Liver Histologies Research Articles

Fructooligosaccharides Supplementation: A Good Choice for the Prevention and Treatment of Non-Alcoholic Fatty Liver Disease?

Background and objectives: Carbohydrates such as fructooligosaccharides (FOSs) are associated with improved gastrointestinal health and the prevention of excess body fat. We evaluated the long-term effects of high amounts of FOS on metabolic parameters, non-alcoholic fatty liver disease (NAFLD) and short-chain fatty acids (SCFAs). Methods: Sixty C57BL/6 mice received the following diets for four months: control (C), normolipid rich in fiber (F), normolipid supplemented with FOS (FOS), high fat (HL), high fat with high fiber (HLF) and high fat with FOS (HLFOS). We analyzed the animal weight; body composition; food intake; fasting blood glucose; serum and liver lipid profiles; liver and intestinal histologies; malondialdehyde (MDA), hepatic retinol and α-tocopherol; and SCFAs in the feces. Results: Supplementation with FOS in a high-fat diet promoted less body weight gain and reduced liver and retroperitoneal adipose tissue weights compared to HL and HF. FOS prevented NASH and decreased alanine aminotransferase and serum cholesterol levels in experimental animal models of obesity and metabolic syndrome (MS). There were statistical differences found in the dosages of the three main SCFAs in feces (acetic, isobutyric and isovaleric acids). Conclusions: Long-term supplementation with high doses of FOS was effective in reducing weight, adiposity, NAFLD and serum cholesterol in C57BL mice with obesity and MS induced by a high-fat diet.

Subtype, Frequency, and Associated Background Histology in Males Diagnosed with Hepatocellular Adenomas: A Single Institution Study

Abstract Introduction/Objective Hepatocellular adenomas (HCAs) are rare, benign hepatic neoplasms arising from noncirrhotic livers, most commonly diagnosed in reproductive-age women using oral contraception with female:male ratio of 10:1. These tumors are also associated with anabolic steroid use, obesity, and diabetes. HCA subtypes and their incidence in the general population are as follows: inflammatory (HCA-I) accounting for approximately 50% of cases, HNF1A- inactivated (HCA-H) in 35-40%, β-catenin-activated (HCA-B) in 10-15%, and unclassified HCA in <5-10%. We aimed to determine HCA subtype frequency in male-only cases and their associated background liver histologies. Methods/Case Report We performed a retrospective review of all HCAs diagnosed in males from 2000–June 2022 at Mount Sinai Hospital using keywords “hepatocellular adenomas” and “hepatic adenomas;” a total of 54 cases fulfilled the criteria. Liver biopsies and resection specimens were included; duplicate cases were eliminated, and only those cases with a definitive diagnosis of HCA were included. Results (if a Case Study enter NA) There were 31 patients (age range 17-73 years) included with a total number of 41 HCAs (17 from resections and 14 from biopsies). HCA-I comprised 23/31 cases (74%), HCA-H and HCA-B each encompassed 2/31 cases (6%, respectively), one HCA-I had coexistent β-catenin mutation (3%, 1/31), and one unclassified HCA had mixed HCA-I and HCA-H features (3%, 1/31). Twelve cases were diagnosed as HCA without further subtype specification (NOS) (38%). Background liver histology was available in 24 cases. Fatty liver disease (steatosis and steatohepatitis) was found in five cases, of which four were classified as HCA-I. Of seven hepatocellular carcinomas diagnosed, four arose from NOS HCAs with either unremarkable or steatohepatitis backgrounds. Three of five HCA-I showed background nonspecific chronic hepatitis while two had glycogen storage disease. The remaining ten cases had unremarkable liver backgrounds and were found involving all HCA subtypes. Conclusion In this cohort of male-only patients, HCA-I was the most common subtype, the frequency of which is significantly higher than is reported in the general population. Only 6% of cases were HNF1A-inactivated and β- catenin-activated, far less than the reported incidence of 35-40% and 10-15%, respectively. Additionally, our findings show that the majority of HCAs had an unremarkable liver background. Further studies are necessary to determine the neoplastic drivers among male patients that develop hepatocellular adenomas.

