Analysis of data from the Haemophilia and Thrombosis Research Society (HTRS) Registry was performed on episodes where doses of ≥250 μg kg−1 were reported. From 2041 rFVIIa-treated bleeds, 172 bleeding episodes were identified in 25 individuals with CHwI who were treated with ≥1 higher doses (≥250 μg kg−1, ≥270 μg kg−1

or ≥300 μg kg−1) of rFVIIa between January 2004 and November 2008. Bleeds occurred in individuals ranging in age from 0.4 to 41.7 years who were predominantly non-Hispanic and white (40%) with haemophilia Doxorubicin A (88%). Bleed types most frequently treated with higher doses of rFVIIa were spontaneous (62–65%) or traumatic (27–32%). Bleed locations most frequently treated with higher doses of rFVIIa were joint (60–68%) or muscle (20–25%). A total of 1521 rFVIIa doses were administered (median, three doses per bleed); 26% were 250 μg kg−1 or higher (initial dose, 82%). Bleeding stopped in 93% (160/172) of bleeds treated with rFVIIa 250 μg kg−1 or higher. No serious adverse drug-related events or thrombotic complications were reported. This data

analysis from the HTRS Registry provides evidence of the safe and effective use of higher doses of rFVIIa (≥250 μg kg−1) in US practice. “
“Summary.  The most problematic complication of haemophilia A treatment is the development of inhibitors Selleck Z-VAD-FMK to FVIII. The highest risk of developing inhibitors is during the first 20 exposure days (EDs). If the patient can be brought through this high risk period without inhibitor development, the subsequent risk is low. Therefore, as a pilot project, we developed a prophylaxis regimen for the first 20–50 EDs specifically designed to induce tolerance to the administered FVIII and to minimize

inhibitor development by avoiding immunological danger signals. Twenty-six consecutive previously untreated patients (PUPs) with severe haemophilia A were treated with the new prophylaxis regimen and the incidence of inhibitor development in this group was compared with that in a historical control group of 30 consecutive PUPs treated N-acetylglucosamine-1-phosphate transferase with a standard joint protection prophylaxis regimen (40–50 IU kg−1, three times a week). There were no significant differences between the study and control groups in patient-related inhibitor risk factors such as ethnicity (all Caucasian), severity of haemophilia (all <1% FVIII), severity of FVIII gene mutation (P < 0.0006) nor in some treatment-related factors such as product type, age at first exposure, vaccination regimen or the need for surgery. 14 of 30 subjects given standard prophylaxis but only one of the 26 subjects given the new regimen developed an inhibitor (P = 0.0003, odds ratio 0.048, 95% CI: 0.001–0.372). Our results indicate that minimizing danger signals during the first 20 EDs with FVIII may reduce the risk of inhibitor formation. These results should be confirmed in a larger prospective clinical study.

The ER stress and cytopathologies seen in the hi559 liver resemble those seen in human NAFLD. Furthermore, Gene Set Enrichment Analysis (GSEA) of microarray data identified selective enrichment of genes involved in ERSR pathway in hi559 larvae; several of these genes are selectively overexpressed in the mutant liver. Together, these data support a model in which disrupted PtdIns synthesis leads to ER stress–mediated intracellular damage resulting in hepatic pathology

similar to that seen in NAFLD. CDIPT, CDP-diacylglycerol-inositol 3-phosphatidyltransferase; dpf, days postfertilization; ER, endoplasmic reticulum; ERSR, endoplasmic reticulum stress response; GI, gastrointestinal; GSEA, Gene Set Enrichment Analysis; ISH, in situ hybridization; mRNA, messenger RNA; NAFLD, nonalcoholic Selleck SCH727965 fatty liver disease; ORO, Oil Red O; PCR, polymerase chain reaction; PI, phosphoinositides; PIS, phosphatidylinositol synthase; RAD001 ic50 PtdIns, phosphatidylinositol; RT-PCR, reverse-transcription PCR;

