These cases highlight the complexity of using phylogenetic analysis to determine the direction or mode of transmission in individual situations when events occurred at unknown times in the past. Among the 12 couples that had concordant (or indeterminant) HCV genotypes or serotypes, 50% were HCV RNA–negative. This rate of spontaneous clearance is similar to that observed among MAPK Inhibitor Library price subjects infected at younger (<30 years) ages (by transfusion of whole blood, receipt of contaminated Rh immune globulin, IDU, or accidental needlestick injuries), and prospectively followed for 20 years.24-26 Although a younger age at infection might explain the high proportion of

anti–HCV-positive, HCV RNA–negative partners in our study, one might speculate that repeated exposures to small “doses” of HCV resulted in an immunization-like effect or facilitated viral clearance once infection occurred. We acknowledge that we have not

genetically proven transmission among the phylogenetically linked partners, but rather have presented strong evidence for such a transmission. The method we used is much more effective for excluding possible transmission than it is for confirming it. The consensus sequence of the virus is heavily dominated by a handful of dominant quasispecies, and it drifts relatively slowly. If the genetic distance is not significantly more similar between the pairs than to the rest of the population, then there is no realistic chance the dominant strains selleck compound came from the same source. Proving (or providing strong evidence for) infection with HCV from a common source is difficult for several reasons. First, HCV passes a bottleneck upon infection (it has been IMP dehydrogenase estimated that only a dozen to <100 infectious particles initiate an infection, and these may not be randomly sampled from the donor quasispecies). Therefore, it is possible even with deep sequencing that finding identical quasispecies

variants shortly after infection may not be possible. Second, HCV rapidly adapts to a new host over the first 1-2 months of infection, leading to a burst of diversity and genetic drift. During the rapid expansion in a new host, there is little constraining adaptive immunity, and consequently novel variants are not selected out as rapidly as in an established infection, and immune escape variants that were selected in the donor often revert to a more-fit sequence. Third, HCV’s mutation rate is far higher than its fixation rate (i.e., the number apparent from population sequencing as we did). Therefore, at a quasispecies level, the viral sequence is essentially “shimmering” from the combined effects of random mutation and its opponent, negative selection.

Conclusions: These results demonstrate a role for Pol in HBV-mediated antagonization of IFN-α signaling and provide a possible molecular mechanism

by which HBV resists the IFN therapy and maintains its persistence. (HEPATOLOGY 2013;) Chronic hepatitis B (CHB) caused by hepatitis B virus (HBV) is a serious health problem worldwide. The mechanism by which chronic infection is established and maintained is unknown but is thought to be due, in part, to a suppressed host immune response. One key component RGFP966 manufacturer of the host antiviral responses is the interferon (IFN) system. Viral infection of the host initiates Sunitinib nmr the synthesis of

type I IFNs, which consist predominantly of IFN-α and IFN-β (IFN-α/β). By binding to type I IFN receptors, IFN-α/β triggers the oligomerization and tyrosine phosphorylation of the two tyrosine kinases of the Janus family, Janus kinase 1 and tyrosine kinase 2, which in turn phosphorylate a single tyrosine residue of signal transducer and activator of transcription (STAT) 1 and 2. The activated STAT1/2 heterodimerize with interferon regulatory factor 9 (IRF9) to form the ISGF3 transcription factor complexes and then translocate into the nucleus, where they bind to the interferon-stimulated response element (ISRE) in the promoter of interferon-stimulated genes (ISGs) to initiate transcription of ISGs.1 IFN-α has been shown to inhibit HBV replication in a variety of systems. However, about 70% of CHB patients respond poorly to exogenous IFN-α treatment.2, 3 Increasing evidence suggests that HBV has developed strategies to counteract the type I IFN system, which may contribute to the ineffectiveness of IFN-α therapy.4, 5 We have shown that HBV polymerase (Pol) is able to inhibit IFN-α–induced MyD88 Adenylyl cyclase induction

and nuclear translocation of STAT1.6 Consistently, it could be demonstrated in a chimeric mice model that HBV infection reduced IFN-α–mediated ISG production and STAT1 translocation.7 However, it is still unknown whether the translocation of STAT1 is impaired in chronic hepatitis B patients as well, and the molecular mechanism by which Pol interferes with the IFN responses remains unclear. In this study, we used cell and mouse models to gain a detailed understanding of how HBV and Pol interferes with IFN-α–induced STAT activation. Furthermore, liver biopsies of CHB patients were used to obtain more information on the blockage of IFN-α–induced STAT nuclear translocation by HBV.

