Collagen type I protein in the supernatant of purified/serum HBV group also increased compared to the control group (408.0 ± 8.0/384.4 ± 6.8

vs 262.7 ± 15.7 ng/mL, P < 0.05). However, the 3 × 107 IU/mL purified/serum HBV increased collagen type I expression similar to that of 3 × 105 IU/mL, while 3 × 103 IU/mL group showed no effect. Human HBV immunoglobulin alleviated HBV-induced collagen I expression, but no evidence of HBV infection was found. Neutralization of transforming growth factor beta, tumor necrosis factor alpha, platelet-derived growth factor, extracellular signal-regulated kinase and TGF-β receptor had no obvious inhibitory effects; only inhibition of p38 Wnt activity mitogen-activated protein kinase decreased collagen type I mRNA expression by 0.5-/0.4- and 0.4-/0.3-fold at 24 and 48 h, respectively.

It reduced collagen type I protein in the purified/serum HBV group for 48 h (252.1 ± 14.1/251.7 ± 18.8 vs 403.9 ± 4.9/385.0 ± 4.2 ng/mL, P < 0.05). Conclusion:  HBV directly promotes collagen type I expression of LX-2 cells without infection in vitro. "
“Hepatocellular mTOR inhibitor carcinoma (HCC) is one of the most common neoplasms worldwide. The recent dramatic increase of HCC cases is associated with chronic hepatitis B and C.[1] Worldwide, there are 300 million people infected with hepatitis B virus and 170 million with hepatitis C virus, respectively. Prothrombin, a 72-kDa plasma protein, is synthesized in hepatocytes as a precursor with 10 glutamic acid (Glu) 上海皓元 residues. Under physiological condition, these Glu residues are converted into gamma-carboxy glutamic acid (Gla) by gamma-glutamylcarboxylase, which uses

vitamin K as a cofactor. However, when gamma-glutamylcarboxylation is impaired, Glu residues of prothrombin remain unconverted, and a des-carboxy prothrombin (DCP) is released into the bloodstream as a result. DCP is elevated in many patients with HCC, as well as those with vitamin K deficiency, so that raised plasma DCP can result from the administration of vitamin K antagonists like warfarin or impaired uptake of vitamin K in cholestasis with hyperbilirubinemia. Plasma level lf DCP is significantly elevated in patients with HCC, and is a clinically established HCC marker. For small tumors, measurement of both alpha fetoprotein (AFP) and DCP is recommended, because DCP is more specific for HCC than AFP. DCP is potentially valuable primarily as a prognostic biomarker, which would be predictive of rapid tumor progression and provide information about a possible poor prognosis.[2] This is because DCP-positive HCCs show aggressive and invasive distinctiveness. A high DCP level implies a poor prognosis, and a slight increase in the DCP concentration after therapy could suggest recurrence.[2] Kiriyama et al. demonstrated significantly poorer prognosis in “triple-positive” HCC patients, who were positive for all three available serum HCC markers, AFP, AFP-L3, and DCP.

6). A few factors may contribute to this phenomenon in fatty liver, as described below. Insulin insensitivity learn more in the fatty liver is detrimental to the hormone’s

inhibitory role in gluconeogenesis, primarily through the inactivation of the phosphatidylinositol 3-kinase/serine/threonine kinase–signaling pathway,15 thereby enfeebling the suppression of key gluconeogenic enzymes PEPCK and glucose-6-phosphatase (G-6-Pase) expression.14 In addition, previous studies utilizing radioisotopic analysis also showed that carboxylation of pyruvate into OAA is up-regulated in the diabetic rat liver, concomitant with dramatic increases in PC,16 PEPCK, and G-6-Pase15 expression. These studies corroborate our finding that both PC and PEPCK enzyme activities

