OBJECTIVE:Non-alcoholic fatty liver disease (NAFLD) is a common debilitating condition in many industrialized countries that increases the risk of cardiovascular disease. The aim of this study was to derive a simple and accurate screening tool for the prediction of NAFLD in the Japanese population.METHODS:A total of 945 participants, 279 men and 666 women living in Hokkaido, Japan, were enrolled among residents who attended a health check-up program from 2010 to 2014. Participants with an alcohol consumption > 20 g/day and/or a chronic liver disease, such as chronic hepatitis B, chronic hepatitis C or autoimmune hepatitis, were excluded from this study. Clinical and laboratory data were examined to identify predictive markers of NAFLD.RESULTS:A new predictive index for NAFLD, the NAFLD index, was constructed for men and for women. The NAFLD index for men = -15.5693+0.3264 [BMI] +0.0134 [triglycerides (mg/dl)], and for women = -31.4686+0.3683 [BMI] +2.5699 [albumin (g/dl)] +4.6740[ALT/AST] -0.0379 [HDL cholesterol (mg/dl)]. The AUROC of the NAFLD index for men and for women was 0.87(95% CI 0.88-1.60) and 0.90 (95% CI 0.66-1.02), respectively. The cut-off point of -5.28 for men predicted NAFLD with an accuracy of 82.8%. For women, the cut-off point of -7.65 predicted NAFLD with an accuracy of 87.7%.CONCLUSION:A new index for the non-invasive prediction of NAFLD, the NAFLD index, was constructed using available clinical and laboratory data. This index is a simple screening tool to predict the presence of NAFLD.

AIM:To evaluate the changes of shear-wave velocity (Vs) by acoustic radiation force impulse after treatment in chronic hepatitis C.METHODS:Eighty-seven patients with chronic hepatitis C were consecutively treated with combinations of interferon (IFN) plus ribavirin (RBV). Vs value (m/s) was measured with acoustic radiation force impulse before treatment, at end of treatment (EOT), 1 year after EOT, and 2 years after EOT.RESULTS:In patients with a sustained virological response (SVR) (n = 41), Vs significantly decreased at EOT [1.19 (1.07-1.37), P = 0.0004], 1 year after EOT [1.10 (1.00-1.22), P = 0.0001], and 2 years after EOT [1.05 (0.95-1.16), P < 0.0001] compared with baseline [1.27 (1.11-1.49)]. In patients with a relapse (n = 26), Vs did not significantly decrease at EOT [1.23 (1.12-1.55)], 1 year after EOT [1.20 (1.12-1.80)], and 2 years after EOT [1.41 (1.08-2.01)] compared with baseline [1.39 (1.15-1.57)]. In patients with a nonvirological response (n = 20), Vs did not significantly decrease at EOT [1.64 (1.43-2.06)], 1 year after EOT [1.66 (1.30-1.95)], and 2 years after EOT [1.61 (1.36-2.37)] compared with baseline [1.80 (1.54-2.01)]. Among genotype 1 patients, baseline Vs was significantly lower in SVR patients [1.28 (1.04-1.40)] than in non-SVR patients [1.56 (1.20-1.83)] (P = 0.0142).CONCLUSION:Reduction of Vs values was shown in SVR patients after IFN-plus-RBV therapy by acoustic radiation force impulse.

The aims of this study were to elucidate the kinetics of Epstein-Barr virus (EBV) DNA load in serially collected peripheral blood mononuclear cells of patients with primary EBV infection, and to determine the correlated host factors. Blood samples were collected from 24 patients with primary EBV infection. EBV DNA copy numbers were measured using real-time polymerase chain reaction. Based on the kinetics of EBV DNA load, the 24 patients were divided into two groups: rapid regression and slow regression. Eighteen of the 24 patients (75%) were included in the slow regression and 6 (25%) in the rapid regression group. No statistically significant differences were observed between the two groups in clinical features and laboratory findings. However, acute phase (3 to 10 days after the onset of the illness) serum samples from six children in the slow regression and four in the rapid regression group revealed significantly higher serum interleukin (IL)-1β (P= 0.018), IL-12 (P= 0.009), tumor necrosis factor-α (P= 0.019), interferon-inducible protein 10, and monokine induced by interferon γ concentrations in the rapid regression than the slow regression group. On the other hand, sera from six children in the slow regression and four in the rapid regression group in the convalescent phase (14 to 21 days after the onset of the illness) showed no statistically significant differences between the two groups in these biomarker concentrations. Based on this, it was concluded that the kinetics of EBV DNA load can be divided to two different patterns after primary EBV infection, and immune response might be associated with viral clearance.

