98 These mechanisms may be additive or synergistic in

treating MHE. Probiotics may represent a safe, effective, long-term therapy for MHE and may be an alternative to lactulose. Clinical studies evaluating the role of LOLA in the treatment of MHE did not show its effectiveness; however, these studies were small and underpowered. A recent study that compared lactulose, a probiotic and LOLA with no treatment, however, showed that LOLA is as effective as lactulose MAPK inhibitor or a probiotic preparation in improving psychometric performance and HRQOL.67 Larger prospective studies are warranted to evaluate the role of LOLA before it can be recommended for the treatment of MHE. The role of antibiotics in

MHE has not been evaluated. Prospective studies with poorly absorbed antibiotics are required to evaluate their efficacy in improving MHE. 45 Lactulose is effective in reducing blood ammonia levels and improving psychometric performance in cirrhotic patients with MHE. (1b) The INASL Working Party recommends that all patients with cirrhosis be screened for the presence of MHE using a standard battery of psychometric tests, PHES, CFF or ICT, depending upon the availability of tests and BMN 673 their validation for local populations from different parts of the world (Fig. 1). Patients whose index psychometric or computerized test results do not indicate pathology should be screened every 6–12 months. Treatment for MHE may be initiated with lactulose; patients should receive 30–60 mL of lactulose in two or three divided doses so that

they pass two to three semi-soft stools per day. Although the appropriate duration of therapy for MHE is unsettled, at least three studies suggest that treatment may be advised for 3–6 months.3,67,95 “
“We read with interest the article by Hongthanakorn et al. published in a recent issue of HEPATOLOGY.1 The authors reported a very high incidence of virological breakthrough (VBT) in patients receiving five different nucleos(t)ide analogues (NUCs) Forskolin mw in clinical practice: 26% (39 patients). They reported that 7% of NUC-naïve patients receiving entecavir (ETV) experienced VBT, and that the cumulative probability of experiencing VBT at 3 years was 13%. The VBT rates reported by Hongthanakorn et al. are higher than described previously. In our population of 69 NUC-naïve chronic HBV patients treated in routine clinical practice with ETV, we found that 100% achieved undetectable HBV DNA after 96 weeks of treatment.2 We did not perform tests to evaluate genetic resistance, but we found no evidence of clinical resistance to treatment or VBT. Other studies in clinical practice have shown high efficacy of treatment with low rates of VBT, around 1%.3-5 In phase 3 randomized clinical trials, VBT rates with ETV treatment were 1.6%.6, 7 Hongthanakorn et al.

395; 95% CI, 0.180-0.896; P = 0.021). Age of onset below 18 was a significant risk factor (HR, 1.963; 95% CI, 1.21-3.20; P = 0.007) for the use of immunosuppressants in CD. Extent of disease was a significant factor associated with surgical resection (p = 0.012) in univariate analysis but not in multivariate analysis. Extensive disease in UC was a significant risk factor in multivariate

cox model (HR, 3.558; 95% CI, 1.32-9.58; P = 0.012) for primary use of immunosuppressants. Conclusion: In CD, colonic disease were associated with decreased risk while stricturing and penetrating behavior were associated with increased risk of surgical resection. In UC, extensive disease was associated with the need for immunosuppressants. Key Word(s): 1. inflammatory bowel disease; 2. bowel resection Presenting Author: GOVIND K MAKHARIA Additional Authors: GOVIND K MAKHARIA, Selleckchem YAP-TEAD Inhibitor 1 ABHISHEK AGNIHOTRI,

GSK1120212 mw SUDIPTO CHAUDHARY, UC GHOSHAL, MANISH K PATHAK, ASHA MISHRA, SIDDHARTHA DATTA GUPTA, RAJU SHARMA, RM PANDEY, VINEET AHUJA, SK SHARMA, BS RAMAKRISHNA Corresponding Author: GOVIND K MAKHARIA Affiliations: All India Institute of Medical Sciences, All India Institute of Medical Sciences, Christian Medical College, Sanjay Gandhi Postgraduate Institute of Medical Sc, All India Institute of Medical Sciences, All India Institute of Medical Sciences, All India Institute of Medical Sciences, All India Institute of Medical Sciences, All India Institute of Medical Sciences,All India Institute of Medical Thymidine kinase Sciences, All India Institute of Medical Sciences, SRM Institute of Medical Sciences Objective: Whether patients with abdominal tuberculosis

