Lentiviral vectors pLKO1 TRC (Addgene 10879) and pWPI1 (Addgene

Lentiviral vectors pLKO.1 TRC (Addgene 10879) and pWPI.1 (Addgene 12254) were used for constructing

recombinant lentiviruses. Oligonucleotides encoding hairpin precursors for si259 (5′-TTTCCAGGAGCAACCA TAATT-3′, corresponding to nt259-279 of PROX1 ORF) and si1646 (5′-GGCTCTCCTTGTCGCTCA TAA-3′, corresponding to nt1646-1666 of PROX1 ORF) were used for generating short interference RNA (siRNA) constructs. A scrambled sequence (Scr) was used as a control. Synonymous mutations were introduced into the target sequence of si1646 (5′-GGCTCTCATTATCACTCATAA-3′, mutations underlined) in PROX1 ORF to generate si1646-resistant PROX1. pEGFP-Prox1 was generated by inserting PROX1 cDNA into pEGFP-C2 (ClonTech, Mountain View, CA). pGL2-HIF-1α selleck chemicals llc contained the promoter (−572/+284) of HIF-1α in pGL2-Basic (Promega, Madison, WI). Cell transfection and luciferase reporter assays were performed as described.[22] The procedures are detailed in the Supporting Methods. Co-IP, GST pulldown, and western blot were performed as described.[22] Antibodies used in western blot experiments are listed in Supporting Table S2. The procedures are detailed in the Supporting

Methods. Anti-PROX1 monoclonal antibody (mAb) (Cell Signaling Technology, Danvers, MA) was used to immunoprecipitate sonicated chromatins prepared from Huh7 or MHCC-97H cells. Preimmune IgG was used for specificity control. Immunoprecipitated DNA was Rucaparib clinical trial quantitated for HIF-1α promoter (Ct[IP]) using qrtPCR (forward, 5′-GGTGAGGCGGGCTT GCGGGAG-3′; reverse, 5′-GAGGAGCTGAGGCAG CGTCAGGG-3′). DNA from 5% input was quantitated for HIF-1α promoter in parallel (Ct[Input]). The relative occupancy was calculated using the equation: 2^(Ct[Input]-Ct[IP]) *100%. The procedures are detailed in the Supporting Methods. Animal experimental protocols were approved by the Animal Ethics Committee of Shanghai Medical College, Fudan University. Eight-week-old nude mice (BALB/c) were randomly divided into groups

(six mice/group) before inoculation or injection. Cells were inoculated into the liver parenchyma of nude mice (BEL-7402 derived cells, 2.0 × 106 cells/mouse) or subcutaneously injected into nude mice (MHCC-97H derived cells, 1.0 × 107 cells/mouse). The mice were sacrificed after 8 weeks and the number of metastatic 上海皓元 tumors was assessed by double-blinded evaluation. Statistical analysis was performed with SPSS v. 13.0. Kaplan-Meier analysis was used for survival analysis and the log-rank test was chosen to compare the difference. Pearson χ2 test or Fisher’s exact test were employed to compare qualitative variables, while Student t test was used for quantitative variables. A Cox proportional hazards model was adopted for multivariate analysis. Receiver operating characteristic (ROC) curve analysis was applied to assess the predictive values of variables. P < 0.05 was considered statistically significant for all tests.

The data described above, together with our previous characteriza

The data described above, together with our previous characterization of the inhibitory effect of supplemental glucose on liver

regeneration,9 suggest Neratinib molecular weight that perturbations in systemic glucose metabolism may contribute to suppressed regeneration in fld mice. The impaired regenerative response associated with dextrose supplementation was characterized by augmented expression of CCAAT/enhancer binding protein alpha (C/EBPα), p21, and p27.9 Therefore, hepatic expression of these factors was compared between fld and control mice. The results showed that C/EBPα and C/EBPβ mRNA and p27 protein expression were comparable in fld and controls (Supporting Fig. 2 and Fig. 5D-F); however, p21 protein was increased in fld liver (Fig. 5D-F). These data raise the possibility that dysregulated p21 expression contributes to impaired regeneration in fld mice. The adipose-derived

