Comparison of the baseline independent predictive factors for fibrosis in the four cohorts of patients with chronic hepatitis C.

<p>Only factors found by univariate analysis to be significant in at least one of the four groups are reported.</p>*<p>Male as reference; § Not included in multivariate model because not significant in univariate analysis.</p><p>HCV, hepatitis C virus; BMI, body mass index; ALT, alanine aminotransferase; GGT, γ-glutamyl transpeptidase; OR, odds ratio; CI, confidence interval.</p

Univariate and multivariate analysis for fibrosis in the whole cohort of patients with chronic hepatitis C.

<p>Only factors found to be significant by univariate analysis are reported.</p>*<p>Male as reference. HCV, hepatitis C virus; BMI, body mass index; ALT, alanine aminotransferase; GGT, γ-glutamyl transpeptidase; OR, odds ratio; CI, confidence interval.</p

Estradiol and Testosterone serum levels and E2/T ratio in men and women divided according to women’s reproductive phases as described in Methods.

<p>Values (reported as mean±SD) were compared by Mann-Whitney test. The levels of significance are reported in the text.</p

A morphological method for ammonia detection in liver

<div><p>Hyperammonemia is a metabolic condition characterized by elevated levels of ammonia and a common event in acute liver injury/failure and chronic liver disease. Even though hepatic ammonia levels are potential predictive factors of patient outcome, easy and inexpensive methods aiming at the detection of liver ammonia accumulation in the clinical setting remain unavailable. Thus, herein we have developed a morphological method, based on the utilization of Nessler´s reagent, to accurately and precisely detect the accumulation of ammonia in biological tissue. We have validated our method against a commercially available kit in mouse tissue samples and, by using this modified method, we have confirmed the hepatic accumulation of ammonia in clinical and animal models of acute and chronic advanced liver injury as well as in the progression of fatty liver disease. Overall, we propose a morphological method for ammonia detection in liver that correlates well with the degree of liver disease severity and therefore can be potentially used to predict patient outcome.</p></div

Hepatic ammonia staining in patients with Non-Alcoholic Fatty Liver Disease (NAFLD).

<p>Representative micrographs of H&E and ammonia staining in liver samples from NAFLD patients with low (<2) and high (≥2) Ammonia Score (AS).</p

Hepatic ammonia staining in mouse models and human samples of acute and chronic liver injury.

<p>Representative micrographs of H&E and ammonia staining in liver samples in bile-duct ligation (BDL) mouse model of liver injury (n = 6 Ctrl, n = 6 BDL 10 days and, n = 4 BDL 14 days, *p<0.05 vs. Ctrl) <b>(A)</b>, in the transgenic mouse model of cholestatic disease, the Mdr2^-/- mouse (n = 6 Mdr2^+/+ and n = 6 Mdr2^-/-, *p<0.05) <b>(B)</b> and, in acetaminophen-induced liver injury in mouse (n = 6 Ctrl, n = 8 Acetaminophen, *p<0.05) <b>(C)</b>. Serum ammonia both in BDL and in the Mdr2 mouse models are shown, *p<0.05 is indicated <b>(D)</b>. Finally, representative micrographs of H&E and ammonia staining in the liver of cirrhotic patients (n = 6 healthy and n = 6 Cirrhosis, *p<0.05) <b>(E)</b> and in idiosyncratic drug-induced liver injury (DILI) patients (n = 5 healthy, n = 5 DILI and n = 3 Acute DILI, *p<0.05 Acute DILI vs. healthy) <b>(F)</b> are shown.</p

Hepatic ammonia staining in mouse models of Non-Alcoholic Fatty Liver Disease (NAFLD).

<p>Representative micrographs of Sudan Red, Sirius Red, F4/80 and ammonia staining in liver samples from high-fat (HF) diet (n = 6 Ctrl vs. n = 7 HF diet), (A) and choline deficient with 0.1% methionine diet (MCDD)-fed rodents (n = 6 Ctrl vs. n = 5 MCDD 2 weeks and n = 5 MCDD 6 weeks) (B). AS was only significantly increased in MCDD 6 weeks vs. Ctrl, p<0.05.</p

Representative micrographs of ammonia staining in healthy mouse tissue.

<p>Representative micrographs of ammonia staining in healthy mouse tissues in samples included in O.C.T <b>(A)</b> and Pearson correlation of the quantification of ammonia in biological tissues between the novel proposed method and the commercial available colorimetric kit <b>(B)</b>. Representative micrographs of ammonia staining in healthy mouse tissues in samples included in paraffin <b>(C)</b>. At least triplicates were used for each sample.</p

Failure rates following the first DAA regimen, by HCV genotype and treatment regimen in patients who completed the 12 weeks post treatment evaluation (n = 3,830 patients).

<p>Failure rates following the first DAA regimen, by HCV genotype and treatment regimen in patients who completed the 12 weeks post treatment evaluation (n = 3,830 patients).</p

Univariate and logistic regression analysis linking failure with independent variables.

<p>Univariate and logistic regression analysis linking failure with independent variables.</p