Digital Pathology Enables Automated and Quantitative Assessment of Inflammatory Activity in Patients with Chronic Liver Disease

Traditional histological evaluation for grading liver disease severity is based on subjective and semi-quantitative scores. We examined the relationship between digital pathology analysis and corresponding scoring systems for the assessment of hepatic necroinflammatory activity. A prospective, multicenter study including 156 patients with chronic liver disease (74% nonalcoholic fatty liver disease-NAFLD, 26% chronic hepatitis-CH etiologies) was performed. Inflammation was graded according to the Nonalcoholic Steatohepatitis (NASH) Clinical Research Network system and METAVIR score. Whole-slide digital image analysis based on quantitative (I-score: inflammation ratio) and morphometric (C-score: proportionate area of staining intensities clusters) measurements were independently performed. Our data show that I-scores and C-scores increase with inflammation grades (p < 0.001). High correlation was seen for CH (ρ = 0.85–0.88), but only moderate for NAFLD (ρ = 0.5–0.53). I-score (p = 0.008) and C-score (p = 0.002) were higher for CH than NAFLD. Our MATLAB algorithm performed better than QuPath software for the diagnosis of low-moderate inflammation (p < 0.05). C-score AUC for classifying NASH was 0.75 (95%CI, 0.65–0.84) and for moderate/severe CH was 0.99 (95%CI, 0.97–1.00). Digital pathology measurements increased with fibrosis stages (p < 0.001). In conclusion, quantitative and morphometric metrics of inflammatory burden obtained by digital pathology correlate well with pathologists’ scores, showing a higher accuracy for the evaluation of CH than NAFLD.

Reliability of Transient Elastography as a non-Invasive Technique for Detection of Fibrosis in Budd Chiari Syndrome Patients after Endovascular Intervention

Aim of the work To test the reliability of fibroscan in detection of fibrosis in patients with Budd Chiari syndrome before and after endovascular intervention (after elimination of hepatic congestion). Background transient elastography (TE) is a noninvasive methodology that has been used to monitor liver stiffness in patients with chronic viral hepatitis. One of the limitations for accurate assessment of liver fibrosis by TE is the liver congestion. Liver congestion can result from Budd Chiari syndrome (BCS).The treatment of BCS is through restoring the flow of the blood between the portal vein to the inferior vena cava, which will lead to decongestion of the liver.TE, will be tested after liver decongestion for proper detection of liver fibrosis. Patients and methods This was a prospective cohort study conducted on 25 Egyptian patients with confirmed diagnosis of primary Budd-Chiari Syndrome (BCS) in the period from June 2017 to September 2019. TE was performed three days before endovascular intervention and three months after it. Liver biopsy was taken during the intervention for assessment of METAVIR score. Comparison was done between TE assessments before and after intervention in detection of the degree of liver fibrosis in comparison to METAVIR score measured in liver biopsy. Results FVLM was the most common hypercoagulable cause in the involved patients. There was significant drop in Liver Stiffness Measurements (LSM) measured three months post-intervention indicating improvement of liver fibrosis after relieving liver congestion but still not correlated to the METAVIR scores measured in the liver biopsy. Conclusion Liver congestion has high impact on Liver stiffness measurement giving overestimation which improves significantly after decongestion of the liver by the endovascular intervention.

Follow-Up of Liver Stiffness with Shear Wave Elastography in Chronic Hepatitis C Patients in Sustained Virological Response Augments Clinical Risk Assessment

This study aimed to observe the effect of the direct-acting antiviral (DAA) therapy on liver stiffness (LS) and serum biomarkers. We prospectively observed 35 patients with chronic hepatitis C infection and attained a sustained virological response (SVR) after antiviral therapy. Shear wave elastography (SWE) measurement was performed at the beginning of DAA treatment and at 48 weeks after the end of treatment (EOT48w). The METAVIR score and the score for varices needing treatment (VNT) were determined based on the LS values; the fibrosis-4 (FIB4) score was calculated from laboratory tests. The baseline LS (mean ± standard deviation = 2.59 ± 0.89 m/s) decreased significantly after successful DAA therapy (1.90 ± 0.50 m/s; p < 0.001). The METAVIR score showed significant improvement at EOT48w (F0/1 = 9, F2 = 2, F3 = 10, F4 = 14) compared to the initial status (F0/1 = 2, F2 = 1, F3 = 7, F4 = 25; p < 0.028). The FIB4 score indicated less fibrosis after therapy (2.04 ± 1.12) than at baseline (3.51 ± 2.24; p < 0.018). Meanwhile, the number of patients with a high-risk of VNT was significantly less at EOT48w (4 vs. 15 at baseline; OR = 0.17 95% confidence interval (CI) = 0.05–0.59, p < 0.007). SWE indicates a significant resolution of liver fibrosis when chronic hepatitis C patients are in SVR, coinciding with a lower risk of VNT.

