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The impact of the HCV genotype 1b core amino acid (aa) 70 mutant on the cumulative rate of hepatocellular carcinoma following eradication of HCV RNA by antiviral therapy was investigated with the Q-Invader assay. Multivariate analysis based on 649 patients indicated that a core aa70 Q-Invader mutant level ≥20% is a predictor of hepatocellular carcinoma.

We present the case of a chronic hepatitis C (CHC) patient who was originally diagnosed with genotype 2a on serogroup 1 CHC genotype testing, but who was subsequently confirmed to have genotype 1b when using the hepatitis C virus (HCV) monitor genotype assay. The genotype 2a diagnosis was attributed to the fact that the type 2a-specific primer used in Okamoto et al.'s method (HCV genotype primer kit) has relatively high homology, which caused the amplification reaction to proceed, rendering a HCV RNA genotype test result of 2a. Genotype testing is important in determining whether a patient is indicated for concomitant dual oral therapy; however, the potential for different diagnoses such as described in this report highlights the importance of serogroup confirmation.

Background The aim of the study was to analyse the predictive host and viral factors of sustained virological response (SVR) in Estonian patients with chronic hepatitis C genotype 1b.

Even when treated with telaprevir-based triple therapy, some patients fail to achieve a sustained virological response. This study identified factors related closely to treatment failure. A total of 146 Japanese genotype 1b chronic hepatitis C patients were enrolled in this prospective, multicenter study and received a 24-week regimen of triple therapy. The end-of-treatment response rate was significantly lower in patients with the interleukin 28B (IL28B) (rs8099917) non-TT genotype (85.2%) than in those with the TT genotype (100%, P = 0.0002). Multiple logistic regression analysis identified high α-fetoprotein levels as an independent factor related to non-end-of-treatment response in patients with the non-TT genotype. A cut-off value of 20 ng/ml was determined for a non-end-of-treatment response; sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy were 75.0%, 95.7%, 75.0%, 75.0%, and 92.6%, respectively. Multiple logistic regression analysis for a sustained virological response identified the IL28B TT genotype, low α-fetoprotein levels, non-responders, and a rapid virological response. The sustained virological response rate was significantly lower in patients with the non-TT genotype (59.3%) than in those with the TT genotype (96.7%, P

Chronic hepatitis C patients display many genotype-specific clinical features of HCV infection. The core proteins encoded by different genotypes dysregulate numerous sets of distinct host genes. In this study we tested the hypothesis that HCV core proteins 1b and 3a would actually act on a limited number of independent cellular players, as well as on several functionally linked gene products. Structural and functional tests identified a core set of host genes dysregulated by HCV core genotypes 1b and 3a. The core proteins of HCV genotypes 1b and 3a target specifically limited sets of functionally related gene products, which may be responsible for the variations in the clinical spectra associated with HCV infection.

The Interferon (IFN) which is the standard treatment for Hepatitis C, may cause a lot of side effects including dermatological anomalies. This paper presents a psoriasis case which occurred in relation with the treatment of acute hepatitis C (AHC) with peginterferon alfa (peg-IFN-α). A 60-year-old male patient came to the hospital with symptoms of high liver enzymes. The patient with history of a recent operation showed anti-HCV (+), HCVRNA 3.5 million IU/mL and HCV genotype 1b in the tests. Without any other etiological factors found in the patient, we started a treatment of peg-IFNα-2b with the diagnosis of AHC. After 3 weeks, psoriatic plaques were observed in various parts of the body. Antiviral treatment of the patient was concluded within 6 months. His psoriasis treatment initially commenced with local agents followed by phototherapy. Permanent viral response was seen in the patient and his lesions recovered rapidly after the antipsoriatic and antiviral treatment. Psoriasis and other autoimmune diseases should be considered even though they are encountered rarely,and the patients should be informed of the possible risks before planning treatment with peg-IFN-α.

Aim Initial hepatitis C virus (HCV) RNA reduction was investigated as a potential index for sustained virological response (SVR) in the treatment of interferon (IFN)-β followed by peginterferon plus ribavirin (PEG IFN/RBV). Methods The treatment course was retrospectively analyzed in 64 genotype 1b patients with a HCV RNA level of 5.0 logIU/mL or higher. IFN-β was administrated twice a day for 2 weeks followed by 24 or 48 weeks of PEG IFN/RBV. The serum HCV RNA level was measured by real-time polymerase chain reaction before administration and at 1, 2 and 4 weeks of therapy. Results By the duration of PEG IFN administration, the SVR rates were 11% (2/18

The prevalence of HCV infection throughout Asia is variable and complex compared with United States and European. Genotype 1 to 6 could be found among different countries in Asia. Before 2000, 1b is the most prevalent genotype and other multi genotypes and subtypes in eastern Asia. The overall profile demonstrated genotype 1b decreased and genotype 2a increased gradually from north to south, from west to east. Since 2000, more genotypes and subtypes were found in Asia, then Asia turned out to be an area with the most complex genotypes distribution, all genotypes, genotype 1 through genotype 6 could be found. We can still find the trend that genotype 1b decreased and genotype 2a increased gradually from north to south, from west to east. In addition, genotype 3 increased from north to south. Meanwhile, genotype 3 occurred in more countries and areas, genotype 4 was mostly in middle east, genotype 5 was reported in Pakistan, India, Syria and Saudi Arab. Genotype 6 was mainly in Thailand and Vietnam. Genotype 1b was associated with high risk for HCC. In Asia, most patients were with favorable host genotype in IL28B.

