Yasushi Seo

ADC Measurement of Abdominal Organs and Lesions Using Parallel Imaging Technique

OBJECTIVE The purpose of our study was to assess the reliability and usefulness of parallel imaging for apparent diffusion coefficient (ADC) measurement of abdominal organs and lesions. MATERIALS AND METHODS Single-shot spin-echo echo-planar diffusion-weighted MRI (TE = 66, b = 0, 600 s/mm2) was performed in phantom and clinical studies. The b value was set to minimize the effects of perfusion in tissue and to maintain signal-to-noise ratio. Bottle phantoms were scanned with and without parallel imaging and with various parallel imaging factors and at various positions to evaluate the effects of parallel imaging on ADCs. In 200 consecutive clinical patients (122 men and 78 women: mean age, 61.9 years), ADCs were calculated for liver (four segments), spleen, pancreas (head, body, tail), gallbladder, renal parenchyma, and back muscle, and then compared to evaluate the reliability of clinical ADC measurements with parallel imaging. ADCs were also calculated for diffuse diseases and focal lesions (94 malignant and 93 benign) of abdominal organs to evaluate the clinical usefulness of ADC. RESULTS Location-dependent changes in water ADCs were minimal with parallel imaging factors first of 3, then of 4, and were small except for measurements at the image periphery. Acetone ADCs were saturated at 4.00 x 10(-3) mm2/s. Degraded image quality prevented ADC measurement of the left hepatic lobe and pancreas in 7-18 patients. There was no significant difference among ADCs of four liver segments (1.50 +/- 0.24 [SD] x 10(-3) mm2/s - 1.56 +/- 0.31 x 10(-3) mm2/s) and between ADCs of the right and left kidneys (2.65 +/- 0.30 x 10(-3) mm2/s, 2.59 +/- 0.33 x 10(-3) mm2/s). ADC of the pancreas tail (1.65 +/- 0.37 x 10(-3) mm2/s) was significantly lower than those of the head (1.81 +/- 0.40 x 10(-3) mm2/s) and body (1.81 +/- 0.41 x 10(-3) mm2/s) (p < 0.005). Renal ADCs were significantly lower in patients with renal failure (right: 2.15 +/- 0.30 x 10(-3) mm2/s; left: 2.11 +/- 0.25 x 10(-3) mm2/s) than in those without disease (right: 2.67 +/- 0.29 x 10(-3) mm2/s; left: 2.60 +/- 0.32 x 10(-3) mm2/s) (p < 0.005). ADC of pancreatic cancer was significantly higher than that of healthy pancreas (p < 0.05). ADC of renal angiomyolipoma was significantly lower than those of renal cell carcinoma and healthy renal parenchyma (p < 0.0005). CONCLUSION Clinical ADC measurements of abdominal organs and lesions using parallel imaging appear to be reliable and useful, and the effect of parallel imaging on calculated values is considered to be minimal.

High Expression of PRL-3 Promotes Cancer Cell Motility and Liver Metastasis in Human Colorectal Cancer: A Predictive Molecular Marker of Metachronous Liver and Lung Metastases

Purpose: Overexpression of PRL-3 has been implicated in colorectal cancer metastases. We investigated the significance of PRL-3 expression in the progression and development of colorectal cancer. Experimental Design: We transfected PRL-3-specific small interfering RNA into human colon cancer DLD-1 cells and analyzed its effect on proliferation, motility, and hepatic colonization. Using an in situ hybridization method, we examined the levels of PRL-3 expression in both primary (177 cases) and metastatic (92 cases) human colorectal cancers and elucidated the relationships with clinicopathological parameters including the incidence of metachronous liver and/or lung metastasis after curative surgery for primary tumor. Results: Transient down-regulation of PRL-3 expression in DLD-1 cells abrogated motility (in vitro) and hepatic colonization (in vivo), but no effect on the proliferation of these cells was observed. In human primary colorectal cancers, the frequency of up-regulated PRL-3 expression in cases with liver (84.4%) or lung (88.9%) metastasis was statistically higher than that in cases without either type of metastasis (liver, 35.9%; lung, 42.3%). In metastatic colorectal cancer lesions, high expression of PRL-3 was frequently detected (liver, 91.3%; lung, 100%). Interestingly, metachronous metastasis was observed more frequently in the cases with high PRL-3 expression (P < 0.0001). Conclusions: These results indicate that PRL-3 expression in colorectal cancers may contribute to the establishment of liver metastasis, particularly at the step in which cancer cells leave the circulation to extravasate into the liver tissue. In addition, PRL-3 is expected to be a promising biomarker for identifying colorectal cancer patients at high risk for distant metastases.

