Background:A chart audit (pre-intervention, PRE) of endoscopic ultrasound (EUS)-fine needle biopsy (FNB) of solid masses identified a diagnostic yield of 75%. To improve diagnostic yield, a quality improvement intervention targeting tissue procurement and processing was developed and trialed (post-intervention, POST). Intervention:Increase needle passes to ≥3/mass, limit the personnel preparing cytology slides, and replace saline with formalin for cell block preparation. Methods:A POST chart audit was undertaken for solid mass EUS-FNB from January 2024-December 2024, with quarterly reviews. Only a definite diagnosis was used to calculate diagnostic yield. ResultsAbout:241 patients underwent 262 EUS-FNBs. Diagnostic yield was 81% in POST vs. 75% in PRE (p = ns). Single passes decreased to 14% in POST vs. 20% in PRE (p = ns), with similar diagnostic yield (57% vs. 56%, respectively). Number of ≥3 passes increased significantly (47% POST vs. 12% PRE, P < .0001). In POST, 86% of single passes were performed in the first half vs. 14% in the second (P = .012). Only 25% of ≥3 passes were performed in the first half vs. 75% in the second (P = .06). Diagnostic yield improved significantly between the first and second halves (72% vs. 90%, P < .0001). Replacing saline with formalin for cell block improved diagnostic yield (57% vs. 75%, P = .062). Limiting the personnel making slides from 25 nurses (PRE) to 3 endoscopists (POST) had no impact on diagnostic yield. Conclusions:Critical practice assessment, regular audit, and continued reinforcement led to a significant improvement in EUS-FNB diagnostic yield to 90% by increasing the number of needle passes to ≥3/mass.

INTRODUCTION:Primary sclerosing cholangitis (PSC) may reoccur following liver transplantation (LT), and the diagnosis established once imaging studies demonstrate the diagnostic cholangiographic appearance. To evaluate whether the development of recurrent PSC (rPSC) is associated with cholestasis soon after LT, we studied whether changes in hepatic biochemistry within the first 12 months were linked with the development of rPSC and graft loss. METHODS:We conducted a retrospective cohort analysis of 158 transplant recipients with PSC in Canada and 549 PSC transplant recipients from the United Kingdom. We evaluated serum liver tests within 12 months after LT and the subsequent development of a cholangiographic diagnosis of rPSC as a time-dependent covariate using Cox regression. Severe cholestasis was defined as either alkaline phosphatase > 3x upper limit of normal or total bilirubin > 100 mu mol/L. RESULTS:Patients who developed rPSC were more likely to have severe cholestasis vs those without at 3 months (20.5% vs 8.2%, P = 0.011), at 6 months (17.9% vs 10.0%, P = 0.026), and 12 months (15.4% vs 7.8%, P = 0.051) in the Canadian cohort and at 12 months in the UK cohort (27.9% vs 12.6%, P < 0.0001). By multivariable analysis, development of severe cholestasis in the Canadian cohort at 3 months (hazard ratio [HR] = 2.41, P = 0.046) and in the UK cohort at 12 months (HR = 3.141, P < 0.0001) was both associated with rPSC. Severe cholestasis at 3 months in the Canadian cohort was predictive of graft loss (HR = 3.88, P = 0.0001). DISCUSSION:The development of cholestasis within 3-12 months following LT was predictive of rPSC and graft loss.

Background:Small bowel bleeding accounts for about 5%-8% of all cases of gastrointestinal bleeding. Suspected small bowel bleeding (SSBB) can be classified into occult, inactive overt, and overt. Most patients with SSBB will undergo balloon-assisted enteroscopy (BAE) for diagnosis and treatment. There are currently no recommendations from practice guidelines on what is the best approach and limited information about diagnostic and therapeutic yields for each subtype of SSBB. Aims and Methods:We aimed to investigate the diagnostic and therapeutic yields of BAE in the 3 subtypes of patients with SSBB by performing a retrospective analysis of all patients that underwent BAE for this diagnosis at the University of Alberta Hospital in a 5-year period. We also aimed to identify other factors that could influence diagnostic and therapeutic yields. Results:The overall diagnostic and therapeutic yields of BAE for SSBB were 66% and 51%, respectively. When stratified by subtypes of SSBB, the diagnostic yield for occult, inactive overt, and active overt SSBB were reported to be 61%, 67%, and 95% (P < .05), respectively. BAE performed within 72 hours of presentation and patients requiring transfusion within the past 12 months had a significantly higher diagnostic yield. Conclusions:Our data showed the clinical differences between the 3 subtypes of patients with SSBB and the usefulness of an appropriate and timely approach to maximize the diagnostic and therapeutic yields.