are increased selleck chemical in the fatty liver, leading to larger 13C-malate, -aspartate, and -OAA signals as well as higher rates of chemical exchange with pyruvate. Indeed, higher hepatic PC activity correlated with increased PEPCK activity (r2 = 0.82; P < 0.0001) (Supporting Fig. 4), further supporting the hypothesis that both PC and PEPCK are important regulators in gluconeogenesis.7 In diabetes, pathological alteration of the precise balance between insulin and glucagon action results in excessive hepatic gluconeogenesis and glycogenolysis, both of which induce hyperglycemia. Moreover, inadequate suppression of postprandial glucagon secretion by insulin in the diabetic state causes hyperglucagonemia and evokes elevated HGP, as observed in HFD mice. We previously reported that combined defects in insulin secretion and signaling were not sufficient to cause hyperglycemia in the absence of dysregulated glucagon secretion in a mouse model with deletion of calcium-sensing protein synaptotagmin-7.17 Indeed, glucagon plays a major role in promoting gluconeogenesis in enhancing G-6-Pase activity and PEPCK transcription in the liver, likely through the protein kinase A–signaling cascade mechanism.18 Thereafter, up-regulated gluconeogenesis increases the demand for OAA. In this work, we demonstrated up-regulated

PC activity in glucagon-stimulated HGP in Chow-fed animals, as detected 上海皓元 in vivo with hyperpolarized 13C MRS, through the biomarker kpyr->asp. Concomitantly, glucagon increases PDH activity.19 This technology appears to possess sufficient sensitivity to detect this phenomenon as well, as evident from the higher kpyr->bic exchange rate. Treatment with a glucagon-receptor antagonist appears to alleviate HGP in the diabetic liver,20 and reducing glucagon signaling is being explored as a potential therapy for diabetes.21 It will be interesting to measure corresponding changes in hepatic metabolism upon therapeutic intervention with a glucagon-receptor antagonist in diabetic animals, and that forms the next phase of our research.

apiculatum and P. rotundifolius on granitic sands; (5) tree savanna dominated by C. mopane on clayey soils formed from shale and mudstone; (6) riparian woodland on alluvial soils fringing the Mphongolo River. Rainfall recorded at Punda Maria

camp averaged 560 mm per year (1960–2007). Rainfall over the seasonal cycle (July–June) was 33% above 3-MA the long-term mean in 2005/6, and 25% below the mean in 2006/7. In 2006, the first spring rains were delayed until early November, whereas in 2007, the first rain of the wet season was received at the end of September. Surface water availability became restricted to pools in the Mphongolo River by mid-August, apart from artificial sources near the western border fence and the tourist camp in the north. In May 2006, GPS/GSM collars (Africa Wildlife Tracking; http://www.awt.co.za) were placed on three adult females representing the sole sable herd of about 20 animals, four female zebra in separate herds of 5–7

animals and two female buffalo present in a single herd of about 400 individuals, later commonly split into two subgroups. In June 2007, collars were replaced on one of the previously collared sable and buffalo, and placed on female zebra representing two new herds, to extend the study period through September 2007. Animal capture was carried out by South African National Parks Angiogenesis inhibitor staff using immobilizing

drugs injected from a helicopter, following their ethical guidelines. No animal fatalities were recorded. Field observations covered two dry seasons (June–October 2006 and May–September 2007). Habitat use through the wet season (December 2006–April 2007) was provided MCE公司 by the GPS tags. GPS collars recorded herd locations routinely every 6 h, at 8:00 and 20:00 representing foraging times during the day, and at 2:00 and 14:00 representing resting times. To facilitate observations at feeding sites, GPS tags on selected herds were temporarily re-set to provide hourly locations. Places where these animals had been present during the morning (6:00–10:00) and late afternoon (16:00–20:00) foraging periods were visited on 2 days per species each week. Feeding sites were identified from fresh hoof prints and signs of recent grazing, generally found within 5 m of the GPS location. Sites with signs of recent use by other grazers were discarded, but represented less than 1% of sites visited. One to five feeding sites were sampled to represent either the morning or afternoon foraging session. In the area surrounding each feeding site, the habitat features recorded included (1) topographic location as lowland, slope or upland; (2) tree (>2.5 m in height) and shrub (<2.

