Physical deconditioning is the common denominator in both obese and overweight subjects with nonalcoholic steatohepatitis.

BACKGROUND: Obesity and decreased physical activity mirror increasing prevalence of nonalcoholic fatty liver disease (NAFLD). AIM: We aimed to investigate associations between aerobic fitness, anthropometrics and disease parameters in patients with nonalcoholic steatohepatitis (NASH). We hypothesised that NASH subjects have lower aerobic power and capacity than untrained, sedentary, normal subjects. METHODS: Forty subjects (60% obese, 40% overweight) with biopsy-confirmed NASH and NAFLD activity score (NAS) ≥4 were enrolled in a clinical trial where anthropometrics, laboratories, liver fat content by MRI, activity, and aerobic fitness by cycle ergometry data were obtained. RESULTS: NASH subjects were significantly deconditioned compared to 148 untrained, sedentary, healthy subjects from our laboratory in aerobic power (VO2peak) (NASH 16.8 ± 6.6 vs control 28.4 ± 10.6 mL/kg/min, P < 0.0001) and capacity (VO2 at lactate threshold [LT]) (NASH 8.3 ± 2.5 vs control 14.1 ± 5.9 mL/kg/min, P < 0.0001). NASH subjects' fitness was comparable to the "least fit" tertile of controls: VO2peak [NASH 16.8 ± 6.6 vs "least fit" 17.3 ± 3.3, P = 0.64]) and VO2 at LT (NASH 8.3 ± 2.5 vs "least fit" 9.3 ± 2.1, P = 0.31). Fitness was similar in obese compared to overweight subjects (adjusted for gender) and was not correlated with visceral adiposity or NAS. Engaging in dedicated cardiovascular activity correlated with higher VO2peak and VO2peak at LT. CONCLUSIONS: Aerobic deconditioning was universally present in NASH subjects. NASH subjects' fitness was similar to our laboratory's "least fit" untrained, sedentary control subjects. Further research investigating NASH patients' ability to improve low baseline aerobic fitness is warranted.

Does hepatocellular carcinoma in non-alcoholic steatohepatitis exist in cirrhotic and non-cirrhotic patients?

Non-alcoholic steatohepatitis (NASH) has been associated with hepatocellular carcinoma (HCC) often arising in histologically advanced disease when steatohepatitis is not active (cryptogenic cirrhosis). Our objective was to characterize patients with HCC and active, histologically defined steatohepatitis. Among 394 patients with HCC detected by ultrasound imaging over 8 years and staged by the Barcelona Clinic Liver Cancer (BCLC) criteria, we identified 7 cases (1.7 percent) with HCC occurring in the setting of active biopsy-proven NASH. All were negative for other liver diseases such as hepatitis C, hepatitis B, autoimmune hepatitis, Wilson disease, and hemochromatosis. The patients (4 males and 3 females, age 63 ± 13 years) were either overweight (4) or obese (3); 57 percent were diabetic and 28.5 percent had dyslipidemia. Cirrhosis was present in 6 of 7 patients, but 1 patient had well-differentiated HCC in the setting of NASH without cirrhosis (fibrosis stage 1) based on repeated liver biopsies, the absence of portal hypertension by clinical and radiographic evaluations and by direct surgical inspection. Among the cirrhotic patients, 71.4 percent were clinically staged as Child A and 14.2 percent as Child B. Tumor size ranged from 1.0 to 5.2 cm and 5 of 7 patients were classified as early stage; 46 percent of all nodules were hyper-echoic and 57 percent were <3 cm. HCC was well differentiated in 1/6 and moderately differentiated in 5/6. Alpha-fetoprotein was <100 ng/mL in all patients. HCC in patients with active steatohepatitis is often multifocal, may precede clinically advanced disease and occurs without diagnostic levels of alpha-fetoprotein. Importantly, HCC may occur in NASH in the absence of cirrhosis. More aggressive screening of NASH patients may be warranted.

Does hepatocellular carcinoma in non-alcoholic steatohepatitis exist in cirrhotic and non-cirrhotic patients?