Hepatitis C is a global health problem with a worldwide prevalence of about 3% (around 170 million people). Hepatitis C virus (HCV) is major concern in the Maghreb countries, Algeria, Libya, Mauritania, Morocco, and Tunisia, but no detailed description of its epidemiology in the region is available. In the present review, a systematic search was undertaken covering HCV data available in peer-reviewed databases as well as institutional reports and regional conference meeting abstracts from the Maghreb countries. Reports written in English and French were included in this analysis. Estimates of national and regional prevalence of HCV infection (based on anti-HCV antibody) and of the size of patient populations were performed. In addition, the molecular features of the circulating viral strains in the region are discussed. A substantial proportion, 1.2-1.9% of the Maghreb inhabitants, have anti-HCV antibodies. Genotype 1b predominates among viral strains in all countries except in Libya, where genotype 4 is dominant as in neighboring Egypt. This epidemiological situation is of significant concern, and requires urgent, broad, and active intervention for the prevention and control of HCV. More specifically, the application of state-of-the-art hygiene procedures and rigorous controls in medical disciplines such as hemodialysis, transfusion, endoscopic procedures, and dentistry is necessary to reduce significantly the number of new infections in the region.

Uyanikoglu, Ahmet/0000-0003-4881-5244 WOS: 000322057500012 PubMed: 23898387 Background/Aims: The aim of this study is to investigate the rate of sustained virologic response (SVR) in chronic hepatitis C patients receiving antiviral treatment. Methods: The files of patients with chronic hepatitis C treated with interferon ribavirin between 1995 and 2009 were reviewed retrospectively. Six months after the end of treatment, patients with negative hepatitis C virus (HCV)-RNA (

A series of symmetrical E-stilbene prolinamides that originated from the library-synthesized lead 3 was studied with respect to HCV genotype 1a (G-1a) and genotype 1b (G-1b) replicon inhibition and selectivity against BVDV and cytotoxicity. SAR emerging from an examination of the prolinamide cap region revealed 11 to be a selective HCV NS5A inhibitor exhibiting submicromolar potency against both G-1a and G-1b replicons. Additional structural refinements resulted in the identification of 30 as a potent, dual G-1a/1b HCV NS5A inhibitor.

A high throughput screen of the Pfizer compound collection was carried out using a hepatitis C virus (HCV) genotype 1b subgenomic replicon cell line. Those confirmed hits that demonstrated broad spectrum activity without overt cytotoxicity were further evaluated, leading to the identification of a series of pyrrolopyridines with excellent antiviral activity in a fully infectious HCV cell-based assay and pharmacokinetic properties.

The presented study had two objectives. The first was to examine distributions of Hepatitis C Virus (HCV) genotypes in Sindh, Pakistan, where HCV is prevalent. The other was to explore clinically relevant relationships between the genotypes, viral load (measured by real-time polymerase chain reaction assays) and biochemical markers. For this, 1471 HCV-infected patients in six cities in Sindh were recruited and sampled. HCV genotype distributions varied among the cities, but genotype 3a was most prevalent, followed by 3b, 1a and 1b (detected in 51.5, 22.7. 9.25 and 3.2% of the cases, respectively). No type-specific sequences were detected in serum samples from 189 (12.8%) of the 1471 patients. Frequencies of low (600,000 IU/mL serum) viral loads were respectively 45.4, 16.5 and 38.1% for patients infected with genotype 3, and 16.9, 36.9 and 46.2%, respectively, for patients with other genotypes. Infection with genotype 1a was associated with significantly higher (p < 0.005) alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase titers than infection with genotype 3a. The results will help in the formulation of treatment strategies.

Objective: Highly variable genome of HCV and high prevalence in many geographical areas made it necessary to conduct local population studies. This study has been conducted to show HCV parameters along with clinical features in the local population of interior Sindh, province of Pakistan. Methods: Present study was conducted in from August 2010 to November 2015 in the rural areas of Sindh, Pakistan. All the 31560 screened samples selected for the study were tested by second Generation Enzyme Linked Immunosorbent Assay (ELISA Biokit 480&96). Results: Total 31560 people were screened for HCV and out of these 13.67% (n= 4314) HCV infected patients. When 4314 samples of patients were examined; the anti-HCV was significantly higher in males 2814 (14.98%) than in females 1500 (11.74%) with P value = 0.06. The age of the patients ranged from 18 to 65 years. Out of 4314 HCV samples, 3020 (70%) were of Genotype 3a, 237(5.5%) of Genotype 2a, 108 (2.5%) of Genotype- 1a, 216 (5%) of Genotype 1b, 237 (5.5%) of Genotype 3b and 43 (1%) of Genotype 4. Additionally, 108 (2.5%) had co-infection and 345 (8%) samples showed no result –designated as untypable by the genotyping. Conclusion: This study showed that HCV is most frequently reported disease with genotype 3a being the most prevalent genotype.