BDL, bile duct ligation; BSA, bovine serum albumin; cAMP, cyclic adenosine monophosphate; CCl4, carbon tetrachloride; ERK1/2, extracellular signal-regulated Cilomilast mw kinase; FACS, fluorescence-activated cell sorting; IBDM, intrahepatic bile duct mass; MEK, mitogen-activated protein kinase kinase;

PCNA, proliferating cell nuclear antigen; PCR, polymerase chain reation; PKA, protein kinase A; SEM, standard error of the mean; SR, secretin receptor; WT, wild-type. Reagents were purchased from Sigma Chemical Co. (St. Louis, MO) unless otherwise stated. The nuclear dye 4′,6-diamidino-2-phenylindole was obtained from Molecular Probes, Inc. (Eugene, OR). Porcine secretin was purchased from Peninsula Laboratories (Belmont, CA). The polyclonal SR antibody (Santa Cruz Biotechnology, Santa Cruz, CA) was raised against a peptide mapping at the C terminus of SR of human origin and cross-reacts with mouse.20 The antibody

against proliferating cell nuclear antigen (PCNA) was purchased from Santa BMS-907351 supplier Cruz Biotechnology. The mouse anti–cytokeratin-19 antibody was purchased from Caltag Laboratories Inc. (Burlingame, CA). Goat phosphorylated ERK1/2 and total ERK1/2 (44-42 kDa) polyclonal affinity purified antibodies were purchased from Santa Cruz Biotechnology. The RIA kits for the determination of intracellular cAMP levels in cholangiocytes were purchased from Perkin Elmer (Shelton, CT). All animal experiments (Table 1) were performed in accordance with a protocol approved by the Scott & White and Texas A&M Health Science Center Institutional Animal Care and Use Committee and conformed to the Guide for the Care and Use of Laboratory Animals published

by the National Institutes of Health (Publication No. 85-23, revised 1996). Our SR+/+ (wild-type [WT]) or SR knockout (SR−/−)21 mice were maintained in a temperature-controlled environment (20-22°C) with a 12:12-hour light/dark cycle. We used adult male WT and SR−/− mice (approximately 25-30 g) of the N5 generation: (1) as normal treated with saline (0.9% NaCl) or secretin 上海皓元医药股份有限公司 (2.5 nmol/kg/day, a dose similar to that used by us for another gastrointestinal hormone, gastrin, in rodents)18 by way of intraperitoneally implanted Alzet osmotic minipumps (Alzet, CA) for 7 days; or (2) for sham operation or BDL (for 3 and 7 days).5, 20, 22 Because our previous studies21 showed that SR−/− mice have a renal defect in water reabsorption and associated polyuria and polydipsia, experiments were performed to determine whether the response of SR−/− mice to BDL was due to the lack of SR rather than severe dehydration. Thus, we evaluated changes in body weight and mortality rate in the experimental groups of Table 1. In addition, both WT and SR−/− mice (after BDL or administration of secretin) received oral hydration therapy, consisting of up to 1 ml of normal saline subcutaneously up to twice daily along with water in gel form on the ground and food supplements.

Results.— Three months of pretreatment prospective diaries were completed by 126 women. ICHD-II menstrually related migraine was diagnosed in 74%, with pure MM in 7%.

