caudata strain DC-LOHABE01 and M. rubrum strain MR-MAL01 to address the status of Dinophysis plastids. Our approach was to experimentally generate D. caudata with “green” plastids and then follow the ingestion and fate of “reddish-brown” prey plastids using light microscopy, time-lapse videography, and single-cell TEM. Our results for D. caudata resolve the apparent Z-VAD-FMK discrepancy between morphological and molecular data by showing that plastids acquired when feeding on M. rubrum are structurally modified and retained as

stellate compound chloroplasts characteristic of Dinophysis species. “
“Department of Microbial Ecophysiology, University of Bremen, Bremen, Germany Unicellular cyanobacteria are now recognized as important to the marine N and C cycles in open ocean gyres, yet there are few direct in situ measurements of their activities. Using a high-resolution nanometer scale secondary ion mass spectrometer (nanoSIMS), single cell N2 and C fixation rates were estimated for unicellular cyanobacteria resembling N2 fixer Crocosphaera watsonii. Crocosphaera watsonii-like cells were observed in the subtropical North Pacific gyre (22°45′ N, 158°0′ W) as 2 different phenotypes: colonial and free-living. Colonies containing 3–242 cells per colony were observed and cell density in colonies increased with

incubation time. Estimated C fixation rates were similarly high in both phenotypes and unexpectedly for unicellular cyanobacteria 85% of the colonial cells incubated during midday were also enriched in 15N above natural abundance. Highest 15N enrichment and

N2 fixation rates were found in cells incubated overnight where up to 64% of the total daily Selleckchem ABT-263 fixed N in the upper surface waters was attributed to both phenotypes. The colonial cells retained newly fixed C in a sulfur-rich matrix surrounding the cells 3-mercaptopyruvate sulfurtransferase and often cells of both phenotypes possessed areas (<1 nm) of enriched 15N and 13C resembling storage granules. The nanoSIMS imaging of the colonial cells also showed evidence for a division of N2 and C fixation activity across the colony where few individual cells (<34%) in a given colony were enriched in both 15N and 13C above the colony average. Our results provide new insights into the ecophysiology of unicellular cyanobacteria. "
“A marine, filamentous, endolithic cyanobacterium, strain BC008, was obtained in pure culture and characterized using a polyphasic approach. BC008 could bore into calcium carbonate minerals (calcite, aragonite) and, weakly, into strontium carbonate (strontianite), but not into other carbonates, phosphates, sulfates, silicates, or oxides, including those of calcium. We describe procedures for its continued cultivation in an actively boring state. BC008 was developmentally complex: it displayed lateral, terminal, and intercalary heterocysts; true branching; trichome tapering; and motile hormogonia. It also displayed considerable morphological plasticity between boring and nonboring modes.

Functionally, previous study indicated that BAF60a stimulates fatty acid β-oxidation and ameliorates hepatic steatosis in vivo.24 We also found that mice have hypoglycemia when BAF60a is knocked down in the liver. Consistently, overexpression of BAF60a in mouse hepatocytes increase the glucose production rate. The positive effects of BAF60a on glucose levels may due to the induction of gluconeogenic genes such as G6Pase and PEPCK (Fig. 5 and data not shown) by BAF60a and the accelerated rate of gluconeogenic process. These observations

collectively extend our recognition of physiological roles of this factor, whose function is traditionally limited in the regulation of chromatin structure. Future study is required to elucidate the functions Mitomycin C molecular weight of BAF60a and other BAF60 proteins, BAF60b and BAF60c, in various metabolic tissues under physiological and pathological status. Similar to the control of the circadian clock on metabolism, food is a very potent synchronizer (zeitgeber) for peripheral clocks. Recent work investigating the respective contributions of feeding and of the circadian clock to hepatic rhythmic EGFR inhibitor gene expression using clock-deficient mice and the temporally restricted feeding paradigm has shown that oscillations of food-induced transcripts, including those from the gluconeogenic and fatty acid

oxidation pathways, are modulated and consolidated by the clock.37 Reciprocally, the robustness of the clock and its immediate downstream targets is increased by temporally restricted feeding. MRIP The expression of BAF60a itself is not altered by food signals,24 but it is a clock target that

