In addition, those studies that report increased prevalence offer no clear explanation and there is no clear evidence of increased obesity in older individuals with haemophilia [23]. Ageing pwh who are HIV positive may also be at higher risk for IHD because of highly active retroviral therapy (HAART). While it is recognized that non-haemophilic individuals on HAART therapy are at increased risk for myocardial infarction, in the absence of specific data it is not clear whether this risk is shared by pwh find more [25]. These studies demonstrate that atherosclerosis and IHD can and do occur in haemophilia. It may be that the severe deficiency of factor

VIII or IX may offer relative protection against the final thrombotic insult in the narrowed arterial lumen that often precipitates the more severe manifestations of IHD. If so, then it may be prudent to exercise caution during intensive replacement therapy such as with major surgical

procedures, particularly in elderly subjects and it may be preferable to use measures such as carefully controlled continuous infusion to avoid peaks of coagulation factor activity in this setting. This may be particularly important during replacement therapy in the setting of acute coronary syndrome [26]. Symptomatic ischaemic heart disease appears to be increasing in haemophilia [27] at least in part because of an ageing population. Acute coronary syndromes (ACS) pose a Ponatinib nmr particular challenge because of the need to consider the risk of bleeding when using antithrombotic therapy. MK-1775 in vivo There is a paucity of data from which to create guidelines for management of

this situation. Most reports are of single cases. In general, the principle of management of these clinical cases is to correct the clotting factor deficiency by using factor replacement and then treating the patient as closely as possible to standard protocols for ACS. Recently, consensus guidelines have been published for this situation and have made recommendations specific for haemophilia such as avoidance of thrombolytic therapy, the use of bare metal stents for percutaneous coronary intervention and the use of prophylaxis during dual anti-platelet therapy [27]. While such guidelines are likely to be useful to guide treatment of individual patients, it must be recognized that such guidelines are largely based on opinion rather than evidence and it is important that they should be reviewed and updated when more robust evidence emerges. Valvular heart disease is also more prevalent in older populations [28] and it is likely that more cardiac surgery will be performed in older persons with haemophilia. Cardiac bypass has been performed safely in haemophilia [29] but requires careful planning and management. Valve prostheses should be of a material that does not necessitate anticoagulation.

Key Word(s): 1. biliary; 2. stenosis; 3. inflammatory; 4. pseudotumor; 5. IPT; 6. benign Presenting Author: CATHARINA TRIWIKATMANI Additional Authors: SARAH MARDHIAH SUDIN, NORZAREEN AZLINDA MOHD ROS, NUR IZZATI ABDUL MALEK, FAHMI INDRARTI, NENENG RATNASARI Corresponding Author: CATHARINA TRIWIKATMANI Affiliations: Gadjah Mada University, Gadjah Mada University, Gadjah Pembrolizumab datasheet Mada University, Faculty of Medicine, Gmu / Dr. Sardjito Hospital, Faculty of Medicine, Gmu / Dr. Sardjito Hospital Objective: Complete removal of all bile duct stones is recommended. The selective use of intraoperative

cholangiography or choledochoscopy exploration with cholecystectomy is still controversial. Usually, liver biochemical tests and transabdominal ultrasound are performed as initial evaluations and risk stratifications. T he aim of this research is to evaluate the difference of bilirubin, alkaline phosphatase (ALP), g amma g lutamyl t ransferase (GGT), aspartate transaminase (AST), and alanine transaminase

(ALT) level s in different Buparlisib research buy gallstone locations. Methods: From 2010 through 2013, 289 adult patients with gallstone disease underwent biliary surgery in Dr. Sardjito Hospital. One hundred and thirty six p atients with appropriate medical record data and criteria were included into this study. The subjects were divided into two groups, i.e. patients with choledocholithiasis and with cholecystolithiasis. The stone locations were determined based on final STK38 surgery reports. The latest liver biochemical test results during one week before surgery were chosen. Results: T abel. Liver biochemical level according to gallstone location Liver biochemical test Gallstone location n Median (min.-max.) p * Direct bilirubin