UPR, unfolded protein response. The zebrafish line hi559 was obtained from a large-scale insertional mutagenesis screen.8 All fish husbandry was performed in accordance with local institutional animal care and use committee protocols. Heterozygous and homozygous fish were confirmed by way of genotyping using multiplex polymerase chain reaction (PCR). CY3-streptavadin (CY3-SA) labeling was performed as illustrated previously.7 For whole-mount in situ hybridization (ISH), embryos were processed as described.5 Probes and their corresponding accession numbers are provided in the Supporting Information. Alkaline phosphatase staining for vasculature and whole-mount Oil Red O (ORO) staining were

performed as described.21, 22 Total RNA was extracted only from 5-dpf wild-type and hi559 larvae using RNAeasy (Qiagen). Oligo dT–primed complementary DNA was then synthesized using SuperScript II RT (Invitrogen) and probed by way of reverse-transcription PCR (RT-PCR). cdipt mRNA was synthesized from a full-length linear DNA template using mMessage (Ambion) and purified by RNA clean (Zymo Research). cdipt and gfp mRNAs were injected into 1-cell stage embryos. For knockdown analyses of Cdipt, two zebrafish cdipt splice-blocking morpholinos were coinjected with tp53 morpholino into wild-type embryos. tp53 morpholino alone was injected as a control morpholino. See Supporting Information for primer and morpholino sequences. The PIS assay was performed essentially as described.23, 24 The assay was conducted in 100 μL total volume containing 0.2 mM CDP-DAG, 0.5 mM myo-[3H]inositol (5,000 cpm/nmol), 2 mM MnCl2, 50 mM Tris-Hcl (pH 8.0), 0.15% Triton X-100, and 50 μg of total protein isolated either from wild-type or hi559 larvae. After 1 hour of incubation at 37°C, the reaction was terminated by adding 0.35 mL chloroform and 0.5 mL 1 M MgCl2.

The 1-year and 3-year posttransplant survival rates in haemophilic recipients, 71% (95% CI:26–92%) and 38% (95% CI:6–72%), were similar to rates in non-haemophilic candidates, 66% (95% CI:44–80%) and 53% (95% CI:32–70%), respectively. The median time to graft loss was also not different between haemophilic and non-haemophilic transplant recipients, 1.29 years vs. 0.73 years, P = 0.80 (Fig. 1b). The 1-year and 3-year cumulative rates of treated rejection in haemophilic transplant recipients were 14% (95% CI:2–67%) and 36% (95% CI:10–85%), Selleckchem Alectinib whereas those in non-haemophilic transplant recipients were 36% (95%CI:21–59%)

and 43% (95%CI:25–66%), respectively. The median time to treated rejection also was not statistically different between haemophilic and non-haemophilic transplant recipients, 0.75 years vs. 0.02 years, P = 0.77 (Fig. 1c). Among transplant candidates who did not undergo transplantation, including 8 of 15 (53.3%) haemophilic and 62 of 89 (69.7%) non-haemophilic candidates, Table 2, significantly fewer haemophilic candidates remain alive, 3 (37.5%) vs. 49 (79.0%), P = 0.03 (Fig 2a). The haemophilic group was more likely than their non-haemophilic Protein Tyrosine Kinase inhibitor counterparts to die before receiving a transplant, 5 of 15 (33.3%) vs.

13 of 89 (14.6%), and more quickly, with a median time to death of 0.07 years in those with haemophilia vs. 0.42 years Pyruvate dehydrogenase in non-haemophilic subjects, P = 0.03, (Fig. 2a). The causes of pretransplant deaths were similar between groups, and included sepsis and multi-organ failure (Table 2). The median time to transplant, as measured by time on the transplant waiting list, was marginally longer in haemophilic as compared with non-haemophilic candidates, 0.15 years vs. 0.03 years, P = 0.15 (Fig. 2b). The median time to MELD = 25, as measured in time on the transplant