(both gastrointestinal and peritoneal) should be treated with six months or nine months is a debatable. There is also a lack of data on the efficacy of short course intermittent therapy in treatment of abdominal tuberculosis. We conducted a multicenter single blinded randomized controlled trial to assess the efficacy of 6 months and 9 months of anti-tuberculous therapy (ATT) in abdominal tuberculosis using Directly Observed Therapy Short Course (DOTS). Methods: Of 499 patients screened, 197 patients with abdominal tuberculosis (gastrointestinal-154, peritoneal-40, mixed-3) were randomized to receive 6-mo (Group A, n = 104) and 9-mo (Group B, n = 93) of ATT using DOTS strategy. All patients were evaluated for primary end point (complete clinical response, partial clinical response, no response, or death) and secondary end point (mucosal healing). Patients were followed up further for one year after completion of treatment to assess recurrence. Results: Both groups had similar baseline characteristics, clinical manifestations, site of disease, proportion of definitive or presumptive diagnosis of tuberculosis. Per protocol analysis showed no difference in complete clinical response (91.5% vs 90.8%, P = 0.882) between group A and group B.

The primary outcome was overall survival at 3 weeks. Secondary outcomes included transplant-free survival and rate of transplantation. RESULTS: A total of 173 patients received NAC (n = 81) or placebo (n = 92). Overall survival at 3 weeks was 70% for patients given NAC and 66% for patients given placebo (1-sided P = .283). Transplant-free survival was significantly better for NAC patients (40%) than for those given placebo (27%; 1-sided P = .043). The benefits of transplant-free survival were confined to the 114 patients with coma grades I-II who received NAC (52% compared

with 30% for placebo; 1-sided P = .010); transplant-free survival for the Selleckchem Silmitasertib 59 patients with coma grades III-IV was 9% in those given NAC and 22% in those given placebo

(1-sided P = .912). The transplantation rate was lower in the NAC group but was not significantly different between groups (32% vs 45%; P = .093). Intravenous NAC generally was well tolerated; only nausea and vomiting occurred significantly more frequently in the NAC group (14% vs 4%; P = .031). CONCLUSIONS: Intravenous NAC improves transplant-free survival in patients with early stage non-acetaminophen-related acute liver failure. Patients with advanced coma grades do not benefit from NAC and typically require emergency liver transplantation. Acute liver failure (ALF) is characterized by loss of liver function without pre-existing chronic liver disease. The grade of hepatic encephalophathy and the international normalized ratio (INR) have been identified as the main prognostic PLX4032 nmr indicators in this disease. The clinical course in severe ALF may be characterized by the rapid development of multiorgan failure, extended intensive care, and liver transplantation may be needed. The definition of ALF has not been clearly established. The clinical presentation of coagulopathy and encephalopathy suggests the diagnosis. However, ALF with less severe Bay 11-7085 liver injury can present with coagulopathy but without encephalopathy. The historical classification of ALF

into fulminant or hyperacute, acute and subacute liver failure depending on the duration of clinical symptoms has no prognostic relevance.1 Causes of ALF with unfavorable prognosis include Wilson’s disease, Budd-Chiari syndrome, and non-A hepatitis B or C. In the United States, approximately 2000 cases of ALF are recorded annually and cause 6% of liver-related deaths overall.2 The most common etiologies of ALF are drug-induced injury, viral hepatitis, and indeterminate causes. Acetaminophen (AAF) toxicity accounts for 39% of the cases of ALF in the United States and Europe.3 Recovery from ALF is determined by the metabolic consequences of reduced liver cell mass, the release of toxic substrates from hepatocytes, and the capability of hepatocyte regeneration.

8 Terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick-end labeling staining and histopathological examination of liver tissue indicated that cell death was necrotic and not apoptotic in nature. Hepatic injury was mediated by intestinally-derived CD4+ T cells during infection and could be mitigated by blocking their entry into the liver. 9 Moreover, transfer of these cells from IL-10 KO mice to recombination activation gene 2 KO animals reproduced the disease, and this suggested that this lymphocyte subset alone was sufficient for inciting injury. Type 2 cytokine production, particularly IL-4 synthesis, was prominent in the infected IL-10 KO liver. 9 Therefore, we hypothesized

that IL-4 promoted inflammation Selleckchem 3MA and necrosis during infection in IL-10 KO mice. The infection of singly and doubly deficient animals revealed that lesion development was dependent on IL-4 in IL-10 KO mice (Fig. 1A). buy Ponatinib While multifocal lesions were grossly