Background Despite the high prevalence of genotype 1b hepatitis C virus (HCV) among patients, a cell culture system that permits entire viral life cycle of genotype 1b isolates is limited. To develop a cell-cultured hepatitis C virus (HCVcc) of genotype 1b, the proper combination of HCV genomic variants and host cells is essential. HCV genomes isolated from patients with distinctive symptoms may provide the variants required to establish an HCVcc of genotype 1b. Results We first established subgenomic replicons in Huh7 cells using HCV cDNAs isolated from two patients: one with fulminant hepatitis after liver transplantation (TPF1) and another with acute hepatitis and moderate symptoms (sAH). Replicons established from TPF1 and sAH showed mutations in NS4B and in NS3 and NS5A, respectively. Using these replication machineries, we constructed HCV genomic RNAs for each isolate. Virus infectivity was evaluated by a focus-forming assay, which is dependent on the intracellular expression of core antigen, and production of virus particles was assessed by density-gradient centrifugation. Infectious virus was only observed in the culture medium of cells transfected with TFP1 HCV RNA. A chimeric genome with the structural segment (5′-untranslated region [UTR] through NS2) from sAH and the replication machinery (NS3 through 3′-UTR) from TPF1 exhibited greater infectivity than did TFP1, despite formation of deficient virus particles in sAH, suggesting that this genomic segment potentiates virus particle formation. To identify the responsible variants, infectious virus formation was assessed in a chimeric genome carrying parts of the sAH structural segment of the TPF1 genome. A variant in NS2 (M170T) was identified that enhanced infectious virus formation. HCVcc carrying an NS2 gene encoding the M170T substitution and adaptive mutations in NS4B (referred to as TPF1-M170T) infected naïve cured Huh7 cells in a CD81-dependent manner. Conclusions We established a novel HCVcc of genotype 1b in Huh7 cells by introducing an amino acid variant in NS2 and adaptive mutations in NS4B from HCV genomic RNA isolated from a patient with fulminant HCV after liver transplantation. Electronic supplementary material The online version of this article (doi:10.1186/s12866-016-0846-9) contains supplementary material, which is available to authorized users.

The aim of this study is to evaluate the prevalence of various genotypes in the population of south-central Sicily (Italy) and to compare recent data with those of 2000/2001. In 2000, the patients tested were 202, all hepatitis C virus (HCV)-RNA and anti-HCV positive. From 2010 to 2014 the patients examined are in total 535, all anti-HCV positive, but 111 with genotype negative and therefore likely HCV-RNA negative. The study showed a clear predominance of genotype 1b for both men and women, however, with a much greater prevalence in the older cohort. In both groups, then, the 3a genotype follows for men, while the 2a/2c follows for women. 1a genotype prevalence rate falls in the most recent group of women. The cases of co-infection of more genotypes remain very content in 2014 as it happened in 2000.

Treatment-experienced chronic hepatitis C (CHC) genotype (GT) 1b represents a major medical burden in China. We evaluate the efficacy, safety and cost-effectiveness of ribavirin (RBV)-free pan-oral direct-acting antivirals (DAAs) in treatment-experienced Chinese with GT1b CHC, including patients with cirrhosis.One hundred forty treatment-experienced GT1b CHC Chinese with and without cirrhosis were included in this study. Ninety-four patients were treated with either daclatasvir (DCV, 60 mg)-sofosbuvir (SOF, 400 mg) (group 1, n = 46) or ledipasvir (LDV, 90 mg)-SOF (400 mg) (group 2, n = 48) for 12 weeks. Forty-six patients treated with pegylated interferon and RBV therapy for 72 weeks were enrolled as the control group (group 3). Patients were followed at 4-weekly intervals till 24 weeks after the end of treatment.All patients in group 1 (46/46, 100 %) and 2 (48/48, 100 %) had achieved sustained virologic response at 24 weeks after the end of treatment (SVR 24), which was significantly higher than that of group 3 (13/46, 28.3 %) (p 0.001). The SVR 24 rates of cirrhotic patients in group 1 (27/27, 100 %) and 2 (27/27, 100 %) were also significantly higher than that of group 3 (3/25, 12 %) (p 0.001). Twelve weeks of RBV-free LDV-SOF and DCV-SOF was either cost-saving or cost-effective. Adverse events were significantly lower in group 1 and 2 compared with group 3 (p 0.001).Compared with standard therapies, 12 weeks of RBV-free DAA therapies is highly effective, well tolerated and cost-effective in treatment-experienced Chinese with GT1b CHC including patients with cirrhosis.

BACKGROUND The efficacy of direct-acting antiviral agents (DAAs) could be attenuated by the presence of resistance-associated variants (RAVs). The aim of this study was to investigate the natural prevalence of RAVs among Chinese HCV genotype 1b patients and analyze the efficacy of pegylated interferon (PegIFN)/ribavirin (RBV) therapy in patients with and without RAVs at baseline. METHODS Direct sequencing of the HCV NS3, NS5A and NS5B regions was performed in baseline serum samples of 117 DAAs-naive subjects infected with HCV genotype 1b. The efficacy of PegIFN/RBV therapy in patients with and without RAVs at baseline was analyzed by comparing the response rates between patients with RAVs and patients with wild type virus. RESULTS The incidence of RAVs was 8.00% (8/100) in the NS3 region (T54S, n = 1, 1.00%; R117H, n = 5, 5.00%; S122T, n = 1, 1.00%; S174F, n = 1, 1.00%), 29.91% (32/107) in the NS5A region (L28M, n = 12, 11.21%; R30Q, n = 10, 9.35%; L31M, n = 1, 0.93%; P58S, n = 4, 3.74%; Y93H, n = 8, 7.48%) and 98.15% (106/108) in the NS5B region (L159F, n = 1, 0.93%; C316N, n = 103, 95.37%; A421V, n = 6, 5.56%). The response rates to PegIFN/RBV treatment did not differ between patients with or without RAVs in the NS5A region. CONCLUSIONS Pre-existing RAVs, including key RAVs, were detected in Chinese DAAs-naive patients infected with HCV genotype 1b. IFN-based therapy could be a good option for patients with RAVs, especially key RAVs, at baseline.