(mg/dL) Choledocholithiasis 16 0.15 (0.02–9.86) 0.000 Chole cysto lithiasis 97 4.40 (0.42–9.39) ALP (U/L) Choledocholithiasis 8 78.5 (45–552) 0.001 Chole cysto lithiasis 36 252 (111–1093) GGT (U/L) Choledocholithiasis 5 103 (14–430) 0.017 Chole cysto lithiasis 17 326 (220–803) SGOT (U/L) Choledocholithiasis 14 23 (8–224) 0.000 Chole cysto lithiasis 104 56 (14–168) SGPT (U/L) Choledocholithiasis 14 27 (7–323) 0.001 Chole cysto lithiasis 103 52.5 (28–248) *Mann-Whitney U test. Conclusion: Serum liver biochemical levels have significant differences in different gallstone location. Key Word(s): 1. liver biochemical level; 2. gallstone location Table 1. Liver Biochemical Level According to Gallstone Location. Liver biochemical test Gallstone location n Median (min.-max.) p* *Mann–Whitney U test.

The main axis and lateral veins were clearly visible. The lateral veins branched off at an angle of less than 90° and most of them were bifurcated. Like the individuals from Brittany, blades were devoid of marginal teeth Autophagy activator or spines.

In Scotland, D. dudresnayi was found in the narrow sea straits between Dunstaffnage and Eilean Mhor (near Oban) on August 20, 2010. The habitat (pebbles and small rocks on a mostly sandy seabed) was different from the localities off Roscoff (underwater cliff faces), but more resembling that where D. dudresnayi was encountered in Galicia – a seabed consisting mostly of gravel at ~15 m below low tide level, with D. dudresnayi thalli growing attached to small pebbles and sea shells. Despite two searches using SCUBA, no D. dudresnayi was found at the same locality in the summer of 2011. Gametophytes of D. dudresnayi from Brittany developed

only in the culture from the unbranched individual collected on July 18, 1999 (Fig. 2c). From Galicia, we obtained gametophytes both from the unbranched and the branched individual. All three cultures gave rise to monoecious gametophytes, selleck screening library indistinguishable from each other. They consisted of branched creeping filaments 10–15 μm in diameter. Germlings became reproductive in 10°C and 15°C and bore antheridia and oogonia on the same thallus. Sporophytes developed from oogonia, without release of eggs (Fig. 2c). Sporophytes of D. dudresnayi grown in our cultures to a length of several centimeters remained unbranched (Fig. 2d). The specimen of D. ligulata collected in Galicia was profusely branched. The maximum width of the main axis was 6 mm (Fig. 3). mTOR inhibitor Gametophytes of D. ligulata from Galicia were monoecious (not shown as they were similar to previously studied isolates, e.g., Peters and Müller 1986, Ramirez and Peters 1992). The time required for gametogenesis in D. dudresnayi and D. ligulata was compared. Vegetative gametophytes of both species from Galicia were simultaneously inoculated at 10°C and the appearance of first young sporophytes (like those illustrated in Fig. 2c) was recorded. D. dudresnayi gametogenesis took 14 d, whereas in D.

ligulata, it required only 10 d. Sequence statistics obtained for the alignments of the five markers used in the present study are summarized in Table 3. Nuclear SSU rDNA and ITS required gaps for multiple sequence alignment, however, no insertion and deletion were found in the three protein markers. SSU was the most conserved gene (98.2% constant positions) and mitochondrial cox1 was the most variable gene (27.2% variable positions) among the five genes used in the present study. The highest P-distance was found in cox1 (0.072 ± 0.01), followed by psaA (0.047 ± 0.005), ITS (0.026 ± 0.004; ligulate Desmarestia species only), rbcL (0.02 ± 0.002), and SSU (0.003). In each alignment, most variable positions were identified as phylogenetically informative sites.

Benign lesions/trauma of the pancreatic neck and proximal body pose an interesting surgical challenge. Central Pancreatectomy is an operation

that allows resection of benign tumours located in the pancreatic isthmus that are not suitable for enucleation. Methods: Between 2006 and 2011, 16 parenchymal sparing conservative pancreatectomies were carried out. There were 6 women and 10 men with a mean age of 35.7 years. In 8 patients who underwent central pancreatectomy, the cephalic pancreatic stump is oversewn and the distal stump is anastomosed end–to–end with a Roux-en-Y jejunal loop see more in two and with the stomach in six patients. The indications for Central pancreatectomy were: non-functional islet cell tumours in two patients, traumatic pancreatic neck transection in two and one each for

insulinoma, solid pseudopapillary tumour, splenic artery pseudoaneurysm and pseudocyst. The indication for Pancreatic head coring (Frey’s procedure) was chronic pancreatitis with benign head mass in 7 GPCR & G Protein inhibitor patients and one underwent spleen preserving distal pancreatectomy for insulinoma in pancreatic tail. Pancreatic exocrine function was evaluated by a questionnaire method. Endocrine function was evaluated by blood glucose level. Results: Morbidity rate was 37.5% with no mortality. Mean postoperative hospital stay was 10.5 days. Neither of the patients developed pancreatic fistula nor required reoperations or interventional radiological procedures. At a mean follow up of

