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Current Issue

July - August, 2013

Vol. 12 Issue 4

On the cover: Magnetic resonance imaging (MRI) showed an intrahepatic tumor involving segment VII. Rigth lobe hepatectomy and histopathology were performed.

EDITORIAL

HEPATOLOGY HIGHLIGHTS

  • Hepatology highlights Jorge A. López-Velázquez, Justo Fernández-Rivero, Norberto C. Chávez-Tapia, Misael Uribe, Nahum Méndez-Sánchez Page 356-358

CONCISE REVIEW

  • Occult hepatitis B in patients on hemodialysis: a review Andrea M.M. Fontenele, Natalino Salgado Filho, Adalgisa S.P. Ferreira Page 359-363

    The hepatitis B virus (HBV) is one of the most frequently transmitted agents in dialysis units. Occult hepatitis B is characterized by HBV infection without detectable surface antigen (HBsAg) in the patient's serum, a positive or negative HBV DNA marker result in the serum and a positive result in the liver tissue, which leads to the potential risk of transmission during renal replacement therapy service. There is variation in occult hepatitis B prevalence rates in this population across various studies that may be related to numerous factors. The presence of occult hepatitis B in individuals undergoing renal replacement therapy is important with regard to both the possibility of transmission and the consequences for the patient, especially the development of chronic liver disease and reactivation of the disease after renal transplantation.

ORIGINAL ARTICLES

Clinical Studies

Viral Hepatitis
  • Efficacy and safety of interferon-based therapy in the treatment of adult thalassemic patients with chronic hepatitis C: a 12 years audit Irene Vafiadis, Panagiotis Trilianos, Jiannis Vlachogiannakos, Markisia Karagiorga, Antonia Hatziliami, Ersi Voskaridou, Spiros D. Ladas Page 364-370

    Background. HCV infection and transfusional iron overload in Thalassemic patients may result in liver disease. HCV treatment in Thalassemia has raised safety concerns. Aim. Estimate effectiveness and tolerability of interferon-based therapy in HCV-infected Thalassemic patients. Material and methods. Over a 12-year period, consecutive patients with β Thalassemia major (TM) and chronic hepatitis C received treatment. Liver biopsy, HCV-RNA and genotyping were performed beforehand. Sustained virological response (SVR) was defined as negative HCV-RNA 6 months post-treatment. Forty eight patients (26 M-22 F, mean age 39.8) were enrolled. Twenty nine patients were treated with conventional interferon alpha (IFNa) for 48 weeks (group A). Nineteen patients (10 naïve-9 previously IFNa experienced) received pegylated interferon (PEGIFN) (group B). Results. HCV-1 was found in 44%, HCV-2 in 14%, HCV-3 in 23% and HCV-4 in 19%. Group A: ten patients (38.5%) achieved SVR, 2 (7.5%) relapsed and 17 (54%) were non responders. Group B: five (28%) achieved SVR, 8 (44%) relapsed and 6 (28%) never responded. High HCV-RNA levels, genotype 1 and advanced liver fibrosis were independently associated with no response. Four patients (3 treated with IFNα, 1 with PEG-IFN) had to discontinue treatment due to complications. Conclusions. The response rate of IFN monotherapy in multi-transfused, HCV-infected Thalassemic patients is not inferior to that in nonmulti- transfused patients. IFNa administration is well-tolerated and should be recommended as initial treatment schedule in this setting.

  • High frequencies of CD158b+ NK cells are associated with persistent hepatitis C virus infections Hui-Fan Ji, Juan Wang, Lei Yu, Jun-Qi Niu, Desalegn Admassu Ayana, Yan-Fang Jiang Page 371-379

