3B, red line). Accordingly, an HBVpreS1-receptor is present on the hepatocytes of mice and rats. We extended this analysis and tested HBVpreS-peptide binding to primary hepatocytes from rabbits, dogs, cynomolgus monkeys, rhesus monkeys, and pigs. As depicted in Fig. 3C, we found specific binding of HBVpreS/2-48myr-K-FITC to rat, rabbit,

and dog hepatocytes but surprisingly not to hepatocytes from cynomolgus and rhesus monkey, despite their closer evolutionary relation to humans. Binding was also not observed on pig hepatocytes. Thus, differentiated hepatocytes from some HBV nonsusceptible species do express the HBVpreS-specific receptor (mouse, rat, dog, and rabbit), while others do not (pig, cynomolgus, and rhesus monkey), indicating that a step downstream specific preS-binding must restrict HBV/HDV-buy RG7204 infection in these species. The susceptibility of HepaRG cells to HBV infection depends on a differentiated state of the cells.7, 30 Moreover, PHH lose their

susceptibility to HBV when DMSO is withdrawn from the medium.31-34 In order to test if this correlates with the presence of the HBVpreS-receptor we analyzed undifferentiated (5 days after seeding) and differentiated HepaRG cells (28 days after seeding, including a 2-week DMSO treatment) for their ability to bind HBVpreS/2-48myr-K-FITC. As seen in Fig. 4A (upper panel, left), HBVpreS/2-48myr-K-FITC did not stain the PM of

undifferentiated cells, whereas binding was seen after differentiation (lower panel, left). This implies a cell state-dependent induction of HBVpreS/2-48myr-receptor expression during HepaRG-differentiation. Accordingly, we investigated whether dedifferentiation of binding competent primary hepatocytes result in an opposite effect. We cultivated PMH in the absence of DMSO and followed their ability to bind HBVpreS/2-48myr-K-FITC Sulfite dehydrogenase over time (Fig. 4A, right panels). While freshly isolated PMH specifically accumulated the peptide at the PM, hepatocytes from the same preparation lost their ability to bind HBVpreS/2-48myr-K-FITC within a few days of cultivation. Loss of binding could be prevented by addition of DMSO (data not shown). This correlates with the fact that PHH lose their susceptibility for HBV during several days of cultivation in the absence of DMSO.28 Thus, expression of an HBVpreS-receptor is linked to pathways controlling the differentiation state of hepatocytes. To investigate whether HuH7 and HepG2 express detectable amounts of the HBVpreS-receptor following DMSO-induced differentiation, we performed binding experiments with these cells under differentiation conditions. As shown in Fig. 4B (upper panel) dividing cultures of HuH7 and HepG2 cells showed no significant binding of HBVpreS/2-48myr-K-FITC. Cultivation in the presence of 0.

Enough time interval after treatment (i.e., 2 years) is necessary to confirm eradication, and it would not be easy to distinguish between recurrence and recrudescence before 2 years without identifying H. pylori

strains. “
“Studies on seroconversion and its reversion rate in Korean adults Ponatinib order with Helicobacter pylori infection are very rare. The purpose of this study was to evaluate the overall seroprevalence, seroconversion rate, and seroreversion rate of H. pylori infection in an adult population. We performed this retrospective cohort study on healthy adults who had visited our health screening center at Asan Medical Center more than twice between January 2000 and December 2010. We reviewed the anti- H. pylori Ab IgG profiles of the enrolled people and their family members and the results of esophagogastroduodenoscopies and a self-reported questionnaire. A total of 67,212 people were enrolled in this study. The mean follow-up duration was 4.6 years, and each participant visited the center for a mean of 3.8 visits.

The overall proportions of participants demonstrating persistent seropositivity, persistent seronegativity, seroconversion, and seroreversion were 53.1%, 32.5%, 4.3%, and 10.1%, respectively. The annual seroconversion rate was 2.79%. The annual crude and spontaneous seroreversion rates of the entire study population were 3.64% and 2.42%, respectively. According to multivariate logistic Stem Cell Compound Library chemical structure regression, old age (HR = 1.015), smoking (HR = 1.216), alcohol consumption more than four times per week (HR = 1.263), marriage (HR = 2.735), and living with H. pylori-infected family members (HR = 1.525) were identified as statistically significant risk factors associated with seroconversion.