Background Evaluating small bowel Crohn’s disease (SBCD) often relies on cross-sectional imaging (eg, computed tomography enterography [CTE]) and small bowel endoscopy (eg, balloon-assisted enteroscopy [BAE]). The accuracy of CTE for evaluating SBCD compared to BAE remains unclear and is assessed in this study. Methods This single-centre retrospective study included patients with SBCD who underwent both CTE and BAE within 6 months. Findings of active inflammation, long-segment disease, skip-segments, and presence of both strictures and high-grade strictures (HGS) were extracted from CTE and BAE reports and analyzed using BAE as the reference standard. Results Sixty-three CTE and BAE pairings were identified. CTE was sensitive for assessing active inflammation (80.0%) and all strictures (92.1%) and specific for long-segment inflammation (95.0%) and HGS (87.2%). Sensitivity was low for HGS (60.9%) and long-segment inflammation (50.0%), with poor specificity for all strictures (68.4%). In surgically naïve bowel, accuracy improved for active inflammation (sensitivity: 83.3%, specificity: 100%) and worsened for HGS (sensitivity: 42.9%, specificity: 84.2%). In postsurgical bowel, CTE sensitivity for HGS improved to 68.8%. Conclusion Computed tomography enterography accurately detected active inflammation and fibrostenotic disease but may not be sufficient to rule out clinically significant findings such as HGS. The accuracy of CTE varied between surgically naïve and postsurgical bowel. CTE remains an important modality for evaluation of SBCD and should be used in combination with BAE when clinical discrepancy arises.

Abstract Background Management of pancreaticobiliary malignancy is complex and multi-disciplinary. Decompression of malignant biliary obstruction (MBO) is preferentially achieved with endoscopic retrograde cholangiopancreatography (ERCP) and biliary stent placement. This may improve the quality of life for patients with unresectable disease and improve outcomes in resectable/borderline-resectable disease. Purpose To assess quality outcomes in patients undergoing biliary stenting for MBO. Method This is a retrospective chart audit of patients referred to the University of Alberta Hospital (UAH) for suspected or confirmed MBO. The primary outcome was clinical success (reduction in bilirubin of >50% at 30 days). Secondary outcomes were technical success, type of stent used, need for re-intervention, adverse events (AEs), time from stent placement to surgery or cancer centre assessment, and survival. Result(s) Between January 2020 and June 2022, 222 patients (102 female, 46%) with a mean age of 70±1 years (range 34-93 years) underwent 290 ERCPs. The cause of MBO was pancreatic cancer in 130 (59%), cholangiocarcinoma in 32 (14%), ampullary cancer in 9 (4%), and others in 51 (23%). Technical success for stent insertion on first ERCP was achieved in 180/222 patients (81%) with only brushings performed in 2. Of the 40 patients (18%) with unsuccessful ERCP, further success was achieved by repeat ERCP (8), percutaneous transhepatic drainage (PTC, 15), PTC with rendezvous ERCP (1), surgical decompression (3), and endoscopic ultrasound-guided biliary drainage (1). No biliary drainage was performed in 12 patients. Overall, ERCP with stent insertion was technically successful in 188/222 patients (85%). A total of 233 biliary stents were inserted (38 plastic, 195 metal). Clinical success was achieved in 20/38 patients (53%) with plastic stents and 151/181 patients (83%) with metal stents (Χ2 17.4 p<0.05). Adverse events were encountered in 33 patients (11%) with stent migration occurring in 8 (3%), cholangitis in 7 (2%), post-ERCP pancreatitis in 7 (2%), post-sphincterotomy bleeding in 4 (1%), and death in 2 (1%). Re-intervention was required in 36 patients (20%) after initial ERCP with successful stent placement. The re-intervention rate was significantly higher with plastic stents (14/27, 52%) than with metal stents (22/153, 14%) after initial ERCP (Χ2 20.1 p<0.001). The overall survival was a mean of 249±16 days (5-967) for patients with plastic stents and 248± 16 days (1-959) for those with metal stents. Survival was significantly worse in patients with unresectable disease vs resectable/borderline-resectable disease (Log rank 38.89, p<0.001), Figure 1. Image Conclusion(s) In MBO, metal stents appear to provide significantly better biliary drainage, with less need for re-intervention, but do not appear to be associated with any survival benefit over plastic stents. We hope that this quality assurance project will help in the development of a regional management pathway for optimizing the care of patients presenting with MBO. Please acknowledge all funding agencies by checking the applicable boxes below None Disclosure of Interest None Declared