26.4 months, no patient had evidence of endocrine or exocrine pancreatic insufficiency, all the patients were alive and well without clinical and imaging evidence of disease recurrence. Conclusion: When technically feasible, Central Pancreatectomy is a safe, conservative pancreatectomy for non-enucleable benign pancreatic pathology confined to pancreatic isthmus that allows for cure of the disease without loss of substantial amount of normal pancreatic parenchyma with preservation of exocrine/endocrine function and without interruption of pancreatico-biliary-enteric continuity appears to have long-term functional Dichloromethane dehalogenase advantages with good quality of life. Central pancreatectomy with pancreaticogastrostomy is also a safe conservative surgical strategy worthy of consideration as an alternative to distal pancreatectomy in the setting of isolated traumatic pancreatic neck transection. Key Word(s): 1. central; 2. segmental; 3. pancreatectomy; 4. Nonstandard; Presenting Author: BING-RONG LIU LIU Additional Authors: JI-TAO SONG SONG, LING-JIAN KONG KONG Corresponding Author: BING-RONG LIU LIU Affiliations: The second affiliated hospital of Harbin medical university Objective: Peritoneal access techniques are among the most important concerns in the clinical application of natural orifice transluminal endoscopy surgery (NOTES).

The effect of miR-409-3p on proliferation and apoptosis

was via directly targeting the transcriptional regulator PHF10 [24], whereas the radixin was the target for the pro-metastatic pathway [16]. MiR-182 expression that was down-regulated in gastric adenocarcinoma tissue samples had an inverse correlation with CREB that was a direct target of this miRNA [25]. An increase in proliferation and colony formation was detected when miR-182 was overexpressed in GC cells [25]. MiR-429, which was down-regulated in GC tissues, acted as inhibitor of cell proliferation and attachment, when it was overexpressed in CGC-7901 cells, and it targeted c-myc [26]. Next-generation sequencing techniques such as ACP-196 research buy exome sequencing have lead to the detection of new molecules and mechanisms that are involved in gastric carcinogenesis. Dysregulation of miRNAs is also evident in GC, and they hold potential as novel diagnostic, prognostic, and predictive markers in this disease. Competing interests: the authors have no competing interests;][#,63]?> “
“Background:  The aim of this study was to produce a recombinant version of the highly antigenic Helicobacter pylori TonB (iron-dependent siderophore X-396 cell line transporter protein HP1341) in

transgenic plants as a candidate oral vaccine antigen. Materials and Methods:  Using Agrobacterium-mediated gene transfer, we introduced three different

constructs of the tonB gene into the genome of the model plant Arabidopsis thaliana. We investigated transgene insertion by PCR, produced TonB antibodies for analysis of the production of the recombinant protein in plants, verified the identity of the protein produced by mass spectrometry analysis, and analyzed the number of genetic inserts in the plants by Southern blotting. Results:  Three different constructs of the expression cassette (full-length tonB, tonB truncated in the 5′ end removing the codons for a transmembrane helix, and the latter construct with codons for the endoplasmic reticulum SEKDEL retention signal added to the 3′ end) were used to find the most effective way to express the TonB antigen. Production of TonB protein was detected in plants transformed with each of the constructs, selleck kinase inhibitor confirmed by both Western blotting and mass spectrometry analysis. No considerable differences in protein expression from the three different constructs were observed. The protein concentration in the plants was at least 0.05% of the total soluble proteins. Conclusions:  The Helicobacter pylori TonB protein can be produced in Arabidopsis thaliana plants in a form that is recognizable by rabbit anti-TonB antiserum. These TonB-expressing plants are highly suitable for animal studies of oral adminstration as a route for immunization against Helicobacter infections.

