01). The expression of Smac protein in the cells increased from 0.097 ± 0.015 to 0.626 ± 0.058 after transfected by si-Livin1 (P < 0.01). The expression of Livin correlated negatively with the expression of Smac in Caco-2 cells (P < 0.05). Conclusion: Livin gene silenced by siRNA induces growth suppression and apoptosis of Caco-2 cells, which could increase the expression of Smac protein in Caco-2 cells. Livin and Smac gene may be the key factors of colorectal carcinoma cell apoptosis signaling pathway. Key Word(s): 1. Livin; 2. Smac; 3. Colorectal carcinoma; 4.

RNA interference; Presenting Author: WEIZHONG YAN Additional Authors: YANQIU LIU, LIHONG JIA, XIANGHUA PIAO, HONGYAN ZHUO Corresponding Author: WEIZHONG YAN Affiliations: Jili center hospital Objective: To study the expression of Ang-1, Ang-2 and receptorTie-2 R788 clinical trial in colorectal cancer tissue, and explore C646 the rela-tionship between the expression of Ang-1,

Ang-2 and receptor Tie-2 with the histological differentiation degree, provide new targets forthe clinical treatment of colorectal cancer1. Methods: The expression of Ang-1, Ang-2 and receptorTie-2 in 64 cases of colorectal cancertissueswere detected with immunohistochemistry SP method, the expression of Ang-1, Ang-2 and receptor Tie-2 mRNA in colorectalcancer tissueswere detected with RT-PCR1. Results: The expression of Ang-1, Ang-2 and receptorTie-2 in colorectal cancer tissuesshowed that the lower the tumor histological differentiation degree, the higher expression of the protein and RNA (P < 0105) 1. Conclusion: The expression degree of Ang-1, Ang-2 and receptor Methane monooxygenase Tie-2 has positive relationwith the progression of colorectal cancer1. Key Word(s): 1. Ang-1; 2. Angiogenesis; 3. Tie-2; 4. Colorectal cancer; Presenting Author: JUN-JI MA Additional Authors: DONG-QIANG ZHAO, JUN-LI SHI, LI-JUAN CHENG, FANG-FANG LI, XIAO-YU JIANG, HUI-QING JIANG Corresponding

Author: HUI-QING JIANG Affiliations: Department of Gastroenterology, The Second Hospital of Hebei Medical University; Department of Gastroenterology, The Second Hospital of Hebei Medical University Objective: Esophageal cancer is a malignant tumor in the world and the common cause of tumor-related death. The development of esophageal cancer is a complex process involving many pathogenic factors, multiple stages, and accumulation of multiple gene mutations and interactions. This study aimed to investigate the effects of Raf kinase inhibitor protein (RKIP) on the proliferation, apoptosis, and invasion of TE-1 cells in esophageal cancer. Methods: The tissues were either fixed in 4% paraformaldehyde solution for hematoxylin-eosin (HE) and immunohistochemical staining. RKIP expression in esophageal tissues was detected by immunohistochemical staining. The esophageal cancer cell line TE-1 was exposed to four different viruses: RKIP-RNAi-AD, NC-RNAi-GFP-AD, RKIP-AD and GFP-AD.

It is an example of a known virus with a pathogenicity that can vary depending on the conditions. It is resistant to solvent/detergent treatment and pasteurization

[84, 85], is only partly removed by nanofiltration [86], and has been shown to be transmissible via blood donations and pdCFCs [80]. The prevalence of B19 parvovirus is higher in patients exposed to pdCFCs than in those exposed to recombinant products or to no replacement factor products at all (Fig. 5). When the independent effect of product exposure on the likelihood of B19 parvovirus positivity was calculated, the odds of positivity of parvovirus B19V were found to be 70% higher among subjects exposed to pdCFCs alone, relative to those unexposed to any type of concentrate [80]. Parvovirus B19 infection typically causes a benign, flu-like illness which occurs most frequently in childhood [80]. It does not normally have any serious implications for the majority of people with haemophilia; BAY 80-6946 price however, cases of aplastic anaemia have been reported in those patients with existing haematological

disease [87], and infection during pregnancy may cause foetal death [80]. Infection in immunocompromised individuals may also lead to severe anaemia and bone marrow alterations [88]. Prions are self-replicating proteins which can cause transmissible spongiform encephalopathies, and several studies have shown that prions can be transmitted through the blood [89]. The most concerning manifestation of transmissible spongiform encephalopathies for humans is vCJD, which is caused by a prion found in lymphoid Alpelisib price tissue and can therefore potentially