Abstract Background Obscure gastrointestinal bleeding (OGIB) is defined as bleeding from an unknown etiology despite initial investigations with upper endoscopy and colonoscopy, of which 75-80% is attributed to a small bowel (SB) source. OGIB poses a significant diagnostic and therapeutic challenge, resulting in high morbidity and mortality with increased utilization of health care resources. Balloon-assisted enteroscopy (BAE) is a useful procedure for the evaluation and management of small bowel bleeding, with reported diagnostic and therapeutic rates up to 87% and 80%, respectively. Purpose This retrospective study aims to evaluate the diagnostic and therapeutic yields in a large cohort of adult patients presenting with different subtypes of OGIB who have undergone BAE, as well as to assess for association between various patient and disease factors, and clinical outcomes. Method We performed a retrospective review of 1057 cases of BAE at a large quaternary referral centre between 2016 to 2021 and 158 OGIB cases were identified. Sex, age, and, preprocedural variables including indication, time from video capsule endoscopy (VCE) to BAE and subclassification of SB bleed were collected. Endoscopic modality, findings, and therapeutic interventions were used to calculate diagnostic and therapeutic yields. The association between the timing of BAE relative to VCE and clinical outcomes including rebleeding rate, diagnostics and therapeutic yields were assessed. Bleeding free-survival was estimated using Kaplan-Meier function. All data analyses were performed with SPSS. Result(s) The overall diagnostic yield of BAE was 74%. Patients with active overt bleeding were found to have a statistically significant higher yield compared to those with inactive overt bleeding (94% vs 67%, P= 0.03). The therapeutic yield was 51%, with a significantly higher rate of injection therapy and hemoclip placement in those with active overt bleeding compared to those with inactive overt and occult bleeding (P< 0.001). BAE performed within 72 hours of overt GI bleeding was found to have a statistically significant higher diagnostic yield when compared to procedures performed after 72 hours (94% vs 67%, p =0.05). Univariable analysis revealed higher diagnostic yield in patients requiring transfusion within the past 12 months (P= 0.02) Finally, rebleeding-free survival in all patients at 1 year and 5 years was 98% and 68%, respectively. Image Conclusion(s) Balloon-assisted enteroscopy continues to be an effective diagnostic and therapeutic modality for the investigation of obscure gastrointestinal bleeding. Our retrospective study shows a higher diagnostic yield in those presenting with active overt GI bleed as well as those who underwent BAE within 72 hours of overt GI bleeding. Furthermore, patients who required transfusion within the past 12 months are more likely to have a positive BAE finding. Please acknowledge all funding agencies by checking the applicable boxes below None Disclosure of Interest None Declared