We manipulate resource acquisition in adults by providing some females with a small mouse carcass (5–10 g) and other females with a large mouse carcass (15–20 g). We found that females breeding on larger carcasses

produced both more and larger offspring than females breeding on smaller carcasses. Furthermore, an increase in brood size had a stronger negative effect on offspring mass in broods produced on smaller carcasses than in broods produced on larger carcasses. We conclude that phenotypic variation in resource acquisition had a strong effect on the number and mass of offspring and the trade-off between the two. Our study contributes to our understanding of phenotypic variation in PD-0332991 ic50 resource acquisition by showing that females with more resources produce both more and larger offspring in situations where such variation is not associated with anatomical or physiological differences between females. “
“The evolution of large body size has often been considered a key trait allowing the evolution of herbivory in lizards. Although many omnivorous lizards appear unspecialized, they typically show high bite forces, allowing them to reduce

tough and fibrous plant matter. In contrast, true herbivores often show a suite of morphological and physiological specializations, allowing them to efficiently NVP-BKM120 price process and assimilate plant material. Moreover, many specialized herbivores have a large body size, thus likely relaxing constraints on bite-force generation given that bite force increases with increasing body mass. In this study, we test whether large herbivorous lizards of the genus Uromastyx have relatively lower bite forces for their body size compared with a medium-sized congener. No differences in bite force or head dimensions were observed between the two species or between both sexes in our sample.

Moreover, bite force scaled with positive allometry relative to jaw length, suggesting that larger animals have disproportionately large bite forces. This suggests that even in the largest species, constraints on bite-force Carnitine palmitoyltransferase II generation are still strong, possibly due to the demands imposed on the jaw system by the mechanical properties of the diet. “
“In species with simultaneous polyandry, male-biased operational sex ratio is expected to increase the risk of sperm competition and thus sperm traits affecting siring success can differ among populations. Here, we test the hypothesis that high male–female ratios will enhance sperm competitiveness of Rana temporaria males. In this species, local populations can show either prolonged or explosive breeding. In a context of sperm competition and in controlled laboratory conditions, prolonged-breeding males sired a higher proportion of eggs than explosive-breeding males, regardless of female origin.

Accordingly, Selleckchem FK506 international guidelines recently published by the European Crohn’s and Colitis Organization (ECCO), recommend screening for latent infections, and immunization of all IBD patients, and in particular those who receive or are scheduled to start an immunosuppressive drug. However, clinical experience has revealed that it is difficult to implement these recommendations in an everyday clinical setting. Aim: To investigate if a program of systematic

information about immunization status and vaccination recommendations will increase adherence to vaccination and screening guidelines in patients with IBD on immunosuppressive therapy. Methods: Methods: The study consists of two parts: 1) An observational retrospective part including patients with IBD in in anti-TNF-α (tumor necrosis factor) therapy, and other immunosuppressive therapy from the IBD Clinic at Herlev Hospital. Patients will be interviewed regarding immunization Tamoxifen nmr and adherence to vaccination guidelines. 2) An interventional prospective study in which healthcare professionals will receive information about international and local guidelines. Likewise patients will receive systematic information about importance of vaccination and screening. Patients

attending the IBD clinic in the study period will be included. Patients will be interviewed after one year using the same questions as in the retrospective study. Results: Endpoints: The proportions of patients in each of the two substudies who Flucloronide are: 1) fully adherent to the screening and vaccination program; 2) partially adherent; 3) completely non-adherent.

Furthermore, the physicians’, nurses’ and patients’ reasons for not following the recommendations for screening and vaccination. Conclusion: In conclusion, the study will show whether systematic information improves adherence to recommended screening and vaccination guidelines, and thereby aid to improve safety. Key Word(s): 1. IBD; 2. Immunosuppressiva; 3. Vaccination; 4. Guidelines; Presenting Author: XIAOFEI ZHANG Additional Authors: WENYU JIANG, WENJIE LI, XIUQIN CHENG, HONGJIE ZHANG Corresponding Author: HONGJIE ZHANG Affiliations: First Affiliated Hospital of Nanjing Medical University Objective: Cytokine signaling (SOCS)-3 plays an important role in autoimmune disorders. High levels of SOCS3 were observed in Crohn’s disease(CD). MicroRNAs(miRNAs) can regulate gene expression during immune responses. The aim of our study was to investigate the contribution of SOCS3 expression-associated miRNA to the regulation of chemokines production in colonic epithelial cells(CEC) in active CD. Methods: Targetscan Human 6.2 was used to screen miRNAs which may be target the 3′ untranslated region(3′ UTR) of SOCS3, and quantitative PCR was used to detect these miRNAs levels in active CD and healthy subjects. The luciferase reporter assay was performed to confirm the target gene.