Among those women diagnosed with ICHD-II MM, 61 completed at least 1 treatment month. Overall change in headache activity was a 46% decrease. The mean percentage of treated menses without migraine occurring during the 6 days of treatment was 71%. The percentage of subjects with 1, 2, and 3 migraine-free menstrual periods (no migraines occurring 2 days before menses through the first 3 days of menstruation) with eletriptan, respectively, were 14%, 19%, and 53%. Among those subjects who remained headache-free during the 6 days of eletriptan treatment, migraine Nutlin-3 mouse occurred during the 3 days immediately after discontinuing eletriptan for 9%. Perimenstrual eletriptan was generally tolerated

and no abnormalities were identified on the 6th day of treatment using either blood pressure this website recording or electrocardiogram. Conclusions.— Among patients with prospectively identified MM, eletriptan 20 mg 3 times daily effectively reduced MM. A significant reduction in headache activity occurred for 53% of patients. (Headache 2010;50:551-562) “
“Objective.— To examine the lifetime comorbidity of migraine with different combinations of mood episodes: (1) manic episodes alone; (2) depressive episodes alone; (3) manic and depressive episodes; (4) controls with no lifetime history of mood episodes, as well as sociodemographic and clinical correlates of migraine for each migraine–mood episode

combination. Background.— Migraine has been found to be comorbid with bipolar disorder and major depressive disorder in clinical and population-based samples. However, variability in findings 上海皓元医药股份有限公司 across studies suggests that examining mood episodes separately may be fruitful in determining which of these mood episodes are specifically associated with migraine. Methods.— Using a cross-sectional, population-based sample from the Canadian Community Health Survey 1.2 (n = 36,984), sociodemographic and clinical correlates of migraine were examined in each combination of mood episodes as well as controls. Logistic regression analyses controlling for age, sex, and education level compared the lifetime prevalence of migraine (1) between controls and each combination of mood episodes, and then (2) among the different combinations of mood episodes. Results.— Migraine comorbidity in all combinations of mood episodes was associated with lower socioeconomic status, earlier onset of affective illness, more anxiety, suicidality and use of mental health resources. Compared with controls, the adjusted odds ratio of having migraine was 2.0 (95% confidence interval [CI] 1.4-2.8) for manic episodes alone, 1.9 (95% CI 1.6-2.1) for depressive episodes alone, and 3.0 (95% CI 2.3-3.

Increased BMI in Primary Biliary Cirrhosis (PBC) is associated with more advanced fibrosis but the effect of BMI in PSC is unknown. Aim: To examine the effect of BMI on fibrosis stage and progression in PSC. Methods: 291 PSC patients were recruited from the Calgary PSC cohort and stratified according to initial BMI at presentation. BMI <25 as normal, 25-30 as overweight and >30 kg/m2 as obese. Fibrosis stage were measured at least once every 12 months by transient elastography using Fibroscan® and classified as F0 to F4 fibrosis. We examined the fibrosis stage at presentation

and the time in months of progression to the next fibrosis selleck products stage. Data from 1368 patient years of follow up were assessed. Patients with existing cirrhosis at their first presentation

or less than 1 year of follow up data were excluded. Results: 247 cases were eligible for the study. 176 individuals had a normal body weight (BMI <25), 57 were overweight (BMI 25-30) and 14 obese (BMI >30). Mean times of progression to the next fibrosis stage were 51 months, 47 months and 13 months for normal body weight, overweight and obese PSC patients respectively. In addition, obese PSC patients were associated with a more advanced fibrosis stage at presentation compared to normal or overweight cases. Conclusion: Significant proportions of patients with PSC can be classified as overweight or obese. Obesity (BMI 30 kg/m2) in PSC is associated with significantly more advanced fibrosis at presentation and more rapid fibrosis progression as measured by non-invasive