Non-alcoholic steatohepatitis (NASH) has been associated with hepatocellular carcinoma (HCC) often arising in histologically advanced disease when steatohepatitis is not active (cryptogenic cirrhosis). Our objective was to characterize patients with HCC and active, histologically defined steatohepatitis. Among 394 patients with HCC detected by ultrasound imaging over 8 years and staged by the Barcelona Clinic Liver Cancer (BCLC) criteria, we identified 7 cases (1.7%) with HCC occurring in the setting of active biopsy-proven NASH. All were negative for other liver diseases such as hepatitis C, hepatitis B, autoimmune hepatitis, Wilson disease, and hemochromatosis. The patients (4 males and 3 females, age 63 +/- 13 years) were either overweight (4) or obese (3); 57% were diabetic and 28.5% had dyslipidemia. Cirrhosis was present in 6 of 7 patients, but 1 patient had well-differentiated HCC in the setting of NASH without cirrhosis (fibrosis stage 1) based on repeated liver biopsies, the absence of portal hypertension by clinical and radiographic evaluations and by direct surgical inspection. Among the cirrhotic patients, 71.4% were clinically staged as Child A and 14.2% as Child B. Tumor size ranged from 1.0 to 5.2 cm and 5 of 7 patients were classified as early stage; 46% of all nodules were hyper-echoic and 57% were <3 cm. HCC was well differentiated in 1/6 and moderately differentiated in 5/6. Alpha-fetoprotein was <100 ng/mL in all patients. HCC in patients with active steatohepatitis is often multifocal, may precede clinically advanced disease and occurs without diagnostic levels of alpha-fetoprotein. Importantly, HCC may occur in NASH in the absence of cirrhosis. More aggressive screening of NASH patients may be warranted.

Enbucrilate for gastric varices: extended experience in 92 patients.

AIM: We assessed N-2-butyl-cyanoacrylate (enbucrilate) in 92 patients with gastric variceal bleeding under an FDA-approved investigation. These results extend our prior report of the first 44 patients. METHOD: Injection was performed with enbucrilate and ethiodol (1:1). Eighty patients had portal hypertension and 12 had splenic vein thrombosis. RESULTS: In the portal hypertensive group, re-bleeding from gastric varices was seen in 4 of 80 (5%) from 0 to 72 h, 5 of 76 (6.5%) from > 72 h to 3 months and 9 of 51 (17%) from > 3 months to 1 year. Re-bleeding and survival were significantly related to the Child-Pugh class. In the splenic vein thrombosis group (n = 12), there was early rebleeding in 2 (17%) patients from 0 to 72 h, 1 (8%) from > 72 h to 3 months and none in the chronic phase (> 3 months to 1 year) although 1-year survival in this group was only 6 (50%) due to the underlying malignancy in most. Serious embolization was suspected in 2 patients (2%). CONCLUSION: Enbucrilate offers an important intervention in gastric variceal bleeding which should be further studied in the US. A randomized trial is warranted to compare this intervention to radiological therapy.

A pilot study of a thiazolidinedione, troglitazone, in nonalcoholic steatohepatitis.

OBJECTIVES: Troglitazone is a thiazolidinedione and peroxisome proliferator-activated receptor gamma (PPARgamma) ligand used to treat diabetes mellitus type II. Because hyperinsulinemia may be a factor in nonalcoholic steatohepatitis (NASH), we postulated that troglitazone could have beneficial effects in this disorder. Our study was initiated before reports of idiosyncratic hepatitis induced by this agent and was completed before its recent withdrawal from the market. METHODS: We studied 10 female patients (age 44 +/- 16) with histological NASH. All but two were obese (mean body mass index, BMI = 38 +/- 6). One had type 2 diabetes, and three had well-compensated cirrhosis with NASH. Troglitazone was given at a dose of 400 mg/day for < or = 6 months. Responders (defined as normal ALT at the end of treatment) were rebiopsied. Paired specimens were compared in blinded fashion. Mitochondria were quantitated using ultrathin electron microscopy. RESULTS: Seven of ten patients responded with normal ALT at the end of treatment. One of three nonresponders initially normalized ALT but returned to pretreatment level at 3 months. In this patient, therapy was stopped, and the ALT has remained at the baseline level with no other clinical or laboratory findings. In the responders, ALT fell from 87 +/- 38 before to 39 +/- 9 at the end of treatment (p = 0.01), and AST decreased from 77 +/- 23 to 30 +/- 8 (p = 0.002). Biopsy comparisons before and after therapy showed persistent steatohepatitis in all cases, although four of seven showed a one-point improvement in the necroinflammatory grade. Electron microscopy revealed elongation of the mitochondria after therapy. CONCLUSIONS: Normal ALT was seen in 70% of NASH patients at the end of treatment, but this biochemical response was associated with only mild histological improvement, and all follow-up biopsies had evidence of NASH. Normalization of the liver enzymes in patients with NASH who are treated with thiazolidinediones should be viewed with reservation. Follow-up biopsy is essential to evaluate the efficacy of these agents, which, at the histological level, appears to be relatively modest.

