However, in addition to DNA damage, there are many other kinds of stimuli in the liver after DEN administration in vivo: inflammatory responses, liver regeneration signals, and toxic metabolites of DEN. Thus, we assessed the role of the ASK1-p38 pathway in p21 induction after DNA damage using UVB irradiation, which is a well-known direct DNA damage-inducer. UVB irradiation to the immortalized human normal hepatocyte line induced strong phosphorylation of p38 and very

weak phosphorylation of JNK, and ASK1 silencing attenuated UVB-induced p38 phosphorylation, especially in the late phase (Fig. 8D, Supporting Fig. 5). Furthermore, UVB-induced p21 up-regulation was attenuated by ASK1 silencing and p38 inhibition (Fig. 8E F). These results suggest that ASK1 is involved in DNA damage-induced p21 up-regulation through p38 buy ICG-001 activation, and an impaired DNA damage response may be one reason for increased hepatocarcinogenesis in ASK1−/− mice. Dysregulation of the balance between cell proliferation

and apoptosis plays a critical role in hepatocarcinogenesis.31 Our results suggest that ASK1 plays only a minor role in cancer cell proliferation and a major role in death receptor-mediated apoptosis in the liver through the JNK pathway. Loss of ASK1 appears to cause an imbalance that accelerates chemical hepatocarcinogenesis in ASK1−/− mice. Furthermore, ASK1 is involved in the DNA damage response, through the p38 pathway. This study provides new insight into the regulation of stress-activated MAPK signaling in hepatocarcinogenesis. JNK (primarily JNK1) has been reported to promote DEN-induced hepatocarcinogenesis by promoting Selleckchem Dabrafenib cancer cell proliferation

and neovascularization.3, Chlormezanone 5 Although JNK activation was attenuated in ASK1−/− mice, the phenotype of ASK1−/− mice and JNK1−/− mice was opposite in hepatocarcinogenesis.3, 5 We suggest that the reasons may be as follows: (1) Although we observed attenuation of JNK activation in ASK1−/− HCC tissues, ASK1 appears to play only a minor role in HCC cell proliferation (Fig. 2). Additionally, vessel density and VEGF expression in ASK1−/− HCC tissues were unaffected (Supporting Fig. 6A,B). Thus, the tumor-enhancing function of JNK1 seems to be preserved in ASK1−/− mice. (2) JNK has also been reported to act as a tumor suppressor by inducing cancer cell apoptosis.32 JNK1 and JNK2 isoforms have distinct or redundant roles in some situations, including apoptosis induction. JNK2, but not JNK1, has been reported to play a major role in TNF-α-mediated hepatocyte apoptosis in vivo.33 In our experiments using a Jo2-induced hepatitis model, the lack of neither JNK1 nor JNK2 resulted in a significant reduction in the ALT elevation or BimEL phosphorylation, unlike ASK1−/− mice or a pan-JNK inhibitor (Supporting Fig. 7A,B). These results suggest that the role of JNK1 and JNK2 in death receptor-mediated hepatocyte apoptosis may be redundant.

0 array, 41,751 probe sets corresponding to the known human genes were selected. Among them, the numbers of probe sets marked as “present” in >70% of samples derived from cancer tissue and adjacent noncancerous tissue were 17,303 and 16,431, respectively. Using cancer tissue samples, 11 probe sets satisfied the criteria by the Cox likelihood ratio test for recurrence free (P < 0.01) and more than 2-fold change between the recurrence and nonrecurrence groups (Supporting Table 1). Among them, the criteria by Wilcoxon signed-rank test (P < Compound Library 0.01) and more than 2-fold change between the paired cancer and noncancerous tissues

were satisfied by two probe sets: immunoglobulin kappa locus and family with sequence similarity 134, member B (Table 1). Using noncancerous tissue samples, five probe sets satisfied the criteria by Cox regression analysis and more than 2-fold change (Supporting Table 1). Among them, three probe sets, cytochrome P450 1A2 (CYP1A2), carnosine dipeptidase 1 (CNDP1), and ornithine aminotransferase (OAT), showed significant down-regulation in paired cancer tissue. It is of interest that both carnosine and ornithine are known as amino acids closely related