Hepatitis C infection is a public health problem. The aim of this retrospective study was to determine the distribution of hepatitis C virus (HCV) genotypes in seven regions of Turkey, by evaluating 7002 patients with chronic HCV in a six-year period. During the 2009–2014 period, serum/plasma samples from 7002 new consecutive HCV RNA positive patients were collected. The female patients were 3867 (55.2%). The genotype distribution of HCV patiens was evaluated by ages and years. Statistical analysis was performed by using the Mann–Whitney test and the ?2 analysis. During the six-year period, genotype 1b was the most common genotype (67.7%) followed by untypeable genotype 1 (7.7%), genotype 4 (7.3%) and genotype 3 (6.7%). In 2014, genotype 3 was the second most common one (11.3%) and genotype 4 was the third most common one (9.8%). In the group with

BACKGROUND: Resistance-associated variants have been related to treatment failure of hepatitis C virus (HCV) therapy with direct-acting antiviral drugs. The aim of our study was to analyze the prevalence of clinically relevant resistance-associated variants within NS3 in patients infected with HCV genotype 1a (GT1a) in Spain. METHODS: We performed a cross-sectional study on 2568 patients from 115 hospitals throughout Spain (2014-2015). The viral NS3 protease gene was amplified by nested polymerase chain reaction and sequenced by Sanger sequencing using an ABI PRISM 377 DNA sequencer. Additionally, clade information for genotype 1a was obtained by using the software geno2pheno (http://hcv.geno2pheno.org/). RESULTS: In total, 875 out of 2568 samples were from human immunodeficiency virus (HIV)/HCV-coinfected patients. Q80K was the main RAV found in our patients (11.1%) and the rest of the resistance-associated variants had a lower frequency, including S122G (6.23%), T54S (3.47%), V55A (2.61%), and V55I (2.15%), which were among the most frequent after Q80K. Overall, 286 samples had the Q80K polymorphism (11.1%) and 614 (23.9%) were GT1a clade I. HIV/HCV-coinfected patients had a higher frequency of Q80K and GT1a clade I than HCV-monoinfected patients (12.9% vs. 9.6% [p = 0.012] and 28.5% vs. 21.4% [p

The aim of the study Was to assess current prevalence of hepatitis C virus (HCV) genotypes in Poland, including their geographic distribution and changes in a given period of time. Material and methods Data were collected with questionnaires from 29 Polish centers and included data of patients diagnosed with HCV infection between 1 January 2013 and 31 March 2016. Results In total, data of 9800 patients were reported. The highest prevalence was estimated for genotype 1b (81.7%), followed by 3 (11.3%), 4 (3.5%), 1a (3.2%) and 2 (0.2%). Genotype 5 or 6 was reported in 6 patients only (0.1%). The highest prevalence of genotype 1 was observed in central (lodzkie, mazowieckie, świetokrzyskie), eastern (lubelskie) and southern (malopolskie, śląskie) Poland. The highest rate for genotype 3 was observed in south-western (dolnośląskie, lubuskie) and eastern (podlaskie, warminsko-mazurskie and podkarpackie) Poland. Compared to historical data, we observed an increasing tendency of G1 prevalence from 72.0% in 2003 to 87.5% in 2016, which was accompanied by a decrease of G3 (17.9% vs. 9.1%) and G4 (9.0% vs. 3.1%). Conclusions Almost 85% of patients with HCV in Poland are infected with genotype 1 (almost exclusively subgenotype 1b), and its prevalence shows an increasing tendency, accompanied by a decrease of genotypes 3 and 4.

A series of macrocyclic compounds containing a cyclic constraint in the P2–P4 linker region have been discovered and shown to exhibit excellent HCV NS3/4a genotype 3a and genotype 1b R155 K, A156T, A156 V, and D168 V mutant activity while maintaining high rat liver exposure. The effect of the constraint is most dramatic against gt 1b A156 mutants where ∼20-fold improvements in potency are achieved by introduction of a variety of ring systems into the P2–P4 linker.