MRI-based delta-radiomic features for prediction of local control in liver lesions treated with stereotactic body radiation therapy

Real-time magnetic resonance image guided stereotactic ablative radiotherapy (MRgSBRT) is used to treat abdominal tumors. Longitudinal data is generated from daily setup images. Our study aimed to identify delta radiomic texture features extracted from these images to predict for local control in patients with liver tumors treated with MRgSBRT. Retrospective analysis of an IRB-approved database identified patients treated with MRgSBRT for primary liver and secondary metastasis histologies. Daily low field strength (0.35 T) images were retrieved, and the gross tumor volume was identified on each image. Next, images’ gray levels were equalized, and 39 s-order texture features were extracted. Delta-radiomics were calculated as the difference between feature values on the initial scan and after delivered biological effective doses (BED, α/β = 10) of 20 Gy and 40 Gy. Then, features were ranked by the Gini Index during training of a random forest model. Finally, the area under the receiver operating characteristic curve (AUC) was estimated using a bootstrapped logistic regression with the top two features. We identified 22 patients for analysis. The median dose delivered was 50 Gy in 5 fractions. The top two features identified after delivery of BED 20 Gy were gray level co-occurrence matrix features energy and gray level size zone matrix based large zone emphasis. The model generated an AUC = 0.9011 (0.752–1.0) during bootstrapped logistic regression. The same two features were selected after delivery of a BED 40 Gy, with an AUC = 0.716 (0.600–0.786). Delta-radiomic features after a single fraction of SBRT predicted local control in this exploratory cohort. If confirmed in larger studies, these features may identify patients with radioresistant disease and provide an opportunity for physicians to alter management much sooner than standard restaging after 3 months. Expansion of the patient database is warranted for further analysis of delta-radiomic features.

Laparoscopic microwave ablation versus percutaneous microwave ablation of hepatic malignancies: Efficacy and recurrence-free survival outcomes in patients

BackgroundHepatic thermal ablation has been found to be effective and equivalent to resection in certain liver histologies. Of the 16,000 annual liver ablations performed in the United States, only 13% (2,080 ablations) are performed laparoscopically. The laparoscopic technique remains underused even with the benefits of improved staging and better access to tumors. The purpose of this study is to compare laparoscopic microwave ablation versus percutaneous microwave ablation in terms of efficacy and recurrence-free survival outcomes in patients with hepatic malignancies. MethodsA comparative analysis was performed on 275 patients (289 ablation procedures) who underwent laparoscopic microwave ablation or percutaneous microwave ablation between February 2011 and May 2021. Ablation success was confirmed postprocedure and recurrence was monitored at follow-up via contrast-enhanced computed tomography/magnetic resonance imaging and/or computed tomography/positron emission tomography. ResultsThe groups were similar for sex, age, body mass index, location of tumor, size of tumor, and number of tumors. Ablation success was 100% in both groups. Local recurrence was significant (5%: laparoscopic microwave ablation vs 22%: percutaneous microwave ablation, P = .002) and same-lobe recurrence (21%: laparoscopic microwave ablation vs 24%: percutaneous microwave ablation) was lower in the laparoscopic microwave ablation group. Median recurrence-free survival was 15.8 months for the laparoscopic microwave ablation group and 5.6 months for the percutaneous microwave ablation group (P = .0002). Overall, 90-day complications were lower in the laparoscopic microwave ablation group (11%) compared with the percutaneous microwave ablation group (21%) (P = .11). ConclusionLaparoscopic surgical ablation is a critical surgical skill that must be taught in fellowship. Laparoscopic microwave ablation leads to better tumor specific outcomes and oncologic outcomes demonstrating clinical efficacy in the treatment of hepatic malignancies compared with percutaneous microwave ablation.

Biliary atresia liver histopathological determinants of early post-Kasai outcome

BackgroundA retrospective chart review of liver histologies in Kasai biliary atresia BA patients operated 1/2017- 7/2019 at our institution was conducted to identify histologic prognostic factors for biliary outcome. MethodsPatients with wedge liver biopsies and portal plate biopsies (n = 85) were categorized into unfavorable and favorable outcome, based on a 3-month serum total bilirubin level of <34 μM or mortality. Hepatocellular histologies, presence of ductal plate malformation (DPM) and of large bile duct of ≥ 150 μm diameter size at the portal plate were evaluated. ResultsTotal Bilirubin levels> 34 μM correlates with worse 1-year survival. Age at surgery, histologic fibrosis or inflammation does not predict outcome. Potential adverse predictors are severe hepatocellular swelling, severe cholestasis, presence of DPM (n = 24), and portal plate bile duct size < 150 µm (n = 28). In multivariate analyses adjusting for age at Kasai and postop cholangitis, bile duct size and severe hepatocellular swelling remain independent histologic prognosticators (OR 3.25, p = 0.039 and OR 3.26, p = 0.006 respectively), but not DPM. ConclusionAdvanced histologic findings of portal plate bile duct size of <150 µm and severe hepatocellular damage predict poor post-Kasai jaundice clearance and short-term survival outcome, irrespective of Kasai timing.Level of Evidence: Level III

Reno-Hepatoprotective and Antidiabetic Properties of Methanol Leaf Extract of Laportea Aestuans in Wistar Rats.

