Methods: Adult treatment-naïve patients were randomly assigned to DCV/peg-alfa-2a/RBV for learn more 12 or 16 weeks or placebo/peg-alfa-2a/RBV for 24 weeks. DCV/peg-alfa-2a/RBV recipients without protocol-defined response (PDR; HCV RNA

(N = 50), DCV 16-week (N = 50) and placebo (N = 51) arms; more patients with GT3 (18/80, 22.5%) than GT2 (1/71, 1.4%) BGJ398 molecular weight were cirrhotic. 78%-88% of DCV recipients achieved PDR. SVR24 rates were higher in GT2 than GT3 with all regimens;

within each genotype, SVR24 rates were similar in DCV arms and higher than placebo/peg-alfa-2a/RBV. In GT2, the SVR24 rate in each DCV arm was 83%; in the placebo/peg-alfa-2a/RBV arm, the SVR24 rate was 63%. In GT3, the SVR24 rate in the 12-week and 16-week DCV arms were 69% and 67%, respectively; in the placebo/peg-alfa-2a/RBV arm, the SVR24 rate was 59%. In DCV arms, one GT2 and 12 GT3 patients relapsed. In GT3, relapse was higher among cirrhotics (3/7, 43%) than non-cirrhotics (3/19, 16%) in the 12-week arm but similar in the 16-week arm (1/4, 25% vs 5/20, 25%). There were 7 on-treatment serious AEs (DCV, 4; placebo, 3); no deaths. AEs were typical of those associated with peg-alfa/RBV. Conclusion: Shorter treatment duration (12 or 16 weeks) with DCV/peg-alfa-2a/RBV demonstrated higher SVR rates than 24 weeks of peg-alfa-2a/RBV in patients with GT2 or GT3 infection, with higher SVR rates in GT2 with all regimens.

These results support further evaluation of DCV-containing regimens for different HCV genotypes. Funding disclosures: This Vorinostat purchase study was funded by Bristol-Myers Squibb (BMS). Editorial support was provided by Articulate Science and BMS and funded by BMS. Resistance analyses were contributed by Fiona McPhee of BMS. T ASSELAH,1 S ZEUZEM,2 V SORIANO,3 J-P BRONOWICKI,4 AW LOHSE,5 B MÜLLHAUPT,6 M SCHUCHMANN,7 M BOURLIERE,8 M BUTI,9 S ROBERTS,10 ED GANE,11 J STERN,12 P BAUM,13 J-P GALLIVAN,14 W BÖCHER,14 F MENSA12 1Hôpital Beaujon, Clichy, France, 2Klinikum der J. W.

However, only 5% ± 2% of hepatocytes were BrdU positive in eNOS−/− mice at 45 hours post-PH, with 82% fold impairment, as compared to WT mice (Fig. 2G,H). At 72 hours post-PH, the percentage of BrdU-positive cells was slightly higher in the eNOS−/− mice (4% versus 5%; nonsignificant) (Fig. 2G,H). At 96 hours

post-PH, BrdU incorporation declined to near basal levels and was comparable between WT and eNOS−/− mice (Fig. 2G,H). Taken together, these results suggest hepatocyte cell-cycle progression (Fig. 2A-F) and proliferation (Fig. 2G,H) in response to PH is impaired in eNOS−/− mice. Based on the established significance of MMP-9 in ECM remodeling in regenerating livers, MMP-9 protein expression was analyzed by western blotting of total liver homogenates