contaminate pdCFCs [89]. In the 1990s, vCJD was identified in blood donated within the UK, and subsequently UK-donated plasma ceased to be used to reduce any potential risk of transmission of vCJD [82]). The 2011 European Medicines Agency position statement on CJD and plasma-derived products also states that ‘there is strong evidence that vCJD may be transmitted through transfusion of blood and plasma products’ [90]. Cases of probable transmission Resveratrol of vCJD by blood from donors who subsequently developed the disorder have been reported [89]. In one case, the vCJD prion was detected in the autopsied spleen of a neurologically asymptomatic 73-year-old patient with severe haemophilia A; UK-sourced pdCFCs were determined to be the most likely source of the infection [81]. Emerging pathogens such as B19 parvovirus and vCJD not only threaten the health of recipients of pdCFCs, but also threaten the security of supply of replacement factor concentrates [91], posing the question of whether more complete blood screenings are required. At the same time, it is simply not practical (or even feasible with current technology) to screen for all circulating viruses in the blood; in the case of emerging pathogens, it is obviously impossible to screen for unknown agents.

Key Word(s): 1. Sphincterotomy; 2. Pancreatic stent; 3. Pancreatitis; 4. ERCP; Table 1. Univariate analysis of EPS and EPS and stent group Group Difficult cannulation   P-value EPS, endoscopic sphincterotomy; ERCP, this website endoscopic retrograde cholagio pacreatography; CBD, common bile duct; GB, Gallbladder; SOD, sphincter of oddi dysfunction; PEP, post ERCP pancreatitis Presenting Author: AHMED OURFALI Corresponding Author: AHMED OURFALI Affiliations: Saudia Arabia Objective: With the advent of fibro-optic technology, an Ultra-Slim Endoscope can be introduced into the common bile duct (CBD) peroral, which enables instant visual diagnosis and targeted treatment

of biliary tree pathologies. This novel technique obviates the need for repeated procedures and the cumbersome “mother-baby endoscopic system” of the Endoscopic Retrograde Cholangio-Pancreatography (ERCP).1 Methods: We performed Y 27632 Peroral Direct Cholangioscopy (PDCS) by upper endoscope GIF-XP260 ‘Slim Sight’ (Olympus) after dilatation of the ampulla by Controlled Radial Expansion Balloon Dilatation

(CRE) (Boston Scientific) under fluoroscopy with the patient in prone position and under deep sedation or general anesthesia. Results: Five patients underwent 6 procedures of PDCS for evaluation of post-dilation, suspected CBD stones. In all but one case (3rd patient), the Ultra-Slim Endoscope was successfully introduced into the common bile duct after dilatation, and all stones and sludge were retrieved without residue (F 1,2). In case 5, with the presence of post-dilation stricture, visual confirmation of the absence of any residual stones or tumor up to the confluence of common hepatic duct was established Conclusion: The Peroral Direct Cholangioscopy (PDCS) by Ultra-Slim Endoscope is feasible. It facilitates complete CBD stone removal and excludes any additional pathology. Key Word(s): 1. Cholangioscopy; 2. ERCP; 3. Biliary tracts; Presenting Author: CHONG WANG Additional Authors: PENG YE, GUO-HUA LI, XIAO-JIANG ZHOU,

YOU-XIANG CHEN, NONG-HUA LV Corresponding Author: GUO-HUA LI Affiliations: The First Affiliated Hospital of Nanchang University Objective: The aim was to investigate the efficacy of raw rhubarb soak in prevention of PEP (post-ERCP Resveratrol pancreatitis). Methods: To investigate the difference between raw rhubarb group and control group on the incidence of PEP, 669 cases with ERCP were randomly divided into two groups, raw rhubarb group and control group, from July 2012 to February 2013. In order to exclude the effect of pancreas disease on PEP, all of patients who had suffered pancreatic disease were excluded. The patients who enrolled in raw rhubarb group took 100 ml raw rhubarb soak every 3 hours after ERCP procedure until they were laxatived (50 g raw rhubarb was immersed in 100 ml boiling water for 10 min. This supernate was the soak).

