member of the INK4 family of CdkIs, is a potential tumor‐suppressor gene product. However, the expression of p18INK4C in hepatocellular carcinoma (HCC) remains unknown. The aim of this study was to examine the expression of p18INK4C in various liver diseases including HCC and to assess its clinical significance in HCC. To that end, we examined the expression of p18INK4C by immunohistochemistry in various liver diseases, including 51 HCCs, and also studied the relationship between p18INK4C expression, the phosphorylation of retinoblastoma protein (pRb), and the activity level of Cdk4 and Cdk6. Immunohistochemical analysis revealed the frequent loss of p18INK4C expression in HCC, especially in poorly differentiated HCC. The loss of p18INK4C expression was shown to be associated with a poor prognosis compared with that associated with p18INK4C‐ positivity. Further, the kinase activity of Cdk4 was found to be higher in p18INK4C‐negative HCCs than in p18INK4C‐ positive HCCs. However, the level of Cdk6 activity was similar in the 2 groups of HCCs. In p18INK4C‐ positive HCCs, p18INK4C dominantly interacted with Cdk4 rather than with Cdk6. pRb phosphorylated at serine(Ser) 780 was detected more frequently in p18INK4C ‐ negative than in p18INK4C ‐ positive HCCs. In conclusion, the loss of p18INK4C expression may play a role in the differentiation and development of HCC through the up‐regulation of Cdk4 activity. (HEPATOLOGY 2004;40:677–686.)

Expression of p33 ING1 in Hepatocellular Carcinoma: Relationships to Tumour Differentiation and Cyclin E Kinase Activity

Background: Inhibitor of growth-1 (ING1) is a new candidate for the tumour suppressor gene that encodes a 33k Da protein (p33 ING1 ). While reduction of p33 ING1 is an important event in some human cancers, the expression of p33 ING1 in human hepatocellular carcinoma (HCC) remains to be examined. We evaluated p33 ING1 expression in various liver diseases including HCC. Methods: Expression of p33 ING1 was evaluated immunohistochemically not only in the normal liver ( n = 5), but also in specimens of chronic hepatitis ( n = 39) and HCC ( n = 86). We also analysed the relationship between p33 ING1 expression and cyclin E kinase activity detected by autoradiography in 29 HCCs. Results: Expression of p33 ING1 was reduced in HCC, especially in moderately and poorly differentiated HCCs, and those at advanced stages. Furthermore, expression of p33 ING1 correlated inversely with cyclin E kinase activity. Conclusions: These data suggest that reduction of p33 ING1 may contribute to the process of malignant transformation, progression and dedifferentiation of HCC via an increase of cyclin E kinase activity.

A precise method for rotating single cells

A precise method to rotate single cells is reported. In this method, the light pressure in the optical axis direction is harnessed as a rotating torque. Two proximal points in each cell are illuminated from different directions using two beams, and a light pressure is created that acts as a rotating torque. Using this proposed method, we could control the rotational direction of a microsphere regardless of the refractive index distribution in a noncontact operation. The microsphere could be rotated using proximal two-beam optical tweezers, and the rotational velocity could be controlled by changing the light intensity.

CD28-negative CD8-positive cytotoxic T lymphocytes mediate hepatocellular damage in hepatitis C virus infection.

The pathogenic mechanism for hepatocellular damage in hepatitis C virus (HCV) infection has not been clearly understood. Analysis of costimulatory molecules on lymphocytes may give us insight into the pathogenic mechanism of hepatocellular damage in HCV infection. Peripheral blood mononuclear cells (PBMCs) and liver infiltrating mononuclear cells (LIMCs) isolated from the HCV-infected patients were analyzed with antibodies directed against a variety of costimulatory molecules by flow cytometry. Blocking experiment against HLA-A24-restricted HCV-specific CTLs and immunohistochemical analysis were also performed. PBMCs expressing CD8, CD28, CD80, or CD154 were significantly reduced in HCV-infected patients compared with the healthy controls. CD28(+)CD8(+) PBMCs in the patients inversely correlated with ALT levels. Conversely, levels of CD28(−)CD8(+) LIMCs correlated with ALT levels. HCV-specific CTL activity was blocked by the treatment with anti-CD8 antibody, but not with anti-CD4 or anti-CD28 antibody. Immunohistochemical analysis revealed the accumulation of CD28(+) cells around the portal area in the liver of a patient with chronic active hepatitis C. These results suggest that CD28(+)CD8(+) T cells leave the circulation, move to the livers, and are activated in the portal area in proportion to the extent of liver diseases. CD28(−)CD8(+) T cells may finally function as effector T cells causing the hepatocellular damage in HCV infection.

Analysis of CD28 and bcl-2 Expression on Peripheral Blood and Liver-Infiltrating Mononuclear Cells in Patients with Autoimmune Hepatitis

Because the underlying mechanism of hepatocellular damages in autoimmune hepatitis (AIH) still remains unclear, analysis of CD28 and bcl-2 molecules, which are critical for T cell activation and survival, was performed in patients with AIH. The number of CD28(+)CD4(+) peripheral blood mononuclear cells (PBMC) in corticosteroid (CS)-treated patients was comparable to normal control individuals but decreased in untreated AIH patients. In contrast, the number of CD28(+)CD8(+) PBMC was decreased in both CS-treated and untreated AIH patients. Analysis of liver-infiltrating mononuclear cells (LIMC) showed that the number of CD28(+)CD4(+) and CD28(−)CD8(+) LIMC were positively correlated with the histology activity index score. Bcl-2(+)CD4(+) LIMC were observed in the portal area of the liver and the numbers fluctuated with disease activity during the time course after CS administration. By contrast, CD8(+) LIMC were shown not to express bcl-2. Taken collectively, these results suggest that bcl-2(+)CD28(+)CD4(+) and bcl-2(−)CD28(−)CD8(+) cells may play critical and distinct roles in hepatocellular damage in AIH.

Translational velocity measurement for single floating cell based on optical Fourier transform theory

This letter reports on translational velocity measurement, which is needed for tracking a low contrast cell. We propose a new optical spatial filtering method that is based on the optical Fourier transform theory. In this method, a pinhole is installed as a spatial filter on the optical Fourier transform plane. By means of this spatial filter, the arbitrary component of the spatial frequency is derived from the random refractive index distribution as the periodic light intensity distribution. By observing the changes of this light intensity, we can obtain the translational velocity of a low-contrast cell by means of a high-response photodiode.

Coexistence of splenic non-Hodgkin's lymphoma with hepatocellular carcinoma in a patient with chronic hepatitis C.

Chronic hepatitis C virus (HCV) infection is believed to play important roles in hepatocarcinogenesis (1), although there is no apparent evidence that HCV has direct oncogenic effects on hepatocytes. On the other hand, HCV involves many kinds of extrahepatic manifestations, including mixed cryoglobulinemia, Sjögren syndrome, interstitial pneumoniae, and chronic thyroiditis (2). Recently, strong association of HCV with non-Hodgkin’s lymphoma (NHL) has been reported (3–9). Therefore, it has been suggested that chronic HCV infection may also contribute to the development of NHL. We report the case of a HCVinfected patient with coexistent splenic NHL with hepatocellular carcinoma (HCC). Fine needle splenic biopsy under the guidance of ultrasound imaging was a useful procedure for the diagnosis of the splenic lesion in the present case.