Functionally, ZNF545 inhibited GC cell proliferation and induced apoptosis. Interestingly, learn more in GC cells, ZNF545 was shown to be localized at the nucleolus and to act as a suppressor of rRNA transcription, by binding directly to the rDNA promoter and recruiting the co-repressor HP1β, and by diminishing the level of H3Lys4 trimethylation (H3K4me3), a promoter-specific histone modification associated with active transcription [39, 41]. Concerning BCL6B, its tumor-suppressor effects on GC cell inhibition

and induction of apoptosis were attributed to upregulation of the proapoptosis genes TNF1RSF1A and CDKN1A, and of the active forms of caspases-3, -7, -8, -9, and PARP, to downregulation of the anti-apoptotic genes S100A4 and VEGFA, and to the induction of tumor-suppressor proteins ATM and p53 [40]. Clinically, promoter methylation Opaganib price of PAX5, ZNF545, BCL6B, and ADAMTS9 was shown to be associated with poor GC patient survival, and the authors of these publications suggest that methylation of these genes may serve as biomarkers for the prognosis of these patients [36, 37, 39, 40]. Data on genetic and epigenetic alteration patterns

that are characteristic of GC are becoming widely available and will certainly constitute an opportunity to improve the clinical management of GC patients. Besides contributing to an increased understanding of the disease, these data may lead to the identification of clinically relevant biomarkers and new therapeutic targets. New biomarkers, in particular, are expected to impact the management of this disease. Assessed both at the time of diagnosis and continuously over disease progression, together with new Racecadotril testing approaches, biomarkers may provide clinically relevant information for early diagnosis, definition of prognosis, therapeutic selection and for identifying

the acquisition of therapy resistance mechanisms. Competing interests: the authors have no competing interests. “
“Helicobacter pylori (H. pylori) infection stimulates the production of proinflammatory cytokines associated with the development of atherosclerosis. Levels of circulating interleukin-18 (IL-18) have been positively correlated with carotid intima-media thickness (IMT) and coronary plaque area and have identified IL-18 levels as important predictors of coronary events and cardiovascular mortality. This study aimed to examine the relationship between serum IL-18 and H. pylori-IgG antibody as a sign of H. pylori infection in patients with carotid atherosclerosis. The carotid IMT, traditional atherosclerotic risk factors, levels of serum H. pylori-IgG and IL-18 were measured in 573 health checkup examinees. Serum IL-18 and H. pylori-IgG levels were significantly increased in subjects with increased IMT in comparison with those with normal IMT. In subjects with increased IMT, serum H. pylori-IgG was positively correlated with serum IL-18 (r = .402, p = .

19 Given its implication as a tumor suppressor in different human cancers, we analyzed the role of mig-6 in human liver cancer cell lines. Importantly, the EGFR and its ligands have been described to be frequently expressed in human liver cancer, thereby contributing to liver tumor development.25–27 In this study, we show that mig-6 is efficiently induced upon EGF stimulation and acts as an endogenous inhibitor of EGFR activity in human liver cancer cell lines. Mig-6 is able to bind to the activated EGFR, thereby most likely regulating receptor activation and stability. Venetoclax manufacturer However, it is important to note that mig-6 could not be induced in primary

hepatocytes upon EGF stimulation (Fig. 1B). This may be because mig-6 levels are already relatively high in unstimulated cells, which may be caused by the activation process that hepatocytes undergo during isolation and culture. Nevertheless, we were able to show that loss of mig-6 in primary hepatocytes leads to increased activation of EGFR signaling (Fig. 1B), suggesting that mig-6 contributes to EGFR regulation. Unexpectedly, we could show that mig-6 is a negative regulator of EGF-induced cell migration in HepG2 cells. Suppression of mig-6 by a specific siRNA led to a marked increase in EGFR-AKT signaling. As a consequence,

mig-6 knockdown cells display increased cell migration toward EGF. This observation was surprising, U0126 because mig-6 was primarily implicated Buspirone HCl in the suppression of EGF-induced cell