hormones adiponectin and leptin have each been identified as regulators of liver regeneration.13, 25–29 To investigate whether deficiency of either hormone might contribute to impaired regeneration in fld mice, plasma levels of each were determined before and after partial hepatectomy in fld and control mice. This analysis showed that circulating adiponectin and leptin levels were significantly lower in fld animals at baseline (Fig. 7A-C). Following partial hepatectomy, leptin levels declined in controls and remained low in fld mice (Fig. 7C). Because leptin deficiency is associated with impaired liver regeneration,13, 28, 30 the effect of leptin supplementation

MCE on regeneration in fld mice was investigated. LDK378 cost This analysis showed that a regimen of leptin supplementation sufficient to rescue impaired regeneration in CCl4-treated ob/ob mice13 did not augment and, in fact, suppressed hepatocellular proliferation 36 hours after partial hepatectomy in fld mice compared to untreated fld mice and leptin-treated controls (Supporting Fig. 3). Adiponectin levels increased after partial hepatectomy in controls (Fig. 7B), but remained almost undetectable in fld mice (Fig. 7A,B). These data suggest that impaired liver regeneration in fld mice may be mechanistically related to that recently described in adiponectin-null mice.26 Diminished activation of signal transducer and activator of transcription 3 (STAT3) and augmented induction of expression of suppressor of cytokine signaling 3 (SOCS3) were observed in liver after partial hepatectomy in those animals.27 Therefore, STAT3 activation and SOCS3 expression, each of which modulate liver regeneration,31, 32 were quantified after partial hepatectomy in fld and control animals. The results showed comparable STAT3 phosphorylation in both groups; however, the ratio of phosphorylated:total STAT3 was reduced. Moreover, in contrast to the analysis of adiponectin-null mice,27 hepatic SOCS3 expression after partial hepatectomy was significantly lower in fld mice than in controls (Fig.

Monitoring the spatial distribution of over 1,000 proteins, we fo

Monitoring the spatial distribution of over 1,000 proteins, we found unexpectedly that all liver metastasis lesions displayed a reproducible, zonally delineated pattern of functional and therapeutic biomarker heterogeneity.

The peritumoral region featured elevated lipid metabolism and protein synthesis, the rim of the metastasis displayed increased cellular growth, movement, and drug metabolism, whereas the center of the lesion was characterized by elevated carbohydrate metabolism and DNA-repair activity. From the aspect of therapeutic targeting, zonal expression of known and novel biomarkers was evident, reinforcing the Selleck NVP-BEZ235 need to select several targets in order to achieve optimal coverage of the lesion. Finally, we highlight two novel antigens, LTBP2 and TGFBI, whose expression is a consistent feature of CRC liver metastasis. We demonstrate their in vivo antibody-based targeting and highlight their potential usefulness for clinical applications. Conclusion: The proteome heterogeneity of human CRC liver metastases has a distinct, organized pattern. This particular hallmark can now be used Opaganib molecular weight as part of

the strategy for developing rational therapies based on multiple sets of targetable antigens. (Hepatology 2014;59:924–934) “
“Aim:  Because polymorphisms of cyclooxygenase-2 (COX-2) and osteopontin (OPN) promoter regions and a promoter/enhancer region of forkhead box protein 3 (FOXP3) gene are known to affect immune responses, we examined whether these polymorphisms can influence susceptibility to hepatitis C virus (HCV) infection and progression of liver disease. Methods:  Peripheral

blood samples were obtained from 104 Japanese patients with chronic HCV infection and 74 healthy Japanese donors. Polymerase chain reaction 上海皓元 single-stranded conformational polymorphism analysis of genomic DNA was performed to determine the polymorphisms. Results:  The risk of persistent HCV infection was decreased in subjects with –1195GG genotype of the COX-2 promoter region. However, in patients with chronic HCV infection, the –1195GG genotype was associated with advanced-stage liver disease. A luciferase reporter assay performed to analyze the effect of single nucleotide polymorphisms (SNP) (–1195A or –1195G) in COX-2 gene on transcriptional activity using the HepG2, Huh7 and HeLa cell lines indicated that the –1195G genotype showed higher transcriptional activity than the –1195A genotype. SNP of OPN and FOXP3 did not differ between patients with chronic HCV infection and controls. However, the –443TT genotype of the OPN promoter region was associated with increased inflammatory activity of the liver.