The annual seroconversion rate was 2.79% in our study population. this website Marriage and living with H. pylori-infected family members were important risk factors affecting seroconversion in our adult population. “
“Peptic ulcer bleeding and recurrence rate are strongly linked to Helicobacter pylori infection even if nonsteroidal anti-inflammatory drugs (NSAIDs) play a relevant role in this setting. Further studies confirm that H. pylori eradication lowers the risk of recurrent peptic ulcer bleeding. Therefore, a test-and-treat strategy appears to be mandatory for patients with a history of ulcer bleeding and NSAIDs and/or aspirin use. Concerning gastroesophageal reflux disease (GERD), evidence clearly shows that H. pylori status has no effect on symptoms and treatment. Therefore, H. pylori treatment is not contraindicated in patients with GERD. The exact role of H. pylori in functional dyspepsia (FD) remains controversial. Novel possible mechanisms by which H. pylori may elicit dyspeptic symptoms include alterations of gastric motility, as well as endocrine and acid-secretory abnormalities.

Conclusion: Transplantation studies indicate that the state of the host microenvironment is critical to the regenerative potential of hepatocytes, and that a change in the extracellular matrix can lead to regeneration and restoration of function by cells derived from livers with end-stage organ failure. (HEPATOLOGY 2011) Expansion and altered composition of the extracellular matrix as a result of collagen deposition is a common response to injury and plays a major role in chronic heart, liver, and kidney

failure. Understanding the extent to which reversal of this process can lead to functional organ recovery is a critical issue, as numerous interventions have been proposed to improve fibrosis and presumably reverse organ failure.1-4 The unique capacity of hepatic parenchymal cells to undergo extensive proliferation in response to injury makes the liver find more an ideal organ to study cellular regeneration in acquired chronic disease. In the liver, expansion of the extracellular matrix with capillarization of the sinusoidal endothelium and loss of fenestrae results in cirrhosis with production Enzalutamide of regenerative hepatic nodules, portal hypertension, loss of hepatocytes, and liver failure.5 Loss of significant hepatocyte mass does not routinely produce hepatic failure, because the liver is capable

of normal function with less than half its normal complement of hepatocytes.6, 7 Thus, the cause of organ failure in cirrhosis is not well understood. Impaired hepatic function results from intrinsic damage to the native liver cells and from the abnormal

microenvironment in which they reside.8-14 Because collagen deposition and vascular changes in cirrhosis can be extensive before there is functional hepatic decompensation, it is not clear to what extent each plays a role or at what point these factors tip parenchymal cell function toward organ failure. Mito et al.15 attempted to address the role of the microenvironment in hepatic failure by transplanting this website hepatocytes from the livers of patients with cirrhosis back into their own spleens to reverse decompensated liver disease. If it is possible to recover the function of parenchymal cells from a cirrhotic liver by changing the microenvironment, it may be possible to restore hepatic function in the cirrhotic liver by reversing hepatic structural abnormalities, and individual cells derived from some cirrhotic livers might prove to be useful as an untapped source of transplantable cells for the treatment of patients with liver-based metabolic disorders, where the liver microenvironment is intact. Here, we demonstrate that primary cells derived from cirrhotic livers with decompensated function exhibit severe alterations in gene expression and defects in proliferative capacity and function directly after isolation, but engraft normally in a noncirrhotic microenvironment.

Conclusion: Transplantation studies indicate that the state of the host microenvironment is critical to the regenerative potential of hepatocytes, and that a change in the extracellular matrix can lead to regeneration and restoration of function by cells derived from livers with end-stage organ failure. (HEPATOLOGY 2011) Expansion and altered composition of the extracellular matrix as a result of collagen deposition is a common response to injury and plays a major role in chronic heart, liver, and kidney

failure. Understanding the extent to which reversal of this process can lead to functional organ recovery is a critical issue, as numerous interventions have been proposed to improve fibrosis and presumably reverse organ failure.1-4 The unique capacity of hepatic parenchymal cells to undergo extensive proliferation in response to injury makes the liver Selleckchem IWR1 an ideal organ to study cellular regeneration in acquired chronic disease. In the liver, expansion of the extracellular matrix with capillarization of the sinusoidal endothelium and loss of fenestrae results in cirrhosis with production Ibrutinib datasheet of regenerative hepatic nodules, portal hypertension, loss of hepatocytes, and liver failure.5 Loss of significant hepatocyte mass does not routinely produce hepatic failure, because the liver is capable