of resected lobes (0 minutes) GPCR Compound Library research buy and remnant livers (0.5-72 hours post-PH).19 PH induced robust MMP-9 protein expression in WT mice. MMP-9 induction was delayed and significantly attenuated in eNOS−/− mice at 30 minutes (52.4%), 1 hour (52.1%), and 45 hours (52%) (Fig. 3A,C). MMP-9 plays a key role in the activation of latent growth factors, such as hepatocyte growth factor (HGF). HGF effects on hepatocyte proliferation are mediated via the phosphorylation and activation of c-Met, a protooncogene essential for liver regeneration.20 Corresponding to MMP-9 activation, eNOS−/− regenerating livers exhibit dysregualtion in HGF signaling, as evidenced by the attenuated induction of c-Met phosphortylation (Tyr1349) at (3 hours, 37%; 24 hours, 36%; 45 hours, 43%) (Fig. 3B,D). Phosphorylation at Ser1177 (activation) and at Thr495 (inhibition) are among the well-characterized AT9283 clinical trial post-translational modifications of eNOS.21 To determine

whether PH regulates eNOS activity in regenerating livers, eNOS phosphorylation at MTMR9 Ser1177 and Thr495 were analyzed by western blotting of total lysates. eNOS phosphorylation at Ser1177 was observed early in liver regeneration (15 minutes to 3 hours post-PH; peak at 30 minutes), and eNOS dephosphorylation at Thr495 was observed later (45-96 hours post-PH) in WT livers (Fig. 4A). Total eNOS expression increased slightly after PH. Additionally, eNOS activity can be regulated by transcriptional regulation. To test whether PH regulates eNOS mRNA expression, qRT-PCR was performed with RNA isolated from liver tissues (resected and remnant livers at post-PH). eNOS gene expression increased several fold from 3 to 24 hours, and the maximal level was observed at 3 hours post-PH (7-fold) (Fig. 4B). To determine whether compensatory iNOS induction plays any role in eNOS−/− regenerating livers, total proteins and RNA isolated from WT and eNOS−/− regenerating livers (15 minutes to 12 hours) were analyzed by western blotting and qRT-PCR for iNOS expression, respectively. Our results suggest that iNOS protein and mRNA expression were comparable between the WT and eNOS−/− livers (Supporting Fig. 1A-C).

Nuclei were stained with 50 ng/mL Hoechst 33258. Cell polarization was determined by counting the number

of bile canaliculi (BC) (identified by dense F-actin staining) per 100 cells (identified by fluorescently labeled nuclei) as described.18 Cells were plated on coverslips in the absence or the presence of soluble rGal-1 (7 μM). After 48 hours, each coverslip was mounted in a chamber placed on the stage of a Nikon TE-200 epifluorescence-inverted microscope. Cells were then loaded with 5-chloromethylfluorescein diacetate.19 Hepatocytes capture and metabolize this compound, generating fluorescent glutathione methylfluorescein, which is actively secreted to the canaliculi by MRP2. A total of 5 × 106 HepG2-M (mock-transfected) or HepG2-G2 (Gal-1–overexpressing) cells were injected subcutaneously into the left Ivacaftor cell line flank of 6-week-old BALB/c nude mice. Tumor volume was calculated as π/6 × length × width2. Liver, lungs, and tumor-draining lymph nodes were serially sectioned and stained with hematoxylin and eosin. Metastases were

examined in size-matched tumors. All animal care and experimentation was conducted in accordance with the National Academy of Sciences Guide for the Care and Use of Laboratory Animals. To examine the effects of Gal-1 on HCC cell physiology, we first assessed its expression and subcellular distribution Y-27632 ic50 in the differentiated HepG2 cell line. We stably transfected HepG2 cells with Gal-1 complementary DNA. Two G418-resistant clones were selected with approximately two-fold (HepG2-G1) and five-fold (HepG2-G2) higher Gal-1 expression compared with nontransfected cells and cells transfected with empty vector (HepG2-M) (Fig. Fluorouracil cell line 1A). Gal-1 immunostaining of HepG2-G2 cells showed clear cytoplasmic localization, whereas no staining was observed on HepG2 (Fig. 1B) and HepG2-M cells (data not shown). However, immunostaining performed on permeabilized HepG2 cells incubated for 48 hours at 37°C in the presence of exogenously added rGal-1 showed positive cytoplasmic localization. Because real-time polymerase chain reaction analysis showed that rGal-1