Heterogeneous expression of claudin-4 in human colorectal cancer: decreased claudin-4 expression at the invasive front correlates cancer invasion and metastasis.

Objective: Claudin-4 plays a key role in constructing the tight junction (TJ), and altered claudin-4 expression has been documented in various human malignancies; however, little is known about the biological significance of claudin-4 in colorectal cancers (CRCs). The aim of this study is to investigate the significance of claudin-4 expression in CRC and its association with clinicopathological factors. Methods: The levels of claudin-4 expression in a total of 129 CRCs and 44 metastatic tumors were examined by immunohistochemistry. A small interfering RNA (siRNA)-mediated claudin-4 knockdown examination was also conducted to assess the biological role(s) of claudin-4 in cultured cells. Results: Expression of claudin-4 at the intercellular membrane was well preserved at the surface of the tumor; however, decreased claudin-4 expression was detected in 57% of CRCs, particularly at the invasive front. Interestingly, decreased claudin-4 expression was detected in metastatic lesions of CRC. The siRNA-mediated claudin-4 knockdown in SW480 claudin-4-positive CRC cells upregulated cell motility, whereas no significant change was detected in cell proliferation. Conclusions: These observations suggested that disruption of claudin-4-mediated TJ construction enhances cancer cell invasion and metastasis in human CRC. Claudin-4 might be a good biomarker for diagnosing the risk of distant metastasis.

Nonalcoholic fatty liver disease in adult hypopituitary patients with GH deficiency and the impact of GH replacement therapy

BACKGROUND Liver dysfunction in adult hypopituitary patients with GH deficiency (GHD) has been reported and an increased prevalence of nonalcoholic fatty liver disease (NAFLD) has been suggested. OBJECTIVE The objective of the present study was to elucidate the pathophysiology of the liver in adult hypopituitary patients with GHD. PATIENTS AND METHODS We recruited 69 consecutive Japanese adult hypopituitary patients with GHD and examined the prevalence of NAFLD by ultrasonography and nonalcoholic steatohepatitis (NASH) by liver biopsy. Patients had been given routine replacement therapy except for GH. We compared these patients with healthy age-, gender-, and BMI-matched controls. We further analyzed the effect of GH replacement therapy on liver function, inflammation and fibrotic markers, and histological changes. RESULTS The prevalence of NAFLD in hypopituitary patients with GHD was significantly higher than in controls (77 vs 12%, P<0.001). Of 16 patients assessed by liver biopsy, 14 (21%) patients were diagnosed with NASH. GH replacement therapy significantly reduced serum liver enzyme concentrations in the patients and improved the histological changes in the liver concomitant with reduction in fibrotic marker concentrations in patients with NASH. CONCLUSIONS Adult hypopituitary patients with GHD demonstrated a high NAFLD prevalence. The effect of GH replacement therapy suggests that the NAFLD is predominantly attributable to GHD.

Clinical significance of serum hyaluronic acid as a fibrosis marker in chronic hepatitis C patients treated with interferon‐α: Histological evaluation by a modified histological activity index scoring system