    Background. During the early phases of a hepatitis C virus (HCV) infection, NK cell activation appears to be critical to the induction of adaptive immune responses that have the potential of clearing the infection. This study aimed to investigate the phenotype and function of NK cells in chronic HCV (CHC) patients, particularly patients who cleared HCV infections spontaneously (SR-HCV). Material and methods. Peripheral blood NK cells were compared between 36 CHC patients, 12 SR-HCV patients, and 14 healthy controls (HC). The phenotype and function of NK cells were characterized by flow cytometry. In addition, the potential associations between the frequency of NK cell subsets and ALT, AST and HCV viral loads were also analyzed. Results. Our data revealed that the population of CD3-CD56+ NK cells was significantly decreased in CHC and SR-HCV patients compared to levels in HC (P = 0.031, P = 0.014). Interestingly, we found that the levels of the CD158b inhibitory receptor were higher in CHC patients compared to levels observed in HC and SR-HCV subjects (P = 0.018, P = 0.036). In addition, the percentages of the activation receptors NKp30 and NKp46 were significantly decreased in CHC and SR-HCV patients compared to their expression levels in HC (P < 0.05). Moreover, the frequencies of inducible CD107a (but not IFN-γ-secreting) NK cells from both CHC and SR-HCV patients were significantly lower than frequencies observed in controls (P = 0.018, P = 0.027). Conclusion. Our data indicated that the higher frequency of inhibitory NK cells combined with fewer activated NK cells may be associated with HCV-related chronic inflammation involved in CHC pathogenesis.

  • Association of plasma visfatin with hepatic and systemic inflammation in nonalcoholic fatty liver disease Halil Genc, Teoman Dogru, Muammer Kara, Serkan Tapan, Cemal Nuri Ercin, Cengizhan Acikel, Yildirim Karslioglu, Sait Bagci Page 380-387

    Background. Visfatin is a proinflammatory and insulin-mimetic adipokine contributing to whole body glucose and lipid metabolism. Studies to date are conflicting regarding the relationship between visfatin and non-alcoholic fatty liver disease (NAFLD). The aim of the present study was to evaluate the relationship of circulating visfatin with NAFLD. Material and methods. The study included 114 NAFLD patients and 60 healthy non-diabetic controls. Plasma visfatin, adiponectin, tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) levels were measured by ELISA. High sensitive C-reactive protein (hsCRP) levels were measured by immunoturbidimetric fixed rate method. Insulin sensitivity determined by homeostasis model assessment (HOMA-IR) index. Results. TNF-α, IL-6 and hsCRP levels were higher and, Adiponectin levels were lower in NAFLD group when compared to healthy controls (p < 0.001, for all). However, no difference was found regarding to visfatin levels between two groups. Different histologic subgroups of NAFLD had a significantly higher TNF-α, IL-6 and hsCRP, and lower adiponectin levels than those with controls (p < 0.001, for all). On the other hand, no statistically significant difference was found regarding to visfatin levels among different histologic groups. Visfatin was found to be negatively correlated with TNF-α (r = -0.236, p = 0.011) in NAFLD group. However, no association was found between visfatin and histological findings. Conclusion. Our findings show that plasma visfatin levels are not altered in the early stages of NAFLD. However, it is inversely associated with TNF-α. These findings suggest a role for visfatin in protection against liver injury in this widespread disease.

Liver transplantation
  • Predicting the prognosis in acute liver failure: results from a retrospective pilot study using the LiMAx test Johan Friso Lock, Amir Nihad Kotobi, Maciej Malinowski, Antje Schulz, Maximilian Jara, Peter Neuhaus, Martin Stockmann Page 388-394

    Background. Acute liver failure (ALF) is a rare but potentially life-threatening condition and liver transplantation (LTX) remains frequently the only effective therapy. Nevertheless, some patients recover without LTX but the individual indication for or against LTX remains difficult. Aim. To evaluate maximal liver function capacity (LiMAx) for predicting the prognosis of ALF. Material and methods. Clinic data of 12 patients was retrospectively analyzed to compare the different liver function test results with the patients' clinical outcome. Patients were assessed by the LiMAx test, a non-invasive breath test determining cytochrome P450 1A2 capacity using intravenous 13C-methacetin. Statistical analysis compared patients with spontaneous recovery versus non-recovery (LTX or death). Results. Twelve patients (6 male, 6 female; 49 [11-72] years) with viral hepatitis (n = 2), toxic liver injury (n = 3), or cryptogenic liver failure (n = 7) were analyzed. Seven patients fully recovered from ALF and were discharged without LTX. Three patients died and two underwent LTX. The King's College Criteria (KCC) was fulfilled in only one out of five patients without recovery. The LiMAx was 19 ± 19 (16-62) for non-recovery vs. 94 ± 119 (39-378) μg/kg/h for recovery (P = 0.018). In contrast, all biochemical parameters [bilirubin (P = 0.106), creatinine (P = 0.343), AST (P = 0.53), ALT (P = 0.876) and INR (P = 0.876) were statistically indistinct. Also the Model for End-Stage Liver Disease (MELD) score did not show a difference [35 ± 4.3 (29-40) vs. 30 ± 11.5 (6-40); P = 0.27]. Conclusions. Maximal liver function capacity determined by LiMAx test is severely impaired in patients with ALF. The LiMAx test might be effective in predicting the individual prognosis and the need for LTX in ALF.