Toxicities due to exposure to arsenic-contaminated water and the occurrence of diabetes mellitus are major health concerns. Treatment of these concerns using therapeutic measures have recorded limited success. Traditionally, Laportea aestuans (LA) has been used in managing various diseases. Hence, we investigated the reno-hepatoprotective/antidiabetic potentials of methanol leaf extract of LA (MeLELA) in male Wistar rats. Thirty rats (100-150 g) were equally distributed into 6 groups: Group I (vehicle-treated); group II received 2.5 mg/kg sodium arsenite (SA) thrice a week for 2 weeks; group III received streptozotocin (STZ, 50 mg/kg once); group IV received 200 mg/kg LA daily for 14 days; group V received SA and LA; group VI received STZ and LA. Sodium arsenite and STZ induced reno-hepatotoxicity and diabetes, respectively. Phytochemical screening, biomarkers/enzyme activities, blood glucose levels, micronucleus assay, kidney, liver and pancreas histologies were determined according to standard procedures. Alkaloids, carotenoids and flavonoids were present in abundance. Both SA-and STZ-treated groups recorded significant (p < 0.05) reductions in serum protein concentrations, while co-treatment with LA significantly restored the levels. The SA-induced significant increase in creatinine/urea levels were significantly reduced by LA. Co-treatment of each of SA-and STZ-treated groups, respectively, with LA significantly decreased the elevated serum alanine and aspartate aminotransferases’ activities. Increased blood glucose level in diabetic group was remarkably lowered by LA. Also, the SA-induced frequency of micronucleated polychromatic erythrocytes was significantly ameliorated by LA. Conclusively, LA is protective against SA-induced toxicity and STZ-induced diabetes in Wistar rats.

Effects of Ecklonia arborea or Silvetia compressa algae intake on serum lipids and hepatic fat accumulation in Wistar rats fed hyperlipidic diets

Seaweeds can generate changes in metabolism of lipids and consequently diminish cholesterol and other serum lipids in mammals. However, the consumption of marine algae may also alter function of other organs. Therefore, the aim of this research was to study the metabolism of lipids as well as the liver steatosis of Wistar male rats fed for two months with hyperlipidic diets composed of 20% of fat supplemented with 2.5% of seaweeds Ecklonia arborea or Silvetia compressa. Blood total cholesterol, lipoproteins, triglycerides and key hepatic enzymes were determined and liver histologies were also assessed. The two algae produced different results, while S. compressa reduced 18% total cholesterol, E. arborea increased it 5.8% compared to the animals fed the positive control. Rats fed the S. compressa supplemented diet comparatively had lower serum low density lipoproteins (54%) and triglycerides (69%) levels compared to counterparts fed the high-fat diet. Livers histologies of rats fed with the high-fat diets showed an evident lipid accumulation. However, the consumption of S. compressa diminished liver steatosis compared to E. arborea. Results herein indicate that S. compressa is a promising resource to decrease serum total cholesterol and low-density lipoproteins and hepatic tissue damage. Further investigations should be made to find out if the ingestion of marine algae extracts would decrease the accumulation of hepatic fat and consequently affects positively liver physiology.

Chinese Herbal Medicine Formula Shenling Baizhu San Ameliorates High-Fat Diet-Induced NAFLD in Rats by Modulating Hepatic MicroRNA Expression Profiles.

Objective The purpose of present study was to investigate the potential mechanism underlying the protective effect of Shenling Baizhu San (SLBZS) on nonalcoholic fatty liver disease (NAFLD) by microRNA (miRNA) sequencing. Methods Thirty male Wistar rats were randomly divided into a normal control (NC) group, a high-fat diet (HFD) group, and an SLBZS group. After 12 weeks, the biochemical parameters and liver histologies of the rats were assessed. The Illumina HiSeq 2500 sequencing platform was used to analyse the hepatic miRNA expression profiles. Representative differentially expressed miRNAs were further validated by qRT-PCR. The functions of the differentially expressed miRNAs were analysed by bioinformatics. Results Our results identified 102 miRNAs that were differentially expressed in the HFD group compared with the NC group. Among those differentially expressed miRNAs, the expression levels of 28 miRNAs were reversed by SLBZS administration, suggesting the modulation effect of SLBZS on hepatic miRNA expression profiles. The qRT-PCR results confirmed that the expression levels of miR-155-5p, miR-146b-5p, miR-132-3p, and miR-34a-5p were consistent with those detected by sequencing. Bioinformatics analyses indicated that the target genes of the differentially expressed miRNAs reversed by SLBZS were mainly related to metabolic pathways. Conclusion This study provides novel insights into the mechanism of SLBZS in protecting against NAFLD; this mechanism may be partly related to the modulation of hepatic miRNA expression and their target pathways.