Nonalcoholic steatohepatitis and cryptogenic cirrhosis within kindreds.

PURPOSE: Familial forms of cryptogenic cirrhosis have been described. We have cared for families in which several members were afflicted with cryptogenic cirrhosis as well as the more recently recognized entity of nonalcoholic steatohepatitis. To examine the familial patterns of these disorders, we reviewed patients with nonalcoholic steatohepatitis, with and without cirrhosis, or cryptogenic cirrhosis to assess how frequently their relatives were afflicted with these disorders. SUBJECTS AND METHODS: Eighteen members of eight kindreds containing 2 or more afflicted members were studied. Diagnoses were based on histology in all but 3 patients (2 elderly women with liver atrophy and severe cirrhotic ascites diagnosed clinically with cryptogenic cirrhosis and 1 adult man with abnormal serum aminotransferase levels and hepatomegaly that was diagnosed as fatty liver by ultrasound). Other forms of liver disease were excluded by extensive serologic testing. RESULTS: There were 8 index patients (1 man, 7 women; 2 with cryptogenic cirrhosis, 4 with nonalcoholic steatohepatitis with cirrhosis, and 2 with nonalcoholic steatohepatitis without cirrhosis) and 10 relatives (4 men, 6 women; 2 with cryptogenic cirrhosis and 8 with nonalcoholic steatohepatitis). Nonalcoholic steatohepatitis and nonalcoholic steatohepatitis with cirrhosis coexisted within four kindreds, one of which also had an afflicted member with cryptogenic cirrhosis. Nonalcoholic steatohepatitis and cryptogenic cirrhosis coexisted within three additional kindreds. Patterns of afflicted patients included mother-daughter, sister-sister, sister-brother, father-daughter, and male-female cousins. Fifteen (83%) of the 18 subjects were obese, and 11 (61%) had type 2 diabetes mellitus. CONCLUSIONS: The coexistence of nonalcoholic steatohepatitis with and without cirrhosis and cryptogenic cirrhosis within these kindreds suggests a common pathogenesis and possible genetic risk. These disorders were frequently but not invariably associated with female sex, obesity, and type 2 diabetes.

Mitochondrial abnormalities in non-alcoholic steatohepatitis.

BACKGROUND/AIMS: We assessed mitochondrial morphology by electron microscopy and the prevalence of a mitochondrial gene deletion in patients with non-alcoholic steatohepatitis (NASH), alcohol-related liver disease and non-fatty liver diseases. Respiratory chain function using a cytoplasmic hybrid (cybrid) assay was further studied in NASH patients and healthy controls. METHODS: Electron microscopy was performed in 26 specimens. Fifteen patients were studied by polymerase chain reaction to detect a 520-bp deletion product of the mitochondrial genome (dmtDNA). Cybrids were created by fusion of platelets with anaerobic neuroblastoma cells in six NASH patients and 12 controls. RESULTS: Eight of ten NASH, one of seven alcoholics and two of nine other patients had linear crystalline inclusions in megamitochondria (p<0.05). Three of five patients with alcohol-related liver disease had dmtDNA compared to one of five NASH patients and one of five non-steatohepatitis controls. Cybrid respiratory chain function in platelets was not different from that of controls. CONCLUSIONS: Respiratory chain dysfunction, if present in NASH, is not expressed in platelet-derived mitochondria. In contrast to alcohol-related liver disease with active drinking, NASH patients do not commonly express the 5-kb mitochondrial DNA gene deletion in liver tissue. As previously described in early alcohol-related liver disease, crystalline inclusions of unknown composition are seen in hepatic mitochondria in NASH. Their presence suggests either an adaptive process or mitochondrial injury.