waiting list with MELD <25, was marginally shorter in haemophilic subjects, 0.01 years vs. 0.7 years, P = 0.06 (Fig. 2c). In univariate proportional hazards models for pretransplant mortality, including haemophilia status and baseline factors (Table 1), having haemophilia, HR = 3.0, P = 0.04, and higher baseline MELD score, HR = 1.2, P < 0.0001, were significantly associated with increased risk of pretransplant death. In the multivariate model, higher baseline MELD score was significantly associated with increased risk of pretransplant death, HR = 1.2 (95% CI:1.1–1.3), P < 0.0001, whereas being haemophilic was marginally associated with increased risk of pretransplant death, HR = 3.6 (95% CI:1.0–13.5), P = 0.06. When the time-to-event and proportional hazards models analyses were rerun using a male-only control, results remain unchanged (data not shown). This study confirms that HIV/HCV co-infected individuals with haemophilia experience poorer pretransplant outcomes than co-infected individuals without haemophilia.

Therapeutic occlusion of tumor feeder vessels is associated with lower local recurrence. “
“Chronic hepatitis C virus (HCV) infection is one of the leading causes of Ceritinib research buy cirrhosis and hepatocellular

carcinoma worldwide. It is highly prevalent among injection drug users (IDUs) but is often undiagnosed because they represent an underprivileged group that faces multiple barriers to medical care. Here, we report the results of the New Life New Liver Project, which provides targeted HCV screening and education for ex-IDUs in the community. Patients were recruited through the social worker networks and referrals by fellow ex-IDUs, and rapid diagnosis was based on point-of-care anti-HCV testing at rehabilitation centers. From 2009 to 2012, we served 234 subjects. One hundred thirty (56%) subjects were anti-HCV positive. The number needed to screen to detect one patient with positive selleck screening library anti-HCV was 1.8 (95% confidence interval, 1.6–2.0). However, only 69 (53%) HCV patients attended subsequent follow-up at regional hospitals, and 26 (20%) received antiviral therapy. Patients who attended follow-up were older, had higher education level and more active disease as evidenced by higher alanine aminotransferase, HCV RNA, and liver stiffness measurement by transient elastography. Targeted

screening in ex-IDUs is effective in identifying patients with HCV infection in the community. Improvement in the referral system and introduction of interferon-free regimens are needed to increase treatment uptake. Chronic hepatitis C is one of the leading causes of end-stage liver disease and hepatocellular carcinoma (HCC) worldwide. Since 2007, hepatitis C virus (HCV) has surpassed stiripentol human immunodeficiency virus as a cause of death in the United States.[1] In the past few years, with the knowledge on the lifecycle

of HCV, there have been exciting developments in direct-acting antivirals that can lead to sustained virologic response in 60–90% of patients.[2] Successful treatment results in regression of cirrhosis and reduces the risk of HCC.[3, 4] Since chronic hepatitis C rarely causes symptoms, at least half of the patients in the community are undiagnosed.[5] The infection is most commonly found in injection drug users (IDUs), with prevalence ranging from 20% to 90%.[6] With proper care, IDUs can have good adherence to treatment and a sustained virologic response rate similar to that of other patients.[7, 8] HCV treatment for IDUs is also cost-effective.[9] Therefore, current guidelines support HCV screening in IDUs.[10, 11] However, there is one missing link. IDUs represent an underprivileged group that faces multiple barriers to medical care.[12] If HCV infection remains undiagnosed, therapeutic efficacy cannot be translated into effectiveness at the population level.[13] In this article, we report a model of targeted HCV screening in ex-IDUs in the community and evaluate the efficacy of the program.