does not induce transcription of its own gene (Supporting Information Fig. 1), this result suggests that rGal-1 may be internalized by HepG2 cells. Furthermore, no membrane-associated Gal-1 was detected in nonpermeabilized cells (data not shown). Moreover, examination of Gal-1 secretion revealed a faint immunoreactive band in concentrated serum-free conditioned medium of HepG2 and HepG2-M cultures, an effect that was considerably increased in Gal-1–transfected cells (Fig. 1C). These results indicate that HepG2-G2 cells express and secrete high levels of Gal-1. To study the influence of Gal-1 on hepatocyte function, we performed cell adhesion assays. When cells were incubated for 1 hour on uncoated plates in the presence of rGal-1 (3.

Nuclei were stained with 50 ng/mL Hoechst 33258. Cell polarization was determined by counting the number

of bile canaliculi (BC) (identified by dense F-actin staining) per 100 cells (identified by fluorescently labeled nuclei) as described.18 Cells were plated on coverslips in the absence or the presence of soluble rGal-1 (7 μM). After 48 hours, each coverslip was mounted in a chamber placed on the stage of a Nikon TE-200 epifluorescence-inverted microscope. Cells were then loaded with 5-chloromethylfluorescein diacetate.19 Hepatocytes capture and metabolize this compound, generating fluorescent glutathione methylfluorescein, which is actively secreted to the canaliculi by MRP2. A total of 5 × 106 HepG2-M (mock-transfected) or HepG2-G2 (Gal-1–overexpressing) cells were injected subcutaneously into the left PLX3397 cell line flank of 6-week-old BALB/c nude mice. Tumor volume was calculated as π/6 × length × width2. Liver, lungs, and tumor-draining lymph nodes were serially sectioned and stained with hematoxylin and eosin. Metastases were

examined in size-matched tumors. All animal care and experimentation was conducted in accordance with the National Academy of Sciences Guide for the Care and Use of Laboratory Animals. To examine the effects of Gal-1 on HCC cell physiology, we first assessed its expression and subcellular distribution Silmitasertib clinical trial in the differentiated HepG2 cell line. We stably transfected HepG2 cells with Gal-1 complementary DNA. Two G418-resistant clones were selected with approximately two-fold (HepG2-G1) and five-fold (HepG2-G2) higher Gal-1 expression compared with nontransfected cells and cells transfected with empty vector (HepG2-M) (Fig. 4-Aminobutyrate aminotransferase 1A). Gal-1 immunostaining of HepG2-G2 cells showed clear cytoplasmic localization, whereas no staining was observed on HepG2 (Fig. 1B) and HepG2-M cells (data not shown). However, immunostaining performed on permeabilized HepG2 cells incubated for 48 hours at 37°C in the presence of exogenously added rGal-1 showed positive cytoplasmic localization. Because real-time polymerase chain reaction analysis showed that rGal-1

does not induce transcription of its own gene (Supporting Information Fig. 1), this result suggests that rGal-1 may be internalized by HepG2 cells. Furthermore, no membrane-associated Gal-1 was detected in nonpermeabilized cells (data not shown). Moreover, examination of Gal-1 secretion revealed a faint immunoreactive band in concentrated serum-free conditioned medium of HepG2 and HepG2-M cultures, an effect that was considerably increased in Gal-1–transfected cells (Fig. 1C). These results indicate that HepG2-G2 cells express and secrete high levels of Gal-1. To study the influence of Gal-1 on hepatocyte function, we performed cell adhesion assays. When cells were incubated for 1 hour on uncoated plates in the presence of rGal-1 (3.