Monitoring the spatial distribution of over 1,000 proteins, we fo

Monitoring the spatial distribution of over 1,000 proteins, we found unexpectedly that all liver metastasis lesions displayed a reproducible, zonally delineated pattern of functional and therapeutic biomarker heterogeneity.

The peritumoral region featured elevated lipid metabolism and protein synthesis, the rim of the metastasis displayed increased cellular growth, movement, and drug metabolism, whereas the center of the lesion was characterized by elevated carbohydrate metabolism and DNA-repair activity. From the aspect of therapeutic targeting, zonal expression of known and novel biomarkers was evident, reinforcing the BMS-907351 datasheet need to select several targets in order to achieve optimal coverage of the lesion. Finally, we highlight two novel antigens, LTBP2 and TGFBI, whose expression is a consistent feature of CRC liver metastasis. We demonstrate their in vivo antibody-based targeting and highlight their potential usefulness for clinical applications. Conclusion: The proteome heterogeneity of human CRC liver metastases has a distinct, organized pattern. This particular hallmark can now be used learn more as part of

the strategy for developing rational therapies based on multiple sets of targetable antigens. (Hepatology 2014;59:924–934) “
“Aim:  Because polymorphisms of cyclooxygenase-2 (COX-2) and osteopontin (OPN) promoter regions and a promoter/enhancer region of forkhead box protein 3 (FOXP3) gene are known to affect immune responses, we examined whether these polymorphisms can influence susceptibility to hepatitis C virus (HCV) infection and progression of liver disease. Methods:  Peripheral

blood samples were obtained from 104 Japanese patients with chronic HCV infection and 74 healthy Japanese donors. Polymerase chain reaction 上海皓元医药股份有限公司 single-stranded conformational polymorphism analysis of genomic DNA was performed to determine the polymorphisms. Results:  The risk of persistent HCV infection was decreased in subjects with –1195GG genotype of the COX-2 promoter region. However, in patients with chronic HCV infection, the –1195GG genotype was associated with advanced-stage liver disease. A luciferase reporter assay performed to analyze the effect of single nucleotide polymorphisms (SNP) (–1195A or –1195G) in COX-2 gene on transcriptional activity using the HepG2, Huh7 and HeLa cell lines indicated that the –1195G genotype showed higher transcriptional activity than the –1195A genotype. SNP of OPN and FOXP3 did not differ between patients with chronic HCV infection and controls. However, the –443TT genotype of the OPN promoter region was associated with increased inflammatory activity of the liver.

2007b) Similarly, a SNP in the cytochrome b gene of Venturia ina

2007b). Similarly, a SNP in the cytochrome b gene of Venturia inaequalis, corresponding to G143A substitution related to strobilurin resistance, was monitored by qPCR and revealed a higher mutation level in populations from conventional chemical control fields as compared to an organic orchard (Michalecka et al. 2011). A field of particular interest is the prediction of epidemics through the

early quantification of the pathogen inoculum during its latent phase. This aspect can be particularly important for plant pathogens that cannot be detected CT99021 by using conventional culturing methods as in the case of cereal rust diseases. The early detection of latent infections of rust on leaves of cereals can be used to estimate infection levels before the appearance of the disease and provides critical information for predicting it. Accurate forecast and prediction systems for stripe rust GW-572016 datasheet (Puccinia striiformis) have been developed for some geographical regions of China and greatly benefit from sensitive and rapid methods to detect rust pathogens