proliferation rather than migration. A previous study, however, showed that mig-6 is a negative regulator of HGF/MET-induced cell migration in neurons and especially in cells of hepatic origin,13 suggesting that mig-6 might be a negative regulator of growth factor–induced cell migration in liver cells. In primary HCCs, mig-6 was found to be down-regulated in a significant number of cases and that correlates with increased EGFR expression. These data suggest that loss of mig-6 in primary human liver tumors might be sufficient to generate increased EGFR signaling, which may lead to tumor formation and progression. Interestingly, mig-6 knockout mice are susceptible to Di-ethyl nitrosamine–induced liver tumor formation, further suggesting that mig-6 is a suppressor of hepatocarcinogenesis (data not shown). It will be the aim of future studies to investigate the exact role and the regulation of mig-6 in HCCs and whether it can serve as a possible marker for EGFR-dependent liver carcinogenesis. In conclusion, we have demonstrated that mig-6 is a negative regulator of EGFR signaling in mouse hepatocytes and have identified mig-6 as a suppressor of EGFR signaling in human liver cancer cell lines. We thank Rüdiger Klein and Sonia Paixao from the Max-Planck Institute of Neurobiology, Martinsried, for providing reagents and for their generous help with hepatocyte isolation. Additional Supporting Information may be found in the online version of this article.

, Debio Pharm, Synageva, Gilead Pharm., Ironwood Pharma, Alnylam Pharm, Tokai Pharmaceuticals, Bristol Myers Squibb, Takeda Pharmaceuticals, Nimbus Discovery, Bristol Myers Squibb, Intermune, Astra Zen-eca, Abbvie, Intermune; Grant/Research Support: Galectin Therapeutics, Tobira Pharm, Vaccinex Therapeutics, Tobira; Stock Shareholder: Angion Biomedica Thomas D. Schiano

– Advisory Committees or Review Panels: vertex, salix, merck, gilead, pfizer; Grant/Research Support: massbiologics, itherx Douglas Dieterich – Advisory Committees or Review Panels: merck, Idenix, Jans-sen ; Consulting: Gilead, BMS Andrea D. Branch – Grant/Research Support: Kadmon, Gilead, Janssen The following people have nothing to disclose: Rachana Yalamanchili, Alicia Sti-vala, Donna Fanelli, Donald Gardenier, Badr Aljarallah, David Sachs, selleck kinase inhibitor Everolimus research buy Michael Linderman, Meena B. Bansal, Priya Grewal, Ritu Agarwal, Gene Y. Im, Lawrence Liu, Nancy Bach, David C. Perlman, Jonathan Yeh, Ponni Perumalswami Background: The combination of SOF+PEG/RBV could become

a treatment option for treatment-experienced (TE) GT1 patients who failed prior treatment with 3- and 4- drug regimens. In this study we evaluated the impact of preexisting resistant-associated variants (RAVs) on treatment outcome and emergence of RAVs at relapse in patients retreated with SOF+PEG/RBV for 12 weeks. Methods: SOF+PEG/RBV was administered for 12 weeks to TE patients chronically infected with genotype 1 HCV who had previously failed prior regimens containing PEG/ RBV and the protease inhibitors GS-9451 or GS-9256 with or without the investigational direct-acting antivirals (DAAs) ledip-asvir and/or tegobuvir. NS3, NS5A and NS5B deep sequencing analysis (cut-off of 1%) was performed for all patients at baseline of the retreatment study

as well as for all patients who did not achieve SVR12. Data was compared to available historical sequencing data from patient’s initial treatment regimens to determine the prevalence and kinetics of RAV burden prior to retreatment with SOF. Results: Overall 37/50 (74%) patients analyzed Dynein in this study achieved SVR12. Patients began retreatment with high RAV burdens with 44/50 patients having one or more class of RAV present, and 22 and 4 patients starting retreatment with 2 or 3 class resistance, respectively. NS3 and NS5A RAVs were highly prevalent with 24 and 38, patients, respectively, having detectable RAVs. Additionally, 20 patients had the Q80K polymorphism, and 6 patients had detectable RBV RAVs at baseline of retreatment. 19/24 (79%) patients with NS3 RAVs and 31/38 (82%) patients with baseline NS5A RAVs achieved SVR12. 12 patients were observed to have NS5B RAVs at baseline, with 10 (83%) achieving SVR12.