A qualitative import risk assessment was undertaken to assess the likelihood of introduction and establishment of viral haemorrhagic septicaemia virus (VHSV) genotype 1a in England and Wales (EW), via the processing of imported rainbow trout (Oncorhynchus mykiss) carcasses from continental Europe. The likelihood was estimated for one import from an infected farm. Four main routes by which susceptible populations could be exposed to VHSV via processing waste were considered: (i) run-off from solid waste to watercourses, (ii) contamination of birds or rodents with VHSV by scavenging solid waste, (iii) discharge of liquid waste to mains drainage, and (iv) discharge of liquid waste directly to watercourses. Data on the biophysical characteristics of VHSV, its epidemiology, fish processing practices and waste management were collected. Likelihoods for each step of the four pathways were estimated. Pathway 4 (discharge of liquid waste to a watercourse) was judged as the most likely to result in infection of susceptible individuals. Levels of virus entering the aquatic environment via pathways 1-3 were judged to be many times lower than pathway 4 due mainly to the treatment of solid waste (pathways 1 and 2) and high levels of dilution (pathways 1, 2 and 3). Thirty-four trout farms process fish, of which seven have imported carcasses for processing. Compared with other processing facilities, on-farm processing results in a higher likelihood of VHSV exposure and establishment via all four pathways. Data availability was an issue; the analysis was particularly constrained by a lack of data on the prevalence of VHSV in Europe, volume of trade of carcasses into the UK and processing practices in EW. It was concluded that the threat of VHSV introduction into EW could be reduced by treatment of liquid effluent from processing plants and by sourcing carcasses for on-farm processing only from approved VHSV free areas.

We evaluated the Abbott RealTime (ART) and Roche Cobas TaqMan Hepatitis C virus (HCV) viral load assays for quantification of HCV genotypes in patient specimens. The ART HCV assay was a more sensitive and precise tool for accurate HCV viral load quantification across the HCV genotypes tested, especially genotype 1b.

NS5A inhibitors are a new class of direct-acting antiviral agents which display very potent anti-HCV activity in vitro and in humans. Rationally designed modifications to the central biphenyl linkage of a known NS5A series led to selection of several compounds that were synthesized and evaluated in a HCV genotype 1b replicon. The straight triphenyl linked compound 11a showed similar anti-HCV activity to the clinical compound BMS-790052 and a superior cytotoxicity profile in three different cell lines, with an EC50 value of 26 pM and a therapeutic index of over four million in an HCV replicon assay. This triphenyl analog warrants further preclinical evaluation as an anti-HCV agent.

Aim: The aim of our study was to investigate the correlation between the presence of plus-/minus-strand HCV RNA in peripheral blood mononuclear cells (PBMCs) and serum following pegylated IFN/ribavirin therapy with response to therapy in HCV-infected patients. Methods: Forty-three HCV-infected patients who completed 48 weeks of IFN/ribavirin therapy, including 25 sustained virologic responders, 12 resistants and six relapsers, comprised the study population. Plus-/minus-strand HCV RNA was detected by reverse transcription PCR in serum and PBMCs. Results: The frequency of plus-strand HCV RNA was significantly higher in PBMC and serum samples of relapsers and resistants, and this might have important implications in clinical practice and patient management. There was no correlation between presence of plus- and minus-strand HCV RNA and genotypes, except the fact that most of the patients who had plus-strand HCV RNA in PBMCs (60%) and in serum (61.53%) belonged to genotype 1a. Conclusion: Presence of plus-strand HCV RNA in PBMCs and serum after termination of therapy is associated with viral relapse and resistance to IFN/ribavirin treatment in HCV-infected patients.

Summary. Background: Telaprevir in combination with peginterferon and ribavirin is a promising advancement in chronic hepatitis C treatment. However, the safety, tolerability, pharmacokinetics and antiviral profiles of telaprevir alone beyond 2 weeks have not been studied. Methods: In a phase 1b study in Japan, 10 treatment-naive patients infected with hepatitis C virus genotype 1b with high viral load (>5 log10 IU/mL) received telaprevir 750 mg every 8 h (q8h) for 12 weeks. We examined the safety, tolerability, pharmacokinetics, hepatitis C virus (HCV) RNA levels and resistant variants of telaprevir. Results: Neither serious adverse events nor discontinuations of study drug owing to an adverse event occurred. The most common adverse drug reactions were rash (80%) and anaemia (70%). Telaprevir concentration reached its steady state within 2 days after the first administration without abnormal accumulation. Telaprevir alone provided potent antiviral activity: a median log10 decrease of 2.325 at 16 h and 5.175 on Day 14. During the treatment, HCV RNA levels at the nadir were below the limit of the quantification in seven patients and undetectable in three of 10 patients. Viral breakthrough associated with mainly Ala156-substituted variants occurred in eight patients, and only one patient showed end-of-treatment response. The selected variants reverted to the wild-type during the 24-week follow-up period. Conclusion: Telaprevir alone was well tolerated at 750 mg q8h for up to 12 weeks. The safety profile and emergence of resistant variants of genotype 1b under telaprevir monotherapy for 12 weeks will become increasingly important in evaluating an oral combination of telaprevir with other direct-acting antiviral agents.

The present study aims to evaluate the anti-HCV activity of hotwater extract from Platycodon grandiflorum (BC703) with HCV genotype 1b subgenomic replicon system and investigate its hepatoprotective activity on carbon tetrachloride (CCl(4))-induced acute liver damage in mice. BC703 produced significant hepatoprotective effects against CCl(4)-induced acute hepatic injury by decreasing the activities of serum enzymes, nitric oxide and lipid peroxidation. Histopathological studies further substantiated the protective effect of BC703. Furthermore, BC703 inhibited the HCV RNA replication with an EC(50) value and selective index (CC(50)/EC(50)) of 2.82 µg/mL and above 35.46, respectively. However, digested BC703 using a simulated gastric juice showed poor protective effect against CCl(4)-induced hepatotoxicity in mice and decreased anti-HCV activity as compared to the intact BC703. Although further studies are necessary, BC703 may be a beneficial agent for the management of acute hepatic injury and chronic HCV infection.