in the dormant stage in young wheat plants (Huang et al. 2011; Yan et al. 2012). Similarly, latent infections play a major role in the development of epidemics of the wheat powdery mildew caused by Blumeria graminis f.sp. tritici, and accurate qualitative and quantitative detection of the pathogen would provide useful information for predicting possible disease development in the coming growing season (Zeng et al. 2010). The detection of fungal latent infections can be very important also for predicting the evolution of a number of diseases of fruit and vegetables that frequently become manifest only after harvest and/or periods of storage and shelf life (Thomidis and Michailides 2010). The most important postharvest

pathogens, including B. cinerea, Monilia spp., Alternaria spp., Colletotrichum spp. and Penicillium medchemexpress spp., commonly cause latent infections in the field on unripe fruits and become active later when fruit ripe and conducive environmental conditions occur. Recently, a very high incidence of latent infection of B. cinerea has been revealed in apparently healthy grape berries and stamens (80 and 65%, respectively) at harvesting time (Sanzani et al. 2012a). Interestingly, the incidence of the latent infections was directly correlated with the actual disease incidence on bunches after cold storage and shelf life. Therefore, the early, rapid and accurate detection of field infections is useful for devising disease prediction models, improving timing and efficacy of preharvest control method applications. Furthermore, it might help in selecting the lowest contaminated parcels to be destined to long-time storage or to be sent to distant markets (Sanzani et al. 2012a).

16 Statistical significance was set to P < 005 and all statistic

16 Statistical significance was set to P < 0.05 and all statistical tests were two-tailed. Selleckchem AZD2014 Statistical analysis was performed using Stata 12.1

(Stata Corp, College Station, TX) together with the user-written OGLM package.15 As mentioned, inclusion in the F4 group (40 patients) derived either from histopathological staging at the time of the study or on clinical, laboratory, sonographic, and endoscopic parameters. In this group, 27 patients out of 40 were classified as Child-Pugh A, whereas 13 were classified as Child-Pugh B. The presence of esophageal varices (OV) was detected in 20 out of 40 patients, eight in the Child-Pugh A group (four OV grade 1, four OV grade 2) and in 12 in the Child-Pugh B group (four OV grade 1, eight OV grade 2). In the absence of previous data precisely indicating the exact time BIBW2992 manufacturer of LS postmeal peak increase, LS measurements

were performed 15, 30, 45, 60, and 120 minutes after the onset of the meal. Figure 1 illustrates the individual changes of stiffness following the onset of the meal test in the whole patient population according to the degree of fibrosis. Although most patients, irrespective of the stage of fibrosis, presented a peak increase after 30 minutes, some variability was observed, with some patients peaking at 15 or 45 minutes. Values returned to baseline levels within 120 minutes in all patients independently of the stage of fibrosis. As illustrated in Table 3, changes in liver stiffness were evaluated by means 上海皓元 of the following continuous indexes: S0 = baseline value of stiffness, S15-60 = values at 15, 30, 45, and 60 minutes during the meal test, respectively; Smin = minimum value of stiffness, Smax = maximum value

of stiffness, Sdelta (kPa) = (maximal stiffness − basal stiffness), Sdelta (%) = (maximal stiffness − basal stiffness) / basal stiffness × 100. With the exception of Sdelta (%), which showed a decreasing trend, all stiffness indexes showed an increasing trend for increasing stages of fibrosis (P < 0.0001 for all, Jonckheere-Terpstra test), as also illustrated in Fig. 2 for Sdelta (kPa). Since most centers do not apply a fasting time before the TE procedure, the probability of detecting fibrosis stage at each timepoint: basal, 15, 30, 45, and 60 minutes postmeal was evaluated (Fig. 3). It is evident from the comparison of the probability curves that no other timepoint was superior than S0 in detecting any stage of fibrosis. The same analysis was applied to the comparison of basal stiffness and delta stiffness based on the peak change irrespective of the postmeal timepoint. Figure 4 illustrates the probability (point estimate and 95% confidence intervals) of fibrosis stage (F0-F1, F2-F3, andF4) on the basis of S0 (kPa) and Sdelta (kPa).