6 Thus, 6-TGN concentration is used as a measure of optimal efficacy (greater than 235 pmol/8 × 108 red cells) and of risk of hematological toxicity and (possibly) nodular regenerative hyperplasia of the liver (>450 pmol/8 × 108 red cells) by identifying those who are under- and overdosed. The second commonly-measured metabolite, 6-methyl

mercaptopurine (6-MMP), has been implicated in cases of hepatic toxicity (>5700 pmol/8 × 108 red cells) and therefore is used as a measure of the risk of adverse hepatic reactions.7,8 Low concentrations of both metabolites also provide evidence for poor compliance. Furthermore, the ratio of 6-MMP to 6-TGN is used to identify ‘shunters’ (ratio > 11), where there is preferential metabolism to

potentially toxic 6-MMP mTOR inhibitor away from the therapeutic 6-TGN.7 This finding has gained new significance in that allopurinol is capable of reversing this metabolic shunt, leading to therapeutic 6-TGN concentrations with efficacy in the disease and without hepatic toxicity.9 With such a story, it is difficult to see why such tests are not more readily available and utilized routinely. However, the routine use of thiopurines metabolite testing has remained controversial for three reasons. First, the quoted therapeutic range has had limited validation. It is based on retrospective analyses of clinical experience. There are methodological difficulties Torin 1 in prospectively validating the therapeutic range, including the delay between dosing and efficacy, the fluctuating course of IBD and the fact that only approximately one half of patients will respond to optimal therapy. There are reports of enhanced efficacy of azathioprine when dosage is increased in response to ‘sub-therapeutic’

6-TGN concentrations in patients not in remission, but again such studies have used retrospective data.7,10 Second, weight-based estimates of dosing in conjunction with regular tests for hematological and hepatic toxicity have been used successfully for many years. The use of surrogate markers of therapeutic dosage, such as a rise in mean corpuscular volume11 and reduced total lymphocyte count, has assisted clinicians by reassuring them that the thiopurines dose is adequate. Unfortunately, the basis for the value of such surrogate markers is limited and there Celecoxib are a number of clinical situations where such an approach might be suboptimal. For example, using this approach in a patient who is not in remission has the disadvantage of having the dose limited by the patient’s weight (no more than 1.5 mg/kg/day for 6-mercaptopurine or 3 mg/kg/day for azathioprine). Clinicians are often timid in pushing the dose of thiopurines on the basis of the patient’s weight, as retrospective and prospective studies of clinical practice have shown,12,13 and weight-based dosage correlates poorly with 6-TGN concentrations.

Although the precise etiological mechanism of DIAIH has not been elucidated yet, we can speculate that the variations in their developing patterns are due to the different metabolic activity and immunological reactions. We think that a wider range of drugs has the potential to cause AIH, and incidence of AIH with a drug-related selleck inhibitor etiology is more frequent than we have previously thought. In cases of DILI, careful follow-up will be needed, keeping in mind that AIH can develop even after normalization of

liver enzymes. Furthermore, establishment of the diagnostic criteria and therapeutic strategy for DIAIH will be needed. Kazushi Sugimoto M.D., Ph.D.*, Takeshi Ito M.D., Ph.D.*, Norihiko Yamamoto M.D., Ph.D.*, Katsuya

Shiraki M.D., Ph.D.*, * Department of Gastroenterology and Hepatology, Mie University School of Medicine, Mie, Japan. “
“Liver fibrogenesis is associated with the transition of quiescent hepatocytes and check details hepatic stellate cells (HSCs) into the cell cycle. Exit from quiescence is controlled by E-type cyclins (cyclin E1 [CcnE1] and cyclin E2 [CcnE2]). Thus, the aim of the current study was to investigate the contribution of E-type cyclins for liver fibrosis in man and mice. Expression of CcnE1, but not of its homolog, CcnE2, was induced in fibrotic and cirrhotic livers from human patients with different etiologies and in murine wild-type (WT) livers after periodical administration of the profibrotic toxin, CCl4. To further evaluate the potential function of E-type cyclins for liver fibrogenesis, we repetitively treated constitutive OSBPL9 CcnE1−/− and CcnE2−/− knock-out mice with CCl4 to induce liver fibrosis. Interestingly, CcnE1−/− mice were protected against CCl4-mediated liver

fibrogenesis, as evidenced by reduced collagen type I α1 expression and the lack of septum formation. In contrast, CcnE2−/− mice showed accelerated fibrogenesis after CCl4 treatment. We isolated primary HSCs from WT, CcnE1−/−, and CcnE2−/− mice and analyzed their activation, proliferation, and survival in vitro. CcnE1 expression in WT HSCs was maximal when they started to proliferate, but decreased after the cells transdifferentiated into myofibroblasts. CcnE1−/− HSCs showed dramatically impaired survival, cell-cycle arrest, and strongly reduced expression of alpha smooth muscle actin, indicating deficient HSC activation. In contrast, CcnE2-deficient HSCs expressed an elevated level of CcnE1 and showed enhanced cell-cycle activity and proliferation, compared to WT cells. Conclusions: CcnE1 and CcnE2 have antagonistic roles in liver fibrosis. CcnE1 is indispensable for the activation, proliferation, and survival of HSCs and thus promotes the synthesis of extracellular matrix and liver fibrogenesis.