Abstract Background Obscure gastrointestinal bleeding (OGIB) accounts for approximately 50% of all GI bleeds. This refers to bleeding from an unknown etiology despite initial investigations with upper endoscopy and colonoscopy, of which 75–80% is attributed to a small bowel source, also termed as small bowel bleeding. Various diagnostic and therapeutic modalities have been used to investigate sources of OGIB. Balloon-assisted enteroscopy (BAE) is a useful procedure for managing small bowel bleeding, with reported diagnostic and therapeutic rates up to 87% and 80%, respectively. Aims The aims of this study are to evaluate the outcomes of patients with OGIB who have undergone BAE, as well as to explore correlations between the timing of BAE and clinical outcomes. Methods We performed a retrospective review of 342 patients who underwent BAE at a tertiary referral centre between 2016 to 2021, of which 116 underwent evaluation for OGIB. Aside from gender and age, preprocedural variables including endoscopy indication, timing from video capsule endoscopy (VCE) and overt GI bleed were collected to allow for further stratification of data. Endoscopic variables including modality, findings, and therapeutic interventions were used to calculate diagnostic and therapeutic yields. Furthermore, the association between timing of BAE after VCE and clinical outcomes including rebleeding rate, diagnostics and therapeutic yields were assessed. All data analyses were performed with SPSS. Results The overall diagnostic yield was 70.3%. Patients with active overt bleeding were found to have a higher yield compared to those with occult and inactive overt bleeding (78.6% vs 72.5% vs 64.6%, respectively). Amongst all OGIB presentations, the most common findings were vascular lesions including angiodysplasia, arteriovenous malformations, and dieulafoy lesions. The majority of lesions were found within the proximal jejunum. Subgroup analysis did not show a statistically significant difference in diagnostic yields between BAE with and without prior VCE (75.9% vs 77.8, P = 0.605). The therapeutic yield was 51.1% with a higher yield found in those presenting with active overt bleeds compared to occult and inactive overt cases. Compared to BAE performed after 30 days of overt GI bleeding, those completed within 30 days was found to have a higher diagnostic and therapeutic rate (80.0% vs 60.0%, 70.0 vs 30.0%, respectively). Conclusions Balloon-assisted enteroscopy continues to be an effective diagnostic and therapeutic modality for the investigation of obscure gastrointestinal bleeding. Our retrospective study shows a higher diagnostic and therapeutic rate in those presenting with active overt GI bleed as well as those who underwent BAE within 30 days of overt GI bleeding Funding Agencies None

Abstract Background Endoscopy teaching is an integral part of gastroenterology (GI) training. Though the number of completed endoscopic procedures does not equate competency, procedure tracking is useful for monitoring an individual’s learning progress. Currently, procedure tracking is typically done on an informal basis using paper or electronic spreadsheets. These methods are non-standardized and may not be shareable between trainees and their programs. Endostation is a smartphone app created by the University of Alberta Therapeutic Endoscopy Program to facilitate the tracking of endoscopic procedures. The app allows trainees to record the number of endoscopies and details such as cecal intubation (CI), ERCP cannulation, and therapeutic interventions. Data can be accessed by users via the app and website (www.endostation.ca), allowing for close monitoring of trainees’ learning progress. Aims Our primary objective was to evaluate the usefulness of the app for tracking the number of endoscopic procedures and therapeutic interventions. Our secondary objective was to evaluate the acquisition of endoscopy skills based on quality endoscopic parameters such as CI rate and ERCP cannulation rate. Methods One therapeutic endoscopy fellow and two GI residents were recruited for the study. Participants were asked to document their procedures over the study period (9-month for therapeutic endoscopy fellow, 12-month for GI residents). Total number of procedures was summed for each trainee. Acquisition of endoscopy skills was tracked by comparing success rates of CI and ERCP cannulation at different points within the study period. Results The therapeutic endoscopy fellow recorded 415 cannulation attempts, 209 sphincterotomies, 282 stone extractions, 71 plastic stent placements, and 37 metal stent placements. There was a significant difference in the cannulation success rate when comparing the 1st trimester and the 3rd trimester of the study period (68% vs 85%; p= 0.0012) (Fig 1). The two GI residents respectively recorded 335 and 170 colonoscopies plus 454 and 305 gastroscopies. Resident 1 recorded 58 polypectomies, 9 esophageal variceal banding, and 16 non-variceal hemostasis. Resident 2 recorded 17 polypectomies, 12 esophageal variceal banding, and 9 non-variceal hemostasis. The CI success rate was significantly higher for both residents when comparing the first 4 months of training vs the last 4 months [24% vs 88% for resident 1 (p=0.00001); 15% vs 42% for resident 2 (p= 0.001)] (Fig 1). Conclusions The smartphone app (Endostation) was a useful tool for endoscopic procedure tracking. Data from the app was useful in demonstrating improvement in CI rate and ERCP cannulation rate over the study period. Funding Agencies None