AL-516 was assessed in vivo in the dog after oral dosing for the

AL-516 was assessed in vivo in the dog after oral dosing for the ability to form the active AL-516 NTP in liver. Results: In the stable genotype (GT) 1b replicon assay, AL-516 exhibits potent antiviral activity with an EC50 of 6.5 nM. In transient chimeric GT-1b replicons with NS5B regions derived from GT 1-4, AL-516 demonstrates pan-genotypic activity with EC50 values < 10 nM. AL-516 retains activity versus replicon mutants resistant to nucleoside Y-27632 solubility dmso and non-nucleoside polymerase, NS3/4A protease and NS5A inhibitors. AL-516 exhibits an excellent selectivity profile with no inhibition of mitochondrial protein synthesis (IC50 >100 μM).

The AL-516 NTP is a potent inhibitor of the HCV NS5B polymerase with an IC50 of 240 nM and a Ki of 22 nM, and acts as a chain-terminator of RNA synthesis. The AL-516 NTP retains potency against the NS5B S282T variant with an IC50 of 180 nM. The AL-516 NTP is not a substrate for human mitochondrial RNA polymerase and demonstrates no inhibition (IC50 >100 μM) of human DNA or RNA polymerases. In primary human hepatocytes, the AL-516 NTP is rapidly formed and demonstrates a 11.4 hr half-life, indicating potential for QD dosing. Following

oral administration to dogs at 5 mg/kg parent nucleoside equivalent, the AL-516 NTP is formed at high levels in liver (AL-516 NTP levels at 4 hrs: LY2157299 in vivo 11.2 μM). Conclusions: AL-516 is a potent guanosine based nucleotide analog that demonstrates a desirable preclin-ical profile. The compound is currently advancing in preclinical studies as a potential treatment for CHC. Disclosures: Kenneth Shaw – Employment: Alios

Biopharma Andreas Jekle – Employment: Alios Biopharma; Stock Shareholder: Alios Bio-pharma Jerome Deval – Employment: Alios BioPharma Zhinan Jin – Employment: Alios BioPharma Inc. Amy Fung – Employment: Alios BioPharma Lawrence M. Blatt 上海皓元 – Management Position: Alios BioPharma Sushmita M. Chanda – Employment: Alios BioPharma Qingling Zhang – Employment: Alios BioPharma Guangyi Wang – Employment: Alios Biopharma, Inc. Julian A. Symons – Employment: Alios BioPharma David B. Smith – Employment: Alios BioPharma The following people have nothing to disclose: Hua Tan, Yuen Tam, Natalia Dyatkina, Leo Beigelman Purpose/Background: Both viral and host proteins are involved during the hepatitis C virus (HCV) life cycle. Direct Acting Anti-virals (DAAs) inhibit HCV infection by targeting viral proteins and are successfully used to treat HCV infections. However, therapy failure caused by the emergence of DAA-resistance associated variants (RAVs) remains a reasonable concern. Presumably, future anti-HCV therapies will consist of drug combinations that target distinct steps of the viral life cycle. The conserved host factors used by the virus for its propagation seem interesting alternative targets for antiviral intervention, complimentary to the action of DAAs.