The software ignored partial nuclei at the borders of each image. Cy3 staining was used to identify further the nuclei of cell lineages of interest. Dabrafenib Using the ScanR analysis software, a mask was incorporated that defined Cy3-positive cells as those where cytoplasmic staining overlapped −5 pixels inside the DAPI-stained nucleus and extended +15 pixels outside the

nucleus. Data were displayed in histograms, so that cells were gated on the basis of intensity of Cy3 staining, preventing incorrect inclusion of Cy3-negative cells adjacent to Cy3- positive cells. This gate was maintained for each cell lineage. To identify telomeres in Cy3-positive cell nuclei, Cy5 Z maximum fluorescence intensity

was used with application of threshold object recognition and a watershed algorithm to outline individual telomeres, enabling maximum separation. These were depicted in histograms. Data obtained Gamma-secretase inhibitor for Cy3-positive nuclei and the subobject of individual telomeres in each nucleus were exported into Microsoft Excel spreadsheets. Telomere area was measured as the mean telomere area in pixels2 per nucleus. Nuclear area and nuclear density for each cell line were measured by the mean DAPI staining pixels2 for each nucleus and mean DAPI Z maximum fluorescence intensity, respectively. Statistical analysis was performed on GRAPH PAD Prism5 software (Graph Pad, San Diego, CA) and SPSS using a linear regression test as data demonstrated a normal distribution (Supporting Fig. 2). A P value of ≤0.05 was considered significant. Analysis of variance and Dunn’s multiple comparison tests were used to compare cell lineages. Five hyperoxalosis liver explants and sixteen age-matched time-zero livers were compared to determine whether organ acquisition, storage, or cold/warm ischemia influenced telomere length. Explanted hyperoxalosis livers were chosen because they had normal histology and PRKD3 were processed immediately with the shortest possible

ischemia time. There was no significant difference in telomere length or telomere number measured by Q-FISH for any cell lineage (Fig. 2). There was no evidence that cold or warm ischemia influenced telomere length. Q-FISH was validated against real-time PCR (the gold standard measure of telomere length) (Fig. 3). Fresh tissue from eight patients with liver disease obtained at liver resection or transplantation was analyzed in tandem using Q-FISH and real-time PCR. Cy5 staining of telomeres using Q-FISH (Fig. 1) provided data on telomere number, area, and intensity. Telomere intensity for all nuclei in liver tissue (not separated by cell lineage) correlated most closely with telomere length in liver homogenate measured by real-time PCR (R2 = 0.659, P = 0.015). Mean Cy5 intensity was therefore used in subsequent experiments as a measure of telomere length.

[48] Less data are available for the utility of serum CEA in CCA diagnosis. The Pittsburgh group reported that serum CEA level of > 5.2 ng/mL had a sensitivity of 68% and specificity of 82%.[44] One study showed that in patients with CCA, the biliary CEA level was about five times that of patients with benign strictures.[50] A combined index of serum CA 19-9 and CEA (CA 19-9 + [CEA × 40]) has been reported to correctly identified

10 of 15 patients with CCA, including 6 of 11 with radiographically occult disease; and without false positive.[46] This corresponded to an accuracy of 86% for CCA detection. A subsequent study however suggested that this score was no better than CA 19-9 alone in predicting the presence of CCA.[51] Afatinib price 6. Cholangiography with tissue acquisition has been the traditional technique to diagnose HCCA. Cholangioscopy may be performed to increase the diagnostic yield. Level of agreement: a—80%, b—20%, c—0%, d—0%, e—0% Quality of evidence: II-2 Classification of recommendation: B Endoscopic tissue acquisition during ERCP can be performed either via papilla under fluoroscopic guidance or via mother–baby cholangioscopy. In general, the diagnostic