of normal function with less than half its normal complement of hepatocytes.6, 7 Thus, the cause of organ failure in cirrhosis is not well understood. Impaired hepatic function results from intrinsic damage to the native liver cells and from the abnormal

microenvironment in which they reside.8-14 Because collagen deposition and vascular changes in cirrhosis can be extensive before there is functional hepatic decompensation, it is not clear to what extent each plays a role or at what point these factors tip parenchymal cell function toward organ failure. Mito et al.15 attempted to address the role of the microenvironment in hepatic failure by transplanting selleck hepatocytes from the livers of patients with cirrhosis back into their own spleens to reverse decompensated liver disease. If it is possible to recover the function of parenchymal cells from a cirrhotic liver by changing the microenvironment, it may be possible to restore hepatic function in the cirrhotic liver by reversing hepatic structural abnormalities, and individual cells derived from some cirrhotic livers might prove to be useful as an untapped source of transplantable cells for the treatment of patients with liver-based metabolic disorders, where the liver microenvironment is intact. Here, we demonstrate that primary cells derived from cirrhotic livers with decompensated function exhibit severe alterations in gene expression and defects in proliferative capacity and function directly after isolation, but engraft normally in a noncirrhotic microenvironment.

Group comparisons were made with the Wilcoxon-Mann-Whitney test. Categorical variables are reported as counts and percentages. Group comparisons were made with the χ2 test or Fisher’s exact test. Survival was assessed with the Kaplan-Meier

nonparametric survivorship Buparlisib in vitro function, and group comparisons were made with the log-rank test. Univariate and multivariate Cox regression analyses were performed to detect the independent predictors of survival. In all survival analyses, the follow-up period ended either on the day of the last visit for nontransplant patients or on the day of transplantation for transplant patients. The multivariate model was built with the backward elimination technique with P < 0.10 for entering the model and P < 0.05 for staying in the model. The results are presented as crude hazard ratios (HRs) with 95% confidence intervals (CIs) in univariate analyses and as adjusted HRs with 95% CIs in multivariate analyses. Crude HRs indicate the relationship between mortality and a single predictor. Adjusted HRs indicate the relationship between mortality and a predictor and take into account the other

independent predictors. A P value < 0.05 was considered significant. Analyses were performed with the PASW statistical package (SPSS version 18.0, SPSS, Chicago, IL). A total of 151 patients were enrolled. Clinical characteristics, FK228 purchase biochemical

values, and treatment at inclusion are summarized in Table 1. One hundred four patients (68.9%) had diuretic-intractable ascites: renal dysfunction was found at entry in 46 patients (30.5%), and hyponatremia was found in 58 patients (38.4%). None of the patients had diuretic-intractable ascites due to abnormal serum potassium levels. Forty-seven patients (31.1%) had diuretic-resistant ascites. All patients were regularly treated with large-volume paracentesis and intravenous albumin. Seventy-seven patients (51%) were treated with nonselective selleck products beta-blockers (propranolol) for the prevention of gastrointestinal hemorrhage. Among these patients, 9 (11.7%) were given 40 mg of propranolol per day, 31 (40.3%) were given 80 mg, 1 (1.3%) was given 120 mg, and 36 (46.7%) were given 160 mg. The median follow-up time was 8 months (1-47 months). The median survival time was 10 months (95% CI = 8-12 months). The probability of survival was 41% at 1 year (95% CI = 33%-49%) and 28% at 2 years (95% CI = 20%-36%; Fig. 1). Ninety-seven patients (64.2%) died. Causes of death were sepsis in 50 patients (spontaneous bacterial peritonitis in 11 cases) and progression of hepatocellular carcinoma in 13. Twenty-five patients died at home of unspecified causes. Twenty-six patients underwent liver transplantation during the study period.