Use of fibrin sealant to reduce bleeding after needle liver biopsy in an anticoagulated canine model: work in progress.

PURPOSE: Needle biopsy of the liver is a common diagnostic procedure. Although relatively safe, bleeding remains a potential complication and may occur more frequently in patients with coagulopathy. The purpose of this study was to evaluate the utility of a fibrin sealant in preventing bleeding after a 15-gauge needle biopsy of the liver in a canine model heparinized to simulate coagulopathy. MATERIALS AND METHODS: Fibrin sealant was delivered to biopsy tract sites in eight dogs anticoagulated with heparin (activated clotting time 387 seconds +/- 94) using the same sheath system that was employed to obtain the biopsy specimen. RESULTS: The results demonstrated complete hemostasis in the sealant-plugged tracts as compared to controls. Continuous bleeding was observed in none of the fibrin sealant-treated sites, compared with all of the control biopsy sites (P = .0078). CONCLUSION: These results demonstrate the high degree of efficacy of fibrin sealant delivered through a sheath system in plugging liver biopsy tracts and eliminating bleeding in a severely coagulopathic animal model. This indicates that fibrin sealant use in cutting needle biopsies can reduce major and minor complications associated with the procedure.

Subumbilical midline vascularity of the abdominal wall in portal hypertension observed at laparoscopy.

BACKGROUND: Large volume paracentesis is a common treatment of ascites. Injury to abdominal wall collateral veins during this procedure can lead to hemoperitoneum. Because of this concern, the midline below the umbilicus is often recommended as a site for paracentesis because of its presumed avascularity. METHODS: We examined the subumbilical peritoneal surface in 20 consecutive patients with liver disease undergoing diagnostic laparoscopy. This area was visualized by table tilting and confirmed by external finger compression. Nineteen patients had cirrhosis of various etiologies, and one had advanced fibrosis with evidence of portal hypertension. RESULTS: In these 20 patients, only 7 had avascular midlines below the umbilicus. Seven had small but definite veins running along the path of the urachus (median umbilical fold), and 6 had more prominent veins in this region. The internal landmarks in this region (median and medial folds) were frequently asymmetric with regard to the external appearance of the midline. CONCLUSION: The subumbilical midline in patients with portal hypertension is commonly vascular. When using this site for paracentesis, care should be exercised to identify venous structures with the narrow-gauge needle used to inject local anesthetic agent before placement of larger paracentesis needles.

Improved clinical outcomes with liver transplantation for hepatitis B-induced chronic liver failure using passive immunization.

OBJECTIVE: The goals were to summarize the results of liver transplantation for chronic hepatitis B disease (HBV) at the University of Virginia, correlate pretransplant viral markers with posttransplant hepatitis B immunoglobulin (HBIg) requirements, and identify the relation between viral protein in the liver and clinical reinfection. SUMMARY BACKGROUND DATA: Liver transplantation is an accepted treatment for end-stage liver disease from chronic HBV infection, although lifelong antiviral treatment (with HBIg or antiviral agents) is still necessary. Patients with evidence of active viral replication (detectable serum HBV-DNA or e antigen) at the time of transplant have a higher rate of allograft infection. Whether clinically stable patients receiving HBIg immunoprophylaxis have detectable viral products in their grafts remains unknown. METHODS: Forty-four transplants performed for HBV disease at the University of Virginia since March 1990 were reviewed. Most patients underwent aggressive passive immunoprophylaxis with HBIg to maintain serum HBV surface antibody (HBsAb) levels > or =500 IU/l for the first 6 months after the transplant, and > or =150 IU/l thereafter. Patients had viral markers quantified, underwent pharmacokinetic analysis of HBsAb levels to adjust dosing, and were biopsied routinely every 3 to 6 months and when indicated. RESULTS: Forty-four transplants were performed in 39 patients. Actual 1-year and 3-year graft survival was 95% and 81%, respectively, and 1-year and 3-year patient survival was 98% and 96%, respectively. After the adoption of indefinite HBIg prophylaxis, nine grafts became infected (all in recipients positive for HBV e antigen). Three occurred within 8 weeks of transplantation and were associated with a short HBsAb half-life and a wild-type virus. Six occurred >8 months after the transplant, and most of these were associated with viral mutation. Quantification of pretransplant markers was an overall poor predictor of HBIg requirements after the transplant. Immunohistochemistry demonstrated transient low-level expression of core protein in the liver in 23% of patients without serum or clinical evidence of recurrent hepatitis. CONCLUSIONS: An excellent outcome is possible after liver transplantation for chronic HBV disease using HBIg dosed by pharmacokinetic parameters. Currently, quantification of pretransplant serum markers of the HBV antigen load does not predict the intensity of posttransplant treatment required for good clinical outcomes. Because HBV is not eradicated from the patient, some form of indefinite antiviral therapy continues to be warranted.