A retrospective case control study identifies peripheral blood mononuclear cell albumin RNA expression as a biomarker for non-alcoholic fatty liver disease.

Non-alcoholic fatty liver disease (NAFLD) improves after bariatric surgery. The aim of this study was to determine whether peripheral blood mononuclear cell albumin gene expression was related to NAFLD and whether albumin (ALB) and alpha fetoprotein (AFP) expression could be detected in whole blood and visceral adipose tissue. Using a retrospective case control study design, RNA isolated from peripheral blood mononuclear cells from patients prior to undergoing bariatric surgery was used for pooled microarray analysis. Quantitative polymerase chain reaction (QPCR) was used to analyze whole blood and visceral adipose tissue. Liver histology was obtained via intra-operative biopsy and clinical data extracted from the electronic health record. The albumin (ALB) gene was the second most up-regulated found in microarray analysis of peripheral blood mononuclear cell RNA from patients with hepatic lobular inflammation versus normal liver histology. Transcript levels of ALB were significantly different across those with normal (n = 50), steatosis (n = 50), lobular inflammation (n = 50), and peri-sinusoidal fibrosis (n = 50) liver histologies, with lobular inflammation 3.9 times higher than those with normal histology (p < 0.017). Albumin expression levels decreased in 11/13 patients in paired samples obtained prior to and at 1year after Roux-en-Y gastric bypass surgery. ALB expression could be detected in 23 visceral adipose tissue samples obtained intra-operatively and in 18/19 available paired whole blood samples. No significant correlation was found between ALB expression in visceral adipose tissue and whole blood RNA samples. Alpha fetoprotein expression as a marker of early hepatocytic differentiation was detected in 17/17 available VAT RNA samples, but in only 2/17 whole blood RNA samples. Albumin RNA expression from blood cells may serve as a biomarker of NAFLD. Albumin and alpha fetoprotein appear to be ubiquitously expressed in visceral adipose tissue in patients with extreme obesity.

Factors Associated With In-field Recurrence in Patients Treated With Liver SBRT

Stereotactic body radiation therapy (SBRT) is an emerging technique for treatment of primary and secondary liver malignancies. Reports in the literature of the impact of set up uncertainties and other factors influencing in-field recurrences are lacking. The aim of this study was to examine the magnitude and consistency of interfractional shifts, their association, and association of other factors with in-field recurrences in patients treated with liver SBRT. All patients treated with liver SBRT at a single institution between 2014 and 2016 with doses 40 – 50 Gy in 5 fractions were included. Tumor histologies included hepatocellular carcinoma (50%), cholangiocarcinoma (4%), liver metastases from colorectal (39%) and other cancers (7%). Median follow-up was 15 months. A bony match was performed using orthogonal kV images. A soft-tissue match was then performed using a cone beam CT (CBCT). The shift data included in this study are the shifts made after the CBCT. Respiratory motion management techniques included abdominal compression, respiratory gating or free breathing, with the target respiratory motion below 1 cm. No dedicated internal fiducial markers were used. Retrospective analysis of prospectively collected data was performed as a part of an institutional Quality Assurance process. A total of 46 patients met the eligibility criteria. Data on interfractional shifts were available for 22 patients. One- and two-year in-field control rates were 64% and 30%, respectively. Mean vector shift length was 0.37 cm (systematic error 0.14 cm, random error 0.29 cm), 86% of patients had no shifts greater than 1 cm, 96% of all the shifts performed were less than 1 cm. Inferior tumor location had lower interfractional shift values compared with other tumor locations (one sided t-test=1.81, p-value=0.047). Despite that, inferior tumor location was weakly associated with in-field recurrence on univariate analysis (HR=2.63, p-value=0.093; 95% CI 0.86, 8.06), with median time to in-field recurrence 11 months as opposed to 21 months in patients with other tumor locations. Liver metastasis histology was also weakly associated with in-field recurrence, compared with primary liver histologies (HR=1.63, p-value=0.087; 95% CI 0.93, 2.87). Radiation dose, motion management technique, planning target volume and interfractional shift magnitude were not associated with in-field recurrence. Set up uncertainty and reproducibility was acceptable and did not impact in-field recurrences. Cone beam CT with soft tissue matching appears to be an appropriate set-up verification method. Inferior tumor location was associated with higher in-field recurrences in this patient cohort, likely due to proximity of critical structures, such as small bowel, resulting in target volume coverage compromises. Detailed evaluation of dose distribution in patients with in-field recurrences is underway.

Elevation of gamma-glutamyl transferase in adult: Should we think about progressive familiar intrahepatic cholestasis?