modulates the expression of food-driven metabolic genes, such as PEPCK and Cpt1a. In this sense, a principle function of BAF60a may be to temporally modulate the food-entrained daily oscillation of components of specific metabolic pathways. On the other hand, we cannot exclude the possibility that BAF60a mediates food-induced reset of the peripheral clock at the posttranslational levels. The rhythmic recruitments of BAF60a to metabolic gene promoters need to be examined. Circadian function is largely based on a program of finely controlled transcriptional regulation at the genome-wide level. The intrinsic nature of its highly specialized, temporally regulated transcription places the cellular clock as a prominent model for the study of dynamic chromatin remodeling.38, 39 Moreover, based on the tight coupling between circadian rhythms and metabolic regulation,18 clock-controlled chromatin reorganization is likely to reveal specific pathways linking histone modifications to cellular metabolism. For example, the central clock protein Clock has histone acetyltransferase (HAT) enzymatic properties.40 It directs acetylation of histone H3 and its dimerization partner Bmal1 at K537, an event essential for circadian function.

“
“To evaluate the quantitative and qualitative changes in amino acids related to internal nitrogen content and growth rate of Ulva ohnoi, the supply of nitrogen to outdoor cultures of the seaweed was manipulated by simultaneously varying water nitrogen concentrations and renewal rate. Both internal nitrogen content and

growth rate varied substantially, and the quantitative and qualitative changes in amino acids were described in the context of three internal nitrogen states: nitrogen-limited, metabolic, and luxury. The nitrogen limited state was defined by increases in all amino acids with increasing nitrogen content and growth up until 1.2% internal nitrogen. The metabolic nitrogen state was defined by increases in all amino acids with increasing internal Selumetinib nitrogen content up to 2.6%, with no increases in growth rate. Luxury state was defined by internal nitrogen

content above 2.6%, which occurred only when nitrogen availability selleck compound was high but growth rates were reduced. In this luxury circumstance, excess nitrogen was accumulated as free amino acids, in two phases. The first phase was distinguished by a small increase in the majority of amino acids up to ≈3.3% internal nitrogen, and the second by a large increase in glutamic acid, glutamine, and arginine up to 4.2% internal nitrogen. These results demonstrate that the relationship between internal nitrogen content and amino acid quality is dynamic but predictable, and could be used for the selective culture of seaweeds. Amino acids are the critical constituent in animal feeds, specifically the essential amino acids methionine and lysine, as these

are the “first” limiting amino acids in plant-based feed formulations (McDonald et al. 2002, Boland et al. 2012). Amino acids are also targeted as a feedstock for biorefineries in the bio-based chemical industry (Scott et al. 2007, Jung et al. 2013). In this scenario, it is the nonessential amino acids that are the preferred primary substrates for bio-based chemicals, specifically glutamic acid, which resembles many industrial intermediates (Lammens et al. 2012). The extraction SB-3CT and concentration of nitrogenous biochemicals is now proposed as a common value-added component of most biofuel conversion and modeling (Ragauskas et al. 2006). Together these applications promote the use of high productivity biological feedstocks for feed and bio-based chemicals before the remaining biomass is converted to a biofuel, for which algae have received much attention (Ragauskas et al. 2006, Rowbotham et al. 2012). However, relatively little is known about the relationship between internal nitrogen content, growth rate and the quantitative and qualitative changes in amino acids for algae (as it is for plants; see Steinlein et al. 1993, Heilmeier et al. 1994, Lipson et al. 1996), and, correspondingly, whether internal nitrogen content can be manipulated to maximize the yields of specific amino acids.

In the c-Myc/Tgfα transgenic model, MHC-1 is down-regulated and Rae1 is up-regulated on dysplastic hepatocytes targeting them for NK surveillance. However, malignant progression still succeeds, presumably due to insufficient NK numbers.49 NKT cells significantly increased in c-Myc/Tgfα dysplastic liver as it progressed toward

malignancy. Distinct NKT cell subsets can either promote (CD4+ NKT) or inhibit (iNKT) liver cancer50 (recently reviewed by Subleski et al.51) and can partially explain this observation. Bridging innate and adaptive immune systems, resident DCs are less functional in liver, Panobinostat but are still capable of priming antiviral T-cell responses sufficient for clearance. Upon viral escape, chronic liver inflammation renders liver DCs suppressed, as observed in chronically infected HCV patients showing a diminished ability to mature and prime T-cell proliferation and induce IFN-γ.52 Interaction between HCV core protein and DCs in culture results in reduced frequency