A Nomogram Predicting the Prognosis of Children With Biliary Atresia After Hepatoportoenterostomy

Background: Although Kasai portoenterostomy (KPE) is performed timely for most children with biliary atresia (BA), the native liver survival (NLS) is still poor due to the progressive liver fibrosis. Many children have to receive liver transplantation (LT) within 2 years after KPE. Early prediction of the prognosis permits the implementation of prophylactic treatments for BA children. However, studies about the prediction are limited.Objective: The purpose of this study is to establish a nomogram to predict the prognosis of BA children within 2 years after KPE.Methods: The follow-up data of 151 BA children were retrospectively reviewed, and were randomly divided into a training cohort for constructing a nomogram (n = 103) and a validation cohort (n = 48). In the training cohort, patients were divided into Group A and Group B according to whether death or LT were observed within 2 years post-KPE. Multivariate Cox regression based on the baseline characteristics, liver function indicators and LSM (liver stiffness measurement) values at KPE and 3 months after KPE was utilized for the establishment of the nomogram in predicting the prognosis of BA within 2 years after KPE. The discrimination and calibration of the nomogram were internally and externally validated.Results: Fifty-six BA children were included in Group A and 47 were included in group B. Age at KPE, METAVIR score F4, LSM at 3 months, first onset of cholangitis within 3 months, and jaundice clearance time were the independent predictors for the prognosis of BA children within 2 years after KPE (all P &lt; 0.05). The developed nomogram based on these independent predictors showed good discrimination and calibration by the internal and external validation. Its performance was better than each predictor in predicting the prognosis (all P &lt; 0.05).Conclusions: The established nomogram based on the indicators from the first 3 months after KPE may be useful for predicting the prognosis of BA children within 2 years post-KPE and helpful for the consideration of LT.

Liver Disease Detection Using Grey Wolf Optimization and Random Forest Classification

Utilizing machine learning approaches as non-obtrusive strategies is an elective technique in organizing perpetual liver infections for staying away from the downsides of biopsy. This chapter assesses diverse machine learning methods in expectation of cutting-edge fibrosis by joining the serum bio-markers and clinical data to build up the order models. An imminent accomplice of patients with incessant hepatitis C was separated into two sets—one classified as gentle to direct fibrosis (F0-F2) and the other ordered as cutting-edge fibrosis (F3-F4) as per METAVIR score. Grey wolf optimization, random forest classifier, and decision tree procedure models for cutting-edge fibrosis chance expectation were created. Recipient working trademark bend investigation was performed to assess the execution of the proposed models.

NOX1 Inhibition Attenuates the Development of a Pro-Tumorigenic Environment in Experimental Hepatocellular Carcinoma

Background The poor prognosis of advanced HCC and limited efficacy of current systemic treatments emphasize the need for new or combined targeted therapies. The development of HCC is a multistage process in which liver injury appears in a complex microenvironment associated with oxidative stress. NOX enzymes are the main source of ROS during hepatocarcinogenesis and NOX1 in particular has shown correlation with poor prognosis of HCC patients. This study evaluates the effect of pharmacological NOX1 inhibition on the development and progression of HCC and its effect on the tumor microenvironment.Methods The in vitro cytotoxic effects of the NOX1 inhibitor GKT771 (Genkyotex) on human Huh7 and Hep3B and murine Hepa1-6 HCC cell lines, and murine macrophages were evaluated via MTT, LDH activity and CaspGlo® assays. In order to induce in vivo HCC, male SV129 wild-type mice received weekly IP injections of diethylnitrosamine (DEN) (35 mg/kg) for 20-25 weeks. Mice were treated with vehicle or GKT771 (30 mg/kg) via oral gavage. Treatment duration and frequency (daily or twice daily) varied in the preventive and therapeutic studies. mRNA transcript levels in the tumor and liver tissue were determined by RT-qPCR. Fibrosis was visualized by Sirius Red staining and quantified by the Metavir​ score. Results A concentration-dependent reduction in cellular activity of the human HCC cell lines without cytotoxicity was observed. GKT771 treatment reduced LPS-induced pro-inflammatory bone-marrow derived macrophage polarization. DEN injections resulted in 100% tumor formation and the induction of HCC markers which could be reduced by twice daily dosing of GKT771 at early onset of advanced HCC. DEN-induced HCC resulted in an upregulation of pro-inflammatory, angiogenic and fibrotic markers which was less pronounced in GKT771 treated mice in all treatment regimens. In line, liver fibrosis was induced in HCC mice and this to a lesser extend upon GKT771 treatment.Conclusion NOX1 inhibition showed to be safe and well tolerated and was able to attenuate the induction of a pro-inflammatory, angiogenic and pro-fibrotic microenvironment suggesting that this might be a promising adjuvant therapeutic strategy in the treatment of advanced HCC.