Introduction: Treatment of chronic hepatitis C (CH-C) with peginterferon α (Peg-IFN) and ribavirin leads to a sustained virological response (SVR) in 50% of patients and depends on HCV and host factors. Polymorphism of the IL28B gene is associated with eradication of HCV and SVR. Aim: The objective of this study was to examine patients from the region of southern Poland and determine whether CH-C therapy depends on the genetic variants of IL28B and whether this polymorphism may predict SVR. Material and methods: One hundred forty-two patients with CH-C and the genotype 1b HCV were treated with PegIFN and ribavirin for 48 weeks. IL28B rs12979860 polymorphisms (C/T) were tested by PCR-RFLP. The HCV-RNA and alanine aminotransferase (ALT) levels were measured before treatment and after 12, 48 and 72 weeks. Results: Viral load before and after 12 weeks of therapy was higher in the genotypes T/C and T/T than in C/C. 71.1% of patients with the genotype C/C achieved SVR vs. 41.4% with the genotype T/C and 23.5% with T/T. Patients with the genotype C/C responded better to treatment as compared to subjects with the genotypes T/C and T/T, and achieved better early response (12 weeks), at the end of treatment (48 weeks) and SVR. Alanine aminotransferase activity was similar among the groups. Tolerance of therapy was similar and independent of the genotypes. Conclusions: The study confirmed the dependence of response to standard treatment of CH-C caused by the genoStreszczenie Wstep: Leczenie przewleklego zapalenia wątroby typu C (PZW-C) pegylowanym interferonem α (Peg-IFN) z ryba wiryną powoduje tzw. trwalą odpowiedź wirusologiczną (sustained virological response – SVR) u blisko 50% chorych. Odpowiedź ta ściśle zalezy od HCV i czynnikow genetycznych chorego. Wykazano, ze polimorfizm genu IL28B wiąze sie z eradykacją wirusa i SVR. Cel: Zbadanie, czy wyniki leczenia PZW-C u chorych z regionu Polski Poludniowej zalezą od wariantow genetycznych IL28B, a polimorfizm moze byc czynnikiem predykcyjnym SVR. Material i metody: Sto czterdzieści dwie osoby z PZW-C spowodowanym genotypem 1b HCV leczono standardowo PegIFN z rybawiryną przez 48 tygodni. Polimorfizmy rs12979860 IL28B (C/T) badano metodą PCR-RFLP. HCV-RNA i aktywnośc aminotransferazy alaninowej (ALT) w surowicy mierzono przed leczeniem i po 12, 48 i 72 tygodniach terapii. Wyniki: Wiremia HCV-RNA przed terapią i po 12-tygodniowym leczeniu byla wyzsza u chorych z genotypem T/C i T/T. Trwalą odpowiedź wirusologiczną osiągnelo istotnie wiecej chorych z genotypem C/C (71,1%) w porownaniu z 41,4% z genotypem T/C i 23,5% T/T. Chorzy z genotypem C/C lepiej reagowali na leczenie w porownaniu z osobami z genotypami T/C i T/T, osiągali wyzsze wskaźniki odpowiedzi wczesnej (12 tygodni), na koniec leczenia (48 tygodni) i SVR. Aktywnośc ALT nie roznila sie istotnie podczas terapii pomiedzy grupami. Tolerancja leczenia byla podobna i nie zalezala od genotypow C/C, T/C i T/T. Przegląd Gastroenterologiczny 2012; 7 (1) type 1b HCV on the patient's genetic predisposition. In contrast to some other reports, poor prognosis was observed in T/T homozygotes of IL28B. Wnioski: Wyniki badania potwierdzily zaleznośc odpowiedzi na standardowe leczenie PZW-C spowodowanego genotypem 1b HCV od predyspozycji genetycznej chorego, jednak w przeciwienstwie do niektorych doniesien zle rokowanie obserwowano u homozygot T/T IL28B.

Aim: To examine the impact of ribavirin dose reduction on the efficacy of pegylated interferon (PEG IFN) plus ribavirin combination therapy for elderly patients infected with genotype 1b and high viral loads. Methods: A total of 72 patients, over 65 years old, were recruited for this study. Patients were divided into groups receiving either 600–800 mg of ribavirin according to bodyweight (Group 1, n = 36) or 400 mg of ribavirin (Group 2, n = 36) plus 1.5 µg/kg (range: 1.3–2.0 µg/kg) of PEG IFN-α-2b for 48 weeks. Results: Total ribavirin doses were administrated at 9.80 ± 2.39 mg/kg per day (3.29 ± 0.80 g/kg) for Group 1 and 5.87 ± 1.82 mg/kg per day (1.97 ± 0.61 g/kg) for Group 2 (P