The use of acupuncture has since received considerable support an

The use of acupuncture has since received considerable support and is discussed in a separate section. More recently, a structured review123 on physical treatments for headache was undertaken, and found only modest support for the use of physical treatments in selected circumstances. Positive recommendations could be made in only a few clinical scenarios.123 For migraine, recommendations were made for physical therapy combined with aerobic exercise, as well as physical therapy combined with relaxation therapy and thermal BFB. For TTH, there was a trend toward benefit from chiropractic manipulation

in TTH, although the evidence was weak. Physical therapy was recommended, especially in high-frequency TTH cases. Cervical spinal manipulative JQ1 mw therapy was found to be as effective as amitriptyline in short-term use for PLX4032 chronic tension-type headache (CTTH), and more effective than massage for cervicogenic headache. Other recent studies127,128 have reported that physical therapy can be effective in reducing headache frequency, intensity and duration in CTTH patients. Overall, these physical treatments are most beneficial when integrated into a multimodal treatment plan including exercise, stretching, and ergonomics training for both the home and the workplace. Patients who express an interest in physical treatments are more likely to

benefit from active strategies such as exercise than passive ones such as massage and heat or cold application.129 Some have suggested that the insufficient evidence supporting or refuting the effect of physical treatments on headache disorders might be related to problems in identifying subgroups of patients who might benefit from the intervention.130 Fernández-de-las-Peñas et al131 thus devised a preliminary clinical prediction rule to identify CTTH patients who experience short-term success with muscle trigger point

therapy, MCE公司 using variables such as headache frequency, duration, bodily pain, and vitality scores. The implementation of clinical decision rules identifying these patients prior to carrying out randomized clinical trials was therefore suggested as a way of attaining stronger effect sizes.131 Although cervical spinal manipulative therapy may provide benefit in some clinical cases as described above, it has been associated with a 6-fold132 increase in the risk of vertebral artery dissection and stroke or transient ischemic attack. As such, one should be cautious when considering a recommendation for this treatment, and patients who express interest in chiropractic maneuvers should be warned of this potential complication.123 Otherwise, the use of physical treatments in headache is unlikely to be harmful in patients who express interest in these modalities.

Primary human hepatocyte cultures were transfected with genomic R

Primary human hepatocyte cultures were transfected with genomic RNAs of HCV genotypes 1a, 1b, and 2a (1 μg/106 cells) using FuGENE6 (Roche). On day 6 postinfection, the small RNA (≤200-nucleotide) fraction was enriched from HCV-infected cell RNA using a mirVana isolation selleckchem kit (Ambion). Four micrograms of each sample together with positive control (synthetic Arabidopsis thaliana mir-157a, which is not present in the human genome) was spiked in and was hybridized to the microarray slide (BioMicro

System). After 16 hours, the hybridized microarray was washed with a standard sodium citrate solution to remove unhybridized probes. After 3 hours of Klenow exonuclease-1 incubation, exo(-) Klenow enzyme was added to extend the miRNAs hybridized to the chip-attached templates in a primer extension step. During this step, biotinylated dATP was

incorporated as a final portion of the extension through the designed polythymidine region. Detection of this template-hybridized miRNA was performed using streptovidin-conjugated Alexa-fluor-555, which binds to the biotinylated stretch of A’s at the 3′-end of the captured miRNA. Fluorescence data sets were collected using GenePix 4000 scanner (Axon). Details of the procedure are described in Yeung et al.14 Primary hepatocytes were transfected with HCV1a genomic RNA (1 μg/106 cells) in triplicate. Parallel cultures were transfected with DLC-1 complementary DNA (cDNA) expression vector (50 ng/106 cells for 6 hours) prior to transfection with HCV 1a genomic RNA. Six days posttransfection, the cells were released with 0.05%

trypsin treatment and were resuspended at 104/100 μL in (phosphate-buffered selleck chemicals llc saline containing 2% fetal bovine serum) processed for Ki67 immunostaining (BD Biosciences) according 上海皓元 to the manufacturer’s instructions. Primary human hepatocytes were transfected with HCV genotypes 1a, 1b, and 2a (1 μg/106 cells) as described.12 Virus released in the culture medium was filtered through 0.25-μm filters from infected cells.12 Viral RNA replication was evaluated at indicated times after infection as outlined above, and the efficiency of virus released in the culture media was validated using the World Health Organization’s HCV standards (Acrometrix, Benicia, CA). Primary human hepatocyte culture was cotransfected with luciferase reporter containing DLC-1 3′ untranslated region (UTR) (50 ng/106 cells), miR-141 (50 nM/106 cells, antagomir) or miR-141 (50 nM/106 cells, Mimic) using Lipofectamine 2000 (Invitrogen). Luciferase assays (Promega) were performed on the third day after transfection according to the manufacturer’s instructions. The results are given as the mean ± SE. Statistical analysis of the data was performed using the Student t test, Fisher’s exact test, or otherwise as described. To assess virus infection-associated changes in host gene expression, we analyzed alterations in miRNAs in primary human hepatocytes infected with HCV genotypes 1a, 1b, and 2a (Supporting Information Fig. 1).