to oxidative stress in the liver.15, 16 The multivariate Cox regression analysis with the forward variable-selection procedure revealed that a R428 model containing only the gene-expression pattern of CYP1A2 (HR, 0.447; 95% CI, 0.249-0.808; P = 0.010) was the best predictive model for the recurrence of HCC (Table 1). We examined the association between clinicopathological Bumetanide factors and recurrence-free survivals of 78 patients with early-stage HCC meeting the Milan criteria (Table 1; lower). Median recurrence-free survival time was 23.7 months. Univariate Cox regression analysis demonstrated that HCV infection (HR, 2.095; 95% CI, 1.048-4.188; P = 0.031) and low serum albumin level (HR, 0.447; 95% CI, 0.248-0.808; P = 0.010) correlated with the cumulative recurrence-free rate. Other clinicopathological

factors were not statistically significant. We further performed the forward variable-selection procedure using the two factors, and the serum albumin level was identified as the only risk factor for recurrence after curative resection for early-stage HCC (Table 1). To evaluate the best predictive model for patients with HCC meeting the Milan criteria, we considered the combined model using the serum albumin level and the gene-expression pattern of CYP1A2 (Table 2). A multiple logistic regression analysis demonstrated that CYP1A2 (OR, 0.534; 95% CI, 0.276-0.916; P = 0.036) was the only factor that contributed to the prediction of the recurrence. Moreover, the variable-selection procedure by logistic regression analysis identified that CYP1A2 was the best predictor for the recurrence of HCC meeting the Milan criteria (OR, 0.493; 95% CI, 0.273-0.787; P = 0.008).

15 Dietary fructose is absorbed into the intestine by way of a saturable, facultative glucose transporter

(GLUT5). Healthy persons are able to absorb up to 25 g. Malabsorption can lead to increased fructose fermentation by gut bacteria.48 Findings regarding endotoxin (lipopolysaccharide [LPS]) HM781-36B datasheet levels in portal blood in human NAFLD have been mixed, in part because portal blood is difficult to sample in human subjects and circulating levels are inconsistent. Normally, endotoxin released from the gut is cleared rapidly on first pass by Kupffer cells. However, a growing body of evidence supports a role for increased gut permeability and endotoxin in human NAFLD. In type II diabetes, endotoxin contributes to the development of the subclinical inflammatory state and insulin resistance by stimulating the innate immune system and inducing release of proinflammatory cytokines from adipose tissue. While HDL is known to neutralize LPS, this antiinflammatory function has been shown to be less effective in patients

with NAFLD.49 If HDL protection of LDL is decreased, that could lead to greater levels of oxidized LDL in NAFLD, which has previously been demonstrated.50, 51 Supporting this, in a small study of children with NAFLD, a low fructose diet resulted in diminished oxidized LDL.51 The relationship of fructose-induced endotoxin to disease in humans is even less well understood than the role of endotoxin Dabrafenib chemical structure in NAFLD; the direct relationships require further exploration. Limited studies suggest an association between fructose consumption and NAFLD. A pediatric study demonstrated increased carbohydrate intake in children with NAFLD identified by ultrasound compared to obese non-NAFLD counterparts.52 Small Cediranib (AZD2171) case-control studies of adults demonstrate higher

fructose and/or soft drink consumption in those with NAFLD.53-55 A study demonstrating excess soft drink consumption predicted NAFLD in a cohort of adults without typical risk factors for NAFLD lends support for a fructose effect independent of obesity.56 Abdelmalek et al.57 evaluated histologic features of a large cohort of adults with NAFLD and correlated this to estimated fructose intake. Although steatosis grade was lower in those with increased fructose intake, the degree of fibrosis was increased. In this same study, serum uric acid was substantially higher in those with increased fructose intake. Uric acid has been proposed as a biologic marker of fructose intake because uric acid levels increase with fructose intake.58, 59 In a large cohort of children with NAFLD, histopathology did not correlate with self-reported sugar consumption; however, uric acid was significantly increased in those with NASH compared to those with steatosis alone.60 It has been proposed that uric acid may mediate some of the abnormalities seen with fructose consumption through induction of retinol binding protein-4 (RBP-4), an adipokine linked to hepatic insulin resistance.