sensitivity of transpapillary biopsy under fluoroscopic guidance for CCA ranges from 40% to 60%.[40, 52-55] Transpapillary biopsy enables the collection of a relatively Idelalisib manufacturer large amount of tissue because of the use of standard biopsy forceps. In contrast, mother–baby or spyglass or percutaneous cholangioscopy-assisted targeted

biopsy cannot always collect a sufficient amount of tissue because of the use of forceps with small jaws. In fact, the diagnostic sensitivity of cholangioscopy-targeted biopsy alone was previously reported to be suboptimal (49%).[56] However, the advantage of cholangioscopy is that it may additionally provide a cholangioscopic impression to better clarify some indeterminate cholagiographies. The pool data demonstrated that mother–baby cholangioscopy plus targeted biopsy can improve the sensitivity to detect biliary malignancy to 89–100% with the specificity of 87–96%.[57-59] 7. Cholangioscopy with image enhancement systems and Celecoxib possible targeted biopsy or probe-based confocal laser endomicroscopy (pCLE) may improve the accuracy of HCCA diagnosis. Level of agreement: a—73%, b—21%, c—6%, d—0%, e—0% Quality of evidence: III Classification of recommendation: C Although peroral cholangioscopy such as conventional mother–baby scope and spyglass system can be useful for detecting CCA, the images’ quality is still suboptimal because of the limitation in resolution of the fiber-optic choledochoscope. With the advent in video endoscope and the image enhancement technology such as narrow band imaging, the vascular pattern of neoplasm in the bile duct can be better characterized by a new video choledochoscope.

12 for all except TIPSS; for MELD ≥15, P > 0.13 for all except ascites). The donor risk index (DRI) provides a quantitative assessment of the risk of donor liver graft failure. Calculation of the DRI provides an objective measure of the quality of organs accepted by transplant centers for deceased donor liver transplantation. We compared donor risk index (DRI)9 for DDLT recipients enrolled in A2ALL and DDLT recipients from the same centers but not enrolled in A2ALL. Median DRI for non-HCC DDLT recipients with MELD <15 at listing enrolled in A2ALL was 1.35

and was 1.40 for 1458 DDLT recipients not enrolled in A2ALL with MELD <15 at listing who were transplanted at the nine participating centers (P = 0.94). For non-HCC DDLT

recipients with MELD ≥15 at listing, the median DRI was 1.33 for A2ALL patients and 1.34 for 2999 non-A2ALL-enrolled DDLT recipients (P = 0.45). Finally, we compared post-DDLT CYC202 manufacturer mortality for non-HCC DDLT recipients. For non-HCC patients with MELD <15 at listing, post-DDLT mortality HR was 0.79 (P = 0.23) for A2ALL patients compared with non-A2ALL-enrolled patients. For non-HCC patients with MELD ≥15 at listing, post-DDLT mortality HR was 1.00 (P = 0.98) for A2ALL patients compared to non-A2ALL-enrolled patients. These analyses were adjusted for recipient age, MELD at transplant, and DRI. One hundred thirty of 868 (15.0%) of the A2ALL transplant candidates carried a diagnosis of HCC at the time of enrollment. Of these, 93 had a laboratory (nonexception) MELD <15 at almost study entry and 37 had MELD ≥15 at study entry. Tumor stages at the time check details of study entry are presented in Table 1 for these two groups of transplant

candidates. Among the 93 transplant candidates in the MELD <15 group, 32 HCC patients received LDLT at a median of 1.6 months after initial living liver donor evaluation, 49 received DDLT at a median of 2.2 months after study entry, and 12 had not undergone any transplant by last follow-up, including seven who died on the waitlist. Among the 37 transplant candidates in the MELD ≥15 group, 17 HCC patients went on to receive LDLT at a median of 1.8 months after initial living donor evaluation, 16 received DDLT at a median of 3.1 months after first living donor evaluation, and four had not undergone any transplant at last follow-up, three of whom died on the waitlist. In an adjusted sequential stratification analysis of time from initial donor evaluation to death for transplant candidates with MELD <15 and HCC at study entry, we were unable to detect a significant survival benefit for LDLT recipients compared to patients who did not receive LDLT (HR = 0.82, 95% CI 0.36-1.89; P = 0.65). In a similar analysis for patients with MELD ≥15 at study entry and HCC, patients who underwent LDLT had significantly lower mortality risk than those who did not receive LDLT (HR = 0.29, 95% CI 0.09-0.96; P = 0.043).