09 [12–68] for CHBV; 47 [11–84] for CHCV,

and 162 [

09 [1.2–6.8] for CHBV; 4.7 [1.1–8.4] for CHCV,

and 16.2 [9.1–24.5] for NAFLD patients respectively) and hepatic steatosis score on biopsy (odds ratio, 95% confidence interval = 30.7 [19.2–42.2] for CHBV; 24.2 [11.5–37.3] for CHCV, and 21.8 [10.1–45.0] for NAFLD patients respectively). Area under the receiver operating characteristics for CAP was 0.683 (0.601–0.757) for steatosis (S) ≥ 6%, 0.793 (0.718–0.856) for S > 33%, and 0.841 (0.771–0.896) for S > 66% respectively for www.selleckchem.com/products/jq1.html CHBV-infected patients. There was no difference in accuracy of CAP for assessing liver fat among CHBV, CHCV, and NAFLD patients. CAP is a novel, non-invasive tool that can detect and quantify steatosis accurately among CHBV, CHCV, and NAFLD patients, the accuracy being similar for all the three groups of patients. “
“TCBOPOP (1,4-bis [2-(3,5-dichaloropyridyloxy)] benzene) an agonist of the constitutive androstane receptor (CAR), produces rapid hepatocyte hyperplasia and hepatomegaly in the absence of hepatic injury. In this study we demonstrate that integrin-linked kinase (ILK), which is involved in transmission of the extracellular matrix (ECM) signaling by way

of integrin receptors, plays an important role in regulating TCPOBOP-induced proliferation of hepatocytes and hepatomegaly. Hepatocyte-specific ILK knockout mice (ILK/liver−/− mice) and wildtype mice (WT) were given a single dose of TCPOBOP (3 mg/kg) by oral gavage. Mice were sacrificed at days 1, 2, 5, and 7 after TCPOBOP administration. WT mice showed maximum proliferation on days 1 and 2, which came back to baseline levels by days 5 and 7 after TCPOBOP administration. The ILK/liver−/− mice, on the other hand, showed a prolonged this website and a sustained proliferative response as evident by an increased number of proliferative cell nuclear antigen assay (PCNA)-positive cells even at days 5 and 7 after TCPOBOP administration. At day 7 the WT mice showed close to a 2.5-fold increase in liver weight, whereas the ILK/liver−/− mice showed a 3.7-fold increase in liver weight. The prolonged proliferative response in the ILK/liver−/− mice seems to be due to sustained induction of CAR leading to sustained induction of c-Myc, which is

known to be a key mediator of TCPOPOP-CAR induced direct liver hyperplasia. Conclusion: The MCE data indicate that ECM-mediated signaling by way of ILK is essential for adjustment of final liver size and proper termination of TCPOBOP-induced proliferation of hepatocytes. (HEPATOLOGY 2011;53:587-595) The liver responds to specific classes of xenobiotics by inducing members of the nuclear hormone receptor superfamily, particularly the pregnane X receptor and the constitutive androstane receptor (CAR).1-3 An ideal candidate for studying xenobiotic metabolism is the halogenated hydrocarbon 1,4-bis [2-(3,5-dichaloropyridyloxy)] benzene (TCPOBOP). TCPOBOP is both a nongenotoxic carcinogen on its own and a potent tumor promoter when combined with genotoxic agents.