  • Sirolimus based immunosuppression is associated with need for early repeat therapeutic ERCP in liver transplant patients with anastomotic biliary stricture James H. Tabibian, Mohit Girotra, Hsin-Chieh Yeh, Dorry L. Segev, Murat T. Gulsen, Guldane Cengiz-Seval, Vikesh K. Singh, Andrew M. Cameron, Ahmet Gurakar Page 395-401

    Introduction. Sirolimus has inhibitory effects on epithelial healing and cholangiocyte regeneration. In liver transplantation (LT) patients, these effects may be greatest at the biliary anastomosis. We therefore investigated whether sirolimus use is associated with need for early or emergent repeat therapeutic endoscopic retrograde cholangiography (ERC) in LT patients with anastomotic biliary stricture (ABS). Material and methods. Medical records of patients who underwent LT from 1998-2009 at Johns Hopkins were reviewed and patients with ABS identified. Primary outcome was early repeat ERC, defined as need for unscheduled (i.e. unplanned) or emergent repeat therapeutic ERC. Univariate and multivariate logistic regression analyses (adjusting for age, sex, LT to ERC time, and stent number) were performed to assess association between sirolimus and early repeat ERC. Results. 45 patients developed ABS and underwent 156 ERCs total. Early (median 26 days) repeat ERC occurred in 14/56 (25%) and 6/100 (6%) ERCs performed with and without concomitant sirolimus-based immunosuppression, respectively (OR 1.22; 95% CI 1.02-1.45; p = 0.03). In multivariate analysis, sirolimus use was associated with early repeat ERC (OR 1.24; 95% CI 1.04-1.47; p = 0.015); this association remained significant when sirolimus dose was modeled as a continuous variable (OR 1.04 for each mg of sirolimus per day; 95% CI 1.02-1.08; p = 0.038). Conclusions. Sirolimus-based immunosuppression appears to be associated with a modest but significantly increased, dose-dependent risk of early repeat ERC in LT patients with ABS. Prospective studies are needed to further investigate these findings and determine if sirolimus use or dose should potentially be reconsidered once ABS is diagnosed.

Miscellaneous
  • Non-invasive assessment of liver fibrosis with transient elastography (FibroScan®): applying the cut-offs of M probe to XL probe Grace Lai-Hung Wong, Julien Vergniol, Peter Lo, Vincent Wai-Sun Wong, Juliette Foucher, Brigitte Le Bail, Paul Cheung-Lung Choi, Faiza Chermak, Kwong-Sak Leung, Wassil Merrouche, Henry Lik-Yuen Chan, Victor de Lédinghen Page 402-412

    Background and rationale for the study. Limited studies have aimed to define the cut-offs of XL probe (XL cut-offs) for different stages of liver fibrosis, whereas those of M probe (M cut-offs) may not be applicable to XL probe. We aimed to derive appropriate XL cut-offs in overweight patients. Patients with liver stiffness measurement (LSM) by both probes were recruited. XL cut-offs probe for corresponding M cut-offs were derived from an exploratory cohort, and subsequently validated in a subgroup patients also underwent liver biopsy. The diagnostic accuracy of XL cut-offs to diagnose advanced fibrosis was evaluated. Results. Total 517 patients (63% male, mean age 58) who had reliable LSM by both probes were included in the exploratory cohort. There was a strong correlation between the LSM by M probe (LSM-M) and LSM by XL probe (LSM-XL) (r² = 0.89, p < 0.001). A decision tree using LSM-XL was learnt to predict the 3 categories of LSM-M (< 6.0kPa, 6.0-11.9kPa and ≥ 12.0kPa), and XL cut-offs at 4.8kPa and 10.7kPa were identified. These cut-offs were subsequently validated in a cohort of 147 patients who underwent liver biopsy. The overall accuracy was 89% among 62 patients whose LSM-XL < 4.8kPa or ≥ 10.7kPa. These cut-offs would have avoided under-staging of fibrosis among patients with body mass index (BMI) > 25-30 kg/m2 but not > 30 kg/m2. Conclusions. XL cut-offs at 4.8kPa and 10.7kPa were the best estimates of 6.0kPa and 12.0kPa of M probe for patients with BMI > 25-30 kg/m2. Patients with BMI > 30 kg/m² might use M probe cut-offs for XL probe.