and histologically visible in IL-10 KO animals, they were completely absent in IL-10/IL-4 KO mice. Lesions were characterized by central necrosis that was surrounded by mononuclear and polymorphonuclear cells. Although liver tissue from infected IL-10/IL-4 KO mice appeared to contain more leukocytes than that from WT animals, areas of hepatocellular necrosis were not detected. Neither WT nor IL-4 KO mice acquired hepatic lesions (Fig. 1B). As we reported previously, serum ALT activity Pembrolizumab in vivo at 12 days post-infection was significantly greater in infected IL-10 KO mice compared to WT mice (Fig. 1C). 9 Infection did not lead to an increase in ALT values in IL-4 KO mice, and this indicated a lack of hepatocyte damage. In contrast, ALT levels in infected IL-10/IL-4 KO mice rose significantly above WT levels but were not different from those in IL-10 KO mice. When considered with

the histological evidence, the results suggested that initial hepatocyte injury occurred in the absence of IL-10, but the evolution of organized necrotic lesions required IL-4. We considered, however, that differences in parasite burden between IL-10 KO and IL-10/IL-4 KO mice might affect lesion development. Accordingly, we counted intestinal worm numbers as an indication of the load that the liver received during the acute phase of infection and found no differences, suggesting that the disparity in hepatic response was due not to parasite burden but rather reflected differences in immunity (data not shown). Immune-mediated hepatic injury is the result of effector leukocyte recruitment and activity, and we find enumeration of hepatic leukocytes to be a sensitive indicator of inflammation. Both infected IL-10 KO and IL-10/IL-4 KO mice had elevated numbers of hepatic leukocytes in comparison with WT mice, implying that IL-10 regulated the total leukocyte content within the liver independently of IL-4 (Fig. 1D).

Gretch, MD, PhD, Minjun Chung Apodaca, BS, ASCP, Rohit Shankar, BC, ASCP, Natalia Antonov, M.Ed. New England Research Institutes, Watertown, MA: (Contract N01-DK-9-2328) Teresa M. Curto, MSW, MPH, Margaret C. Bell, selleck chemicals llc MS, MPH. Inova Fairfax Hospital, Falls Church, VA: Zachary D. Goodman, MD, PhD, Fanny Monge, Michelle Parks. Data and Safety Monitoring Board Members: (Chair) Gary L. Davis, MD, Guadalupe Garcia-Tsao, MD, Michael Kutner, PhD, Stanley M. Lemon, MD, Robert P. Perrillo, MD. “
“Here, we identify (−)-epigallocatechin-3-gallate (EGCG) as a new inhibitor of hepatitis C virus (HCV) entry. EGCG is a flavonoid present in green tea extract belonging to the subclass of catechins, which has many properties.

Particularly, EGCG possesses antiviral activity and impairs cellular lipid metabolism. Because of close links between HCV life cycle and lipid metabolism, we postulated that EGCG may interfere with HCV infection. We demonstrate that a concentration of 50 μM of EGCG inhibits HCV infectivity by more than 90% at an early step of the viral life cycle, most likely the entry step. This inhibition was not observed with other members of the Flaviviridae family tested. The antiviral activity of EGCG on HCV entry was confirmed with pseudoparticles https://www.selleckchem.com/products/Roscovitine.html expressing HCV envelope glycoproteins E1 and E2 from six different genotypes. In

addition, using binding assays at 4°C, we demonstrate that EGCG prevents attachment of the virus to the cell surface, probably by acting directly on the particle. We also show that EGCG has no effect on viral replication and virion secretion. By inhibiting cell-free virus transmission using agarose or neutralizing antibodies, we show that EGCG inhibits HCV cell-to-cell spread. Finally, by successive inoculation of

naïve cells with supernatant of HCV-infected cells in the presence of EGCG, we observed that EGCG leads to undetectable levels of infection after four passages. Conclusion: EGCG is a new, interesting anti-HCV molecule that could be used in combination with other direct-acting antivirals. Furthermore, it is a novel tool to further dissect the mechanisms of HCV entry into the hepatocyte. (HEPATOLOGY 2012;) Hepatitis C virus (HCV) is a major cause of chronic liver disease. It is estimated that 3% of the world population is currently infected and thus is at high risk of developing cirrhosis and hepatocellular Tenoxicam carcinoma. 1 No vaccine is available, and the current standard-of-care therapy with pegylated interferon-alpha (IFN-α) and ribavirin has a limited efficacy and significant side effects. 2 Very recently, an addition to the therapy of new direct-acting antivirals (DAAs) targeting HCV nonstructural protein (NS)3-4A protease, telaprevir, and boceprevir was shown to increase the sustained virological response in patients infected with HCV genotype 1 by up to 70%. 3 Efforts are currently being made to identify new DAAs with additive potency. The majority of these molecules target the replication step.