Group comparisons were made with the Wilcoxon-Mann-Whitney test. Categorical variables are reported as counts and percentages. Group comparisons were made with the χ2 test or Fisher’s exact test. Survival was assessed with the Kaplan-Meier

nonparametric survivorship PD0325901 research buy function, and group comparisons were made with the log-rank test. Univariate and multivariate Cox regression analyses were performed to detect the independent predictors of survival. In all survival analyses, the follow-up period ended either on the day of the last visit for nontransplant patients or on the day of transplantation for transplant patients. The multivariate model was built with the backward elimination technique with P < 0.10 for entering the model and P < 0.05 for staying in the model. The results are presented as crude hazard ratios (HRs) with 95% confidence intervals (CIs) in univariate analyses and as adjusted HRs with 95% CIs in multivariate analyses. Crude HRs indicate the relationship between mortality and a single predictor. Adjusted HRs indicate the relationship between mortality and a predictor and take into account the other

independent predictors. A P value < 0.05 was considered significant. Analyses were performed with the PASW statistical package (SPSS version 18.0, SPSS, Chicago, IL). A total of 151 patients were enrolled. Clinical characteristics, Ku-0059436 mw biochemical

values, and treatment at inclusion are summarized in Table 1. One hundred four patients (68.9%) had diuretic-intractable ascites: renal dysfunction was found at entry in 46 patients (30.5%), and hyponatremia was found in 58 patients (38.4%). None of the patients had diuretic-intractable ascites due to abnormal serum potassium levels. Forty-seven patients (31.1%) had diuretic-resistant ascites. All patients were regularly treated with large-volume paracentesis and intravenous albumin. Seventy-seven patients (51%) were treated with nonselective see more beta-blockers (propranolol) for the prevention of gastrointestinal hemorrhage. Among these patients, 9 (11.7%) were given 40 mg of propranolol per day, 31 (40.3%) were given 80 mg, 1 (1.3%) was given 120 mg, and 36 (46.7%) were given 160 mg. The median follow-up time was 8 months (1-47 months). The median survival time was 10 months (95% CI = 8-12 months). The probability of survival was 41% at 1 year (95% CI = 33%-49%) and 28% at 2 years (95% CI = 20%-36%; Fig. 1). Ninety-seven patients (64.2%) died. Causes of death were sepsis in 50 patients (spontaneous bacterial peritonitis in 11 cases) and progression of hepatocellular carcinoma in 13. Twenty-five patients died at home of unspecified causes. Twenty-six patients underwent liver transplantation during the study period.

The VWF monomer is heavily glycosylated with 20% by mass being composed of carbohydrate. The N-linked glycan structures of plasma VWF are complex type chains, with the majority being bi-antennary (78%) or tri-antennary (12%). The O-linked glycan structures of plasma VWF are short mucin-type carbohydrates and sialylated T antigen structures. Plasma VWF expresses terminal ABO(H) group antigens. In contrast, platelet VWF does not express A or B blood group antigens

[25], but it does express precursor H antigen (Fig. 3 [26]). Sialic acid expression on platelet VWF is significantly Dinaciclib price reduced, by >50% compared to plasma VWF. It is known that expression of terminal ABO blood group and sialic acid determinants on the N-linked glycans of VWF influence susceptibility to ADAMTS13 proteolysis [27, 28]. Recent work in the laboratory of O’Donnell et al. has demonstrated that due to differences in its glycosylation profile, platelet VWF is resistant to learn more proteolysis by ADAMTS13. Interestingly, other functional differences between platelet and plasma VWF have also been described. For example, GpIb binding is reduced five-fold for platelet VWF compared to plasma VWF [29]. In contrast, platelet VWF demonstrates significantly enhanced GpIIbIIIa binding and heparin binding compared to plasma VWF. Crossed bone marrow transplantation

studies in both pigs and mice have shown that platelet VWF is important in regulating normal haemostasis in vivo. In pigs with plasma VWF only, bleeding time was largely but not completely corrected and all pigs developed thrombosis in an arterial injury model. In contrast, selleck compound in pigs with platelet VWF only, the bleeding time was not corrected and none of the

pigs developed thrombosis [30]. A similar study in mice showed that platelet VWF plays a role in platelet adhesion to collagen under shear [31]. Human studies have demonstrated significant heterogeneity in platelet VWF:Ag and VWF:RCo among patients with type I VWD [32]. In addition, in vitro parallel plate flow studies suggest that platelet VWF may be important in regulating the adhesion of human platelets to collagen under shear stress [33]. Notwithstanding these data, the importance of platelet VWF in managing patients with VWD/pathological bleeding disorders remains poorly defined. This section focused on type 1 patients; type 1 is the most common form of the disease. Patients with type 1 VWD do not produce enough VWF, resulting in heavy bleeding during a bad injury or surgery. Although much is known about type 1 VWD, there are still areas where knowledge is lacking. A diagnosis of type 1 VWD is likely in patients with previous bleeding history (at least two bleeds at different sites) and reduced VWF:RCo in plasma (