Persistent hepatitis C virus (HCV) infection causes chronic liver diseases and is a global health problem. HuH-7 hepatoma-derived cells are widely used as the only cell-based HCV replication system for HCV research, including drug assays. Recently, using different hepatoma Li23-derived cells, we developed an HCV drug assay system (ORL8), in which the genome-length HCV RNA (O strain of genotype 1b) encoding renilla luciferase replicates efficiently. In this study, using the HuH-7-derived OR6 assay system that we developed previously and the ORL8 assay system, we evaluated 26 anti-HCV reagents, which other groups had reported as anti-HCV candidates using HuH-7-derived assay systems other than ORB. The results revealed that more than half of the reagents showed different anti-HCV activities from those in the previous studies, and that anti-HCV activities evaluated by the ORB and ORL8 assays were also frequently different. In further evaluation using the HuH-7-derived AH1R assay system, which was developed using the AH1 strain of genotype 1b, several reagents showed different anti-HCV activities in comparison with those evaluated by the OR6 and ORL8 assays. These results suggest that the different activities of anti-HCV reagents are caused by the differences in cell lines or HCV strains used for the development of assay systems. Therefore, we conclude that plural HCV assay systems developed using different cell lines or HCV strains are required for the objective evaluation of anti-HCV reagents.

Background: A retrospective analysis was carried out in all specimens from subjects with chronic hepatitis C sent for testing to our laboratory in Hospital Universitario Rio Hortega (Valladolid, Spain) over the period 1999-2009. The reason for this study was to examine the suggestion of other authors on the spread of genotype 4 strains. The objective was to describe the distribution of VHC genotypes in our geographical area and compare it with other state-wide reports. Methods: A total of 1074 patients were studied. Specimen preparation to isolate HCV RNA was carried out with the COBAS AmpliPrep system (Roche). Reverse transcription, amplification and reverse hybridization were performed with Qiagen and Siemens kits. Results: The most frequent HCV genotype was 1 (69%), followed by 3 (19,6%) and 4 (8.2%). The most frequent HCV subtype was 1b (41.3%). Most infections with genotype 4 (58%) were found among HCV-HIV-coinfected patients. From our previous report in 2002 to date (end December 2010), both an increase in the prevalence of genotypes 4 (from 7.3 to 8.8%) and 1 and 1a (from 25.9 to 29.4%) and a decrease in the prevalence of genotype 1b (from 44% to 39,5%) has been observed over time. Patients with genotype 4 were, mostly, men and with HIV-HCV coinfection. Conclusions: The prevalence of genotype 4 in our geographical environment was significantly different than the national average which leads to the conclusion that the spread of this genotype was much slower than suggested.

Aim: Elevation of alanine aminotransferase (ALT) levels during pegylated-interferon (peg-IFN) plus ribavirin therapy in patients with chronic hepatitis C [CHC] is a problem that cannot be disregarded. The aim of this study is to assess the frequency and to characterize clinical parameters of this phenomenon. Methods: Two hundred and thirty-five (235) CHC patients with genotype 1b receiving peg-IFN α-2b plus ribavirin therapy were analyzed. Clinical parameters that may be associated with abnormal ALT values during treatment and therapy outcomes were evaluated statistically. One hundred and sixteen (116) patients treated with peg-IFN α-2a plus ribavirin were also included for partial analysis. Results: Abnormal ALT values during treatment were observed in 23.0% of patients. It was observed in 14.5% of those with sustained virological response (SVR) and 17.8% of those with relapse, in whom viral clearance was observed during therapy. Multivariate logistic regression analysis revealed that pretreatment ALT values, therapy outcome, and body mass index (BMI) were significant factors related to abnormal ALT values during treatment. Abnormal ALT values during treatment became normal in SVR patients at 6 months after the completion of treatment, but not in NR (non-response) patients. Mean ALT values were significantly higher at some time points during treatment in patients treated with α-2a when compared to those treated with α-2b. Conclusion: Abnormal ALT values during peg-IFN plus ribavirin treatment are observed relatively frequently, even in patients without detectable HCV RNA. Direct or indirect involvement of drugs is considered as one possible cause.

Background and Objectives A collaborative study was undertaken to evaluate a replacement World Health Organization International Standard for hepatitis C virus (HCV) RNA for nucleic acid amplification technology (NAT)-based assays. The candidate preparations were calibrated in International Units (IUs). Materials and Methods Three new candidate preparations were produced from a single bulk containing anti-HCV-negative, genotype 1a HCV RNA–positive plasma. Two samples were lyophilized (coded Sample 2 and Sample 3), whilst a third (Sample 4) contained liquid/frozen material. The samples were distributed together with the 2nd International Standard (Sample 1, NIBSC code 96/798) for evaluation by thirty-three laboratories, from fourteen countries. The panel of samples were assayed on four separate occasions. Stability studies were performed for the lyophilized samples by accelerated thermal degradation. Results Participants returned data from a wide range of commercial and in-house quantitative and qualitative assays. Twenty-five data sets were returned for quantitative assays and fourteen for qualitative assays. Excellent agreement was observed between laboratories and assay methods. The mean relative potencies of Samples 2–4 were 5·19, 5·41 and 5·70 log10 IU/ml, respectively, when compared against the 2nd International Standard. Samples 2 and 3 demonstrated stability of a similar order to the previous standards. Conclusions Based upon the results of the collaborative study, Sample 2 (code number 06/100) was established as the 3rd International Standard for HCV RNA with an assigned unitage of 5·19 log10 IU/ml. Each vial contains the equivalent of 0·5 ml of material; each vial contains 4·89 log10 IU of HCV RNA.