Hepatoprotective effect of Linagliptin against liver fibrosis induced by carbon tetrachloride in mice

The current study aimed to investigate linagliptin for its potential role in the prevention of liver fibrosis progression. Balb-C mice were randomly allocated into 5 groups (10 each), (1) control, (2) mice were injected intraperitoneally with 50 μl carbon tetrachloride (CCl4) in corn oil in a dose of 0.6 μl /g 3 times a week for 4 weeks, (3) linagliptin was orally administered in a dose of 10 mg/kg/day simultaneously with CCl4, (4) silymarin was orally in a dose of 200 mg/kg/day concomitantly with CCl4, (5) linagliptin only. Hepatic injury was manifested in CCl4 group by elevation of biochemical parameters; alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), hepatic fibrosis was evident histopathologically by increased METAVIR score and immunostaining expression of α-smooth muscle actin (α-SMA), also, increased liver tissue oxidative stress parameters, transforming growth factor-β1 (TGF-β1) and mammalian target of rapamycin (mTOR). Linagliptin was able to stop the progression of liver fibrosis; evident histopathologically with reduced METAVIR score and α-SMA expression. The possible mechanism may be via suppression of oxidative stress, TGF-β1, and mTOR, this was associated with improvement of serum biochemical parameters; ALT and AST. In conclusion, linagliptin might help to protect the liver against persistent injury-related consequences.

The Enhanced Liver Fibrosis Index Predicts Hepatic Fibrosis Superior to FIB4 and APRI in HIV/HCV Infected Patients

Background Accurate noninvasive biomarkers of fibrotic progression are important for hepatitis C virus (HCV) management, but commonly used modalities may have decreased efficacy in human immunodeficiency virus (HIV)/HCV-coinfected persons. The enhanced liver fibrosis (ELF) index is a highly sensitive noninvasive marker of hepatic fibrosis that has had limited assessment in the HIV/HCV population. We compared ELF index performance to FIB4 and aspartate to platelet ratio index (APRI) at different stages of liver fibrosis as determined by liver histology, and validated the efficacy of the three noninvasive biomarkers in HIV/HCV-coinfected versus HCV-monoinfected. Methods The ELF index was determined in 147 HIV/HCV-coinfected and 98 HCV-monoinfected persons using commercial ELISA assays for the component elements of the index. Area under the receiver-operator curve was used to validate ELF and to compare its performance to liver histology as well as to other noninvasive biomarkers of liver fibrosis, FIB4, and APRI. Results The ELF index increased with histological stage of liver fibrosis and exhibited a linear relationship with Metavir score in all subjects. ELF performance was comparable between HIV/HCV and HCV with advanced liver fibrosis/cirrhosis. In the HIV/HCV cohort ELF cutoffs of 8.45 and 9.23 predicted mild and moderate fibrosis with 85% sensitivity, whereas the ELF cutoff of 9.8 had the highest specificity for advanced fibrosis and the cutoff of 10.4 was 99% specific for cirrhosis. ELF performance was superior to FIB4 and APRI in all subjects regardless of HIV status. Conclusions ELF index demonstrated excellent characteristics toward accurate prediction of liver fibrosis and cirrhosis with superior performance to APRI and FIB4 in HIV/HCV coinfection. Applying this noninvasive biomarker index for diagnosis of liver fibrosis and progression in HIV/HCV is warranted.

Liver Fibrosis Assessment in a Cohort of Greek HIV Mono-Infected Patients by Non-Invasive Biomarkers

Background: Non-alcoholic Fatty Liver Disease (NAFLD) is common in HIV-infected individuals. Liver biopsy remains the gold-standard procedure for the diagnosis of liver fibrosis, but both Transient Elastography (TE) and Non-invasive Biomarkers (NIBMs) have emerged as alternatives. Objectives: Our study’s aim was to validate commonly used NIBMs for the assessment of liver fibrosis in a cohort of Greek HIV-mono-infected patients. Methods: Inclusion criteria were confirmed HIV-infection and age>18 years and exclusion criteria HBV or HCV seropositivity, liver disease other than NAFLD, alcohol abuse, ascites, transaminases levels>4xULN(upper limit of normal) and Body-Mass index(BMI)>40. Liver stiffness (LS) measurement with TE and thorough laboratory work up and medical history were acquired at study entry. FIB-4, APRI, NFS, BARD, Forns and Lok scores were calculated for each patient. Results: A total of 157 patients were eligible for this study. Significant liver fibrosis, compatible with Metavir score of F3-F4, was found in only 11(7%) patients. These findings were in accordance with those of the NIBMs; the BARD score constituting the only exception, allocating 102(65%) patients as having significant liver fibrosis. In order to obtain a balance between sensitivity and specificity new cut-offs for each NIBM were calculated; FIB-4 score yielded the best results, since by changing the cut-off to 1.49 a sensitivity and specificity balanced for both close to 85% was achieved. Conclusions: Our findings suggest that NIBMs can be used for the evaluation of liver fibrosis in HIV mono-infected patients. New cut-offs for NIBMs should probably be calculated, to help distinguishing patients with significant from those with mild/no fibrosis.