The VWF monomer is heavily glycosylated with 20% by mass being composed of carbohydrate. The N-linked glycan structures of plasma VWF are complex type chains, with the majority being bi-antennary (78%) or tri-antennary (12%). The O-linked glycan structures of plasma VWF are short mucin-type carbohydrates and sialylated T antigen structures. Plasma VWF expresses terminal ABO(H) group antigens. In contrast, platelet VWF does not express A or B blood group antigens

[25], but it does express precursor H antigen (Fig. 3 [26]). Sialic acid expression on platelet VWF is significantly selleck kinase inhibitor reduced, by >50% compared to plasma VWF. It is known that expression of terminal ABO blood group and sialic acid determinants on the N-linked glycans of VWF influence susceptibility to ADAMTS13 proteolysis [27, 28]. Recent work in the laboratory of O’Donnell et al. has demonstrated that due to differences in its glycosylation profile, platelet VWF is resistant to www.selleckchem.com/products/GDC-0941.html proteolysis by ADAMTS13. Interestingly, other functional differences between platelet and plasma VWF have also been described. For example, GpIb binding is reduced five-fold for platelet VWF compared to plasma VWF [29]. In contrast, platelet VWF demonstrates significantly enhanced GpIIbIIIa binding and heparin binding compared to plasma VWF. Crossed bone marrow transplantation

studies in both pigs and mice have shown that platelet VWF is important in regulating normal haemostasis in vivo. In pigs with plasma VWF only, bleeding time was largely but not completely corrected and all pigs developed thrombosis in an arterial injury model. In contrast, selleck inhibitor in pigs with platelet VWF only, the bleeding time was not corrected and none of the

pigs developed thrombosis [30]. A similar study in mice showed that platelet VWF plays a role in platelet adhesion to collagen under shear [31]. Human studies have demonstrated significant heterogeneity in platelet VWF:Ag and VWF:RCo among patients with type I VWD [32]. In addition, in vitro parallel plate flow studies suggest that platelet VWF may be important in regulating the adhesion of human platelets to collagen under shear stress [33]. Notwithstanding these data, the importance of platelet VWF in managing patients with VWD/pathological bleeding disorders remains poorly defined. This section focused on type 1 patients; type 1 is the most common form of the disease. Patients with type 1 VWD do not produce enough VWF, resulting in heavy bleeding during a bad injury or surgery. Although much is known about type 1 VWD, there are still areas where knowledge is lacking. A diagnosis of type 1 VWD is likely in patients with previous bleeding history (at least two bleeds at different sites) and reduced VWF:RCo in plasma (

Of note, CcnE1−/− livers revealed a normal frequency of resident

HSCs (Supporting Fig. 4A). Primary analysis of HSCs was performed by fluorescence-activated cell-sorting (FACS) analysis of DNA content and immunofluorescence staining of Ki67 and α-SMA serving as markers for cell-cycle activation and myofibroblast differentiation, respectively. As expected, the total number of living WT HSCs increased continuously within the observation period of 10 days, whereas the number of CcnE1−/− HSC PXD101 chemical structure remained constant at low levels (Supporting Fig. 4B,C). In agreement with these findings, WT HSCs revealed the marked occurrence of a 4n cell population after 10 days, indicating continuous cell-cycle progression (G2/M phase; Fig. 6A) with a tendency to form polyploid cells, which is in agreement with earlier observations.12 These cells were characterized by the expression of α-SMA and Ki-67 (Fig. 6A and Supporting Fig. 5A), indicating that they proliferate and transdifferentiate into myofibroblasts. In sharp contrast, CcnE1−/− HSCs did not show 2n/4n conversion throughout the 10-day observation time, demonstrating G1 cell-cycle arrest of these cells. Instead, we observed a large sub-G1 population of apoptotic cells with reduced