transient elastography. selleck kinase inhibitor Disclosures: The following people have nothing to disclose: Aliya Gulamhusein, Danielle Reid, Bertus Eksteen BACKGROUND: The pathogenesis of primary sclerosing cholangitis (PSC) is largely unknown due to lack of an ideal animal model. The association of PSC with inflammatory bowel disease suggests a critical role of gut-derived factors in its pathogenesis. Our aim was to investigate the role of small bowel bacterial overgrowth (SBBO) in the development of PSC-like cholangiopathy. medchemexpress METHODS: We surgically created a jejunal self-filling blind loop (SFBL) to induce SBBO in a genetically susceptible mouse strain (NOD.B6Abd3), developed by introgression of a 1-Mb non-MHC insulin-dependent diabetes locus from B6 onto NOD background. Control mice underwent laparotomy (sham). Bacterial 16s rRNA gene sequencing was used to analyze bacterial populations of jejunal lumen content. H/E and Trichrome staining were used to assess hepatic inflammation and fibrosis. Flow cytometry was utilized to assess liver immune cell profiles. Chemokine expression was assessed by ELISA (serum) and by RT-PCR (liver tissue). RESULTS: Creation of SFBL led to dramatic increase in bacterial counts in jejunal lumen content, compared to sham mice.

Thereafter,

effects of ADR with or without PPZ pretreatment were compared by determining the tumor size and weight, apoptotic cells in tumor tissues were detected by TUNEL assay. Results: At concentrations greater than 20 μg/ml, PPZ pretreatment reduced ADR releasing index and significantly enhanced intracellular ADR concentration of SGC7901 (P < 0.01). Similarly, PPZ pretreatment Angiogenesis antagonist significantly decreased ADR releasing index of SGC7901/ADR dose-dependently (P < 0.01). PPZ pretreatment also decreased cell viabilities of SGG7901 and SGC7901/ADR dose-dependently. After 24-h PPZ pretreatment, administration of chemotherapeutic agents demonstrated maximal Cilomilast cytotoxic effects on SGC7901 and SGC7901/ADR cells (P < 0.05). The resistance index in PPZ pretreatment group was significantly lower than that in non-PPZ pretreatment

group (3.71 vs. 14.80). PPZ at concentration >10 μg/ml significantly decreased pHi in SGC7901 and SGC7901/ADR cells and diminished or reversed transmembrane pH gradient (P < 0.05). PPZ pretreatment also significantly inhibited protein expressions of V-ATPases, mTOR, HIF-1α, P-gp, and MRP1, and alter intracellular expressions in parent and ADR-resistant cells (P < 0.05). In vivo experiments further confirmed that PPZ pretreatment could enhance anti-tumor effects of ADR on xenografted tumor of nude mice and also improve the apoptotic index in xenografted tumor tissues. Conclusion: PPZ pretreatment enhances the cytotoxic

effects of anti-tumor drugs on SGC7901 and reverse MDR of SGC7901/ADR by downregulating the V-ATPases/mTOR/HIF-1α/P-gp and MRP1 signaling pathway. Key Word(s): 1. pantoprazole; 2. V-ATPases; 3. multidrug resistance; Presenting MCE Author: YING XING Additional Authors: YING HAN, ZHIHONG WANG Corresponding Author: YING HAN Affiliations: Doctor’s Degree of 2010 session in Medical School of Chinese PLA; General Hospital of Beijing Military Command, Beijing Objective: Recent studies indicated that FTY720, a novel immunosuppressant, could inhibit proliferation and induce apoptosis in many cancer cells. However, the effects and mechanisms of FTY720 on inducing growth inhibition and potentiating cytotoxicity of anticancer drugs in human colon cancer cell lines were little and controversial. Methods: Cell viability and apoptosis after treatment with FTY720 alone or in combination with chemotherapeutic drugs (e.g.