1) lead to protein intolerance and massive accumulation of ammonia, with most catastrophic presentations in full-term infants in the first week of life.1 There is wide genotypic and phenotypic heterogeneity such that milder forms of these diseases may present in childhood or in adults manifesting as dietary protein aversion and encephalopathy

that may be misdiagnosed for years. Treatment entails restriction of dietary protein intake and pharmacological selleck chemicals llc activation of alternate pathways of waste nitrogen synthesis and excretion. Despite these therapeutic strategies, there remains significant unmet need, and even in apparently well-controlled patients, there is suboptimal control of ammonia, subclinical neurocognitive dysfunction, and impaired quality of life.2 It is instructive to trace the evolution of therapies for urea cycle disorders (UCDs). It illustrates daring efforts to circumvent formidable hurdles facing investigators attempting to develop therapies for rare orphan diseases that generally exhibit

extreme genotype and phenotype heterogeneity.3 The history is rich with triumphs that have been uniformly propelled by powerful advocacy of individual treating physicians in collaboration with patient support groups. Pioneering efforts by Saul Brusilow and Mark Batshaw at Johns Hopkins spanning more than a quarter selleck screening library of a century have dramatically improved prospects for patients and families affected by UCDs.4 Following on from observations in animal studies of nitrogen metabolism, these investigators developed a scientific rationale for the first-generation alternate pathway therapy in the form of oral Na benzoate

and Na phenylacetate combined with dietary protein restriction. This alone was sufficient to have approval of this regimen; what a far cry from current exacting requirements for demonstrating Dimethyl sulfoxide efficacy for therapies for rare diseases. However, this treatment resulted in incapacitating body odor, prompting Brusilow to conceive of a prodrug in the form of Na Phenylbutyric Acid (NaPBA), which was efficiently cleaved to phenylacetate and, subsequently, to phenyacetylglutamine (Fig. 1). Early studies indicated good bioavailability and it was clearly more tolerable. However, the drug could not be approved by the U.S. Food and Drug Administration (FDA) because of the lack of any efficacy and safety data, nor was there any federal support to carry out these studies. It was philanthropy and patient advocacy that advanced the field beyond the gridlock. An ad-hoc dispensary was set up at Johns Hopkins to ensure the supply of the drug to families affected by UCDs. Again, without a clinical trial, but based on real-world experience and advocacy of patient groups, the FDA approved the use of NaPBA in 1996.

2003a, Dehn et al. 2006a). Conclusions on Hg biomagnification are mixed. Dehn et al. (2006b) found little DNA Damage inhibitor evidence for Hg biomagnification, whereas data from Atwell et al. (1998) suggest a significant, positive log relationship between [Hg] and δ15N values in arctic food webs. Theory and empirical studies show

that [Pb] is lowest in top trophic level consumers (Michaels and Flegal 1990), because Pb is biodepleted relative to its biogeochemical analogue calcium. The combination of [Pb] and stable Pb isotopes (207Pb/206Pb) have been especially useful in documenting historical shifts in the source(s) and sometimes concentrations of Pb between preindustrial and modern times (Smith

et al. 1990, Outridge et al. 1997, Caurant et al. 2006). Most of the studies on organochemical contaminants evaluate exposure at the community or ecosystem level and present data from multiple trophic levels RG7204 solubility dmso that often include one or more marine mammal species. Significant, positive correlations among PCB, DDT, and FOC concentrations and trophic level, as derived from δ15N values, are strong evidence for bioaccumulation of these compounds in marine food webs (Jarman et al. 1996, Fisk et al. 2001, Hobson et al. 2002, Tomy et al. 2004). Coupled contaminant and δ13C analysis also suggest differences in FOC contaminant loads among marine mammal species that occupy nearshore versus offshore habitats (Van de Vijver et al. 2003). The blending of contaminant and SIA also yields information on population structure and niche variation at the individual, population, or species level. At first, contaminant concentrations alone were used in this capacity (see review by Aguilar 1987). More recently, however, researchers have begun to integrate toxicological and isotopic proxies. In essence, geographic

variability in natural elements (i.e., food web isotope values) or anthropogenic compounds (i.e., contaminants) provides independent but complimentary chemical tracers that can have signatures unique to the region(s) in which an organism forages. This strategy has been applied to small Interleukin-3 receptor cetacean populations in the southwestern Mediterranean Sea, the northeast Atlantic Ocean, and Black Sea (Das et al. 2000, 2004b; Borrell and Aguilar 2005; Borrell et al. 2006); ringed seal populations in the Canadian Arctic (Fisk et al. 2002a); minke whales in the North Atlantic (Born et al. 2003); and killer whale ecotypes in the North Pacific Ocean (Herman et al. 2005; Krahn et al. 2007, 2008). As with most ecological applications of stable isotope analysis, diet and habitat preferences are the primary pieces of information acquired through study of population structure and niche separation.

