Expression Levels of the Uridine-Cytidine Kinase Like-1 Protein As a Novel Prognostic Factor for Hepatitis C Virus-Associated Hepatocellular Carcinomas.

The expression levels of the two novel oncoproteins uridine-cytidine kinase like-1 (UCKL-1) and mitochondrial ribosomal protein S18-2 (MRPS18-2) were assessed in samples of hepatitis C virus (HCV)-associated hepatocellular carcinoma (HCC) using immunohistochemistry. Tissue microarray (TMA) paraffin blocks were prepared from 42 HCC tumor samples with the corresponding peri-tumor tissues and from 11 tissues of a liver with HCV-induced cirrhosis. We found that the UCKL-1 signal in the liver tissues of the peri-tumor zone in the HCC samples was stronger than that in cirrhosis (50 ± 49.44 vs. 24.27 ± 14.53; p = 0.014). The MRPS18-2 expression was weak, and there was no differences between the groups (p = 0.26). Noteworthy, the UCKL-1 protein was expressed at higher levels in peri-tumor tissues in the cases of HCC recurrence; this was confirmed for 27 older patients (63.78 ± 9.22 vs. 53.53 ± 4.07 years, p < 0.001), in parallel with enhanced UCKL-1 staining in former HCC nodules (62.69 ± 50.4 vs. 26.0 ± 30.19, p = 0.006) and microvascular invasion (p = 0.02). A multivariate analysis of prognostic factors for HCC recurrence showed that the best predictive factors for these conditions were UCKL-1 expression in tumor, vascular invasion, and HCC treatment modality, other than liver transplantation (odds ratios: 1.029, 18.143 and 11.984, R2 = 0.633, p = 0.002). In conclusion, the high UCKL-1 expression might be a prognostic factor for HCC relapse, in combination with age and microvascular invasion. MRPS18-2 protein expression has no prognostic significance in the cases of HCV-associated HCC.

Strategies to manage hepatitis C virus infection disease burden - volume 3.

The hepatitis C virus (HCV) epidemic was forecasted through 2030 for 15 countries in Europe, the Middle East and Asia, and the relative impact of two scenarios was considered: increased treatment efficacy while holding the annual number of treated patients constant and increased treatment efficacy and an increased annual number of treated patients. Increasing levels of diagnosis and treatment, in combination with improved treatment efficacy, were critical for achieving substantial reductions in disease burden. A 90% reduction in total HCV infections within 15 years is feasible in most countries studied, but it required a coordinated effort to introduce harm reduction programmes to reduce new infections, screening to identify those already infected and treatment with high cure rate therapies. This suggests that increased capacity for screening and treatment will be critical in many countries. Birth cohort screening is a helpful tool for maximizing resources. Among European countries, the majority of patients were born between 1940 and 1985. A wider range of birth cohorts was seen in the Middle East and Asia (between 1925 and 1995).

Association of the prevalence and grade of steatosis in patients with chronic hepatitis C with the host and viral factors.

AIM: The aim of this study was to investigate the prevalence of hepatosteatosis in chronic hepatitis C patients, evaluate the potential impact of some host- and virus-related factors on its occurrence and possible influence of steatosis on the consequences of hepatitis. PATIENTS AND METHODS: The case records of 387 patients with hepatitis C and cirrhosis were studied. The prevalence and grade of steatosis were investigated and evaluated by logistic regression analysis as dependent variable to age, gender, alcohol consumption, body mass index, hepatitis C virus (HCV) genotypes, liver enzymes activity, histological activity index and fibrosis. RESULTS: Steatosis was found in 47.3% of the patients. It was more prevalent in males, alcohol abusers, overweight and obese patients, and in those with HCV genotypes 3 and 2. Multivariate analysis confirmed body mass index as an independent risk factor for steatosis in the overall patient cohort and in those with genotypel without any correlation with the steatosis grade. The prevalence and grade of steatosis were associated with alcohol consumption and higher fibrosis stage. The age of the patients showed converse association. CONCLUSIONS: The male gender, body mass index, alcohol consumption, genotype 2 and 3 were confirmed as risk factors for hepatosteatosis. Older patients had a lesser steatosis grade. The correlation of histological activity index and fibrosis scores with the prevalence and higher grade of steatosis suggested a possibility to worsen the course of hepatitis C and to accelerate disease progression.

Acetazolamide inhibits stimulated feline liver and gallbladder bicarbonate secretion.