Dual infection was defined as the co-existence of sequences distributed into two or more geno/subtypes. DS results showed the most prevalent genotype in hemophiliac patients was genotype 1b (52.3%), followed by genotype 1a (23.8%) and undetermined (19.0%). All genotype 1a cases were co-infected with HIV. Genotype analyses of NGS consensus sequences yielded consistent results with those of DS, and additionally revealed the genotypes of those undetermined samples to be 1a (4.8%), 2a (9.5%) and 2b (4.8%). Moreover, haplotype reconstruction of HCV hypervariable region (HVR) and NS3 region indicated Selleckchem GSK1120212 that 42.9% of the patients

had dual/triple geno/subtype infections. Focusing on NS3 region, categorical analyses revealed the association between HCV mono-infection and 1b as a dominant genotype (p = 0.008), HIV co-infection and multiple genotypes (p = 0.009), and, histories of blood transfusion (BTF) and multiple genotypes (p = 0.012). Furthermore, the existence of non-1b sequence was tightly associated with HIV co-infection (p = 0.0002), and BTF histories (p = 0.003).

This pilot study demonstrated that multi-geno/subtypic multiple infections may occur more frequently than previously expected, especially in patients having HIV co-infection and BTF histories. Also, our NGS-based haplotype reconstruction approach click here is useful for detecting low-abundant haplotypes undetectable from DS. Whether those harbored viral populations may affect the outcome of DAA therapy should be clarified in forthcoming studies. Disclosures: The following people have nothing to disclose: Masato Ogishi, PFKL Hiroshi Yotsuyanagi, Takeya Tsutsumi, Hiroyuki Gatanaga, Kyoji Moriya, Kazuhiko Koike BACKGROUND MK-5172, a potent HCV NS3/4A protease inhibitor, is being assessed in two phase 2 studies in combination with MK-8742 (a NS5a inhibitor)+/−RBV (C-WORTHy, PN035) and IFN/RBV (PN038) for 12 weeks.

The aim is to characterize the impact of IFN/RBV-free therapy on HRQOL. METHODS HRQOL is assessed using the SF-36v2® Health Survey Acute at baseline, therapy week 4 (TW4), end of therapy (EOT), and follow-up weeks 12 and 24. Means (standard deviations(SD)) are used to describe change from baseline during therapy in the health domain scores and mental component summary (MCS) and physical component summary (PCS) scores. Wilcoxon signed-rank test is used to compare changes from baseline in MCS and PCS scores within each treatment group at TW4 and EOT. RESULTS 123 subjects (24% HIV co-infected, 35% cirrhotic, 27% null-responders to IFN/ RBV) received MK-5172/MK-8742; 125 subjects (23% HIV co-infected, 34% cirrhotic, 26% null-responders to IFN/RBV) received MK-5172/MK-8742/RBV; and 58 mono-infected, treatment-naïve, non-cirrhotic subjects with GT 1 infections received MK-5172 (50 or 100 mg) + IFN/RBV. The SVR rates are high for all treatment groups and subpopulations (86%-100%).

Dual infection was defined as the co-existence of sequences distributed into two or more geno/subtypes. DS results showed the most prevalent genotype in hemophiliac patients was genotype 1b (52.3%), followed by genotype 1a (23.8%) and undetermined (19.0%). All genotype 1a cases were co-infected with HIV. Genotype analyses of NGS consensus sequences yielded consistent results with those of DS, and additionally revealed the genotypes of those undetermined samples to be 1a (4.8%), 2a (9.5%) and 2b (4.8%). Moreover, haplotype reconstruction of HCV hypervariable region (HVR) and NS3 region indicated Tamoxifen in vitro that 42.9% of the patients

had dual/triple geno/subtype infections. Focusing on NS3 region, categorical analyses revealed the association between HCV mono-infection and 1b as a dominant genotype (p = 0.008), HIV co-infection and multiple genotypes (p = 0.009), and, histories of blood transfusion (BTF) and multiple genotypes (p = 0.012). Furthermore, the existence of non-1b sequence was tightly associated with HIV co-infection (p = 0.0002), and BTF histories (p = 0.003).