30 In the second part of our study we showed that αVEGFR2 treatment prevents the progression of NASH by attenuating steatosis and inflammation, both in a preventive and therapeutic setting, thereby confirming our hypothesis that angiogenic factors play an early role in the disease progression from steatosis to NASH. The grade of steatosis and inflammation was significantly less in the liver of αVEGFR2-treated mice in a preventive setting. Moreover, αVEGFR2 treatment was able to block the progression to NASH in mice with steatosis and inflammation. This could indicate that αVEGFR2 treatment could temper the effect of angiogenic stimuli. Moreover, in vitro we

found that αVEGFR2 therapy significantly

decreased lipid accumulation in fat-laden primary hepatocytes. This is in line with previous studies that https://www.selleckchem.com/products/Cisplatin.html the VEGF/VEGFR2 pathway is critical for both angiogenesis and adipogenesis during de novo adipose tissue formation from preadipocytes.31 Moreover, previous studies EPZ-6438 showed that angiogenic, inflammatory, and lipogenic processes are tightly crosslinked and are capable of sustaining each other.32 The reason why αPlGF treatment did not have a significant effect on the liver histology of mice with NASH compared to untreated mice with NASH could probably lie in the fact that PlGF signals only through the VEGFR1 pathway and not by way of VEGFR2. To obtain a clear insight into the molecular events, we examined the transcript levels of lipogenic genes (Scd1 and L-fabp1). These experiments showed that αVEGFR2 treatment increased Scd1 gene expression

in both a preventive and therapeutic setting. Scd1 plays a key role in the prevention of steatohepatitis by partitioning excess lipid into monounsaturated fatty acids that can be safely stored.33 The literature confirms our results that MCD feeding causes a significant decrease in hepatic Scd1 gene expression as well as the down-regulation of L-fabp1 gene expression compared to mice fed a control diet.34, 35 Our study showed that αVEGFR2 increases Scd1 gene expression to a more normal Scd1 expression in the liver of mice treated in a therapeutic and preventive setting compared to untreated mice fed an Celecoxib MCD diet. This could suggest that αVEGFR2 therapy improves lipid metabolism in the liver. However, the exact molecular mechanism responsible for MCD diet-induced down-regulation of Scd1 as well as changes in Scd1 expression in human NAFLD, and its relation to liver damage and disease progression, remain unknown and will require further investigation. In summary, we demonstrated the role for VEGF in the pathophysiology in two mouse models for NASH. This study shows for the first time that αVEGFR2 treatment attenuates steatosis and inflammation in the liver of mice with NASH both in a preventive and therapeutic setting.

We produced VSVg pseudotyped lentiviral vectors expressing the murine Fah complementary DNA (cDNA)

from an internal SFFV promoter. The enhanced green fluorescence protein (eGFP) was coexpressed either by an internal ribosomal entry site (IRES) or by a 2A proteolytic cleveage site (RRL.PPT.SFFV.eGFP.pre*, RRL.PPT.SFFV.Fah.ires.eGFP.pre*, RRL.PPT.SFFV.Fah.2a.eGFP.pre*).29 The vector supernatants were produced by 293T cells, concentrated with Centricon Plus-70 filter units ITF2357 concentration (Millipore, Schwalbach, Germany) and titrated on HepA1.6 cells.30 Viral titers were in the range of 107 to 108 IU/mL. Hepatocytes were isolated from 3 to 6-month-old mice as described.6 For further enrichment of hepatocytes we used discontinuous Percoll (GE Healthcare) gradients. A 25% phase prevented dead cells and debris from entering the gradient. A lower 50% phase facilitated the enrichment of small, highly viable, and robust hepatocytes. For depletion of nonparenchymal liver cells, we used PE (Phycoerythrin)-labeled anti-CD45 and anti-CD31 antibodies (BD Pharmingen), anti-PE MicroBeads on an automated MACS Separator (Miltenyi Biotech, Bergisch Gladbach). Analysis of eGFP expression was performed on day 5 after in vitro transduction of primary murine hepatocytes

or directly after hepatocyte isolation during serial transplantations by FACS. For in vitro experiments, hepatocytes were cultured on Primaria dishes (1.1 × 106 cells/35 cm2) in HCM medium (Lonza). Fah(-/-) hepatocytes isolated from C57Bl/6-Fahtm1Mgo were cultured in the presence of NTBC (9.6 ng/mL) selleck chemicals (Swedish Orphan). Transduction (multiplicity of infection [MOI] 10) was performed as previously described by our group.6 RNA from hepatocytes was hybridized on the Affymetrix Mouse 430 2.0 GeneChip.