Thereafter,

effects of ADR with or without PPZ pretreatment were compared by determining the tumor size and weight, apoptotic cells in tumor tissues were detected by TUNEL assay. Results: At concentrations greater than 20 μg/ml, PPZ pretreatment reduced ADR releasing index and significantly enhanced intracellular ADR concentration of SGC7901 (P < 0.01). Similarly, PPZ pretreatment GSK2126458 solubility dmso significantly decreased ADR releasing index of SGC7901/ADR dose-dependently (P < 0.01). PPZ pretreatment also decreased cell viabilities of SGG7901 and SGC7901/ADR dose-dependently. After 24-h PPZ pretreatment, administration of chemotherapeutic agents demonstrated maximal http://www.selleckchem.com/products/ly2606368.html cytotoxic effects on SGC7901 and SGC7901/ADR cells (P < 0.05). The resistance index in PPZ pretreatment group was significantly lower than that in non-PPZ pretreatment

group (3.71 vs. 14.80). PPZ at concentration >10 μg/ml significantly decreased pHi in SGC7901 and SGC7901/ADR cells and diminished or reversed transmembrane pH gradient (P < 0.05). PPZ pretreatment also significantly inhibited protein expressions of V-ATPases, mTOR, HIF-1α, P-gp, and MRP1, and alter intracellular expressions in parent and ADR-resistant cells (P < 0.05). In vivo experiments further confirmed that PPZ pretreatment could enhance anti-tumor effects of ADR on xenografted tumor of nude mice and also improve the apoptotic index in xenografted tumor tissues. Conclusion: PPZ pretreatment enhances the cytotoxic

effects of anti-tumor drugs on SGC7901 and reverse MDR of SGC7901/ADR by downregulating the V-ATPases/mTOR/HIF-1α/P-gp and MRP1 signaling pathway. Key Word(s): 1. pantoprazole; 2. V-ATPases; 3. multidrug resistance; Presenting MCE公司 Author: YING XING Additional Authors: YING HAN, ZHIHONG WANG Corresponding Author: YING HAN Affiliations: Doctor’s Degree of 2010 session in Medical School of Chinese PLA; General Hospital of Beijing Military Command, Beijing Objective: Recent studies indicated that FTY720, a novel immunosuppressant, could inhibit proliferation and induce apoptosis in many cancer cells. However, the effects and mechanisms of FTY720 on inducing growth inhibition and potentiating cytotoxicity of anticancer drugs in human colon cancer cell lines were little and controversial. Methods: Cell viability and apoptosis after treatment with FTY720 alone or in combination with chemotherapeutic drugs (e.g.

4 We evaluated, by IF, whether small control vector- or CaMK I short hairpin RNA (shRNA)-transfected cholangiocytes express GABA receptors. Then, we performed studies to demonstrate that GDC-0449 mw (1) GABA increases IP3 levels, mRNA, and/or protein expression for CaMK I and AC8 in small cholangiocytes4 and (2) silencing of CaMK I in small cholangiocytes prevents GABA-induced differentiation of small into large cholangiocytes and AC8 activation. The primers (from SABiosciences) used are described in the Supporting Materials. Knockdown

(∼70%)4 of the CaMK I gene in small cholangiocytes was established by a SureSilencing shRNA (SABiosciences) plasmid for mouse CaMK I, containing the gene for neomycin (geneticin) resistance for selection of transfected cells.4 Control or

CaMK I shRNA-transfected small cholangiocytes were incubated at 37°C with GABA (1 μM) for 3 days before evaluating (1) expression of GABA receptors by IF, (2) PCNA protein expression by immunoblottings, (3) expression of SR, CFTR, and Cl−/HCO3− AE2 by IF in a coded fashion, and (4) basal and secretin-stimulated cAMP levels by RIA.3, 22 Because AC8 regulates the function of large cholangiocytes,9 we proposed to demonstrated that IP3/Ca2+/CaMK high throughput screening compounds I-dependent, GABA-induced differentiation of small into large cholangiocytes are dependent on the presence or activation of AC8. Thus, we studied: (1) biliary expression of AC8 (by IHC) in liver sections and small cholangiocytes from BDL mice treated with saline