  • Serum cholesterol is a significant and independent mortality predictor in liver cirrhosis patients Martin Janicko, Eduard Veselíny, Dušan Leško, Peter Jarcuška Page 413-419

    Background and Aim. Accurate assessment of cirrhotic patient's prognosis is essential for decisions regarding the course of treatment. Therefore we aimed to confirm and quantify the predictive value of serum cholesterol and serum triglycerides in liver cirrhosis patients. Material and methods. We performed a retrospective observational cohort study on consecutive patients with liver cirrhosis (n = 191). Relevant clinical and laboratory variables were obtained from patients' charts and patients were followed for two months. Mortality was the main outcome. Results. Thirty-eight patients died in the follow-up period. Significant difference was observed in the level of total serum cholesterol between surviving and deceased patients (2.27 ± 1.02 mmol/L vs. 2.97 ± 1.00 mmol/L, P < 0.0001 respectively). Cholesterol was confirmed as a significant predictor of mortality in univariate logistic regression analysis, and independent predictor beside bilirubin, creatinine and MELD score in multivariate logistic regression analysis. Addition of serum cholesterol level to a prognostic model based on total bilirubin, creatinine and INR increased its accuracy by 4%. Adding cholesterol to the MELD score improved prediction accuracy by 3%. There was no significant difference in serum levels of triglycerides between surviving and deceased patients. Conclusion. Serum cholesterol is a routinely measured parameter, which has independent prognostic value in patients with liver cirrhosis.

  • Predicting portal hypertension and variceal bleeding using non-invasive measurements of metabolic variables Mohammed Eslam, Javier Ampuero, Maria Jover, Hesham Abd-Elhalim, Diego Rincon, Mohammed Shatat, Ines Camacho, Amal Kamal, Oreste Lo Iacono, Zainb Nasr, Lourdes Grande, Rafael Banares, Mahmoud A. Khattab, Manuel Romero-Gomez Page 420-430

    Background & aim. This study assessed the involvement of metabolic factors (anthropometric indices, insulin resistance (IR) and adipocytokines) in the prediction of portal hypertension, esophageal varices and risk of variceal bleeding in cirrhotic patients. Material and methods. Two prospective and retrospective cohorts of cirrhotic patients were selected (n = 357). The first prospective cohort (n = 280) enrolled consecutively in three centers, underwent upper gastrointestinal endoscopy, seeking evidence of esophageal varices. Clinical, anthropometric, liver function tests, ultrasonographic, and metabolic features were recorded at the time of endoscopy, patients were followed-up every 6 months until death, liver transplantation or variceal bleeding. The second retrospective cohort (n = 48 patients) had measurements of the hepatic venous pressure gradient (HVPG). Statistical analyses of the data were with the SPSS package. Results. The presence of esophageal varices was independently associated with lower platelet count, raised HOMA index and adiponectin levels. This relationship extended to subset analysis in patients with Child A cirrhosis. HOMA index and adiponectin levels significantly correlated with HVPG. Beside Child-Pugh class, variceal size and glucagonemia, HOMA index but not adiponectin and leptin plasma levels were associated with higher risk of variceal bleeding. Conclusion. In patients with cirrhosis, HOMA score correlates with HVPG and independently predict clinical outcomes. Three simple markers i.e. platelet count, IR assessed by HOMA-IR and adiponectin significantly predict the presence of esophageal varices in cirrhotic patients.