Bile acidification is a key factor in preventing calcium carbonate precipitation and gallstone formation. Carbonic anhydrase II (CA II), that is inhibited by acetazolamide, plays a role in regulation of the acid-base balance in many tissues. This study examines the effect of acetazolamide on secretin- and vasoactive intestinal peptide (VIP)-stimulated gallbladder mucosal bicarbonate and acid secretion. Gallbladders in anaesthetized cats were perfused with a bicarbonate buffer bubbled with CO2 in air. In 20 experiments VIP (10 microg kg(-1) h(-1)) and in 10 experiments secretin (4 microg kg(-1) h(-1)) were infused continuously intravenous (i.v.). Hepatic bile and samples from the buffer before and after perfusion of the gallbladder were collected for calculation of ion and fluid transport. During basal conditions a continuous secretion of H+ by the gallbladder mucosa was seen. Intravenous infusion of vasoactive intestinal peptide (VIP) and secretin caused a secretion of bicarbonate from the gallbladder mucosa (P < 0.01). This secretion was reduced by intraluminal (i.l.) acetazolamide (P < 0.01). Bile flow was enhanced by infusion of VIP and secretin (P < 0.01) but this stimulated outflow was not affected by i.v. acetazolamide. The presence of CA II in the gallbladder was demonstrated by immunoblotting. Biliary CA activity has an important function in the regulation of VIP- and secretin-stimulated bicarbonate secretion across the gallbladder mucosa.

Multispecificity of Na+-dependent taurocholate uptake in basolateral (sinusoidal) rat liver plasma membrane vesicles.

To test the hypothesis of broad specificity of the hepatocellular bile acid uptake system(s) we investigated the kinetics and substrate specificity of Na+-dependent taurocholate uptake in basolateral (sinusoidal) rat liver plasma membrane vesicles in the presence and absence of bovine serum albumin. Bovine serum albumin selectively stimulated the Na+-dependent portion of taurocholate uptake and decreased its apparent Km from 46 +/- 6 to 17 +/- 3 microM, whereas it had no effect on Vmax (4.2 +/- 0.2 nmol.mg-1.min-1). Based on complementary analysis by Dixon- and Cornish-Bowden-plots the following compounds were identified as competitive inhibitors of Na+-dependent taurocholate uptake: cholate (Ki = 140 +/- 30 microM); taurochenodeoxycholate (Ki = 9 +/- 3 microM); chenodeoxycholate (Ki = 53 +/- 6 microM); progesterone (Ki = 110 +/- 30 microM); 17-beta-estradiol-3-sulfate (Ki = 28 +/- 4 microM); bumetanide (Ki = 440 +/- 85 microns); furosemide (Ki = 460 +/- 140 microM); verapamil (Ki = 65 +/- 35 microM); and phalloidin (Ki = 850 +/- 350 microM). In contrast, noncompetitive inhibition was found with bromosulfophthalein (Ki = 12 +/- 2 microM), cyclosporin A (Ki = 3 +/- 1 microM) and 4,4'-diisothiocyano-2,2'-disulfonic acid stilbene (Ki = 45 +/- 7 microM). These results support the concept of multispecificity of the Na+-dependent basolateral bile acid uptake system with respect to different bile acids and drugs. In addition, the findings provide further evidence for bile acids and bromosulfophthalein being taken up into rat hepatocytes by different transport systems, thus supporting the assumption of multiple basolateral organic anion "carriers" with distinct, yet partially overlapping substrate specificities.

Bicarbonate sulfate exchange in canalicular rat liver plasma membrane vesicles.

The mechanism(s) and driving forces for biliary excretion of sulfate were investigated in canalicular rat liver plasma membrane vesicles (cLPM). Incubation of cLPM vesicles in the presence of an inside-to-outside (in, out) bicarbonate gradient (50 mM in, 5 mM out, pH 8.0 in and out), but not pH (pH 8.0 in, 6.0 out) or out-to-in sodium gradients, stimulated sulfate uptake 10-fold compared with the absence of bicarbonate and approximately 2-fold above sulfate equilibrium ("overshoot"). Initial rates of this bicarbonate gradient-driven sulfate uptake were saturable with increasing concentrations of sulfate (apparent Km, approximately 0.3 mM) and could be inhibited by probenecid, N-(4-azido-2-nitrophenyl)-2-aminoethylsulfonate, acetazolamide, furosemide,4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid, and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (IC50, approximately 40 microM). Cisinhibition of initial bicarbonate gradient-stimulated sulfate uptake and transstimulation of sulfate uptake in the absence of bicarbonate were observed with sulfate, thiosulfate, and oxalate but not with chloride, nitrate, phosphate, acetate, lactate, glutamate, aspartate, cholate, taurocholate, dehydrocholate, taurodehydrocholate, and reduced or oxidized glutathione. These findings indicate the presence of a sulfate (oxalate)-bicarbonate anion exchange system in canalicular rat liver plasma membranes. In conjunction with the previously reported chloride-bicarbonate exchanger (J. Clin. Invest. 75: 1256-1263, 1985), these findings support the concept that bicarbonate-sensitive transport system might play an important role in bile acid-independent canalicular bile formation.