of pDC and direct inhibition of IFNα production.53 Core protein can also inhibit IL-12 production by DCs through an intracellular mechanism dependent on a combination of TLR4 signaling and cross-linking of the complement receptor,54 thereby contributing to a Th2 skewed microenvironment. HBV-infected patients have diminished pDC functions resulting in part from a specific HBV antigen (HBeAg).55 These findings suggest persistent viral infection and chronic inflammation deprive DC’s ability to prime T-cell surveillance, augmenting hepatocellular carcinogenesis. VX 809 Progesterone Circulating B cells from cirrhosis patients have been reported to be hyporesponsive to ex vivo CD40/TLR9 stimulation, as characterized by LTα secretion, IgG production, and T-cell allostimulation. A reduction in CD27+IgM+ memory B cells was also observed in cirrhosis patients.56 These changes support a reduced B-cell-mediated antiviral response, allowing persistent viral infection, associated inflammation, and HCC development. In contrast, results from an inflammatory skin model of HPV16 squamous cell carcinomas (SCC) suggest a more direct, tumor-promoting role for B cells, possibly by way of immunoglobulin

accumulation.57 Although controversial, increased levels of immunoglobulin in murine HCC models, serum from cirrhotic individuals, and HCC lesions58 all suggest a possible cause-and-effect linkage between the presence of immunoglobulin and HCC development. Previously, we established a murine de novo liver tumor model of adenoma and carcinoma by hydrodynamic injection of transposons containing myrAKT (AKT) and Δ90 β-catenin (β-CAT).59 We observed hepatocellular tumor development/progression to be largely dependent on B cells (authors’ unpubl. obs.). We also found that tumor infiltrating B cells express elevated levels of TNF-α (authors’ unpubl. obs.), suggesting that B cell-derived cytokines could be instrumental in tumor development/progression.

5 years. Patients in whom HCV RNA was undetectable at week 20 were categorized as responders and continued full-dose combination therapy for up to 48 weeks. Initial responders

were eligible for randomization into the trial if virologic breakthrough occurred during extended therapy or relapse followed 48 weeks of therapy. In addition, patients who were treated with peginterferon and ribavirin outside the lead-in phase of the HALT-C Trial were also eligible for randomization (“express” group) if they met criteria for nonresponse, breakthrough, or relapse. This approach to enrollment ensured that all patients had received optimal therapy with peginterferon and ribavirin8, 9 before they were enrolled into this long-term buy Carfilzomib trial, during which they might not be treated.5 After randomization, patients in both groups were seen at 3-month intervals for 3.5 years, at which point peginterferon was discontinued in the treatment group.

Nine patients assigned to the control group were treated “off-protocol” by nonstudy physicians, but they were included as controls in find more our intention-to-treat analysis. Thereafter, all patients remained untreated and were seen at 6-month intervals. At each visit the occurrence of clinical outcomes (which had been established prospectively) was noted, including clinical events and laboratory markers of hepatic decompensation, HCC, or death. Although not a primary clinical outcome in the

HALT-C Trial, liver transplantation was included in this mortality analysis because these patients were likely to have died in the absence of liver transplantation. Most deaths were identified by study coordinators interacting with family members by way of telephone. In addition, periodic on-line searches were performed of the U.S. Social Security Death Index (SSDI) (http://ssdi.rootsweb.com/), which is generated from the U.S. Social Security Administration’s Death Master File. The SSDI was queried for any participant with whom the study site had no contact for at least 6 months. The last search of the SSDI was conducted in October, 2009. To account for the potential lag between date of death and mafosfamide report to the SSDI, we included in our analysis deaths occurring on or before December 31, 2008. All deaths were reviewed by a seven-person, central review committee consisting of HALT-C Trial investigators blinded to the identity of the subject, study site, fibrosis versus cirrhosis stratum, but not randomization allocation (treatment or control), this information being required to assess treatment relatedness. The committee classified the primary cause of death into one of 15 categories (Supporting Table 1).