  • Acute phase proteins for the diagnosis of bacterial infection and prediction of mortality in acute complications of cirrhosis César Lazzarotto, Marcelo Fernando Ronsoni, Leonardo Fayad, Christiane Lourenço Nogueira, Maria Luiza Bazzo, Janaína Luz Narciso-Schiavon, Leonardo de Lucca Schiavon, Esther Buzaglo Dantas-Corrêa Page 431-439

    Introduction. Bacterial infection is a frequent complication in patients with decompensated liver cirrhosis and is related to high mortality rates during follow-up of these individuals. We sought to evaluate the diagnostic value of C-reactive protein (CRP) and procalcitonin (PCT) in diagnosing infection and to investigate the relationship between these biomarkers and mortality after hospital admission. Material and methods. Prospective study that included cirrhotic patients admitted to the hospital due to complications of the disease. The diagnostic accuracy of CRP and PCT for the diagnosis of infection was evaluated by estimating the sensitivity and specificity and by measuring the area under the receiver operating characteristics curve (AUROC). Results. A total of 64 patients and 81 hospitalizations were analyzed during the study. The mean age was 54.31 ± 11.87 years with male predominance (68.8%). Significantly higher median CRP and PCT levels were observed among infected patients (P < 0.001). The AUROC of CRP and PCT for the diagnosis of infection were 0.835 ± 0.052 and 0.860 ± 0.047, respectively (P = 0.273). CRP levels > 29.5 exhibited sensitivity of 82% and specificity of 81% for the diagnosis of bacterial infection. Similarly, PCT levels > 1.10 showed sensitivity of 67% and specificity of 90%. Significantly higher levels of CRP (P = 0.026) and PCT (P = 0.001) were observed among those who died within three months after admission. Conclusion. CRP and PCT were reliable markers of bacterial infection in subjects admitted due to complications of liver cirrhosis and higher levels of these tests are related to short-term mortality in those patients.

  • Non-invasive assessment of liver fibrosis using ARFI with pathological correlation, a prospective study Wan Wan Yap, Rathy Kirke, Eric M. Yoshida, David Owen, Alison C. Harris Page 440-447

    Introduction.ARFI is a new technique that uses acoustic push pulse to generate tissue displacement resulting in shear wave propagation, can be used to measure elasticity of tissue. We aim to assess feasibility of ARFI as a non-invasive method to measure liver fibrosis compared to histological fibrosis scores and to compare our results with the published pooled-meta-analysis cut off values. Material and methods. Prospective study to compare median velocities of ARFI shear wave measurements (Virtual Touch Imaging™ ACUSON S2000, Siemens, Mountain View CA) with Batts and Ludwig liver fibrosis scoring system F0-F4. Results. 70 patients (mean = 49 years) were included. Etiologies were chronic hepatitis C (n = 43), chronic hepatitis B (n = 7) and others (n = 20). Median ARFI values (m/sec) for fibrosis stages and inflammatory stages measured were F0: 1.52, 1.42; F1: 1.50, 1.37; F3: 2.36, 2.41 and F4: 2.61. Areas under the curve for grade 3 = 0.875, stage 3 = 0.867; grade 2 = 0.4, stage 2 = 0.3.Using the cut-off ARFI value of 1.34 m/s for F ≥ 2 suggested in the meta-analysis, we found sensitivity of detecting true F ≥ 2 is 68%, specificity 66%, PPV 74% and NPV 59%. For F ≥ 3 using the cut-off ARFI value of 1.55 m/s, we found sensitivity of 95%, specificity 86%, PPV 74% and NPV 98%. No stage 4 was compared due to insufficient cases. Conclusion. ARFI has strong correlation with higher fibrosis scores compared to lower. When compared to the pooled meta-analysis

Basic studies

  • Pegylated interferon-α2b and ribavirin decrease claudin-1 and E-cadherin expression in HepG2 and Huh-7.5 cells Erika P. Rendón-Huerta, Ana Torres-Martínez, Claudia Charles-Niño, Ana M. Rivas-Estilla, A. Paez, Teresa I. Fortoul, Luis F. Montaño Page 448-457