Results: Of the 162 accredited GI fellowship programs, 70 GI PDs completed the survey. Eighty three percent of respondents were from a University-Affiliated Medical Center with 67% having 8 or more GI fellows in their program. The majority of GI PDs (87%) were in favor of fellow participation in the pilot if the fellow was on a trajectory of competence in GI. Concerns about the pilot noted in this section included: competence in endoscopic training, compromised

GI workload of other fellows Tyrosine Kinase Inhibitor Library ic50 in the program, and penalization of fellows training at non-transplant centers. In the area of coverage of non-transplant services, 52% of participants thought there would not be difficulty. Comments in this section focused on coverage issues at smaller programs. On the issue of competency, only 58% of GI PDs believed that graduates of the pilot program would be as competent in GI as those who completed the traditional program. Overall, 65% believed that there would be increased Selleckchem SB203580 interest and participation

in pursing TH fellowship by incorporating the training into the 3-year model and 55% believed the current shortage of transplant hepatologists is likely to improve with implementation of the pilot. Conclusion: The majority of GI PDs embrace competency based fellowship education and sub-specialty training in TH during the designated three-year fellowship. GI PDs concerns about the pilot are mainly about coverage of non-transplant services and endoscopic competency. Future studies

will be needed to re-evaluate GI PDS beliefs after several years of the pilot enrollment. Disclosures: Steven K. Herrine – Grant/Research Support: BMS, Merck, Schering, Vertex The following people have nothing to disclose: Dina Halegoua-De Marzio Background Patients with cirrhosis receive low rates of recommended liver care such as varices surveillance and hepatoma screening, and many are diagnosed too late to benefit from preventive management strategies. Nationally, >50% of cirrhosis patients are followed exclusively by Primary Care Providers (PCPs) as opposed to liver specialists. A growing national dipyridamole shortage of specialists will increasingly shift cirrhosis management towards the primary care setting. We conducted a qualitative analysis to determine PCPs’ attitudes toward patients with cirrhosis, their self-reported roles in caring for them, and perceived barriers to care. Methods We recruited PCPs from 7 Veterans Affairs facilities in the Pacific Northwest during in-service trainings and via direct email from March- October 2012. Trained staff administered 20–30 minute semi-structured telephone interviews covering 4 domains (general attitudes, roles and practices, barriers and supports, and suggestions for enhancing cirrhosis management). We used an Editing analysis approach to thematically code interview responses. Two trained, independent coders reviewed each interview.

Conclusion: Oral tacrolimus is a safe and effective therapy for the treatment of moderate to severe UC, although still more longer follow-up of patients and compilation of further clinical data will be necessary. Key Word(s): 1.

ulcerative colitis; 2. tacrolimus Presenting Author: MANABU SHIRAKI Additional Authors: Fluorouracil concentration TAKAYUKI YAMAMOTO, KOICHI MATSUMOTO Corresponding Author: MANABU SHIRAKI Affiliations: Yokkaichi Hazu Medical Cener, Yokkaichi Hazu Medical Cener Objective: It has been reported that CT can be used for the evaluation of inflammatory bowel disease; nevertheless, there have been few reports on the efficacy of low dose CT for ulcerative colitis. We report here the efficacy of low dose CT for ulcerative colitis.

Methods: The patients with relapsing ulcerative colitis between https://www.selleckchem.com/products/cetuximab.html July 2013 and April 2014 were included in this study. All patients had undergone sigmoidoscopy and low dose CT scan. The colon CT image was divided into six segments, and then we evaluated wall thickening, stratification, contrast enhancement and mesenteric vascular engorgement, assigning a CT score to each segment. We calculated a total CT score by the sum of CT scores of 6 segments. To assess endoscopic severity, Ulcerative Colitis Colonoscopic Index of Severity (UCCIS) was used. The clinical severity was assessed by Mayo partial score. We investigated the correlation between those CT scores and UCCIS. The correlation between partial Mayo score and total CT score also investigated. Results: Twenty three cases of ulcerative colitis were included in this study. We achieved a 57% reduction of effective dose by adjusting the scan conditions and the reconstruction conditions (P = 0.00326). We observed a high degree of correlation between the sum of the CT scores of the rectum and sigmoid colon and the sum of the UCCIS of the rectum and sigmoid colon (ρ = 0.629). Although the UCCIS of the rectum and sigmoid colon segment calculated by sigmoidoscopy and partial Mayo scores correlate (ρ = 0.456, R2 = 0.267), the correlation analysis between the total CT score and the partial Mayo score indicated a higher coefficient of determination