For DNA analysis, 20-25 μL of viral samples were treated according to Qiagen QIAamp DNA blood kit protocol. Viral DNA, 2 μL, was BTK animal study amplified by PCR with specific HBV primers. pGEM-1.3xHBV was used for standard calibration. Analysis of HBV DNA replication from cells was performed as described.15 dNTP extraction

is based on19 and dNTP level was quantified by DNA polymerase fill-in reaction as described.20 Nondividing cells have minimal amounts of dNTPs that are produced by de novo synthesis. We hypothesized that HBV induces de novo dNTP synthesis in nondividing cells, to ensure sufficient levels of dNTPs for the synthesis of progeny DNA. HBV does not readily infect cells in tissue culture; thus, a commonly used tool for the study of HBV is the hepatic HepG2 cell line stably-tranfected with HBV,21 known as HepG2.2.15, that is active in HBV gene expression and virion production.22 To investigate HBV production in resting cells, we treated HepG2 and HepG2.2.15 cells with DMSO to induce G0/G1 Selleck SAHA HDAC arrest.3, 13 Cells were

arrested in a gradual manner and a complete growth arrest was obtained after about 5 days of treatment (Fig. 1A). FACS analysis revealed that both HepG2 and HepG2.2.15 DMSO-treated cells did not incorporate BrdU, indicating that both stopped proliferating (Fig. 1B). Growth arrest was also confirmed by the [3H]thymidine-incorporation assay (Fig. 1C). Finally, in DMSO-treated cells, Ki67 expression, a cell cycle marker, was markedly attenuated over time (Fig. 1D), Thymidylate synthase confirming the quiescent state (G0) of DMSO-treated cells. HBV replication and virion production was examined in quiescent, DMSO-treated HepG2.2.15 cells. We examined whether RNR, the key enzyme for dNTP synthesis, is required for HBV replication in nondividing cells, by using the specific RNR inhibitor HU.23 Remarkably, the level of HBV replication was dramatically attenuated in HU-treated quiescent HepG2.2.15 cells, as examined by monitoring the intracellular viral DNA in the cytoplasm (Fig. 2A). Next, we quantified the level of secreted virions

and revealed that it was higher in the DMSO-treated HepG2.2.15 cells, compared to the nontreated cells (Fig. 2B, lower panel), demonstrating that sufficient levels of dNTPs were available in HepG2.2.15 nondividing cells. In addition, the amount of viral particles released to the medium was sharply reduced as determined by western blot analysis of HBV core protein (Fig. 2C) and PCR-based quantification of viral DNA (Fig. 2B), suggesting that RNR inhibition blocks viral replication and secretion. The level of RNR activity is determined by R2 expression, because the R1 protein level is almost constant, while the R2 protein has a short half-life of 3 hours and its gene is not expressed in quiescent cells.10 To examine the effect of HBV on R2 expression, we quantified R2 level in HepG2 and HepG2.2.15 quiescent cells.

The primary outcome was a difference in the improvement of steatosis, hepatocellular inflammation, or fibrosis between treatment groups. A minimum 1-point improvement in each quartiled graded parameter was required to meet the primary end-point. Secondary outcomes included overall changes in steatosis, hepatocellular inflammation, hepatocyte ballooning degeneration, fibrosis, NAS, insulin,

and alanine aminotransferase (ALT), as all three groups received rosiglitazone therapy. Changes in weight, other metabolic parameters, and other liver enzymes were additional secondary end-points. The primary analysis was a per-protocol analysis. Comparisons for primary and secondary Erlotinib outcomes were made using a two-factor analysis of variance (treatment, time), with repeated measures on one factor (time). Correlations were determined by linear regression analysis with backward elimination.

Sample size was derived using a look-up table, based on employing the methods of Kraemer and Thiemann (1988), to obtain an initial sample size. The sample size was adjusted with 1,000 iterations of a Monte Carlo simulation until the power was between 80% and 85%, with a level of confidence of 95%. With 45 subjects per group, an 0.8 standard deviation would be detected between groups. An additional 5 patients were added to allow for dropouts. In the fall of 2010, the U.S. Food and Drug Administration (FDA) restricted rosiglitazone to type II diabetics, prematurely halting

the study at 137 patients enrolled. Of the 135 subjects that underwent randomization, learn more 41 were assigned to receive rosiglitazone alone, 49 were assigned to receive rosiglitazone and metformin, and 45 were assigned to receive rosiglitazone and losartan (Fig. 2). Baseline characteristics were well matched between groups with respect to age, percent of diabetic subjects, gender, PIK3C2G race, biochemical markers, metabolic factors, and histologic findings (Table 1). The difference in baseline NAS was significantly different (P = 0.014), with rosiglitazone alone having a higher baseline NAS, compared to the other two study groups. After a planned blinded, independent expert pathologist review at the completion of the study, 19 subjects were excluded based on the absence of stringent criteria for the diagnosis of NASH on their initial liver biopsy: 5 subjects (6%) in the rosiglitazone-alone arm, 9 subjects (19%) in the rosiglitazone and metformin arm, and 5 subjects (5%) in the rosiglitazone and losartan arm. A total 108 subjects underwent an end-of-treatment liver biopsy. There was no statistically significant difference between rosiglitazone, rosiglitazone and metformin, and rosiglitazone and losartan with respect to improvement in steatosis (25%, 28%, and 25%; P = 0.