BackgroundThere are three types of progressive familial intrahepatic cholestasis (PFIC). Type 3 is characterized by elevated gamma-glutamyl transferase (γ-GT) and it can be diagnosed in adolescence/adulthood. The genetic defect of PFIC 3 appears to explain the pathogenesis of intrahepatic cholestasis of pregnancy (ICP). AimsDraw attention to this rare disease, especially in adulthood, and clarify the association between ICP and PFIC 3. ResultsWe describe a series of cases from a Portuguese northern family with two brothers presenting chronic cholestasis since adolescence. Brother 1: since 15-years-old with pruritus and elevated γ-GT ∼6x. Brother 2: pre-term, due to severe maternal pruritus and jaundice, since 13-years-old with pruritus, jaundice and ∼8x γ-GT elevation. Common causes of cholestasis were excluded and liver histologies were nonspecific. Research for mutation on ABCB4 gene showed mutations in both alleles. ConclusionDisease and mechanisms that determine cholestasis are complex and their understanding may provide new therapeutics.

Growth, survival and the histology of the digestive tract of juvenile Osteoglossum bicirrhosum (Cuvier, 1829) fed three diets containing different protein and lipid levels

The influence of three commercial diets with different protein and lipid contents (40 : 14, 45 : 8, 48 : 8 protein : lipid – P : L, in % dry weight of diet) on growth performance, survival and the histology of the liver and intestine was analysed in early juveniles of silver arowana, Osteoglossum bicirrhosum, reared in captivity. Individuals (initial wet weight–Wi = 1.07 ± 0.04 g; total length–TL = 6.0 ± 0.17 cm) were raised in triplicate (n = 12 fish per tank) at 26.2 ± 0.1°C and fed one of three commercial diets over 60 days. At the end of the trial, survival (90.5 ± 3.3%) and condition factor (0.5 ± 0.02) were not affected by dietary treatments, whereas specific growth rate, TL and W gain were lower in animals fed the 40P : 14L diet. Histological analysis revealed a larger accumulation of lipid droplets in the intestines of the group supplied with the 40P : 14L diet, as well as a larger surface of hepatic lipid vacuoles compared to the other treatments, although no signs of steatosis were observed. Juveniles fed the 48P : 8L diet displayed the lowest liver lipid accumulation; however, a protein content higher than 45% did not improve growth performance. The intestine and liver lipid accumulation of individuals fed the 45P : 8L diet might indicate a more balanced lipidic metabolism. Intestine and liver histologies proved to be useful markers to identify the nutritional condition in juveniles of O. bicirrhosum, even when differences in nutritional composition were subtle (i.e. protein variation of 3%).

Outcomes including liver histology after liver transplantation for chronic hepatitis B using oral antiviral therapy alone.

The outcomes of hepatitis B virus (HBV)-related hepatitis after liver transplantation (LT) without hepatitis B immune globulin (HBIG) is not well documented. This study aims to determine the outcomes of chronic hepatitis B (CHB) patients using an HBIG-free regimen. All biopsies performed 3 months or more after LT in consecutive CHB patients transplanted from 2003 to 2012 were reviewed. None of the patients received HBIG. Results of all liver histologies were reviewed to determine the cause of graft dysfunction. Of the 435 patients transplanted during this period, 263 liver biopsies were performed in 144 patients. Thirty-six patients were positive for hepatitis B surface antigen (HBsAg) with undetectable HBV DNA at the time of biopsy, and none had histological evidence of HBV infection. Of the 263 biopsies, 44 (17%) had evidence of fibrosis. There was a significantly higher rate of fibrosis in those with large duct obstruction compared to those without (51% versus 9%, respectively; P < 0.001). Of the 291 patients without a liver biopsy during the same period, 43 were HBsAg+. Seven patients had evidence of virological rebound, of whom 6 had evidence of rtM204V/I mutation and 1 had recurrence of hepatocellular carcinoma with low-level rebound and wild-type virus. In conclusion, for patients without virological rebound, positive serum HBsAg was not associated with histological evidence of HBV-related hepatitis after LT. To prevent virological rebound, nucleos(t)ide analogues with higher barriers to resistance should be used.

