5 ��g/kg body weight once weekly+RBV (Rebetol; Schering-Plough, Kenilworth, NJ, USA): 1000�C1200 mg daily, according to body weight (1000 mg ��75 kg, 1200 mg >75 kg). The total duration of the antiviral therapy was defined Vismodegib medulloblastoma by the type of antiviral response within the first 12�C24 weeks of therapy. Patients with HCV RNA decrease <2 log from baseline level at week 12 were classified as null responders and their antiviral therapy was terminated at week 12. Patients with HCV RNA decrease ��2 log or with undetectable HCV RNA at week 12 were treated to week 24. If HCV RNA was detectable at week 24, patients were classified as slow responders and their therapy was terminated at week 24. Only patients with undetectable HCV RNA at week 24 were treated up to week 48.

Based on the treatment response, patients were classified into two groups. Responders were defined as patients with sustained virologic response (SVR, undetectable HCV RNA at weeks 24 after completion of antiviral therapy, n=38). Treatment-failure patients (non-SVR, n=20) included those who did not achieved SVR (n=15) and patients who relapsed (n=5). For PBMC and PBL studies, 55 healthy volunteers (blood donors or employees of General Faculty Hospital and 1st Faculty of Medicine, Charles University in Prague) were used as control subjects. The study was registered under ID: NCT 00842250 (www.clinicaltrial.gov). The study protocol conformed to all ethical guidelines of the 1975 Declaration of Helsinki, reflected in the a priori approval by the institution��s Ethics Committee.

Additionally, all subjects in this study had provided written informed consent. Methods Material sampling and storage Patients�� blood samples were collected on the day before treatment initiation (day 0, n=58), as well as at 12 (n=37), 24 (n=31), 36 (n=27) and 48 (n=16) weeks after initiation of the standard antiviral treatment. Samples for gene expression analyses were collected into PAXgene Blood RNA Tubes (PreAnalytix, Hombrechtikon, Switzerland) and stored at ?80��C. Samples for determination of total HMOX activity in PBMC were collected in BD Vacutainer Blood collection tubes with heparin (BD Diagnostics-Preanalytical Systems, Franklin Lakes, NJ, USA). PBMC were isolated through a Ficoll-density gradient within 5 hours, and stored in potassium phosphate buffer at ?80��C.

The blood samples for analysis of interleukin 28B (IL28B, OMIM*607402) gene polymorphism were collected in BD Vacutainer Blood collection tubes with EDTA (BD Diagnostics-Preanalytical Systems, Batimastat Franklin Lakes, NJ, USA) and stored at ?80��C. All liver samples were immediately placed into RNAlater (Ambion Diagnostics, Austin, TX, USA) and stored at ?80��C until total RNA isolation. Reagents Hemin, nicotinamide adenine dinucleotide phosphate (NADPH) and sulfosalicylic acid were purchased from Sigma (St. Louis, MO, USA).