This pilot study demonstrated that multi-geno/subtypic multiple infections may occur more frequently than previously expected, especially in patients having HIV co-infection and BTF histories. Also, our NGS-based haplotype reconstruction approach www.selleckchem.com/EGFR(HER).html is useful for detecting low-abundant haplotypes undetectable from DS. Whether those harbored viral populations may affect the outcome of DAA therapy should be clarified in forthcoming studies. Disclosures: The following people have nothing to disclose: Masato Ogishi, ioxilan Hiroshi Yotsuyanagi, Takeya Tsutsumi, Hiroyuki Gatanaga, Kyoji Moriya, Kazuhiko Koike BACKGROUND MK-5172, a potent HCV NS3/4A protease inhibitor, is being assessed in two phase 2 studies in combination with MK-8742 (a NS5a inhibitor)+/−RBV (C-WORTHy, PN035) and IFN/RBV (PN038) for 12 weeks.

The aim is to characterize the impact of IFN/RBV-free therapy on HRQOL. METHODS HRQOL is assessed using the SF-36v2® Health Survey Acute at baseline, therapy week 4 (TW4), end of therapy (EOT), and follow-up weeks 12 and 24. Means (standard deviations(SD)) are used to describe change from baseline during therapy in the health domain scores and mental component summary (MCS) and physical component summary (PCS) scores. Wilcoxon signed-rank test is used to compare changes from baseline in MCS and PCS scores within each treatment group at TW4 and EOT. RESULTS 123 subjects (24% HIV co-infected, 35% cirrhotic, 27% null-responders to IFN/ RBV) received MK-5172/MK-8742; 125 subjects (23% HIV co-infected, 34% cirrhotic, 26% null-responders to IFN/RBV) received MK-5172/MK-8742/RBV; and 58 mono-infected, treatment-naïve, non-cirrhotic subjects with GT 1 infections received MK-5172 (50 or 100 mg) + IFN/RBV. The SVR rates are high for all treatment groups and subpopulations (86%-100%).

Generally, 15–25% of cases have colorectal liver metastasis (CRLM) at diagnosis.[1, 2] Furthermore, CRLM occurs in 25–50% of cases with the resection of primary colorectal tumor over 3 years.[3-5] Hepatic resection is accepted as the only treatment contributing to the long-term survival and cure of patients with CRLM.[6] However, only 15–20% of patients with CRLM are considered candidates for hepatic resection at the time of presentation.[7-10] The significance of other tumor destruction modalities, such as radiofrequency ablation, remains controversial.[11] Of those patients who

undergo hepatic resection, there are at least two categories of patients with CRLM. The first category is clearly or potentially resectable at the time

of presentation. selleckchem The second category is initially unresectable, but convertible to be resectable after treatment with anticancer agents including molecular targeted agents, which we refer to as “conversion surgery”. The purpose of neoadjuvant chemotherapy for resectable CRLM is to downsize CRLM lesions and maximize the remnant liver as well as to reduce the residual micrometastasis, while less extensive resections can be carried out in keeping with the curative intent. However, until now, the role of neoadjuvant therapy prior to the resection of CRLM is not yet proven and remains controversial. click here The largest prospective trial consisted of 364 patients with less than five initially resectable CRLM (European Organization for Research and Treatment of Cancer Intergroup 40983 trial) randomized with perioperative chemotherapy (four to six preoperative and six postoperative cycles of FOLFOX4) or surgery alone, and showed a clinical benefit in 3-year progression-free survival (36.2% vs 28.1%) but not in 5-year overall survival.[12] The FOLFOX regimen may reduce the risk of events in terms of progression-free survival but not necessarily improve long-term survival compared with surgery alone in eligible and initially resectable patients. On the other hand, regarding initially unresectable CRLM, the “conversion surgery” strategy

has been widely used and accepted. Actually, 5-fluorouracil (5-FU)/leucovorin (LV) plus oxaliplatin (L-OHP); FOLFOX or irinotecan (Iri); FOLFIRI or combination Idelalisib order of both; FOLFOXIRI with or without molecular-targeted agents as preoperative strategy have recently achieved higher conversion rates and better clinical outcomes.[13-20] Particularly in L-OHP-based chemotherapy, the conversion rate ranged 7–51% in patients with unresectable CRLM. The effectiveness of triple combination chemotherapy, FOLFOXIRI, for patients with initially unresectable CRLM has been reported to have an improved response rate (60% vs 34%) and higher R0 resection rate among patients with CRLM only compared with the FOLFIRI regimen (36% vs 12%).