BACKGROUND:Gastrointestinal neurofibromas are commonly found in patients diagnosed with neurofibromatosis type 1. However, isolated gastrointestinal neurofibromas are a rare entity and only fourteen cases of isolated colorectal neurofibromas have been documented in literature. Isolated gastrointestinal neurofibromas have not been associated with Lynch syndrome (LS). Patients with LS are at an increased risk of colorectal cancer, and are recommended to undergo screening colonoscopy.CASE SUMMARY:A 33-year-old healthy female with a family history of LS was found to have unresectable polyp in the ascending colon on screening colonoscopy suspicious for malignancy. The patient was asymptomatic and had no stigmata of neurofibromatosis. A staging workup for colorectal cancer revealed no evidence of metastatic disease. A discussion with the patient resulted in the decision to undergo a segmental resection with ongoing surveillance. The patient underwent a laparoscopic right hemicolectomy. Histopathology was consistent with a gastrointestinal neurofibroma. Post-operatively, the patient recovered well. She will not require further treatment with regards to her colonic neurofibroma, but will continue to follow-up for ongoing surveillance of her LS.CONCLUSION:We present the first case of an isolated colonic neurofibroma in a patient with LS. This case explores considerations for the management of isolated gastrointestinal neurofibromas given the lack of guidelines in literature.

Inflammatory bowel diseases (IBD), Crohn's disease and ulcerative colitis, are poorly understood disorders affecting the intestinal tract. The current model for disease suggests that genetically susceptible patients develop intolerance to gut microflora, and chronic inflammation develops as a result of environmental insults. Although interest has mainly focused on studying genetic variants and gut bacterial flora, little is known about the potential of viral infection to contribute to disease. Accordingly, we conducted a metagenomic analysis to document the baseline virome in colonic biopsy samples from patients with IBD in order to assess the contribution of viral infection to IBD. Libraries were generated from colon RNA to create approximately 2 GB sequence data per library. Using a bioinformatic pipeline designed to detect viral sequences, more than 1000 viral reads were derived directly from tissue without any coculture or isolation procedure. Herein, we describe the complexity and abundance of viruses, bacteria/bacteriophage, and human endogenous retroviral sequences from 10 patients with IBD and 5 healthy subjects undergoing surveillance colonoscopy. Differences in gut microflora and the abundance of mammalian viruses and human endogenous retroviruses were readily detected in the metagenomic analyses. Specifically, patients with herpesviridae sequences in their colon demonstrated increased expression of human endogenous viral sequences and differences in the diversity of their microbiome. This study provides a promising metagenomic approach to describe the colonic microbiome that can be used to better understand virus-host and phage-bacteria interactions in IBD.

Our laboratory has characterized and cloned a human betaretrovirus resembling the mouse mammary tumor virus (MMTV) from biliary epithelium and lymph nodes of patients with the autoimmune biliary disease, primary biliary cirrhosis (PBC). Evidence for betaretrovirus infection has been detected mainly in patients’ lymphoid tissue by RT-PCR and immunohistochemistry in approximately 75% of PBC patients. However, the viral burden was observed to be far less in the liver and we could only identify viral sequence in a third of patients using RT-PCR. Other labs have been unable to confirm our data using PCR of hepatic DNA. Not surprisingly, the betaretroviral association with PBC has become a controversial issue. In order to provide more robust evidence of infection in patients with liver disease, we have isolated an infectious betaretrovirus from lymph nodes. We cloned and sequenced the proviral isolates and confirmed viral particles by electron microscopy. We have also conducted ligation-mediated PCR studies with next generation sequencing and found proviral integration sites both in vivo and in vitro. To date, we have identified more than 3,000 integration sites in perihepatic lymph nodes, liver and isolated biliary epithelium in the majority of patients with PBC and seldom in patients with other liver diseases. These data confirm the presence of an MMTV-like virus in patients with autoimmune liver disease. Ongoing studies using biliary disease mouse models with MMTV infection and randomized controlled anti-retroviral trials for patients with PBC will be required to further characterize the association of human betaretrovirus infection with PBC.