Accuracy for monitoring of the concentration of hepatitis C virus (HCV) RNA represents a major challenge throughout the management of patients with chronic hepatitis C. To investigate the genotype-independent efficiency and the accuracy of two real-time detection reverse transcription-polymerase chain reaction (RT-PCR) assays; the Cobas Ampliprep/Cobas TaqMan (CAP/CTM); and the Abbott RealTime HCV (ART), a total of 184 samples with different HCV subtypes were examined; 1b (n=58), 2a (n=39), 2b (n=26), 3a (n=20), and 4 (n=41). A robust linear correlation was observed between the two assays applied to genotypes 1b, 2a, 2b, and 3a [the correlation coefficient (R) ranged from 0.99 to 0.98], but not to genotype 4 specimens (R=0.78). A significant difference in measurements of HCV RNA using CAP/CTM and ART in serum samples with genotypes 1b and 4 was observed (0.72, -0.53 log IU/ml, P

HCV infection is a leading cause of chronic liver disease, including cirrhosis of the liver. There are at least six major genotypes and more than 50 subtypes of HCV. The prevalence and distribution of HCV genotypes depend on geographical location. The aim of this study was to identify and compare the HCV genotypes in HCV infected blood donors and patients. In this cross-sectional study, 167 serum samples from 103 blood donors and 64 patients with hepatitis C were investigated for HCV genotypes. HCV genotyping was carried out using type-specific primers from the core region of the viral genome. The highest frequency was for genotype 1a, with 53 and 34 (51.5% versus 53.1%) of subjects in blood donors and patients respectively. Genotype 3a and 1b were the other frequent genotypes with 4 and 16 (3.9% versus 25%) and 39 and 10 (37.9% versus 15.6%) subjects, respectively. There was not any statistical significant association between the place of infection of the patients and genotype. The results of this study indicate that the distribution of genotypes in the two populations was similar. The dominant HCV genotypes between blood donors and patients were 1a, 3a and 1b respectively.

Interferon (IFN) response rate in hepatitis C virus (HCV) patients has been varied with genotypes. In this study, we investigated the effects of HCV NS5A protein on IFN resistance and compared the genotypic differences of NS5A. We showed that IFN-α-, poly I:C-, and Sendai virus-induced ISRE transcriptional activities were inhibited by both genotype 1b and 2a NS5A protein. We demonstrated that not only genotype 1b but also genotype 2a NS5A exerted the similar extent of IFN-α-induced antiviral activity. We showed that NS5A derived from both genotype 1b and 2a showed no significant differential IFN responses as seen in HCV patients. These data imply that some other host factor may be involved in genotypic differences of IFN antagonism in HCV patients.

Acute pancreatitis, an uncommon side effect of pegylated interferon α (PEG-IFN α) and ribavirin (RBV) combination therapy, has rarely been reported in the English language literature. Here, acute pancreatitis associated with PEG-IFN plus RBV treatment is described in three patients with chronic hepatitis C, genotype 1b with high serum hepatitis C virus RNA levels. The patients had been started on weekly subcutaneous injections of PEG-IFN α (60, 80, and 90 μg) plus a daily oral dose of RBV (600 mg). The therapy was discontinued, however, because of the onset of acute pancreatitis (after 15 weeks, 48 weeks, and 3 weeks respectively). The drug-induced pancreatitis was diagnosed on the basis of elevated levels of amylase and lipase and the absence of other identifiable causes. High tumor necrosis factor-α was found in one patient and high interleukin-6 in the other two. The immune system stimulated by PEG-IFN and RBV combination therapy might have caused the acute pancreatitis. Further study is needed to clarify the mechanism of the onset of drug-induced pancreatitis by PEG-IFN and RBV combination therapy.

Substitution of amino acid (aa) 70 and 91 in the core region of HCV genotype 1b is a useful pretreatment predictor of efficacy of 48-week peginterferon (PEG-IFN) plus ribavirin (RBV) therapy. Here, we determined the efficacy of 72-week PEG-IFN/RBV and the predictive factors to such therapy in a case–control study matched for sex, age, and periods from the start of treatment to initial point of HCV RNA-negative. We compared the treatment efficacy of 72-week regimen in 65 patients with that of 48-week in 130 patients, who were infected with HCV genotype 1b and treated with PEG-IFN/RBV. They consisted mainly of late virological responders (LVR) (HCV RNA-positive at 12 weeks and negative at 24 weeks after start of treatment). Sustained virological response (SVR) was achieved by 61.5% and 32.3% of patients of the 72- and 48-week groups, respectively, while non-virological response was noted in 9.2% and 29.2% of the respective groups. Multivariate analysis identified substitution of aa 70 and 91 (Arg70 and/or Leu91) and duration of treatment (72-week) as independent parameters that significantly influenced SVR. For Arg70 and/or Leu91 of core region, SVR rate was significantly higher in 72- (68.0%) than 48-week group (37.8%). For wild-type of ISDR, SVR rate was significantly higher in 72- (61.2%) than in 48-week group (29.3%). We conclude that 72-week PEG-IFN/RBV improves SVR rate for LVR, especially those with Arg70 and/or Leu91 of core region or wild-type of ISDR. Substitution of aa 70 and 91 is also a useful pretreatment predictor of response to 72-week PEG-IFN/RBV. J. Med. Virol. 81:452–458, 2009. © 2009 Wiley-Liss, Inc.