alJour. Nutr. Bioch. 2013). The aim of the present study was to understand the effects of increased oxidative stress on AMPK signaling and activity in vitro as well as in a murine model of chronic ethanol consumption. Methods: Using recombinant protein, an in vitro human hepatocyte HepaRG cell culture system

or a murine model of ALD, the direct effects of lipid peroxidation were examined with respect to AMPK phosphorylation, carbonyla-tion, enzymatic activity and mTOR inhibitor phosphorylation of ACC. Results: In HepaRG cells, incubation with increasing concentrations of 4-HNE resulted in decreased phosphorylation of AMPK and ACC. Pretreatment of 4-HNE inhibited both hydrogen peroxide and adiponectin induced phosphorylation of AMPK and subsequent phosphorylation of ACC. Using biotin hydrazide capture, it was confirmed that exposure of HepaRG cells to 4-HNE resulted in carbonylation of AMPKα/β which was not observed

in untreated control cells. Based on this data, mass spectral analysis of 4-HNE treated DAPT recombinant AMPKα identified Michael addition adducts of 4-HNE on Cys130, Cys174, Cys227 and Cys309. Global computational based molecular modeling analysis of AMPK following 4-HNE modification revealed no significant changes in secondary or tertiary structure. Molecular modeling analysis of 4-HNE adducted to Cys174 AMPKα

suggest inhibition of AMPK activity by steric hindrance of the active site pocket. Using a murine model of alcoholic liver disease, chronic ethanol consumption resulted in an increase in carbonylated AMPKα despite increased phosphorylation of Thr172AMPK. There was no significant change in phosphorylation of ACC. Conclusions: Combined these data demonstrate for the first time that AMPK is a direct target of reactive aldehyde during conditions PI-1840 of increased oxidative stress in the liver. The inhibition of AMPK by reactive aldehydes provides a novel mechanism for defective AMPK signaling and β-oxidation in ALD. This work was funded by NIH 5F32 AA018613-03 (C.T.S.) and 5R37 AA009300-16 (D.R.P.). Disclosures: The following people have nothing to disclose: Colin T. Shearn, David J. Orlicky, Dennis R. Petersen Alcoholic hepatitis (AH) is the most severe form of alcoholic liver disease and is one of the most deadly conditions in hepa-tology. There is a clear need to identify non-invasive biomarkers and molecular drivers in order to develop novel targeted therapies. We recently showed that progenitor cell expansion is a hallmark of severe AH and correlates with disease severity. Here, we performed a translational study including human and experimental data using a systems biology approach to identify new molecular biomarkers of AH.

8 to 2.5 times increased odds of having no surveillance. Patients

with complete surveillance had a significantly (p<0.05) greater number of physician visits during follow-up when stratified by hepatic decompensation status: 1) no hepatic decompensation: mean-1.8 visits (complete surveillance), 1.1 (incomplete), and 0.6 (none); 2) prior hepatic decompensation: mean-2.8 visits (complete), 1.5 (incomplete), and 1.1 (none). In linear regression models, non-GI provider, non-PPO/POS health insurance, and increasing age were associated with decreased PUTDS, while a history of a hepatic decompensation, presence of any component of the metabolic syndrome, and diagnosis of hepatitis B www.selleckchem.com/products/gsk1120212-jtp-74057.html or C were associated with increased PUTDS. Conclusions: HCC surveillance rates in commercially insured at-risk patients remain poor despite Small Molecule Compound Library formalized guidelines. Improving access to appropriate specialized care should