or GABA for 1 week and (2) message expression of AC8 by real-time PCR4 in control vector- or CaMK I shRNA-transfected small cholangiocytes treated with 0.2% BSA MCE or GABA (1 μM) for 3 days. We studied the effect of in vitro GABA treatment (1 μM, 3 days) in the absence or presence of preincubation (2 hours) with the AC8 inhibitor, 2′-deoxyadenosine 3′-monophosphate (10 mM),23 on the differentiation of small into large cholangiocytes by measuring the semiquantitative expression of SR, CFTR, and Cl−/HCO3− AE2 by IF. The primers used are shown in the Supporting Materials. Data are expressed as mean ± SEM. Differences between groups were analyzed by the Student unpaired t test when two groups were analyzed and by analysis of variance when more than two groups were analyzed, followed by an appropriate post-hoc test. Mann-Whitney’s U test was used to determine ultrastructural differences between cells treated with BSA or GABA. For SEM, statistical analyses were performed using SPSS statistical software (SPSS, Inc., Chicago, IL). Both small (yellow arrows) and large (red arrows) bile ducts from normal (not shown) and BDL (treated with vehicle or GABA) mice express GABAA, GABAB, and GABAC receptors (Fig. 1A). By real-time PCR and IF (Fig.

4 We evaluated, by IF, whether small control vector- or CaMK I short hairpin RNA (shRNA)-transfected cholangiocytes express GABA receptors. Then, we performed studies to demonstrate that Nivolumab clinical trial (1) GABA increases IP3 levels, mRNA, and/or protein expression for CaMK I and AC8 in small cholangiocytes4 and (2) silencing of CaMK I in small cholangiocytes prevents GABA-induced differentiation of small into large cholangiocytes and AC8 activation. The primers (from SABiosciences) used are described in the Supporting Materials. Knockdown

(∼70%)4 of the CaMK I gene in small cholangiocytes was established by a SureSilencing shRNA (SABiosciences) plasmid for mouse CaMK I, containing the gene for neomycin (geneticin) resistance for selection of transfected cells.4 Control or

CaMK I shRNA-transfected small cholangiocytes were incubated at 37°C with GABA (1 μM) for 3 days before evaluating (1) expression of GABA receptors by IF, (2) PCNA protein expression by immunoblottings, (3) expression of SR, CFTR, and Cl−/HCO3− AE2 by IF in a coded fashion, and (4) basal and secretin-stimulated cAMP levels by RIA.3, 22 Because AC8 regulates the function of large cholangiocytes,9 we proposed to demonstrated that IP3/Ca2+/CaMK VX-770 order I-dependent, GABA-induced differentiation of small into large cholangiocytes are dependent on the presence or activation of AC8. Thus, we studied: (1) biliary expression of AC8 (by IHC) in liver sections and small cholangiocytes from BDL mice treated with saline

or GABA for 1 week and (2) message expression of AC8 by real-time PCR4 in control vector- or CaMK I shRNA-transfected small cholangiocytes treated with 0.2% BSA medchemexpress or GABA (1 μM) for 3 days. We studied the effect of in vitro GABA treatment (1 μM, 3 days) in the absence or presence of preincubation (2 hours) with the AC8 inhibitor, 2′-deoxyadenosine 3′-monophosphate (10 mM),23 on the differentiation of small into large cholangiocytes by measuring the semiquantitative expression of SR, CFTR, and Cl−/HCO3− AE2 by IF. The primers used are shown in the Supporting Materials. Data are expressed as mean ± SEM. Differences between groups were analyzed by the Student unpaired t test when two groups were analyzed and by analysis of variance when more than two groups were analyzed, followed by an appropriate post-hoc test. Mann-Whitney’s U test was used to determine ultrastructural differences between cells treated with BSA or GABA. For SEM, statistical analyses were performed using SPSS statistical software (SPSS, Inc., Chicago, IL). Both small (yellow arrows) and large (red arrows) bile ducts from normal (not shown) and BDL (treated with vehicle or GABA) mice express GABAA, GABAB, and GABAC receptors (Fig. 1A). By real-time PCR and IF (Fig.