The centre did not have a specialized team to properly administer prophylaxis and to ensure proper conduct of the study. The treating physicians were not sure about the benefits of prophylaxis. Patients/parents doubting the benefit of prophylaxis and discontinuing on their own. Patients/parents disliking the need for repeated injection and/or frequent Rucaparib visits to hospitals and discontinuing on their own. Statistics Package for Social Science

(spss, IBM Corporation, Armonk, NY, USA) 13.0 for Windows was used for data analysis. We used Rank-test to determine the significance of data difference between the observation and prophylaxis period. P < 0.05 is considered statistically significant. Fifteen centres participated click here in this trial enrolling a total of 191 patients. All 66 patients (66/191, 34.6%) from three centres (3/15, 20%) completed 6–12 weeks prophylaxis (compliant group), while none of the 125 patients (125/191,

65.4%) from the remaining 12 centres completed at least 6 weeks of prophylaxis (non-compliant group). All the 66 patients (age 2–17.5 years, mean 8.6) were analysable. These include: 28 (42.4%) from the Nanfang Hospital Center, 22 (33.3%) from the Shandong Center, Jinan and 16 (24.2%) from the BCH Center. Haemophilia A was severe in 34 patients (51.5%), moderate in 32 (48.5%). Compared with the observation period, prophylaxis resulted in significant reduction in bleeding (joint bleeding and severe bleeding; P < 0.01; 4-Aminobutyrate aminotransferase Table 1 and Fig. 1). The reduction in joint bleeding was more significant in patients who (i) had undergone prophylaxis for more than 10 weeks (P = 0.001), (ii) had severe disease (P = 0.005), and (iii) were older than age 12 (P = 0.024; Table 2). Sixteen cases were assessed by the BCH score and 27 cases were assessed by FISH. The remaining 23 patients were not assessed. After prophylaxis, the BCH assessment scale was upgraded in 31% (5/16; Table 3) while FISH assessment showed 60% improvement from a score range 7–28 (mean 15.81 ± 4.99) to 14–28 (mean 25.15 ± 4.01), (P < 0.001; Table 4). Consumption

of factors for on-demand therapy (and breakthrough bleeding) varies according to the regimen (A1 and A2) used (Table 5). For those using the ‘optimal-dose’ (A1), the mean consumption during the observation and prophylaxis period were similar (103.2 vs. 102.9 IU kg−1 per month). The consumption during the prophylaxis period was as expected higher when ‘suboptimal-dose’ (A2) was used (Table 5). One hundred and twenty-five patients (age 2–18 years, mean 8.4. Age between the non-compliant and compliant groups were not significantly different, P = 0.229) enrolled to 12 centres were in the non-compliant group. Their duration of prophylaxis was 2–5 weeks (mean 2.7). All the 15 centers were able to provide paediatric haemophilia treatment, but not all have the professional components of a comprehensive haemophilia team.

The centre did not have a specialized team to properly administer prophylaxis and to ensure proper conduct of the study. The treating physicians were not sure about the benefits of prophylaxis. Patients/parents doubting the benefit of prophylaxis and discontinuing on their own. Patients/parents disliking the need for repeated injection and/or frequent ICG-001 cost visits to hospitals and discontinuing on their own. Statistics Package for Social Science

(spss, IBM Corporation, Armonk, NY, USA) 13.0 for Windows was used for data analysis. We used Rank-test to determine the significance of data difference between the observation and prophylaxis period. P < 0.05 is considered statistically significant. Fifteen centres participated Angiogenesis inhibitor in this trial enrolling a total of 191 patients. All 66 patients (66/191, 34.6%) from three centres (3/15, 20%) completed 6–12 weeks prophylaxis (compliant group), while none of the 125 patients (125/191,