Background & AimsThe NOD.c3c4 mouse model develops autoimmune biliary disease characterized by spontaneous granulomatous cholangitis, antimitochondrial antibodies and liver failure. This model for primary biliary cirrhosis (PBC) has evidence of biliary infection with mouse mammary tumour virus (MMTV), suggesting that the virus may have a role in cholangitis development and progression of liver disease in this mouse model. We tested the hypothesis that MMTV infection is associated with cholangitis in the NOD.c3c4 mouse model by investigating whether antiretroviral therapy impacts on viral levels and liver disease.MethodsNOD.c3c4 mice were treated with combination antiretroviral therapy. Response to treatment was studied by measuring MMTV RNA in the liver, liver enzyme levels in serum and liver histology using a modified Ishak score.ResultsCombination therapy with the reverse transcriptase inhibitors, tenofovir and emtricitabine, resulted in a significant reduction in serum liver enzyme levels, attenuation of cholangitis and decreased MMTV levels in the livers of NOD.c3c4 mice. Furthermore, treatment with the retroviral protease inhibitors, lopinavir and ritonavir, in addition to the reverse transcriptase inhibitors, resulted in further decrease in MMTV levels and attenuation of liver disease in this model.ConclusionsThe attenuation of cholangitis with regimens containing the reverse transcriptase inhibitors, tenofovir and emtricitabine, and the protease inhibitors, lopinavir and ritonavir, suggests that retroviral infection may play a role in the development of cholangitis in this model.

Autoimmune hepatitis (AIH) and primary biliary cirrhosis (PBC) are poorly understood autoimmune liver diseases. Immunosuppression is used to treat AIH and ursodeoxycholic acid is used to slow the progression of PBC. Nevertheless, a proportion of patients with both disorders progress to liver failure. Following liver transplantation, up to a third of patients with PBC experience recurrent disease. Moreover a syndrome referred to as "de novo AIH" occurs in a proportion of patients regardless of maintenance immunosuppression, who have been transplanted for disorders unrelated to AIH. Of note, the use of cyclosporine A appears to protect against the development of recurrent PBC and de novo AIH even though it is a less potent immunosuppressive compared to tacrolimus. The reason why cyclosporine A is protective has not been determined. However, a virus resembling mouse mammary tumor virus (MMTV) has been characterized in patients with PBC and AIH. Accordingly, we hypothesized that the protective effect of cyclosporine A in liver transplant recipients may be mediated by the antiviral activity of this cyclophilin inhibitor. Treatment of the MMTV producing MM5MT cells with different antivirals and immunosuppressive agents showed that both cyclosporine A and the analogue NIM811 inhibited MMTV production from the producer cells. Herein, we discuss the evidence supporting the role of MMTV-like human betaretrovirus in the development of PBC and de novo AIH and speculate on the possibility that the agent may be associated with disease following transplantation. We also review the mechanisms of how both cyclosporine A and NIM811 may inhibit betaretrovirus production in vitro.

Background) We have previously reported the efficacy of dual treatment, which is consisted of reductive hepatectomy and percutaneous isolated hepatic perfusion (PIHP), for patients with advanced HCC.However these patients are frequently complicated with Vp4 portal vein tumor thrombus (PVTT), and conventional en bloc resection is not always feasible.To overcome this situation, we have developed back flow thrombectomy (BFT) technique.For right hemihepatectomy with Vp4 PVTT, which reached to the contralateral left second portal branch, the portal trunk should be at first clamped at the superior border of the pancreas.After one transverse venotomy at an appropriate site of the right portal branch, tumor thrombus is extracted by forceps and scissors using suction devices.Of particular note, the vascular clamp at the left first portal branch should be avoided because it may split PVTT and enhance portal vein embolization with fragmented tumor thrombus.Instead, back flow pressure in the portal system generated by BFT technique should be kept throughout the thrombectomy procedure.This pressure eases effective extraction of both micro-and macroscopic cancer nests liberated to the blood stream and avoid the migration into the future remnant liver.(Methods) Until the end of 2011, 43 multiple bilobular HCC patients with Vp4 were performed reductive hepatectomy with tumor thrombectomy.In 22 of 43 patients, BFT techniques were used.Sixteen of 23 patients had PVTT in the contralateral second portal branch.Seventeen of 43 patients were not performed PIHP because of economical reason, extrahepatic metastases, aggressive tumor progression, hepatic dysfunction, infectious complications or unfavorable conditions after surgery.(Results) Patency of portal vein at thrombectomy site of all/BFT patients 3 and 6 months after hepatectomy were 92%/90% and 87%/86%, respectively.The median OS of all 43 patients was 14 months and the 1 and 3-year OS rate was 55.5% and 19.1% respectively.In 26 patients who could undergo PIHP as second treatment, the median OS was 17 months and the 1 and 3-year OS rate was 69.2% and 23.1% respectively.(Conclusions) Tumor thrombectomy by BFT technique allows multi-disciplinary treatment for patients with PVTT.An impressively increased survival rate achieved by additional PIHP supports the dual treatment strategy for multiple bilobular HCC patients with Vp4 PVTT.