Abstract A04: Systemic agonistic anti-CD40 treatment of tumor bearing mice modulates hepatic myeloid suppressive cells and causes immune-mediated liver damage

Abstract Immune stimulatory monoclonal antibodies are currently evaluated as antitumor agents in clinical trials. Although overall toxicity appears to be moderate, liver toxicities have been reported and are not completely understood. Therefore, we decided to study the effect of systemic CD40 antibody treatment on the liver immune microenvironment. Agonistic CD40 antibody or isotype control was injected intraperitoneally into naïve and tumor-bearing mice. Liver enzymes, histologies and immune infiltrates were analyzed after antibody administration. The effect of agonistic CD40 administration was evaluated using bone marrow chimeras and genetically modified mouse strains Cd40-/- and Nox2-/-. Mouse and human tumor-induced myeloid suppressive cells were cultured to address changes in their immune function upon CD40 ligation. Agonistic CD40 antibody caused liver damage within 24 hours after injection in two unrelated tumor models and mice strains. Using bone marrow chimeras we could demonstrate that CD40 antibody-induced hepatitis in tumor-bearing mice was dependent on the presence of CD40+ immune cells. Agonistic CD40 ligation-dependent liver damage was induced by the generation of reactive oxygen species. Furthermore, agonistic CD40 antibody resulted in increased CD80 and CD40 positive liver CD11b+Gr-1+ immature myeloid cells. CD40 ligation on tumor-induced hepatic myeloid cells or CD14+HLA-DRlow PBMC from cancer patients reduced their immune suppressor function. Collectively, agonistic CD40 antibody activates the tumor-induced liver myeloid suppressive compartment, enhancing acute inflammation and thus liver damage. Our data in murine and human myeloid suppressive cells point towards a pivotal role for CD40 as a key molecule in modulating myeloid suppressive cell plasticity. Citation Format: Jose Medina-Echeverz, Chi Ma, Tobias Eggert, Tim F. Greten. Systemic agonistic anti-CD40 treatment of tumor bearing mice modulates hepatic myeloid suppressive cells and causes immune-mediated liver damage. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A04.

The influence of dietary restriction on hepatic mononuclear cell numbers and functions in mice

BackgroundSurgical patients with gastrointestinal malignancies are at increased risk for malnutrition. However, the mechanism by which dietary restriction (DR), one form of malnutrition, impairs hepatic immunity remains to be clarified. The present study was designed to examine the influence of DR on hepatic mononuclear cell (MNC) numbers, subpopulations, and cytokine productions (tumor necrosis factor α [TNF-α], interferon gamma [IFN-γ], and interleukin 10 [IL-10]) in response to lipopolysaccharide (LPS) in mice. Immunoglobulin (Ig) A levels in the gallbladder and histopathologic changes in the liver were also assessed. Material and methodsMale Institute of Cancer Research mice were randomly assigned to three dietary groups: ad libitum (AD), mild restriction (MR), and severe restriction (SR). The AD, MR, and SR groups received daily mouse chow in amounts of 190, 133, and 76 g/kg, respectively, for 7 d. After the mice had been fed for 7 d, hepatic MNCs were isolated. Total hepatic MNCs were counted and subpopulations were determined by flow cytometry. Cytokine productions (TNF-α, IFN-γ, and IL-10) by hepatic MNCs in response to LPS were measured. Blood samples were analyzed for hepatobiliary biochemical parameters. IgA levels in gallbladder bile were measured with enzyme-linked immunosorbent assay. In addition, liver histologies were examined. ResultsHepatic MNC numbers were significantly lower in the MR and SR groups than in the AD group, with no significant difference between the MR and SR groups. The percentage of B cells was significantly lower in the SR group than in the MR and AD groups, whereas the T-cell percentage was higher in the SR group than in the MR and AD groups. The percentage of Kupffer cells was significantly lower in the SR group than in the AD group, whereas that in the MR group was midway between those in the SR and AD groups. TNF-α and IL-10 levels in hepatic MNC culture supernatants were increased LPS-dose dependently in the AD group. However, the increase was slight in the MR group and absent in the SR group. The IgA levels in gallbladder bile were significantly lower in the SR and MR groups than in the AD group. On the basis of hematoxylin and eosin staining of hepatic sections, livers from the SR group showed atrophic hepatocytes and sinusoidal dilatation, whereas these changes were absent in the AD group. ConclusionsDR decreases hepatic MNC number with subpopulation changes, reduces IgA levels in gallbladder bile, blunts cytokine production by hepatic MNCs, and induces pathologic damage in the liver, which may be an important mechanism underlying the impaired host defense associated with malnutrition.

Systemic Agonistic Anti-CD40 Treatment of Tumor-Bearing Mice Modulates Hepatic Myeloid-Suppressive Cells and Causes Immune-Mediated Liver Damage.