65.4%) from the remaining 12 centres completed at least 6 weeks of prophylaxis (non-compliant group). All the 66 patients (age 2–17.5 years, mean 8.6) were analysable. These include: 28 (42.4%) from the Nanfang Hospital Center, 22 (33.3%) from the Shandong Center, Jinan and 16 (24.2%) from the BCH Center. Haemophilia A was severe in 34 patients (51.5%), moderate in 32 (48.5%). Compared with the observation period, prophylaxis resulted in significant reduction in bleeding (joint bleeding and severe bleeding; P < 0.01; Dichloromethane dehalogenase Table 1 and Fig. 1). The reduction in joint bleeding was more significant in patients who (i) had undergone prophylaxis for more than 10 weeks (P = 0.001), (ii) had severe disease (P = 0.005), and (iii) were older than age 12 (P = 0.024; Table 2). Sixteen cases were assessed by the BCH score and 27 cases were assessed by FISH. The remaining 23 patients were not assessed. After prophylaxis, the BCH assessment scale was upgraded in 31% (5/16; Table 3) while FISH assessment showed 60% improvement from a score range 7–28 (mean 15.81 ± 4.99) to 14–28 (mean 25.15 ± 4.01), (P < 0.001; Table 4). Consumption

of factors for on-demand therapy (and breakthrough bleeding) varies according to the regimen (A1 and A2) used (Table 5). For those using the ‘optimal-dose’ (A1), the mean consumption during the observation and prophylaxis period were similar (103.2 vs. 102.9 IU kg−1 per month). The consumption during the prophylaxis period was as expected higher when ‘suboptimal-dose’ (A2) was used (Table 5). One hundred and twenty-five patients (age 2–18 years, mean 8.4. Age between the non-compliant and compliant groups were not significantly different, P = 0.229) enrolled to 12 centres were in the non-compliant group. Their duration of prophylaxis was 2–5 weeks (mean 2.7). All the 15 centers were able to provide paediatric haemophilia treatment, but not all have the professional components of a comprehensive haemophilia team.

Regarding BA metabolism, Cyp7A1, NTCP, BSEP, and OATP2 were lower in patients with adiponectin levels below the cutoff (Fig. 4D). In contrast, death receptor expression was increased in patients with DMXAA cell line lower adiponectin levels (Fig. 4E). Various growth factors, regulatory proteins, and (nuclear) receptors were analyzed for mRNA expression (Fig. 4F), although differences were observed for few targets (MET, KLF6/KLF6SV1,

and LXRa). The principal findings of this study relate BA transporters to hepatocyte apoptosis in NAFLD and uncover a potential role for adiponectin in BA homeostasis. The observations demonstrate a marked induction of genes involved in hepatocellular BA uptake and synthesis, which are repressed by SHP under physiological selleck chemicals conditions, in our cohort of superobese individuals. Treatment of hepatoma cells with FFA induces the same BA uptake and synthesis-related genes in a similar fashion. Adiponectin is inversely correlated with serum BAs and hepatocellular injury,

and low adiponectin levels predict simple steatosis as opposed to NASH in obese individuals. Patients with adiponectin levels below 29.16 ng/mL have significantly greater histological features of NASH, higher BA levels, and a lower expression of BA metabolism-related genes, uncovering a novel role for adiponectin and FFA in bile salt metabolism (Fig. 6). The pathogenesis of NAFLD is widely known to be associated with hepatocyte steatosis and FFA-induced lipotoxicity followed by the secretion of proinflammatory cytokines and stellate cell (HSC) activation, which in Mephenoxalone the end results in disease progression and fibrosis.21, 22 Since our group and others observed increasing BA concentrations

in NASH, in addition to lipotoxicity, BAs, as products of endogenous hepatic synthesis, may themselves contribute to liver injury in NAFLD.5 In this context, accumulation of BAs in hepatocytes causes hepatocyte death, giant cell hepatitis, and progressive liver damage in hereditary disorders requiring liver transplantation at a young age.23 The mutagenic potential of BAs may even explain the early development of hepatocellular carcinoma in children with hereditary BSEP deficiency.24 Hepatobiliary transport systems are regulated at a transcriptional and posttranscriptional level.9, 25 Nuclear receptors have been identified to function as regulators for positive and negative feedback pathways orchestrating bile formation under different clinical conditions.26 The nuclear BA receptor FXR plays a central role in BA homeostasis and regulates Na+-dependent (NTCP) BA uptake, apart from canalicular excretion (BSEP), as well as the rate-limiting step of BA formation (CYP7A1).27–30 Upon activation by BAs, FXR represses BA uptake and synthesis (NTCP, CYP7A1) by way of SHP and simultaneously activates BA efflux (BSEP).