5 ± 4.5% vs 1.8 ± 1.6%, NAFLD vs no NAFLD, P < 0.001). Total liver volume was 29% higher in subjects with NAFLD (1.91 ± 0.45 L) than in those with no NAFLD (1.49 ± 0.31 L, P < 0.001). In multiple linear regression analysis, the percentage of liver fat and bodyweight independently explained variation in total liver volume (r2 = 0.42, P < 0.001). The r-values for the relationship between metabolic selleck kinase inhibitor parameters and the total liver fat volume were not significantly better than those between metabolic parameters and

the percentage of liver fat. Both bodyweight and NAFLD increase liver volume independent of each other. Measurement of liver fat by 1H-MRS allows accurate quantification of NAFLD and calculation

of total liver volume. “
“A powerful way to identify driver genes with causal roles in carcinogenesis is to detect genomic regions that undergo frequent alterations in cancers. Here we identified 1,241 regions of somatic copy number alterations in Autophagy Compound Library 58 paired hepatocellular carcinoma (HCC) tumors and adjacent nontumor tissues using genome-wide single nucleotide polymorphism (SNP) 6.0 arrays. Subsequently, by integrating copy number profiles with gene expression signatures derived from the same HCC patients, we identified 362 differentially expressed genes within the aberrant regions. Among these, 20 candidate genes were chosen for further functional assessments. One novel tumor suppressor (tripartite motif-containing 35 [TRIM35]) and two putative oncogenes (hairy/enhancer-of-split related with YRPW motif 1 [HEY1] and small nuclear ribonucleoprotein polypeptide E [SNRPE]) were discovered by various in vitro and in vivo tumorigenicity experiments. Importantly, it was demonstrated that decreases of TRIM35 expression are a frequent event in HCC and the expression level of TRIM35 was negatively correlated with tumor size, histological grade, and serum alpha-fetoprotein concentration.

Conclusion: These results showed that integration of genomic and transcriptional data offers powerful potential for identifying novel cancer genes in HCC pathogenesis. (HEPATOLOGY 2011;) © 147. Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related mortality worldwide. New insights into Dichloromethane dehalogenase the pathogenesis of this lethal disease are urgently needed. Chromosomal copy number alterations (CNAs) can lead to activation of oncogenes and inactivation of tumor suppressors in human cancers.1 Thus, identification of cancer-specific CNAs will not only provide new insight into understanding the molecular basis of tumorigenesis but also facilitate the discovery of new cancer genes.2, 3 Using traditional methodologies, frequent DNA copy number gains at 1q, 8q, 7q, 17q, and 20q and losses at 4q, 8p, 13q, 16q, and 17p have been identified in HCC.

The final part explores the clinical value of miRNA as prognostic and diagnostic markers. Hepatocellular carcinoma is a highly aggressive tumor that currently ranks the fifth most prevalent cancer worldwide. Although few studies have reported on promising treatment strategies for HCC, the dismal outcome remains unchanged; the median survival of most patients is still 6–9 months from diagnosis.15 Epidemiological studies have firmly established a number of etiologic risk factors in the development of HCC. These can be broadly divided into host factors and environmental factors. Host factors include male gender and genetic metabolic defects contributing to obesity,

whereas environmental factors entail viral hepatitis (types B and C) infections, excessive U0126 research buy chronic alcohol intake,

dietary aflatoxin B1 exposure, and cigarette smoking.15 Complex interactions among these factors ultimately lead to chronic liver disease and/or liver cirrhosis, and the increased risk of HCC development. Several studies have begun to examine for specific miRNA deregulation in hepatitis B virus (HBV)-related and hepatitis C virus (HCV)-related HCCs. While a microarray profiling study in 25 HCC specimens showed no significant difference in miRNA expression between HBV and HCV-associated cases,16 another complementary study measuring the expression of 188 miRNAs in 12 HBV-related and 14 HCV-related HCCs identified 19 miRNAs that could differentiate the HBV and HCV groups.17 Thirteen miRNAs exhibited a decreased expression in the HCV group (including miR-190, miR-134, miR-151), and six showed specific reduced expression in the HBV group (including AP24534 supplier miR-23a, miR-142-5p, miR-34c).17 Concordantly, several studies have also identified miRNAs that were differentially regulated by HBV or HCV. Transfection of the HCV genome resulted in downregulation of 10 miRNAs and upregulation of 23 miRNAs, amongst which elevated miR-193b expression was apparent.18 On the other hand, miR-96 was reported to be