(ρ = 0.643, R2 = 0.315). Conclusion: This study Parvulin suggested that low dose CT could provide more effective images to assess the disease activity of ulcerative colitis less invasively compared with sigmoidoscopy. Key Word(s): 1. low dose CT; 2. sigmoidsocpy; 3. ulcerative colitis Presenting Author: DAIMON SHIROSE Additional Authors: KOJI MATSUDA, DAISUKE SUENAGA, YUICHI KINOSHITA, DAISUKE KUMON, MIKIHITO HAYASHI, KAYO ADACHI, TOSHIYA ISHII, AKIRA SATO Corresponding Author: DAIMON SHIROSE Affiliations: St. Marianna Univ. Yokohama-City Seibu Hopsital, St. Marianna Univ. Yokohama-City Seibu Hopsital, St. Marianna Univ. Yokohama-City Seibu Hopsital, St. Marianna Univ. Yokohama-City Seibu Hopsital, St. Marianna Univ. Yokohama-City Seibu Hopsital, St. Marianna Univ.

(HEPATOLOGY 2010;52:1758-1768) Chronic cholestatic liver diseases such as primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC) are characterized by impaired biliary secretion of bile acids

and other potentially harmful cholephiles. Intrahepatic accumulation of endogenous hydrophobic bile acids, together with cytokine- and chemokine-mediated inflammatory bile ductular and liver parenchymal BMS 354825 injury, may contribute to development of fibrosis and cirrhosis in chronic cholestatic disorders. The ratio of toxic to less toxic bile acids correlates with severity of liver injury.1 The hydrophilic C24 bile acid, ursodeoxycholic acid (UDCA) improves biochemical and histological features in PBC, halts progression to cirrhosis in the majority of patients with PBC, Silmitasertib ic50 normalizes life expectancy in two-thirds of patients with PBC2-5 and currently represents the only approved therapy for PBC.6 In PSC, its therapeutic long-term benefit remains so far unclear although serum liver tests and surrogate markers of long-term prognosis improved during UDCA treatment in pilot trials.6 Mechanisms of action of UDCA in cholestatic disorders have not yet been fully resolved.7 Stimulation of impaired hepatocellular secretion by mainly

posttranscriptional mechanisms, detoxification of bile, antiapoptotic effects in hepatocytes and cholangiocytes, and stimulation of cholangiocyte HCO secretion may contribute to the beneficial effect observed during UDCA treatment in cholestatic disorders.7 NorUDCA is a C23 homolog of UDCA. C23 bile acids are found only in trace amounts in human bile.8 They are poorly conjugated with taurine and glycine in liver, and their pharmacokinetic properties differ from their naturally occurring C24 homologs.8NorUDCA has recently been shown to exert therapeutic effects superior to those of UDCA in

multidrug resistance protein 2 (Mdr2)/ATP-binding cassette b4 (Abcb4) knockout mice, a murine model of nonsuppurative, sclerosing cholangitis.9, 10 The mechanisms of action of norUDCA Selleckchem Ibrutinib in Mdr2−/− mice remain obscure.10NorUDCA is a potent (hyper-)choleretic bile acid as a result of cholehepatic shunting.8, 10 Whether this property may explain, at least in part, the anticholestatic, anti-inflammatory, and antifibrotic effects of norUDCA in Mdr2−/− mice9, 10 still remains unresolved. Most importantly, it is unclear whether norUDCA, like UDCA, may exert anticholestatic effects at the level of the hepatocyte.7, 11 Lithocholic acid conjugates are the most potent cholestatic agents among the major human bile acids.12 Taurolithocholic acid (TLCA)-induced cholestasis is an established experimental model of hepatocellular cholestasis.

In this line, in vitro neutralization of IL-10 in PBMC from HCV-infected patients recovered the activity of low-responsive T-cells.3, 31 Although the mechanism responsible for this recovery is not characterized, restoration of functional properties of DC and concomitantly of T-cells might explain these results. Thus, IL-10 inhibition in HCV infection might enhance T-cell immunity facilitating viral clearance. An important finding obtained using peptide inhibitors of IL-10 is that they not only inhibit IL-10

released in response to HCV proteins, but also IL-10 induced by maturation stimuli. Indeed, activation of mDC with CD40L in the presence of p9 enhanced IL-12 production. Thus, we hypothesized that inhibiting an endogenous brake, like IL-10, synthesized upon CD40L stimulation, may be useful to improve Selleckchem LY294002 the functional properties of DC. This buy Ibrutinib strategy may increase