Abstract The aim of the present study was to investigate the histological changes effected by interferon (IFN) treatment and to evaluate the clinical significance of serum hyaluronic acid (HA) as a marker of fibrosis. Forty‐nine patients with chronic hepatitis C treated with IFN‐α were divided into three groups according to the existence of viraemia: sustained complete responders (CR), complete responders with relapse (PR) and non‐responders (NR). Needle biopsy sections of the liver taken before and at the end of IFN treatment were assessed according to the modified histological activitindex (HAI) scoring system. Serum fibrosis markers, including HA, were measured at needle biopsies. Biopsies of CR at the end of treatment showed a significant improvement in fibrosis and necroinflammatory scores. More significant correlation was observed between fibrosis scores and serum levels of HA before IFN treatment (r= 0.607, P < 0.0001) than those between fibrosis scores, on the one hand, and pepride of type III procollagen (PIIIP; r= 0.531, P= 0.0004) or type IV collagen 7S domain (type IV‐C; r= 0.241, P= 0.1062) on the other. Moreover, serum HA levels fell significantly in patients in whom fibrosis improved (P= 0.011). This is the first paper describing the advantages of the modified HAI scoring system over others in estimating the effect of IFN‐α; the results also indicate that serum HA can be useful in monitoring liver fibrosis in chronic hepatitis C patients treated with IFN‐α.

Activation of the aryl hydrocarbon receptor induces hepatic steatosis via the upregulation of fatty acid transport

The aryl hydrocarbon receptor (AHR) is a basic helix-loop-helix/Per-ARNT-Sim domain transcription factor, which is activated by various xenobiotic ligands. AHR is known to be abundant in liver tissue and to be associated with hepatic steatosis. However, it has not yet been elucidated how the activation of AHR promotes hepatic steatosis. The aim of this study is to clarify the role of AHR in hepatic steatosis. The intraperitoneal injection of 3-methylcholanthrene (3MC), a potent AHR ligand, into C57BL/6J mice significantly increased the levels of triglycerides and six long-chain monounsaturated fatty acids in the livers of mice, resulting in hepatic microvesicular steatosis. 3MC significantly enhanced the expression level of fatty acid translocase (FAT), a factor regulating the uptake of long-chain fatty acids into hepatocytes, in the liver. In an in vitro experiment using human hepatoma HepG2 cells, 3MC increased the expression level of FAT, and the downregulation of AHR by AHR siRNA led to the suppression of 3MC-induced FAT expression. In addition, the mRNA level of peroxisome proliferator-activated receptor (PPAR) α, an upstream factor of FAT, was increased in the livers of 3MC-treated mice. Taking together, AHR activation induces hepatic microvesicular steatosis by increasing the expression level of FAT.

Perfusion measurement of the whole upper abdomen of patients with and without liver diseases: Initial experience with 320-detector row CT

OBJECTIVES To report initial experience of upper abdominal perfusion measurement with 320-detector row CT (CTP) for assessment of liver diseases and therapeutic effects. MATERIALS AND METHODS Thirty-eight patients who were suspected of having a liver disease underwent CTP. There were two patients with liver metastases, two with hemangiomas, and four with cirrhosis (disease group). CTP was repeated for four patients with cirrhosis or hepatocellular carcinoma (HCC) after therapy. Hepatic arterial and portal perfusion (HAP and HPP) and arterial perfusion fraction (APF), and arterial perfusion (AP) of pancreas, spleen, stomach, and intra-portal HCC were calculated. For disease-free patients (normal group), the values were compared among liver segments and among pancreatic and gastric parts. The values were compared between groups and before and after therapy. RESULTS No significant differences were found in the normal group except between APFs for liver segments 3 and 5, and fundus and antrum. Mean HAP and APF for the disease group were significantly higher than for the normal group. APF increased after partial splenic embolization or creation of a transjugular intrahepatic portosystemic shunt. HPP increased and AP of intra-portal HCC decreased after successful radiotherapy. CONCLUSIONS 320-Detector row CT makes it possible to conduct perfusion measurements of the whole upper abdomen. Our preliminary results suggested that estimated perfusion values have the potential to be used for evaluation of hepatic diseases and therapeutic effects.