Finally, the three animals with proven envelope mutations always experienced a delay in the appearance of viral RNA in the plasma, comparable to that of four H06-treated animals in which the viral amino acid sequence Vadimezan remained conserved. This again points towards a failure of neutralization rather than viral escape. Despite this seemingly low cross-genotype neutralizing efficiency, our results are nevertheless encouraging for the quest for a potent cocktail of broad neutralizing monoclonal antibodies that could be used in a clinical setting for the prevention of graft reinfection after liver transplantation and holds promise for the development

of a successful prophylactic HCV vaccine. First, only a minor fraction of the polyclonal H06-antibody pool is directed against HCV envelope proteins, and definitely not all of these antibodies will have neutralizing activity. Second, Zhang et al.29 recently showed that plasma from Patient H contains antibodies against epitope II (residues 434-446 of E2) that interfere with the action of neutralizing antibodies. Depletion of these interfering antibodies from our H06-pool could possibly improve the protection rate. For this reason, the use of a pool consisting of well-defined monoclonal antibodies selected for their neutralizing

ability would be more efficacious for prophylaxis than pooled nonfractionated polyclonal plasma.30-32 “
“Dyspepsia is perhaps the most common gastrointestinal

disease universally. The prevalence Selleckchem Fulvestrant of dyspepsia ranges from 7–40% in population based studies worldwide. These figures vary with definition of dyspepsia Thalidomide used and also with the survey methodology. As with Western studies, functional dyspepsia (FD) predominates in Asia. With a decline in peptic ulcer disease and gastric cancer, the proportion of FD is set to increase further. Studies have shown FD to account for 50–70% of cases of uninvestigated dyspepsia. In Malaysia dyspepsia has been reported in up to 15% of a rural and 25% of an urban population. No racial differences were seen in the rural survey. In the urban survey, Malays and Indians were found to have significantly more dyspepsia than Chinese. No clear explanation can be found for these racial differences. In clinical practice, Malays seem to complain a lot of wind and bloating in the “stomach.” This is interesting to note when you compare it with the prevalence of H. pylori which is distinctly less common amongst Malays compared to the Indians and Chinese. As with many Asian populations, many Malaysians do not consult for complains of dyspepsia. Many will self medicate and others may even bear with their complains. This is probably true in the rural population.

There was no difference in the proportion of deficiencies between elderly who reported a dental visit in the preceding year or not having seen a dentist. A quarter of the prostheses required replacement. The findings from this and the NHANES studies demonstrate that an engaged and recognized prosthodontic dental school faculty continues to be as important now as it was a generation ago. “
“Purpose: The purpose of this study was to assess the performance of an intraoral dental colorimeter. Materials and Methods: In Dasatinib chemical structure vivo repeatability of an intraoral colorimeter was assessed by performing color measurements of 30 individuals’ right maxillary central incisor.

Three consecutive measurements from each individual were made. In the in vitro part of the study, 25 metal-ceramic and 25 all-ceramic specimens were prepared. Five shades of metal-ceramic

and all-ceramic specimens were selected for color determination. A widely recognized in vitro colorimeter was used as the control group for the in vitro performance assessment of the in vivo colorimeter. The color differentiation capability of two colorimeters was compared with the readings obtained from ceramic specimens. ΔE values between shade groups of ceramic specimens were calculated and statistically analyzed with Student’s t-test. The repeatability of the intraoral instrument was evaluated statistically with Intraclass correlation coefficient. Results: The in vivo evaluation results showed that the overall repeatability coefficient values of L*, a*, and b* notations of the intraoral buy BGB324 colorimeter were “excellent.” The color differences (ΔE) calculated between the colorimeters were significant only between shades A1-B1 for metal-ceramic specimens (p= 0.002); however, from 5 of 10 shade couples of all-ceramic specimens, the color differences obtained from the readings of the in vivo colorimeter were significantly different from that of the in vitro