the immunogenicity of DC, leading to higher efficacy of DC-based vaccination procedures. Using human MoDC (DC population commonly used in vaccination), we found that inhibition of endogenous IL-10 with p13 enhanced their immunogenicity in vitro, increasing lymphocyte proliferation and IFN-γ production, the prototypical Th1 cytokine. Similar results were obtained with murine DC, in agreement with the ability of these peptides to bind and inhibit murine IL-10, which has more than 70% homology with hIL-10. More important, immunization with p13-treated DC in different antigenic models, including mice expressing a secretable version of HCV core as well as transgenic mice expressing the full HCV polyprotein, led to the induction of stronger anti-NS3 T-cell responses, measured as IFN-γ production. Thus, these peptides may have important applications in HCV infection not only in vivo, to inhibit IL-10, thus modulating immune responses, but also ex vivo, in clinical protocols based on the use of DC loaded with HCV antigens for further

administration in therapeutic vaccination. An interesting result regarding the activity of p13 and p9 is their selective effect on their ability ADAMTS5 to restore cytokine production by different DC populations. We do not have a clear explanation, but it might be speculated that these DC populations and their functions have a different sensitivity to be inhibited by IL-10. This might be related to differences in the site of binding to IL-10 by p9 and p13, resulting in specific effects on both types of DC populations. Similarly, binding of the anti-IL-10 antibody to another site on the cytokine may also explain differences in its effect. Finally, because IL-10 plays an immunosuppressive role32 in other diseases (infections by HBV, human immunodeficiency virus [HIV], Epstein-Barr virus [EBV], or cancer), we believe that these peptides might be also useful in these settings.

Acute UGIB is a serious medical problem in cirrhotic patients. In published literature, most reports focus on variceal bleeding while data on acute non-variceal upper GI bleeding in cirrhosis are limited. This has meant that many physicians over the years assume only variceal bleeding in cirrhosis. Moreover, there are very few reports in which the characteristics of variceal and non-variceal bleeding are analyzed together. Despite the fact that variceal bleeding is a life-threatening complication in cirrhosis with consistently high morbidity and mortality, non-variceal bleeding may also decompensate

cirrhotic patients and even may be fatal. Therefore, we conducted this prospective study in our endoscopy center in TUH to assess the magnitude of the problem as well as its different causes among

cirrhotic patients in the region of the middle of Nile Delta. Methods: In the period from March 2013 LDK378 to September 2013, a total of 650 patients underwent emergency upper GI endoscopy for acute UGIB in the endoscopy center in TUH. Out of these patients, 550 (84.6%) patients proved to have cirrhosis, who were the subject of the present study. All patients included in the selleck chemicals llc study were subjected to full history taking, clinical examination, with special emphasis on stigmata of chronic liver disease, and emergency upper gastrointestinal endoscopy after initial assessment and resuscitation in the emergency department searching for the source of bleeding. A lesion was considered the source of bleeding, if there is stigmata of recent hemorrhage or if it’s the only lesion detected in the presence of fresh or altered blood in the upper GI tract. After identification of the bleeding lesion, the appropriate endoscopic hemostatic procedure was done to control bleeding whenever indicated. Endoscopic hemostasis was obtained by injection, thermal and mechanical

methods or combination of these modalities. The outcome of these modalities was not included in the present analysis. Different endoscopic findings were recorded & ratio of non-variceal in relation to the total number of cases was calculated. Results: Our results showed that UGIB in cirrhotic patients was much more common in males and patients from rural Unoprostone areas. Bleeding varices were detected in 75.5% while non-variceal sources of bleeding were detected in 24.5% of the patients. Regarding age, the bleeding variceal group was younger than the bleeding non-variceal group & the difference was statistically significant. Bleeding variceal group was more commonly presented with hemodynamic instability than the bleeding non-variceal group. 22% of the studied cirrhotic patients had negative viral markers while 78% had positive viral markers. 99.1% of patients with positive viral markers were HCV positive, (0.2%) were HBV positive and (0.7%) had mixed viral etiology. Within bleeding variceal patients, bleeding esophageal varices were predominant (90.

The first is on Mechanisms of Gastrointestinal and Liver Diseases, the second is about Advances in Clinical Practice. The first review articles in these new series are both about irritable bowel syndrome. That in the Mechanisms of Disease series written by Ghoshal and colleagues canvasses the potential roles of gut infections and microbiotica,2 whereas that by Gibson and Shepherd discusses the Advances in Clinical Practice afforded by consideration of food (specifically FODMAP) sensitivity.3 Hereafter, one article from each series will appear in most issues of the Journal. While the Editors take responsibility for developing suitable

topics and inviting authors to write these reviews, we would be interested to hear from you, our readers, on your ideas for topics that should be covered in the Asia–Pacific

region. In addition to making the Journal more efficient, more readable and more effective as a vehicle for promoting Asia–Pacific science BIBW2992 research buy and the practice of gastroenterology and hepatology, we are also making it more accessible. Specifically, most subscriptions to JGH are now electronic ones, submission and review are entirely electronic processes, and we have recently taken the decision to increase the content available for download free of charge. This now includes all editorial content (Table of Contents, Editorials, In this Issue, Images of Interest and Education), and all review articles, including meta-analyses, miniseries reviews and the selleck inhibitor new series mentioned earlier. From this issue, it will also include a selected number