Immune-stimulatory mAbs are currently being evaluated as antitumor agents. Although overall toxicity from these agents appears to be moderate, liver toxicities have been reported and are not completely understood. We studied the effect of systemic CD40 antibody treatment on myeloid cells in the spleen and liver. Naïve and tumor-bearing mice were treated systemically with agonistic anti-CD40 antibody. Immune cell subsets in the liver and spleen, serum transaminases, and liver histologies were analyzed after antibody administration. Nox2(-/-), Cd40(-/-), and bone marrow chimeric mice were used to study the mechanism by which agonistic anti-CD40 mediates its effects in vivo. Suppressor function of murine and human tumor-induced myeloid-derived suppressor cells (MDSC) was studied upon CD40 ligation. Agonistic CD40 antibody caused liver damage within 24 hours after injection in two unrelated tumor models and mice strains. Using bone marrow chimeras, we demonstrate that CD40 antibody-induced hepatitis in tumor-bearing mice was dependent on the presence of CD40-expressing hematopoietic cells. Agonistic CD40 ligation-dependent liver damage was induced by the generation of reactive oxygen species. Furthermore, agonistic CD40 antibody resulted in increased CD80-positive and CD40-positive liver CD11b(+)Gr-1(+) immature myeloid cells. CD40 ligation on tumor-induced murine and human CD14(+)HLA-DR(low) peripheral blood mononuclear cells from patients with cancer reduced their immune suppressor function. Collectively, agonistic CD40 antibody treatment activated tumor-induced myeloid cells, caused myeloid-dependent hepatotoxicity, and ameliorated the suppressor function of murine and human MDSC. Collectively, our data suggest that CD40 may mature immunosuppressive myeloid cells and thereby cause liver damage in mice with an accumulation of tumor-induced hepatic MDSC.

Relationship Between Changes in Serum Levels of Keratin 18 and Changes in Liver Histology in Children and Adults With Nonalcoholic Fatty Liver Disease

Cross-sectional studies have associated serum levels of the keratin 18 (K18) fragment with histologic features of liver in individuals with nonalcoholic fatty liver disease (NAFLD). We investigated the relationship between changes in serum levels of K18 and changes in liver histology in adults and children with NAFLD. We measured levels of K18 in stored serum samples collected at baseline and various time points from 231 adults with nonalcoholic steatohepatitis and 152 children with NAFLD who participated in 2 separate prospective randomized clinical trials. Liver biopsy specimens collected at baseline and week 96 were reviewed centrally. There were greater decreases in serum levels of K18 in adults with histologic improvement at week 96 than in those without histologic improvement at week 16 (decrease, 193 ± 293 vs 139 ± 467 U/L; P < .001), week 48 (decrease, 232 ± 360 vs 113 ± 425 U/L; P < .001), or week 96 (decrease, 269 ± 368 vs 97 ± 400 U/L; P < .001). There were greater decreases in serum levels of K18 in children with histologic improvements than in those without histologic improvements at week 48 (decrease, 197 ± 467 vs 47 ± 350 U/L; P = .005) and week 96 (decrease, 206 ± 432 vs 2 ± 474 U/L; P < .001). However, reductions in serum levels of K18 were not better than reductions in levels of alanine aminotransferase in identifying adults with histologic improvement (area under the receiver operator characteristic [AUROC], 0.71; 95% confidence interval [CI], 0.63-0.80; vs AUROC, 0.68; 95% CI, 0.61-0.79; P = .34) or children (AUROC, 0.72; 95% CI, 0.63-0.81; vs AUROC, 0.79; 95% CI, 0.70-0.87; P = .42). Decreases in serum levels of K18 are associated strongly with improved liver histologies in adults or children with NAFLD. However, reductions in K18 do not perform better than those in alanine aminotransferase level in identifying histologic changes in NAFLD.

Influence of fish to soybean oil ratio on hepatic mononuclear cell function and survival after intraportal bacterial challenge in parenterally fed mice