be targeted for quality improvement interventions. Disclosures: David S. Goldberg – Grant/Research Support: Bayer Healthcare Adriana Valderama – Employment: Bayer Sujit S. Sansgiry – Consulting: Bayer Pharmaceuticals James D. Lewis – Grant/Research Support: Bayer The following people have nothing to disclose: Rajesh Kamalakar, Svetlana Babajanyan Background: The United Network for Organ Sharing (UNOS) provides patients (pts) with HCC who are listed for LT with exemption MELD points that can place them at an advantage for earlier LT compared to pts listed for non-malignant indications. Aim: Identify Avelestat (AZD9668) a scoring system that achieves survival benefit equity among pts with and without HCC who are listed for LT. Methods: We defined LT survival benefit as the difference between life expectancy if transplanted and life expectancy if the subject remains on the waiting list (WL). Adult pts listed for LT in the United States between 2003 and 2012 were identified from the UNOS database, including HCC pts meeting exemption policy 3.6.4.4 (stage T2). A univariable analysis was performed to assess differences between HCC and non-HCC pts; this was done for WL and LT populations. Pre-LT survival

was modeled using competing risks analysis and post-LT survival was assessed using Cox regression. Using these models, life expectancy on WL and after LT was estimated for each patient by calculating the area under the survival curve up to 5 years using the trapezoidal rule. Linear regression was used to obtain a regression model defining 5-year LT survival benefit based on MELD score for non-HCC pts and based on MELD and AFP for HCC pts. These 2 models were equated to obtain an adjusted HCC-MELD score which matches the LT survival benefit of HCC pts to non-HCC pts having the same biochemical MELD. Results: 101,458 pts were included in the analysis. Average age at time of listing was 53 ± 10 years, 65% were male and 13% had HCC. 69% of HCC pts underwent LT compared to 47% of non-HCC pts.

6C). Anti–β1-integrin antibody and echistatin promoted cellular rounding in both the

Huh7 and HepG2 cells cultured on collagen-I–coated 12 kPa (stiff) supports. Huh7 cell proliferation was reduced by treatment with both 6S6 antibody (38% reduction, P < 0.05) and echistatin (29% reduction, P = 0.07) relative to relevant controls. Similarly, in HepG2 cells, cell proliferation was reduced by treatment with both 6S6 antibody (92% reduction, P < 0.001) and echistatin (21% reduction, P < 0.01). The effect of FAK activation on HCC cell proliferation was investigated in experiments with the small molecular FAK inhibitor PF573228 (Fig. 6B,C). Treatment with PF573228 (5 μM) was associated with a reduction in the proliferation of both Huh7 (42% reduction, P < 0.01) and HepG2 cells (45% reduction, P < 0.001) cultured on collagen-I–coated 12 kPa polyacrylamide gels. Furthermore,

inhibition of β1-integrin or FAK expression in HepG2 cells with siRNA click here resulted in a significant reduction in cellular proliferation relative to control siRNA transfection (Supporting Fig. 8). A similar trend in respect to cellular proliferation was observed following siRNA-dependent inhibition of β1-integrin or FAK expression GSK1120212 in vivo in Huh7 cells, although in this case the reduction was not statistically significant. HCC is resistant to treatment with conventional chemotherapeutic agents. We therefore investigated whether the stiffness of the cancer cell niche regulates the susceptibility of HCC cells to chemotherapy-induced apoptosis. In both cell lines, there was decreased apoptosis in cells cultured on stiff supports, as indicated by reduced poly-ADP-ribose polymerase (PARP) cleavage (Fig. 7A). There was a nonsignificant trend toward increased numbers of surviving cells on stiff supports (data not shown). We also performed a series of clonogenic assays to investigate whether changes in matrix stiffness modulate the survival and behavior of tumor-initiating cells after chemotherapy. Following cisplatin treatment, the surviving cell population included an increased frequency of clone-initiating also cells for both HepG2 (2.4-fold, P < 0.001) and Huh7 cells (2.2-fold,

P < 0.05) cultured on soft (1 kPa) versus stiff (12 kPa) supports (Fig. 7B). In addition, there was a nonsignificant trend toward an absolute increase in the total number of clone-forming cells from soft supports (data not shown). To assess the validity of this finding, experiments were repeated using a second, unrelated chemotherapeutic agent, 5-fluorouracil (5-FU). Consistent with our findings with cisplatin, following 5-FU chemotherapy there was an increased frequency of clone-initiating cells from HepG2 (3.6-fold, P < 0.001) and Huh7 cells (1.9-fold, P < 0.05) cultured on soft versus stiff supports. There was no difference in the frequency of clone-initiating cells in untreated HepG2 or Huh7 cells after culture on soft or stiff supports in the absence of chemotherapy.