Liver stiffness (LS) has been reported to correlate with fibrosis stage (F). The correlation between LS and fibrosis stage and the reduction of LS by antiviral therapy were examined in patients with hepatitis B infection.

AIM:To construct and evaluate a new non-invasive fibrosis index for assessment of the stage of liver fibrosis.METHODS:A new fibrosis index (Fibro-Stiffness index) was developed in 165 of 285 patients with chronic hepatitis C, and was validated in the other 120 patients where liver biopsy was performed. Its usefulness was compared with liver stiffness (LS) measured by FibroScan, the aminotransferase-to-platelet ratio index, the Forns index and the FibroIndex.RESULTS:The Fibro-Stiffness index consists of LS, platelet count and prothrombin time. The values of the Fibro-Stiffness index differed significantly between neighboring fibrosis stages except F0-F1. The area under the receiver operating characteristics curves of the Fibro-Stiffness index for prediction of F ≥ 2 (0.90), F ≥ 3 (0.90) and F = 4 (0.92) in the estimation group and those for F ≥ 3 (0.93) and F = 4 (0.97) in the validation group were the highest among the 5 methods examined. The accuracy of the Fibro-Stiffness index had highest values for F ≥ 2, F ≥ 3 and F = 4 in both the estimation and validation groups. The diagnostic performance for F = 4 was improved by a combination of the Fibro-Stiffness index with serum hyaluronic acid level.CONCLUSION:The Fibro-Stiffness index was constructed and validated. It showed superior diagnostic performance to other indices for F ≥ 2, 3 and 4.

A variant specific direct loop-mediated isothermal amplification (LAMP) method was developed to detect human herpesvirus-6 (HHV-6) variants in serum samples. Specific primers were designed against HHV-6 U86 gene. Initial validation analysis confirmed high specificity and sensitivity of the method. This method was shown to be highly reliable for monitoring active HHV-6 infection in hematopoietic stem cell transplant recipients.

The etiology of Behcet’s disease (BD) has been linked to viral infection, autoimmune disease, streptococcal-related antigens, specific major histocompatibility complex alleles, and hazardous chemicals. Because Herpes simplex virus (HSV) can cause stomatitis, many investigators have searched for an association between HSV and BD. Eglin et al. demonstrated HSV-1 gene fragments in the peripheral blood mononuclear cells of BD patients using in situ DNA-RNA hybridization. Moreover, serum anti-HSV-1 antibodies were found in a higher percentage of BD patients than in controls, and other groups have reported circulating immune complexes to HSV-1 antigen. In addition to the human studies, 30% of ICR mice inoculated with HSV were reported to exhibit Behcet’s disease-like symptoms, including genital ulcers and skin and eye lesions. Viral infection alone, however, cannot sufficiently explain BD pathogenesis. BD has a strong genetic component with familiar aggregation. Among the various genetic markers, a correlation with human leukocyte antigen (HLA) class 1 antigens B5 and the subclass B51 is the most commonly reported, and linkage of the HLA-B locus during BD has also been shown. Here, we describe a patient with the HLA-B51 haplotype with Behcet’s disease-like symptoms concurrent with HSV-1 reactivation.