Parenteral nutrition (PN) is indispensable for meeting the caloric and substrate needs of patients who cannot receive adequate amounts of enteral nutrition. However, PN decreases hepatic mononuclear cell (MNC) numbers and impairs their functions. We examined the effects of various ratios of ω-3 to ω-6 polyunsaturated fatty acids on hepatic MNC number and function in a murine model. We focused on serum liver enzymes, lipid metabolism, cytokine production, histopathology, and the outcomes of an intraportal bacterial challenge. In experiment 1, male Institute of Cancer Research mice were randomized to CHOW, 67%, 33%, 16%, 8%, 4%, and 0% fish oil (FO)-PN groups. After receiving their respective diets for 5 days, 1.0 × 10(7) Pseudomonas aeruginosa were delivered by intraportal injection. Survival was observed ≤ 7 days after injection. Liver histologies after intraportal bacterial challenge were examined in the CHOW, 33%, 8%, and 0% FO-PN groups. In experiment 2, the mice were divided into 4 groups: CHOW, 33%, 8%, and 0% FO-PN. After the mice had been fed for 5 days, MNC were isolated. Hepatic MNC were counted and cytokine productions (tumor necrosis factor [TNF]-α and interleukin [IL]-10) by MNC in response to lipopolysaccharide (LPS) were measured. Blood samples were analyzed for lipid metabolism and hepatobiliary biochemical parameters. Liver histologies were also examined. In experiment 1, survival times were significantly shorter in the 4% and 0% FO-PN groups than in the CHOW group. Survival rates at 168 hours were 100%, 64%, 86%, 73%, 67%, 11%, and 13% in the CHOW, 67%, 33%, 16%, 8%, 4%, and 0% FO-PN groups, respectively. At 72 hours after intraportal bacterial challenge, the 0% FO-PN group showed severe tissue damage, whereas such damage was reduced in the 8% and 33% FO-PN groups. In experiment 2, the CHOW, 33%, 8%, and 0% FO-PN groups showed LPS dose-dependent increases in TNF-α levels. IL-10 levels were also LPS dose-dependently increased in the CHOW and 33% FO-PN groups. However, no marked changes were observed in response to LPS stimulation in either the 8% or the 0% FO-PN group. There were no differences in serum levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, or total bilirubin among these 4 groups. In the 0% FO-PN group, serum total cholesterol levels were greater than those in the 8% and 33% FO-PN groups. Without bacterial challenge, livers from the 0% FO-PN group showed steatosis, but these changes were attenuated in the 8% and 33% FO-PN groups. The 30-40% ratio of FO to soybean oil with 20% of total calories supplied by lipid seems to be the best PN for preservation of hepatic MNC number and function.

480 ANTIVIRAL THERAPY CAN LEAD TO FIBROSIS REGRESSION IN ANTIHBE-POSITIVE CHRONIC HEPATITIS B PATIENTS TREATED IN CLINICAL PRACTICE

of patients according to various guidelines and liver histology of these patients. Patients andMethods: 574 HBV patients (353; 61.5% male – median age 46; range 19–78) without clinical or laboratory evidence of cirrhosis were included in the study. Patients were grouped into treatment, follow-up (no treatment) and borderline sub-groups according to age, gender, HBeAG status, serum ALT, HBVDNA levels defined in HBV guidelines published by AASLD, EASL, APASL and WGO. Liver histologies of all patients were evaluated and were grouped as treatment and follow-up groups according to the degree of histologic activity or stage of fibrosis. Results: Of the 574 patients 54(9.4%) had no inflammation, 206(35.9%) had minimal (HAI: 1–4), 141 (24.6%) had mild, 138 (24.1%) had moderate and 35 (6.1%) had severe inflammation. Fibrosis stages were 0 (no fibrosis) in 211 (36.8%), 1–2 in 303 (52.7%) and >2 in 60 (10.45%) patients according to modified Ishak’s method. Treatment was indicated in 157 (27.4%) of the patients according to liver biopsy. Distributions of patients according to treatment guidelines and histopathologic findings (Treatment indicated according to biopsy/treatment indicated according to serum markers) in liver biopsy is shown in table 1. Among the 347 patients with minimal-mild histologic activity 76 (21.9%) had HBVDNA between 2000–20,000 IU/ml and 134 (38.6%) had HBVDNA >20,000 IU/ml.

Defining high-fat-diet rat models: metabolic and molecular effects of different fat types

High-fat (HF)-diet rodent models have contributed significantly to the analysis of the pathophysiology of the insulin resistance syndrome, but their phenotype varies distinctly between different studies. Here, we have systematically compared the metabolic and molecular effects of different HF with varying fatty acid compositions. Male Wistar rats were fed HF diets (42% energy; fat sources: HF-L - lard; HF-O - olive oil; HF-C - coconut fat; HF-F - fish oil). Weight, food intake, whole-body insulin tolerance and plasma parameters of glucose and lipid metabolism were measured during a 12-week diet course. Liver histologies and hepatic gene expression profiles, using Affymetrix GeneChips, were obtained. HF-L and HF-O fed rats showed the most pronounced obesity and insulin resistance; insulin sensitivity in HF-C and HF-F was close to normal. Plasma omega-3 polyunsaturated fatty acid (omega-3-PUFA) and saturated fatty acid (C(12)-C(14), SFA) levels were elevated in HF-F and HF-C animals respectively. The liver histologies showed hepatic steatosis in HF-L, HF-O and HF-C without major inflammation. Hepatic SREBP1c-dependent genes were upregulated in these diets, whereas PPARalpha-dependent genes were predominantly upregulated in HF-F fed rats. We detected classical HF effects only in diets based on lard and olive oil (mainly long-chain, saturated (LC-SFA) and monounsaturated fatty acids (MUFA)). PUFA- or MC-SFA-rich diets did not induce insulin resistance. Diets based on LC-SFA and MUFA induced hepatic steatosis with SREBP1c activation. This points to an intact transcriptional hepatic insulin effect despite resistance to insulin's metabolic actions.