Regarding BA metabolism, Cyp7A1, NTCP, BSEP, and OATP2 were lower in patients with adiponectin levels below the cutoff (Fig. 4D). In contrast, death receptor expression was increased in patients with MI-503 ic50 lower adiponectin levels (Fig. 4E). Various growth factors, regulatory proteins, and (nuclear) receptors were analyzed for mRNA expression (Fig. 4F), although differences were observed for few targets (MET, KLF6/KLF6SV1,

and LXRa). The principal findings of this study relate BA transporters to hepatocyte apoptosis in NAFLD and uncover a potential role for adiponectin in BA homeostasis. The observations demonstrate a marked induction of genes involved in hepatocellular BA uptake and synthesis, which are repressed by SHP under physiological selleck screening library conditions, in our cohort of superobese individuals. Treatment of hepatoma cells with FFA induces the same BA uptake and synthesis-related genes in a similar fashion. Adiponectin is inversely correlated with serum BAs and hepatocellular injury,

and low adiponectin levels predict simple steatosis as opposed to NASH in obese individuals. Patients with adiponectin levels below 29.16 ng/mL have significantly greater histological features of NASH, higher BA levels, and a lower expression of BA metabolism-related genes, uncovering a novel role for adiponectin and FFA in bile salt metabolism (Fig. 6). The pathogenesis of NAFLD is widely known to be associated with hepatocyte steatosis and FFA-induced lipotoxicity followed by the secretion of proinflammatory cytokines and stellate cell (HSC) activation, which in before the end results in disease progression and fibrosis.21, 22 Since our group and others observed increasing BA concentrations

in NASH, in addition to lipotoxicity, BAs, as products of endogenous hepatic synthesis, may themselves contribute to liver injury in NAFLD.5 In this context, accumulation of BAs in hepatocytes causes hepatocyte death, giant cell hepatitis, and progressive liver damage in hereditary disorders requiring liver transplantation at a young age.23 The mutagenic potential of BAs may even explain the early development of hepatocellular carcinoma in children with hereditary BSEP deficiency.24 Hepatobiliary transport systems are regulated at a transcriptional and posttranscriptional level.9, 25 Nuclear receptors have been identified to function as regulators for positive and negative feedback pathways orchestrating bile formation under different clinical conditions.26 The nuclear BA receptor FXR plays a central role in BA homeostasis and regulates Na+-dependent (NTCP) BA uptake, apart from canalicular excretion (BSEP), as well as the rate-limiting step of BA formation (CYP7A1).27–30 Upon activation by BAs, FXR represses BA uptake and synthesis (NTCP, CYP7A1) by way of SHP and simultaneously activates BA efflux (BSEP).

6C; Supporting Table 3). Acute reduction of PHB1 for 24 hours resulted in a 50% increase in cell proliferation (Fig. LY294002 supplier 6D). To see if the effect of PHB1 knockdown on cyclin D1 expression may be exerted at the level of E2F binding to its consensus sites on the cyclin D1 promoter, we performed ChIP analysis comparing E2F binding to different regions of the promoter that contain E2F binding sites. E2F binding increased (Fig. 6E), particularly in region −513 to −697 (500 ± 12% of scrambled control from three experiments, P < 0.05) of the cyclin D1 promoter in cells where PHB1 expression was reduced. E2F binding to the other regions also increased

significantly but to a much lesser degree (150% to 200%). Overexpression of PHB1 in AML12 cells reduced proliferation (Fig. 7B,D). However, whereas overexpression in Huh-7 cells