We prospectively recorded accompanying visual and auditory symptoms in a large cohort of patients with VS and correlated these symptoms with comorbid migraine and typical migraine aura. To assess potential pathophysiological correlates, we further studied brain metabolism in patients with the hypothesis that VS is associated with regional hypermetabolism distinct from previous findings in migraine.[8, 9] Clinical data

of a subgroup of the study population have been previously presented in a report on the detailed phenotype[5] and in beta-catenin inhibitor preliminary form.[10, 11] The study was approved by the Institutional Review Board (# 11-07270 and # 11-07431) and the radiation safety committee (58605-RU-04-URH) of the University of California, San Francisco. Patients were recruited via advertisements in social media

Deforolimus order with the support of a self-help group on VS (Eye on Vision Foundation; http://www.eyeonvision.org/). After being contacted by the patient, eligibility was assessed during telephone interviews. After being approached by the patient, verbal consent was obtained and subjects with self-suspected VS underwent a semi-structured telephone interview. The following items were covered during the interview: Demographics (age, gender) and handedness. Patients were asked to describe their current visual symptoms in their own words. Based on that information and additional open questions, a diagnosis of VS was made and associated visual symptoms were recorded as described recently.[10] In brief, VS was defined as dynamic, continuous, tiny dots in the entire visual field (similar to “TV static” or “TV snow”) lasting longer than 3 months (criterion A).[5] Other Loperamide symptoms were palinopsia (“afterimages” and “trailing” of moving objects),

entopic phenomena (phenomena arising from the structure of the visual system itself including (1) excessive floaters in both eyes; (2) excessive blue field entoptic phenomenon, ie, uncountable little gray/white/black dots or rings shooting over the visual field in both eyes when looking at homogeneous bright surfaces, such as the blue sky; (3) self-light of the eye, ie, colored waves or clouds when closing the eyes in the dark; and (4) spontaneous photopsia, ie, bright flashes of light),[7] photophobia, and nyctalopia (impaired night vision). Due to its high prevalence in subjects with VS,[5] the presence or history of tinnitus was also covered during the interview despite being a non-visual symptom. Headache history was assessed according to the International Classification of Headache Disorders – 2nd edition.[6] Migraine aura was only diagnosed when typical features were present, which are unilaterality (homonymous), development over 5 minutes, duration for less than 60 minutes, reversibility, zigzag lines, and scotoma.[4, 6] SPSS (v20, IBM Corp.

021) was significantly associated with preS/S mutant infection. Of note, infection with the preS/S HBV mutants was positively correlated with cirrhosis (r = 0.386; P = 0.014), and this correlation persisted find more after adjustment for age, viral loads, HBeAg status, and presence of basal core promoter mutations. Sequencing of the basal core promoter (BCP)/precore (PC) region of the HBV isolates obtained from the 40 patients revealed the presence of the G1896A mutation (PC mutation) combined with the

A1762T and/or G1764A mutations (BCP mutations) in nine patients, the presence of BCP mutations alone in three patients, and of PC mutation alone in 18 patients. In 10 patients, the HBV DNA sequence was WT at both BCP and PC sites. Statistical analysis applied to all variants revealed that BCP/PC mutations—either in combination or alone—were not associated with preS/S mutations and, when present, BCP/PC mutations had no impact on HBsAg production and replication capacity of preS/S HBV mutants (data not shown). Three different HBV full-length genomes were cloned and functionally tested by three independent transient selleck inhibitor transfection experiments of HepG2 cells. pHBV-mtpreS1, which was isolated from patient 14 (HBV DNA, 2 × 107 IU/mL; HBsAg, 1.6 × 103 IU/mL) showed an in-frame deletion of 183 nucleotides in the preS1