Data can be found at Geo link http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE34731. LM-PCR was performed and analyzed as described.31, 32 (See also Supporting Material and Methods.) C57Bl/6-Fahtm1Mgo (Fah(-/-)) mice are defective for the Fah gene, which encodes the enzyme fumaryacetoacetate hydrolase (Fah).25 Mice were maintained by supplementation of drinking water with NTBC (4 mg mL-1, Swedish Orphan International).26, Niclosamide 27 To allow engraftment of hepatocytes the medication was discontinued For in vivo gene transfer, lentiviral particles were injected in 250 μL phosphate-buffered saline (PBS) intrasplenically (∼1 infectious particle / parenchymal liver cell). For ex vivo gene therapy 1 × 106 cultured Fah(-/-) hepatocytes were transduced overnight (MOI 10). For serial hepatocyte transplantation the livers from gene-corrected mice were perfused with collagenase. Aliquots of 0.5 × 106 liver cells were injected in 250 μL PBS into the next generation of Fah(-/-) mice.

Volume 01: New South Wales drawings (‘The Lambert Drawings’), Acknowledgements. Mitchell Library, State Dabrafenib Library of New South Wales. Table S1. List of pre-1900 CE dingo specimens used in analyses. Table S2. Dates of previously undated dingo cave specimens. “
“The male reproductive tract of most Australian hopping mice in the genus Notomys has a suite of highly derived features that differ markedly from those of other Australian rodents. These include, among others,

extremely small testes, a reduced complement of accessory sex glands and a spiny penis. Here we ask the question – what are the coevolved features of the female reproductive tract? To answer this question, we used histology and resin casts to compare the reproductive tract of the Australian plains mouse (Pseudomys australis) with that of the Spinifex hopping mouse (Notomys alexis). In P. australis, the cervix is highly fibrous and has learn more two small canals whereas the vagina has prominent fornices, a large lumen and a folded epithelial lining. By contrast,

in N. alexis the cervix is not prominent and is far more cellular. It has a very small, single lumen with the boundary between it and the vagina not being readily evident. The vagina has minute fornices and is surrounded by a comparatively thick muscle coat. Shortly after ejaculation, N. alexis had many uterine sperm that associated with coagulated material but, unlike in P.australis, no large vaginal plug occurs after ejaculation. These observations support Lumacaftor cell line the conclusion

that N. alexis has a highly derived distal region of the female reproductive tract which has coevolved with that of the male. It appears to facilitate rapid sperm transport postcoitum without the need for a large copulatory plug. “
“The distribution of ovulation patterns and penile ornamentation in mammals is thought to be shaped by sexual selection. Alternatively, ovulation patterns have been linked to factors such as phylogeny, social system and ecological constraints but no conclusive pattern has emerged. African mole-rats exhibit a unique range of social organizations and experience diverse ecological conditions (i.e. rainfall patterns), with various species exhibiting either induced or spontaneous ovulation in addition to a corresponding variation of penile ornamentation. Investigations of members of this family conducted so far do not allow conclusions to be drawn about the importance of phylogenetic versus ecological constraints for the evolution of ovulation patterns because all species of the genus Cryptomys studied occur in mesic habitats and exhibit induced ovulation. In contrast, the one representative of the genus Fukomys is a spontaneous ovulator that occurs in arid habitats.

The internal structure of both lineages was different from the isolectotype of Lessonia nigrescens. It is therefore concluded that the name Lessonia nigrescens should not be used for the Chilean material. Chordaria spicata Suhr appears as the oldest available name for the central lineage, while Lessonia

berteroana Montagne is the oldest name for the northern lineage. In both cases, the type material consisted of small-sized, apical branches of larger plants. The new combination Lessonia spicata (Suhr) Santelices is proposed Selleck Vemurafenib for the central lineage and we reinstate Lessonia berteroana for the northern lineage. Laminaria scissa Suhr is reduced to synonym of L. spicata. Representative specimens of Lessonia nigrescens were not found during new visits to its type locality in Cape Horn and along Chile. Future studies should verify the status of this species. “
“In the Ross Sea, the prymnesiophyte Phaeocystis antarctica G. Karst. dominates deeply mixed water columns, while diatoms dominate shallower mixed layers. Understanding what controls the dynamics of these two phytoplankton taxa is essential because they dominate virtually all coastal polar waters, have different Maraviroc ic50 nutrient