Predictive value of intraoperative biopsies and liver function tests for preservation injury in orthotopic liver transplantation.

Eighty liver allografts were studied to determine the predictive value of intraoperative biopsies and postoperative liver function tests for the development of preservation injury (PI). Peak transaminase (aspartate transaminase [AST] and alanine transaminase [ALT]) and prothrombin time (PT) values achieved by each patient during postoperative days (POD) 1 through 7 were determined. PI in day 0 preperfusion biopsies (0Pre) (obtained immediately before implantation) and postperfusion biopsies (0Post) (obtained immediately after revascularization) was categorized by histological criteria as present or absent. PI in biopsies taken during POD 2 through 14 was histologically graded as either moderate-to-severe, mild, or absent. Of the 80 allografts, 8 were omitted because of primary nonfunction or postoperative complications. 0Pre and 0Post biopsies were available on 25 of 72 (35%) and 69 of 72 (96%) allografts, respectively. Only 2 (8%) of the 0Pre biopsies showed histological PI compared with 48 (70%) of the 0Post biopsies. Fifty-nine patients were biopsied between POD 2 through 14. Of these, 15, 28, and 16 patients developed moderate-to-severe, mild, or no evidence of PI, respectively. The presence of PI in the 0Post biopsy strongly correlated with the development of PI during POD 2 through 14 (P < .0005). Peak AST and ALT values in patients with moderate-to-severe PI on POD 2 through 14 were significantly elevated compared with those patients with either mild (P = .01 and .03) or no PI (P = .02 and .006). Because of extensive overlap in AST and ALT values between the three groups, however, transaminase values were not useful in predicting the presence or absence of PI in the individual case. The development of PI during POD 2 through 14 correlated with advanced donor age (P = .06) but was unassociated with 0Pre biopsy findings, cold ischemia time, or peak PT values. We conclude that the 0Post biopsy is a valuable tool for the prediction of subsequent PI in the early postoperative period. In contrast, 0Pre biopsy findings and peak AST and ALT values are not useful in the assessment of PI.

Accuracy and significance of pretransplant liver volume measured by magnetic resonance imaging.

Measurement of liver volume in patients with advanced liver disease is used to gauge the appropriate size of donor organs and may have prognostic value. We sought to determine the accuracy of magnetic resonance imaging (MRI) in measuring liver volume in 19 adult patients under consideration for liver transplantation. We also correlated the liver volume determination to the clinical severity of disease. Liver volume was measured at MRI by averaging the calculated volumes from coronal and transverse breath-hold T1-weighted images. These results were compared to the explanted liver volume measured by fluid displacement and the explant mass. The correlation coefficient for MRI liver volume and the explant displacement volume was 0.90. The mean liver volume for Child-Pugh class AB by MRI was 1986 +/- 568 mL (1002-2470 mL) compared to 1433 +/- 379 mL (540-1889 mL) in Child-Pugh class C patients (p = .02). We conclude that MRI offers an anatomically accurate means of determining adult liver volume in vivo. Lower mean liver volumes were observed in Child-Pugh class C patients. In addition to its ability to provide tumor screening and vascular assessment, MRI is able to provide accurate determinations of liver volume in patients undergoing liver transplant evaluations.

Intrahepatic biliary endoscopy in sclerosing cholangitis.

Sclerosing cholangitis is usually diagnosed by clinical findings coupled with radiographic imaging of the bile ducts by ERCP. Direct imaging of both the intra- and extrahepatic biliary tree provides an opportunity to further study this disorder and its potential complications such as biliary malignancy. However, endoscopic visualization of the intrahepatic bile ducts in sclerosing cholangitis is potentially limited by the size of available cholangioscopes and the presence of strictures. Below, we report our initial results using a 0.8-mm fiberoptic endoscope placed through a partially steerable 1.8-mm guide catheter. The system allows visualization of the intrahepatic biliary tree beyond areas of stricture in the more distal ducts.