    Background. Hepatitis C virus (HCV) infection usually results in long-term viremia. Entry of HCV into the hepatocyte requires claudin-1, -6, -9 and occludin. The efficacy of Pegylated interferon-α (PEG-IFN) treatment against HCV infection increased when ribavirin (RBV) was added to the therapeutic scheme. Our aim was to investigate if PEG-IFN plus RBV regulate claudin expression. Material and methods. HepG2, Huh-7 and Huh-7.5 cells were treated with PEG-IFN-α2a or α2b and/or RBV at different times before obtaining the cytosolic, membrane and cytoskeletal fractions. Claudin-1, 3, 4, 6, and 9, E-cadherin and occludin expression was evaluated by Western blot analysis. Transepithelial electrical resistance (TER) was also determined. Results. Claudin-1, 3, 4, 6, E-cadherin and occludin are constitutively expressed mainly in HepG2 cell membrane. Claudin-1 and E-cadherin cell membrane expression diminished after exposure to PEGIFNα2b (50 ng) + RBV(50 μg); the maximal decrease was observed with 200 ng of PEG-IFNα2b + 200 μg of RBV. The effect was less intense with PEG-IFNα2a. The inhibition of claudin-1 and E-cadherin expression in Huh-7 and Huh-7.5 cells was only observed with 200 ng of PEG-IFNα2b + 200 μg of RBV. TER diminished marginally in the HCV containing hepatoma cells with 200 ng of PEG-IFNα2b + 200 μg of RBV. Claudin-1 mRNA expression level was not affected by the combined treatment. Conclusion. The increased therapeutic efficacy of the PEG-IFNα2b plus RBV treatment could be secondary to the inhibition of claudin-1 and E-cadherin cell membrane expression.

IMAGES IN HEPATOLOGY

  • Hepatocellular carcinoma arising in a telangiectatic hepatocellular adenoma María Fernanda González-Lara, Juan-Carlos Córdova-Ramón, Armando Gamboa-Domínguez, Juan Cosme-Labarthe, Diego Luis Carrillo-Pérez Page 458-460

    Telangiectatic hepatocellular adenoma is a rare, recently recognized subtype of benign liver tumor that may very rarely undergo transformation into hepatocellular carcinoma. We report an unusual case of a 75- year-old woman with no history of oral contraceptive use that underwent malignant transformation of a telangiectactic hepatocellular adenoma. No risk factors for adenoma development were identified in this otherwise healthy woman. Radiological characteristics, gross features and histopathology are herein described. In conclusion, telangiectatic hepatocellular adenoma can undergo malignant transformation. Further studies are needed to better clarify the factors associated with malignant progression.

CASE REPORTS

  • Pulmonary complications of treatment with pegylated interferon for hepatitis C infection-Two case reports Vinod S. Hegade, Ruchit Sood, Dinesh Saralaya, Sulleman Moreea Page 461-465

    Pegylated interferon (Peg-IFN) in combination with ribavirin is the standard of care in the treatment of chronic hepatitis C (HCV). Peg-IFN is known to have a number of side effects but severe respiratory complications are uncommon. We report two cases, one of Peg-IFN induced interstitial pneumonitis (IP) and the other of bronchiolitis obliterans organising pneumonia (BOOP) in patients with chronic hepatitis C infection. In general, respiratory complications of Peg-IFN are mild and resolve with withdrawal of Peg-IFN. However, as illustrated in our first case fatal interstitial pneumonitis can occur. We present a review of the available literature on Peg-IFN induced lung toxicity. In conclusion, pulmonary toxicity with Peg-IFN is rare but fatality can occur. We highlight the importance of maintaining a high index of suspicion for early diagnosis and prompt treatment, which includes withdrawal of Peg-IFN and consideration of corticosteroid treatment.

  • Unexpected discovery of massive liver echinococcosis. A clinical, morphological, and functional diagnosis L. Bonfrate, F. Giuliante, G. Palasciano, J.T. LaMont, P. Portincasa Page 466-473

    18 cm), clinical presentation, geographical area, and natural history of echinococcosis, we estimate that the initial infection should have occurred 9-20 yrs before. Presenting symptoms were those of typical mass effect with RUQ pain, pruritus, malaise, and recent weight loss. Abdominal ultrasound diagnosis of probable echinococcal cyst was subsequentely confirmed by positive serology and further detailed by radiological imaging. The cyst was massively occupying subdiaphragmatic liver segments and extending to the omentum and the stomach. The characteristics of the lesion were compatible with the WHO 2003 classification type CE2l, indicating a large active fertile cyst with daughter cysts. The cyst was successfully treated with medical therapy followed by surgery. The prevalence, diagnostic workup, management, and costs of echinococcosis are discussed in this case presentation.

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