(four to six per issue) of what we perceive to be our most exciting original articles, corresponding to those selected for comment in In This Issue. The occasion of our 25th Anniversary in December 2010 will also be marked by a Silver Jubilee supplement; this will accompany the first issue of 2011 (Volume 26 : 1). We have invited 20 or so of our most successful authors of the last 25 years, whose articles rank at the top of our most cited ever, to write thought-provoking reviews of past, present and future developments in their field. We anticipate that this will allow us to compile an incredibly stimulating and readable Supplement. With all these improvements and exciting developments, we hope not to be two, well before we turn Casein kinase 1 30! But only you, the authors, can determine how much our reputation, utility and impact factor can improve by sending us some of your best articles to publish in JGH. Working with a panel of editors who are regional leaders in their countries and fields, together with our expanded panel of approximately100 Editorial Board Members, we hope to promote the further growth and development of our great twin specialities in this important Asia–Pacific part of the world. “
“We read with great interest the article by Tanaka et al.1 showing high serum levels of tauro-β-muricholic and taurocholic acid in two animal models of nonalcoholic steatohepatitis (NASH).

The

study was a prospective investigation of the changes in immunoregulatory markers in the blood, bone marrow, and liver allograft in recipients converted from TAC monotherapy to SRL monotherapy for clinical indications (e.g., TAC toxicity). Inclusion criteria were as follows: age ≥18 years; ≥6 months post-LT; TAC monotherapy ≥1 month before SRL monotherapy conversion for nephrotoxicity (glomular filtration rate [GFR] 30-60 cc/min by modified diet in renal disease [MDRD]) or other indication; ≥6 months without a rejection episode; Tanespimycin mouse no lymphocyte depletion therapy for ≥1 year; normal liver-function tests; and no rejection or fibrosis on preconversion liver biopsy. Exclusion criteria were as follows: previous liver or multiorgan transplant; previous immune or viral liver disease unless hepatitis C virus (HCV) RNA was undetectable; proteinuria (≥0.5 g/day); estimated glomerular filtration rate (eGFR) ≤30 cc/min; ≥2 rejections

post-LT; history of hepatic artery thrombosis; hematological abnormalities or severe hypertriglyceridemia; active infection or malignancy; and inadequacy for follow-up. All patients signed informed consent and were followed for 7 months after SRL conversion. The protocol conformed to the Declaration of Helsinki guidelines and was approved by the Northwestern Institutional this website Review Board (Northwestern University Feinberg School of Medicine, Chicago, IL). History and physical exams, complete blood counts, comprehensive metabolic panels, fasting lipids, hemoglobin A1C tests (HgA1C), and spot urine protein:creatinine ratios were performed before and 3 and 6 months after conversion. Bone marrow aspirations and percutaneous liver biopsies were performed once before and 6 months after conversion. For conversion, SRL at 2 or 3 mg (< or ≥100 kg body weight) daily was initiated with subsequent weekly SRL trough-level monitoring. When these reached ≥5 ng/mL, TAC was discontinued followed by weekly laboratory tests and SRL trough levels (goal, 5-8 ng/mL) for 1 month,

then monthly. Prospective liver- and renal-function tests, lipid levels, urine protein:creatinine Smoothened ratios, and any new SRL toxicities were recorded. Treg immunophenotyping (twice before conversion and 3, 4, 6, and 7 months after conversion): Peripheral blood mononuclear cells (PBMCs) were isolated from heparinized samples on Ficoll-Hypaque gradients. Tregs were enumerated utilizing extracellular immunofluorescent staining with CD3-FITC (fluorescein isothiocyanate), CD4-PerCP (peridinin-chlorophyll protein complex), CD8-PerCP, CD25-APC, and CD127-FITC (BD Biosciences, San Diego, CA). After fixation and permeabilization, the cells were washed and incubated with anti-human FOXP3-PE (phycoerythrin) or rat immunoglobulin G2a-PE isotype control (eBioscience, San Diego, CA) (21, 22).