Variation between centers in technique and guidelines for liver biopsy.

Hospitals have few published guidelines to follow when performing a liver biopsy. In 1992, we began revising our protocol in an effort to institute new guidelines for our teaching hospitals. To assess the current practice of liver biopsy, we sent 500 multilingual questionnaires to international academic centers, and 85 U.S. centers were surveyed by telephone. The survey assessed: 1) patient preparation, 2) technical aspects of the biopsy, and 3) post-procedural care. One hundred and eighty international centers and 85 U.S. centers responded (total = 265). We found a wide variation in the practice of this surgical procedure at both national and international centers. Many Asian centers (73%) performed a bleeding time prior to liver biopsy. This practice was seen in only 36% of the U.S. centers. Most centers preferred platelet counts of 50,000/mm3 and above. The aspiration needle was more widely used in the U.S. (74%) and in many international centers, but Asian centers (61%) preferred a cutting needle. Thirty percent of Japanese centers performed more than 50% of their liver biopsies laparoscopically. Few laparoscopies were done at other centers. While about a quarter of the reported U.S., European, Asian, and South American centers observed patients for 4-6 hours after a biopsy, the majority of centers observed patients 10 hours or more. In addition to the wide variation seen, this survey provided us with an academic view of the contemporary practice of liver biopsy and an insight into how to redefine our present guidelines.

Hepatitis C virus in body fluids after liver transplantation.

Recurrence of hepatitis C virus (HCV) after liver transplantation is common and is associated with high blood levels of HCV RNA. Higher blood levels of HCV may promote body fluid expression of the virus. We tested 152 body fluid specimens from 33 patients with chronic hepatitis C, 21 of whom had undergone prior liver transplantation. All patients had hepatitis C viremia, as determined by a reverse-transcription polymerase chain reaction (PCR) to the 5' noncoding region. The virus was quantitated in serum by the branched chain DNA assay (bDNA). Body fluids (33 sputum, 33 saliva, 33 urine, 32 tear, 9 vaginal, and 12 semen samples) were analyzed using PCR for HCV RNA. Serum HCV RNA by bDNA in the posttransplantation group was 255 +/- 229 x 10(5) compared with 50 +/- 56 x 10(5) eq/mL in the patients who did not undergo transplantation (P = .01). All urine, tear, and semen specimens were negative for HCV RNA. Five of 21 (24%) posttransplantation patients had detectable HCV RNA using PCR in oral secretions compared with 0 of 12 patients who did not undergo transplantation (not statistically significant). However, 5 of 11 patients with serum HCV RNA by bDNA results greater than 150 x 10(5) eq/mL had positive RNA in oral secretions compared to 0 of 22 patients with bDNA less than 150 x 10(5) eq/mL (P = .01). Posttransplantation patients were more likely to have bDNA levels exceeding 150 x 10(5) eq/mL (11 of 21 v 1 of 12, P = .03). Patients within the first year of transplantation were particularly prone to viral RNA levels exceeding 150 x 10(5) eq/mL (8 of 9 v 3 of 12, P = .01). We conclude that HCV RNA can occasionally be detected using PCR in oral secretions after liver transplantation. This is more likely during the first year posttransplantation when blood levels of HCV RNA often exceed 150 x 10(5) eq/mL by the branched chain DNA assay. Whether or not these observations represent an increased risk of transmission of infection during the early posttransplantation period is not certain.

Diagnostic laparoscopy of the liver and peritoneum using conscious sedation.

Laparoscopic study of the liver is now common in many U.S. medical centers. With proper preparation, the procedure can be performed safely in the endoscopy suite using conscious sedation. Multiple sites in the liver can be biopsied in a directed and rapid sequence. Postbiopsy bleeding can be detected early and stopped effectively should it occur. Restrictions after the procedure are similar to those for other outpatient surgical procedures along with the typical post-liver biopsy instructions. A successful laparoscopy requires careful nursing evaluation, monitoring, and assistance. In this article, the authors discuss the practical aspects of diagnostic laparoscopy performed in the endoscopy suite, the nurse's role during and after the procedure, and management of the instruments.