region (a 61-aa deletion [Δ47-108 aa] within the L protein). pHBV-mtpreS2, which was isolated from patient 4 (HBV DNA, 5.7 × 107 IU/mL; HBsAg, 1.9 × 103 IU/mL) showed the deletion of the preS2 start codon. pHBV-mtS, which was isolated from patient 8 (HBV DNA, 2.4 × 108 IU/mL; HBsAg, 9 × 102 IU/mL) showed a G1035A mutation introducing a stop signal at codon 182 within the S gene (sW182*) (Fig. 1A). As a note, pHBV-mtpreS1 and pHBV-mtpreS2 cloned genomes carried

the nucleotide mutation G1896A, introducing a stop signal at codon 28 within the ROS1 precore region and preventing HBeAg synthesis. A plasmid-free HBV transfection cell–based replication assay relying on the generation of transcriptionally active nuclear cccDNA to replicate HBV was used.28, 29, 30 Because the three mtHBV genomes were genotype D, a standard WT HBV genome of the same genotype was used as a control. Two days after transfection, HBV DNA from intracellular replicative intermediates and extracellular viral particles were analyzed by way of Southern blotting. As shown in Figs. 3A and 3B, all three HBV genomes were replication-competent and were able to release viral particles into the cell culture medium. However, whereas the levels of intracellular replicative intermediates were comparable between cells transfected with mutated HBV genomes and cells replicating WT HBV, the HBV DNA level in the supernatant of cells transfected with mutant viruses was 30%-50% lower than in cells transfected with WT virus.

1C). These data thus suggested that HBx may promote expansion of HPCs in DDC-treated mice. It has been shown that the EpCAM+ cells isolated from DDC liver have adult progenitor potential that possess the capacity for unlimited proliferation and bidirectional differentiation.18 Flow cytometric analysis of

nonparenchymal cells prepared from the liver of HBx mice fed with DDC for 1 month showed a higher percentage of EpCAM+CD45− HPCs (Fig. 1D). When cultured at low density, EpCAM+CD45− HPCs isolated from HBx mice were able to form more and larger colonies than EpCAM+CD45− Alectinib clinical trial cells from WT mice (Fig. 2A), suggesting that these cells may have a stronger stem-like property. To further determine the characteristics of HPCs, the gene expression profile of magnetic sorted primary EpCAM+CD45− cells (Supporting Figs. S1, S2) was examined by quantitative reverse-transcription polymerase chain reaction (RT-PCR). As shown in Fig. 2B, HPCs from HBx mice demonstrated stronger expression of stem/progenitor cell markers (EpCAM, CD133, CD90, ABCG2, AFP, and CK19) than those from WT controls. However,

there were no significant differences in the expression of the oncogenes (K-ras, N-ras) between HPCs from HBx and control WT mice at 1 and 2 months DDC treatment (Fig. 2B; Fig. S3). To evaluate the self-renewal ability of HPCs, we performed a sphere formation assay. Compared with WT control, more spheres were observed RG7420 in EpCAM+CD45− HPCs isolated from HBx mice (Fig. 2C). Recent evidence shows that DDC not only causes liver damage along with the proliferation

Coproporphyrinogen III oxidase of HPCs, but also facilitates some oncogene-induced tumorigenesis in the adult liver.19 After consecutive administration of 0.1% DDC for 4 months, both HBx and WT mice showed an increase in liver size and toughness on the liver surface compared with livers from mice treated with DDC for 1 month (Fig. S4A,B). Histological and flow cytometric analysis revealed persistent HPC expansion in both groups, but more robust in HBx mice (Fig. 1C,D). HPCs from HBx mice also showed increased expression of stem/progenitor cell markers and stronger sphere formation ability compared with those from WT mice (Fig. 2B,C). Furthermore, the expression of the oncogenes (K-ras, N-ras) was up-regulated in HPCs from HBx mice (Fig. 2B), suggesting that HBx may also promote the HPCs to obtain transformed characteristics after long-term DDC treatment. To further observe whether long-term exposure to a DDC diet could induce tumors, HBx and WT mice were continuously fed a DDC diet for 5-7 months. As the results in Fig. 3A show, all HBx mice (n = 15) developed liver tumors after 7 months, whereas none of WT mice (n = 15) had any liver tumors. Interestingly, after 5 months DDC treatment, although some HBx did not develop liver tumors, GSTpi and GPC3-positive dysplastic nodules were detectable in their liver tissues (Fig. S4C,D).