utilization characteristics, and support dissimilar food webs. We cultured two strains of P. antarctica and one strain of the diatom Fragilariopsis cylindrus (Grunow) Willi Krieg under three dynamic Calpain irradiance regimes that simulated different mixed-layer depths and measured their photosynthetic characteristics, cellular pigment concentrations, and cellular carbon and nitrogen content. In both species, chl a–normalized maximum carbon uptake rate (Pm* ) and specific growth rate were highest in the deeply mixed treatment that had a dark period.

In all irradiance treatments, both (Pm* ) and photosynthetic efficiency (α*) were greater for the two P. antarctica strains than for the F. cylindrus strain. In contrast, P. antarctica strains were more susceptible to photoinhibition (β*) than the F. cylindrus strain. When photosynthetic rates of each phytoplankton taxon were normalized by cellular particulate organic carbon (POC), the difference in the maximal photosynthetic rate () was generally reduced. In the dynamic irradiance treatment that simulated the shallowest mixed-layer irradiance, all three phytoplankton had similar ; however, the diatom had a 2-fold higher POC-normalized photosynthetic efficiency (αC). Finally, we performed calculations using the measured POC-normalized photosynthetic parameters to show that αC and can play a greater role than βC in determining the competitive outcome between P. antarctica and F. cylindrus in both shallow and deep mixed-layer environments of the Ross Sea. “
“The chl-specific short-term 14C-based production (Pb) measurement is a widely used tool to understand phytoplankton responses to environmental stresses.

We also establish a web-based tool, HNF4 Motif Finder, that can be used to identify potential HNF4α-binding sites in any sequence. (HEPATOLOGY 2009.) Hepatocyte nuclear factor 4α, HNF4α (HNF4A), is a member of the nuclear receptor superfamily of ligand-dependent transcription factors (NR2A1) and a liver-enriched transcription factor (TF) that is also expressed in the kidney, pancreas, intestine, colon, and

stomach.1 Originally identified based on its ability to bind DNA response elements in the human apolipoprotein C3 (APOC3) and mouse transthyretin (Ttr) promoters,2 FK506 chemical structure HNF4α has since been shown to play a critical role in both the development of the embryo and the adult liver.3, 4 Mutations in the HNF4A coding sequence and promoter regions are linked to Maturity Onset Diabetes of the Young 1 (MODY1),5 and mutations in HNF4α response elements have been directly linked to disease, most notably in genes encoding blood coagulation factors in hemophilia and in HNF1α in MODY3.6–8 Through classical promoter analysis, functional HNF4α-binding sites have been identified in >140 genes, including those involved in the metabolism of glucose, lipids, and amino acids, as well as xenobiotics and drugs1, 4, 9 (see Supporting Table 1A for a listing of those genes). Recent genome-wide location analyses suggest

that the number of HNF4α targets may be much greater (>1000) based on widespread binding of HNF4α to promoter regions,10–12 although it is not known how many of those are functional targets. A more comprehensive Selleck Sorafenib list of direct HNF4α targets was recently made even more critical with our finding that HNF4α binds an exchangeable ligand and hence may be a potential drug target.13 HNF4α binds DNA exclusively Buspirone HCl as a homodimer.14, 15 The canonical HNF4α consensus sequence consists of the half site AGGTCA with one nucleotide spacer (referred to as a DR1, AGGTCAxAGGTCA).16 Whereas the number of experimentally verified HNF4α binding sequences is sizable (>217) (Supporting Tables 1A and 1B), they were derived in a biased

fashion building on the first HNF4α-binding sites,2 and subsequently on the direct repeat rules for nuclear receptor DNA binding.16 Furthermore, the total number of 13-base oligomer (13-mer) permutations is much greater than 217 (413 ∼ 67 million), and whereas HNF4α will certainly not bind all potential 13-mers, the total number of DNA sequences that will bind HNF4α is anticipated to be in the tens of thousands. Because the presence of one or more HNF4α response elements in the promoter region of a gene is a prerequisite for classification as a direct HNF4α target, it is desirable to accurately predict all the HNF4α-binding sites throughout the genome in an unbiased fashion. Recent genome-wide technologies, most notably genome-wide location analysis (i.e.