colorimeter (p < 0.001). For all specimens, the differences between nearly ΔE values were within clinically acceptable limits (<3.5). Conclusions: Within the limitations of this study, the intraoral colorimeter exhibited successful in vivo repeatability; however, the color difference detection performance of the device varied depending on the translucency of the specimens. "
“When a screw fracture occurs on a cement-retained, implant-supported restoration, the abutment and restoration are completely separated from the implant’s internal connection. Traditionally, an access hole is drilled through the crown to retrieve the broken screw, and the restoration can be placed again as a screw-retained restoration. This clinical report documents a patient whose broken abutment screw was retrieved from the restoration by burning off the cement and separating from the abutment without drilling an access hole.

nubiscan.unibas.ch). The SHP1-Luc-pGL3-basic vector was prepared as described11 containing the −571 to +1 bp fragment of the 5′-UTR of SHP1 (NR0B2)

gene inserted in the luciferase reporter containing vector pGL3-basic (Promega, Madison, WI). The BSEP promoter construct containing a 140-bp fragment of the 5′-UTR BSEP (ABCB11) has been described.12 Detailed outline of primers and subcloning strategies for the promoter fragments of the 5′-UTR of the SLCO1B1 from genomic DNA is summarized in Supporting Table 1. HepG2 cells were plated in 24-well plates. After 24 hours, the cells were transfected with 250 ng of the reporter vector (pGL3 basic variants), 25 ng of pRL-CMV (Promega) to normalize transfection efficiency, and 250 ng of the human nuclear receptors expression plasmid (LXRα- or FXR-pEF6) or vector control in 200 μL Opti-MEM Epacadostat datasheet (Invitrogen) using Lipofectin (Invitrogen). Cells were incubated for 16 hours with the transfection mixture, then treated

for 24 beta-catenin activation hours with 1 μM of a tested compound. Luminescence was quantified using a plate reader (Fluoroskan Ascent FL, Thermo-Fisher, Waltham, MA). Luciferase activities in the presence of the nuclear receptor were expressed as the percentage of cells transfected with blank vector. Huh-7 cells or primary hepatocytes were grown in 12-well plates and pretreated with agonists of LXRα or FXR or vehicle control, respectively, for

12 hours. Afterward, the cells were briefly washed with OptiMEM and then incubated with tritium-labeled taurocholate (0.4 μM) or rosuvastatin (1 μM). After 10 minutes of incubation at 37°C, the cells were washed twice with ice-cold PBS and lysed in the presence of 1% sodium dodecyl sulfate. Cellular uptake Glycogen branching enzyme was determined using a liquid scintillation counter (Tri-Carb 2900TR, PerkinElmer Life Sciences). After treatment of human hepatocytes, the cells were harvested and lysed by way of repeated thawing and freezing in 5 mM Tris HCl in the presence of Protease Inhibitors. Protein content was determined using bicinchoninic acid. Cell lysates were separated by way of sodium dodecyl sulfate–polyacrylamide gel electrophoresis and electrotransferred to a nitrocellulose membrane in a tank-blotting system (Bio-Rad, Munich, Germany). For detection of OATP1B1, a specific antiserum was used as described.4 For DNA cross-linking and chromatin immunoprecipitation, the EZ ChIP Assay (Millipore, Billerica, MA) was used according tot the manufacturer’s instructions. Briefly, Huh-7 cells were cultured in 10-cm dishes and treated for 24 hours with dimethyl sulfoxide, chenodeoxycholic acid (CDCA) (1 μM), fexaramine (1 μM), GW4065 (1 μM), TO-901317 (1 μM), or GW3965 (1 μM). DNA was cross-linked, sheared by sonication (Virsonic 100, Virtis, Gardiner, NY), then incubated with 5 μg antibody overnight at 4°C.