distinctively upregulated in HBV-associated HCC tumors.19 Interactions between host miRNAs and HCV have also been studied. MiR-122, a liver-specific miRNA, is abundantly expressed all in liver tissues and accounts for 70% of the total liver miRNA population.20 Host miR-122 was found to accelerate ribosome binding to HCV RNA, which in turn stimulated viral translation.21 Functional inactivation of miR-122 could lead to 80% reduction of HCV RNA replication in HCC cell lines.22 In this connection, repression of miR-122 in HCC might represent a compensatory mechanism that confers resistance to HCV replication in HCC cells.22 Hepatocellular carcinoma predominantly affects men, with an incidence typically two to four times higher than in women.23 In an attempt to elucidate the role of miRNAs in this gender disparity, the expression profile of a panel of 17 frequently deregulated miRNAs was compared between male and female HCC patients.

Because tea Cytoskeletal Signaling inhibitor is the most widely consumed beverage in the world, future studies designed to comprehensively elucidate the association of tea intake and liver diseases are encouraged. Liang Shen Ph.D.*, * Shandong Provincial Research Center for Bioinformatic Engineering and Technique, Shandong University of Technology, Zibo, People’s Republic of China. “
“I read with interest the article by Serste et al.,1 who showed that beta-blockers are deleterious to the survival of patients with cirrhosis and refractory ascites. Because the study was performed with two groups of patients with cirrhosis with striking differences in their characteristics, it should be

interpreted with caution. As indicated in the editorial by Wong and Salerno,2 the patients on beta-blockers were Lorlatinib mw indeed sicker: they had higher bilirubin levels, lower albumin levels, lower arterial pressures, and lower serum sodium levels, and more patients had hepatic encephalopathy, hepatocellular carcinoma, and Child-Pugh class C cirrhosis. In addition, the patients receiving beta-blockers were taking them for the prevention of variceal hemorrhaging, but only 4% of the patients not receiving beta-blockers had esophageal varices. I do not know whether the patients received beta-blockers mainly for primary

or secondary prevention of variceal bleeding. If patients had episodes of variceal bleeding with concomitant refractory ascites and hepatocellular carcinoma, the outcome was naturally very dismal. Moreover, Serste et al. did not disclose the proportions of (1) hepatitis B patients receiving antiviral treatment, (2) alcoholic patients who were abstinent, and (3) patients whose refractory ascites was managed with transjugular intrahepatic portosystemic stent shunts in the two groups. All these factors may partly influence the outcomes of patients with advanced cirrhosis.3, 4 The causes of death in the two groups were not revealed in detail. As many as 25 patients (17%) died at home of unspecified causes. It is generally believed that the use of beta-blockers is contraindicated

in patients with hypotension or bradycardia. However, some patients with hypotension or bradycardia were still enrolled in the beta-blocker group. This appears to be against the principle of using cAMP propranolol to prevent gastrointestinal hemorrhaging.5 My group has previously shown that patients receiving medical therapy have improved survival in comparison with those receiving endoscopic ligation to prevent variceal rebleeding.6 For patients with cirrhosis, refractory ascites, and a high risk of variceal bleeding, controlled trials are still required to compare the efficacy and safety of beta-blockers and endoscopic ligation; otherwise, liver transplantation will be needed. Gin-Ho Lo M.D.*, * Department of Medical Nutrition, I-Shou University, Kaohsiung, Taiwan. “
“We read with great interest the work by Kohly et al.

Disaccharides, such as lactulose, are absorbed through the paracellular junction complex, which corresponds to the permeability of larger molecules.13 The L/M (lactulose/mannitol) ratio thus comprises an index to appraise intestinal permeability (IP); this ratio has been reported to be elevated in patients with liver cirrhosis, like those with