Cyclopamine supplier tended to lower proliferation, it was not statistically significant (Fig. 7A,C). To see if PHB1 expression in liver cancer cells can affect sensitivity to sorafenib, Huh-7 cells were treated with siRNA against PHB1 or overexpression vector to raise PHB1. This was then followed by sorafenib treatment. Apoptosis and proliferation were measured thereafter. PHB1 knockdown did not sensitize Huh-7 cells to sorafenib-induced apoptosis or inhibition in proliferation (Fig. 8). Overexpression of PHB1 also had no influence on sorafenib-induced apoptosis or inhibition of proliferation (data not shown). MAT is an essential enzyme for survival as it is responsible for the biosynthesis of SAMe, the principal biological methyl donor and, in mammalian liver, a precursor of GSH.13MAT1A is one of two MAT genes that encode for the catalytic subunit of MAT that

is largely expressed in normal differentiated Fossariinae mammalian liver.13 The expression and activity of hepatic MAT falls in patients with liver disease due to lower MAT1A mRNA level and inactivation of the MAT1A-encoded isoenzymes.13 This work was originally prompted by our observation that Mat1a KO mice have reduced PHB1 protein level from birth that persisted up to 8 months of age.10 Because PHB1 is known to stabilize mitochondrial proteins, we speculated that reduced PHB1 might have led to impaired mitochondrial function, oxidative stress, and susceptibility to many liver injuries in Mat1a KO mice.10–12Mat1a KO mice also develop HCC spontaneously.11 Whether reduced PHB1 could have contributed to this was unclear because there is tremendous controversy with regard to PHB1′s role as a tumor suppressor.1 Although the functional role of the PHB complex as a mitochondrial chaperone is well characterized, particularly in yeast,1, 3 whether it plays a similar role in mammals in vivo has been unclear because Phb1 and Phb2 knockout mice are lethal embryonically (www.informatics.jax.org/external/ko/lexicon/2210.html).

The aim of this study was to determine if baseline analysis of the NS3 viral region using ultra-deep pyrosequencing (UDPS) could help to predict SVR to triple therapy. Methods: Forty genotype 1 patients failing to achieve a SVR with Peg-IFNa + Ribavirin combination

(null responders: n=18; partial responders: n=14, relapsers: n=8 and retreated with triple therapy adding BOC or TPV were included. Their main characteristics were: mean age 55+/-8 years, 47.5% subtype 1a, 77.5% F3-F4. Baseline UDPS of the NS3-protease viral gene was performed on plasma and peripheral blood mononuclear cells (PBMC). Sequences obtained were analyzed in terms of resistance mutations with a threshold of 1% determined by using a control High Content Screening transcript. Heterogeneity of quasispecies was evaluated by the calculation of Shannon Entropy (SE). Results: Baseline mutations were found in 4 patients who achieved SVR with triple Proteasome inhibitor therapy and in 4 patients who did not. For these last patients, mutations were already major in three patients and persisted

until viral breakthrough. In the fourth patient, the mutated population accounted for only 1.4% of the total viral population at baseline but dramatically rose upon failure. In two patients, minor mutations were found in PBMC while not in plasma, and corresponded to mutations observed at the viral rebound. Compartmentalization between plasma and PBMC was confirmed with the analysis of the obtained sequences. More broadly, the NS3 quasipecies heterogeneity expressed

as SE was significantly lower at baseline in patients achieving SVR compared BCKDHB to nonSVR (SE= 26.98016.64 x 10-3 vs 44.93 ± 19.58 x 10-3, p=0, 0049). By multivariate analysis, independent predictors of SVR were F0F2 fibrosis stage (OR =13.3, CI95% 1.25141.096, p<0.03) and SE below median value (oR=5.4, CI95% 1.22-23.87, p<0.03). Conclusion: More than the presence of baseline minor mutations in plasma or in PBMC, NS3 viral heterogeneity determined by UDPS is an independent factor of SVR in previously treated patients receiving a triple therapy with an anti-protease drug. This parameter could be included in a score predicting response to therapy. Disclosures: Jean-Pierre H.