AIM:Liver stiffness (LS) measured by transient elastography (TE) has been reported to correlate with liver fibrosis, which is usually semiquantitatively assessed. In the present study, the fibrosis area was measured by image analysis software in liver biopsy specimens and its correlation with LS was assessed. METHODS:LS was measured by TE in all 165 patients with chronic hepatitis C virus (HCV) infection who underwent liver biopsy consecutively in Fujita Health University Hospital from July 2004 to September 2007. RESULTS:Fibrosis area was significantly correlated with fibrosis stage as assessed by the Metavir score (rho = 0.733, P < 0.0001). The optimal cut-off value of fibrosis area was 1.6% for F > or = 2, 3.1% for F > or = 3, and 3.8-6.4% for F4. LS was significantly correlated with fibrosis stage (rho = 0.734, P < 0.0001). The optimal cut-off value of LS was 7.1 kPa for F > or = 2, 9.6 kPa for F > or = 3 and 11.6-16.9 kPa for F4. Multiple linear regression analysis selected fibrosis area (P = 0.0002), alanine aminotransferase (ALT) (P = 0.0237), gamma-glutamyltransferase (gamma-GTP) (P = 0.0114), prothrombin time (P = 0.0114) and hyaluronic acid (P < 0.0001) as factors correlating with LS. CONCLUSION:The correlation between LS and liver fibrosis was confirmed by the objective measurement of fibrosis area. ALT was significantly correlated with LS, suggesting that inflammatory activity also affects LS values. Despite some limitation, LS measurement is a useful method for the diagnosis of liver fibrosis.

Although it has been demonstrated that human herpesvirus 6 (HHV-6) reactivation generally occurs approximately 2–3 weeks after transplantation in the hematopoietic stem cell transplantation (HSCT) recipients, the mechanism of viral reactivation remains unclear. To explore the relationship between HHV-6 reactivation and plasma cytokine levels, 24 HSCT recipients underwent measurements of plasma proinflammatory cytokine levels (IL-6, TNF-α, IL-1β, and IFN-γ), viral isolation, and serological assays. Of these patients, 14 developed an HHV-6 reactivation, and 9 developed HHV-6 viremia approximately 2–3 weeks after transplantation. IL-6 levels were significantly higher in the recipients with an HHV-6 reactivation than in the subjects without an HHV-6 reactivation at 1 week, 2 weeks, and 4 weeks after transplantation. In addition, the level of TNF-α was significantly higher in recipients with an HHV-6 reactivation than in those without an HHV-6 reactivation at 2 weeks post-transplantation. Low levels of IL-1β and IFN-γ were detected in a small number of the plasma samples, although there were no significant differences between the two groups in the levels of these cytokines. These results imply that proinflammatory cytokines, in particular IL-6 and TNF-α, play a role in the pathogenesis of HHV-6 reactivation after HSCT.

This study compares herpes simplex virus (HSV) type‐specific loop‐mediated isothermal amplification (LAMP) with virus isolation and real‐time PCR. Genital tract specimens were obtained from 25 patients with genital lesions; two swab samples were collected from the vulva and cervix of each patient, for a total of 50 specimens. After culturing, 10 of 50 (20%) samples were positive for HSV‐1 and 12 of 50 (24%) samples were positive for HSV‐2. None of the patients excreted both HSV‐1 and HSV‐2 virus. An original HSV type‐specific LAMP assay (30 min reaction) was compared with virus isolation and HSV type‐specific real‐time PCR. Viral DNA was detected by LAMP in 9 of 10 HSV‐1 isolated samples and 11 of 12 HSV‐2 isolated samples. No viral DNA was detected in samples without virus isolation. Thus, if virus isolation was used as the standard method, the LAMP protocol was highly sensitive and specific. In comparing LAMP to real‐time PCR, viral DNA was detected by the LAMP method in 9 of 12 HSV‐1 DNA positive samples and 11 of 18 HSV‐2 DNA positive samples. If real‐time PCR was used as the standard method, then, sensitivity of the LAMP method (in particular, for HSV‐2) was low. Taking this into consideration, the LAMP reaction was extended to 60 min. This led to an increase in sensitivity, resulting in an additional one and three samples testing positive for HSV‐1 LAMP and HSV‐2 LAMP, respectively, compared to the original LAMP protocol. Therefore, the sensitivity of the LAMP method increased to about 80%. J. Med. Virol. 75:583–587, 2005. © 2005 Wiley‐Liss, Inc.