Crohn’s disease.13 Elevation of the L/M ratio is marked in end-stage cirrhosis.8,9 Although the results by 51Cr-EDTA, the most frequently used isotope probe, have been conflicting,6,7,11 a recent study by Wnt inhibitor Scarpellini et al.6 showed that impairment of instestinal permeability was significantly associated with Child-Pugh status. Parlesak et al.14 found also that permeability of polyethylene glycol (PEG) with high molecular mass (PEG 1500 and PEG 4000) was increased in patients with alcoholic liver diseases.14 They discussed PEG as an appropriate probe for the assessment of endotoxin translocation on the basis of its homogeneous chemical properties, appropriately adaptable molecular mass and linear, chain-like shape mimicking the structure of endotoxin.14 These demands cannot be met by other commonly used permeability RAD001 nmr marker compounds

described above.15 Lee et al.15 reported that intestinal permeability determined by PEG 400 and 3500 was significantly high in cirrhotics with ascites. In this issue of the Journal of Gastroenterology and Hepatology, Kim et al.16 report that the intestinal permeability index, the percentage of permeability of PEG 3350 to that of PEG 400, was significantly increased on admission for active GI hemorrhage in patients with liver cirrhosis and proven or possible infections. This study is especially interesting on the point that the authors described a strong correlation between the increased

intestinal permeability and the serum level of endotoxin in their discussion, although the precise data were not shown in the text. In this study, the most frequent etiology of liver cirrhosis was alcoholism. There is now accumulating evidence that alcohol misuse in patients with liver disease is associated with increased intestinal permeability and endotoxemia. Thus, significant correlation between the plasma endotoxin levels and intestinal permeability Thymidylate synthase determined by PEG 4000 has been reported in patients with alcoholic liver disease.14 Although the mechanism of increased intestinal permeability in patients with alcoholic cirrhosis is still undetermined, genetic factors and/or environmental factors may be involved. These include the generation of acetaldehyde in the colonic lumen, the status of the intestinal flora,17 nitric oxide and superoxide anion in the intestinal barrier,18 and so on. It is not known if these or other factors especially affect intestinal permeability in patients with liver cirrhosis and gastrointestinal hemorrhage.

Louis, MO) containing 1 mg/mL of Mycobacterium tuberculosis strain H37RA, and was subsequently boosted every 2 weeks with 2OA-BSA in incomplete Freund’s adjuvant (Sigma-Aldrich). Additionally, mice received 100 ng of pertussis toxin find more (List Biological Laboratories, Campbell, CA) at the time of initial immunization with 2OA-BSA in Complete Freund’s Adjuvant. Peripheral blood samples from individual mice were obtained from the tail vein prior to the initiation of treatment with mAbs (baseline) and then at 2-week intervals. Sera was collected prior to mAb treatment, 1 week afterward, and thereafter every 4 weeks, and stored at −70°C until

use. Animals were sacrificed at 15 weeks of age. Serum titers of anti–PDC-E2 autoantibodies were measured by way of enzyme-linked immunosorbent assay using our well-standardized recombinant autoantigens.32 Peripheral blood mononuclear cells were isolated from heparinized murine blood using Accupaque (Accurate Chemical & Scientific Company, Westbury, CT) to assess levels of B cells. Cells were preincubated with anti-mouse FcR blocking reagent and then incubated at 4°C with a predetermined optimum concentration of antigen-presenting cell (APC)-conjugated anti–T cell receptor β (BioLegend), phycoerythrin-conjugated anti-mouse IgM

(Caltag), and fluorescein isothiocyanate–conjugated anti-CD19 https://www.selleckchem.com/products/ch5424802.html (BioLegend); B cell frequency was then determined by way of flow cytometry. The liver and spleens were collected from mice immediately following sacrifice, and single-cell mononuclear cell suspensions

were prepared for multicolor flow analysis as described.23 Immediately after sacrifice, liver and spleen tissues were harvested and fixed in 10% buffered formalin, embedded in paraffin, and cut into 4-μm sections for routine hematoxylin (DakoCytomation, Carpinteria, CHIR-99021 concentration CA) and eosin (American Master Tech Scientific, Lodi, CA) staining. Evaluation under light microscopy and scoring of liver inflammation was performed on coded hematoxylin and eosin–stained sections of liver using a set of three indices by a blinded pathologist (K. T.); indices included degrees of portal inflammation, parenchymal inflammation, and bile duct damage.33 Phenotypic analysis of bile duct damage was performed as described.34 Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphate (ALP) levels were measured using the Roche Diagnostics COBAS INTEGRA 400 Plus (Indianapolis, IN). Serum levels of the proinflammatory cytokines interleukin (IL)-6, IL-10, monocyte chemotactic protein-1 (MCP-1), interferon-γ (IFN-γ), tumor necrosis factor α, and IL-12p70 were quantified using the BD Cytometric Bead Array Mouse Inflammatory Kit (BD Biosciences) as described.35 The serum samples were loaded onto the plate neat.