Significance of Serum Matrix Metalloproteinases and Their Inhibitors on the Antifibrogenetic Effect of Interferon-Alfa in Chronic Hepatitis C Patients

Objective and Methods: The imbalance between matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) is considered to be an important determination of deposition and breakdown of the extracellular matrix. To investigate the antifibrogenetic effect of interferon-α (IFN-α) treatment on factors regulating hepatic fibrosis, serum MMP-1, MMP-2, TIMP-1 and TIMP-2 levels were measured by the one-step sandwich enzyme immunoassay in 27 patients with chronic hepatitis C and compared with the histological status of the patients before and at the end of treatment. Results: After 6 months of IFN-α treatment, the histological status of liver fibrosis showed improvement in 9 patients (IF group) and no change or a worsening in 18 patients (NIF group). Compared with pretreatment levels, in the IF group, IFN treatment caused a significant increase in the MMP-1/TIMP-1 ratio. In the NIF group, however, the MMP-1/TIMP-1 ratio tended towards a decrease; moreover, there was not only a significant increase in TIMP-2 levels but also a tendency towards an increase in TIMP-1 levels. Conclusion: These results suggested that an elevated MMP-1/TIMP-1 ratio may ameliorate liver fibrosis by interferon in cases of chronic hepatitis C, whereas elevated levels of TIMP-1 and TIMP-2 might impede improvement.

Histological grading and staging in chronic hepatitis: Its practical correlation

Although the histological features of various causes of chronic liver disease have been well described, usually the inflammatory activity of the disease is important after the cause has been established. Some patients have co‐infection or concomitant liver disease and on occasion it is difficult to decide the treatment. In order to clarify the histological differences, we investigated the inflammatory activity among autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC), chronic hepatitis C (CHC) and chronic hepatitis B (CHB) in a standardized way using the modified histological activity index (HAI). According to the modified HAI, inflammatory activity is divided into four categories; categories A/D explains portal/periportal inflammation and categories B/C explains lobular activity. The inflammatory score of AIH tended to be greater in all categories from the early stage of fibrosis, whereas scores of PBC were lower, except for portal inflammation. Chronic hepatitis C patients had portal or periportal inflammation, and their inflammatory scores were linked to the development of fibrosis. Chronic hepatitis B patients tended to have severe lobular injury, but did not have a relationship between the inflammatory score and their stage. To know the distribution of inflammation using the modified HAI scoring system may be helpful and convenient in evaluating patients with chronic inflammatory liver disease.

Sequence Heterogeneity in NS5A of Hepatitis C Virus Genotypes 2a and 2b and Clinical Outcome of Pegylated-Interferon/Ribavirin Therapy

Pegylated-interferon plus ribavirin (PEG-IFN/RBV) therapy is a current standard treatment for chronic hepatitis C. We previously reported that the viral sequence heterogeneity of part of NS5A, referred to as the IFN/RBV resistance-determining region (IRRDR), and a mutation at position 70 of the core protein of hepatitis C virus genotype 1b (HCV-1b) are significantly correlated with the outcome of PEG-IFN/RBV treatment. Here, we aimed to investigate the impact of viral genetic variations within the NS5A and core regions of other genotypes, HCV-2a and HCV-2b, on PEG-IFN/RBV treatment outcome. Pretreatment sequences of NS5A and core regions were analyzed in 112 patients infected with HCV-2a or HCV-2b, who were treated with PEG-IFN/RBV for 24 weeks and followed up for another 24 weeks. The results demonstrated that HCV-2a isolates with 4 or more mutations in IRRDR (IRRDR[2a]≥4) was significantly associated with rapid virological response at week 4 (RVR) and sustained virological response (SVR). Also, another region of NS5A that corresponds to part of the IFN sensitivity-determining region (ISDR) plus its carboxy-flanking region, which we referred to as ISDR/+C[2a], was significantly associated with SVR in patients infected with HCV-2a. Multivariate analysis revealed that IRRDR[2a]≥4 was the only independent predictive factor for SVR. As for HCV-2b infection, an N-terminal half of IRRDR having two or more mutations (IRRDR[2b]/N≥2) was significantly associated with RVR, but not with SVR. No significant correlation was observed between core protein polymorphism and PEG-IFN/RBV treatment outcome in HCV-2a or HCV-2b infection. Conclusion: The present results suggest that sequence heterogeneity of NS5A of HCV-2a (IRRDR[2a]≥4 and ISDR/+C[2a]), and that of HCV-2b (IRRDR[2b]/N≥2) to a lesser extent, is involved in determining the viral sensitivity to PEG-IFN/RBV therapy.