DNA content (<2n) resulting from DNA degradation (Fig. 6B) and low total cell numbers throughout the observation period. Thus, quiescent ex vivo isolated CcnE1−/− HSCs have a defect in entering the cell cycle and are prone to excessive cell death. Using CcnE2−/− Daporinad HSCs, completely opposite effects were observed, showing already highly polyploid cells after isolation undergoing a further, time-dependent

increase in DNA synthesis and polyploidization (Fig. 6C). The complete data, including all investigated time points, selleck chemical are shown in Supporting Fig. 6 and demonstrates that in WT cells, Ki-67 expression started at day 4 after seeding, whereas transdifferentiated (i.e., α-SMA-positive) myofibroblasts were first detected after 7 days. After 10 days, the majority of HSCs were activated and reached confluence (Supporting Fig. 5A). CcnE2−/− HSCs showed accelerated transactivation, starting day 3 after seeding, with overall stronger Ki-67 expression pointing at an enhanced cell-cycle activity of these cells (Supporting Fig. 6C,F). mRNA quantification revealed substantial α-SMA induction—and thus transactivation—after 7 days in WT HSCs, but already after 3 days in CcnE2−/− cells (Fig. 6D). Importantly, overall α-SMA expression in CcnE2−/− HSCs significantly exceeded WT levels at all time points investigated. In contrast, overall α-SMA levels in CcnE1−/− HSCs were lower, compared to WT cells, and especially lacked induction after 7 days. These findings suggested that CcnE1 is essential for HSC transactivation. To further test our hypothesis, we measured the expression of platelet-derived growth factor receptor beta (PDGF-Rβ), which is usually induced when HSCs are activated and start to transdifferentiate into myofibroblasts.

GW4064 was provided by the University of Kansas (Kansas City, KS). Other reagents, unless mentioned, were obtained from Sigma-Aldrich (St. Louis, MO). Whole body (WB) Fxr knockout (KO) mice (Fxr WB KO) were reported on and were on a pure C57BL/6J genetic HTS assay background.16, 17 The generation of tissue-specific Fxr KO mice on a mixed genetic background has been described previously using loxP/Cre technology with specific disruption of the Nr1h4 gene in hepatocytes (Fxr Liv KO) or in enterocytes (Fxr Int KO).18 Specifically, Fxr Liv KO and Fxr Int KO mice were generated by cross-breeding Fxr floxed/floxed mice with albumin cre (+) or villin cre (+) mice. But, these mice were on a mixed genetic background with variable

basal expression of bile-acid synthetic genes. So, in the current study, congenic Fxr Liv KO and Fxr Int KO mice in the C57BL/6J genetic background were produced. Shp KO mice and hepatocyte-specific Shp transgenic (Tg) mice (albumin promoter derived, Shp Tg) have been reported on.19, 20 Fxr WB KO mice with hepatocyte-specific Shp overexpression (Fxr WB KO/Shp Tg) were generated by crossing Fxr Maraviroc clinical trial WB KO mice with Shp Tg mice, with all three strains on the pure C57BL/6J genetic background. Fgfr4 KO mice on a mixed C57/129SvJ background were provided by Dr. Curtis Klaassen (University of Kansas Medical Center). Fgfr4/Shp double-KO (Fgfr4/Shp DKO) mice were generated

by cross-breeding Fgfr4 KO and Shp KO mice. Egr1 KO mice on a C57BL/6 genetic background were obtained from Taconic (Hudson, NY). C57BL/6J mice bred in the same animal

facility were used as wild-type (WT) controls for KO mice on the C57BL/6J background. If KO mice were on a mixed genetic background (Fgfr4 KO and Fgfr4/Shp DKO), littermates were used as controls. Mice were bred and maintained in the laboratory animal research facility at the University of Kansas Medical Center in rooms under a 12-hour light-dark cycle. All protocols were approved by the institutional animal care and use committee. All experiments used 10-16-week-old male mice, and all mice were sacrificed within a 30-minute period in the morning. In addition, all treatments were repeated twice. The activation of Fxr in vivo was achieved by treatment with an Fxr synthetic selleck kinase inhibitor agonist (GW4064) at 150 mg/kg. GW4064 or vehicle was administered by oral gavage at 6 p.m., followed by a second administration at 8 a.m. the next morning. Two hours later, the liver and ileum were harvested. The generation of purified Fgf15 has been reported on previously.15 Fgf15 protein was injected into mice through the tail vein at a dosage of 10 μg/kg. Two hours or at indicated time points (for time-course study) after injection, livers were collected. Total bile-acid pool size was determined by measuring bile acids of the small intestine, gallbladder, liver, and their contents. Ten 16-week-old mice were fed a chow diet with 2% cholestyramine for 10 days.