We conducted an unbiased metagenomics survey using plasma from patients with chronic hepatitis B, chronic hepatitis C, autoimmune hepatitis (AIH), non-alcoholic steatohepatitis (NASH), and patients without liver disease (control). RNA and DNA libraries were sequenced from plasma filtrates enriched in viral particles to catalog virus populations. Hepatitis viruses were readily detected at high coverage in patients with chronic viral hepatitis B and C, but only a limited number of sequences resembling other viruses were found. The exception was a library from a patient diagnosed with hepatitis C virus (HCV) infection that contained multiple sequences matching GB virus C (GBV-C). Abundant GBV-C reads were also found in plasma from patients with AIH, whereas Torque teno virus (TTV) was found at high frequency in samples from patients with AIH and NASH. After taxonomic classification of sequences by BLASTn, a substantial fraction in each library, ranging from 35% to 76%, remained unclassified. These unknown sequences were assembled into scaffolds along with virus, phage and endogenous retrovirus sequences and then analyzed by BLASTx against the non-redundant protein database. Nearly the full genome of a heretofore-unknown circovirus was assembled and many scaffolds that encoded proteins with similarity to plant, insect and mammalian viruses. The presence of this novel circovirus was confirmed by PCR. BLASTx also identified many polypeptides resembling nucleo-cytoplasmic large DNA viruses (NCLDV) proteins. We re-evaluated these alignments with a profile hidden Markov method, HHblits, and observed inconsistencies in the target proteins reported by the different algorithms. This suggests that sequence alignments are insufficient to identify NCLDV proteins, especially when these alignments are only to small portions of the target protein. Nevertheless, we have now established a reliable protocol for the identification of viruses in plasma that can also be adapted to other patient samples such as urine, bile, saliva and other body fluids.

Background & Aims: A human betaretrovirus resembling the mouse mammary tumor virus (MMTV) has been characterized in primary biliary cirrhosis (PBC) and associated with aberrant pyruvate dehydrogenase complex (PDC)-E2-like expression. As MMTV is prevalent in mice as either an exogenous or endogenous infection, we tested the hypothesis that MMTV is linked with anti-mitochondrial antibody (AMA) production in models with severe immune dysfunction.Methods: Evidence for MMTV was assessed in the liver and spleen of mice by PCR and immunochemistry and PDC-E2-like protein by immunochemistry. ELISA and Western blot were used to investigate AMA and anti-MMTV antibody production.Results: Increased MMTV gag or env expression was detected in the livers of AMA producing mice including NOD.c3c4, CD4 directed dominant negative TGF-beta receptor II and IL-2 receptor alpha knockout mice as well as the NOD parental strain when compared to healthy strains and biliary disease control mice. The NOD.c3c4 mice expressed MMTV surface and capsid proteins and aberrant PDC-E2-like protein in the bile ducts, whereas IL-2 receptor alpha knockout mice, NOD.c3c4 and the NOD mice expressed MMTV proteins and aberrant PDC-E2-like protein in the spleen. A significant correlation between anti-MMTV antibody production and AMA production was observed in the sera of NOD and NOD.c3c4 mice (p < 0.0001).Conclusions: The association of betaretroviral protein production and aberrant PDC-E2-like protein expression in the NOD.c3c4, NOD, and the IL-2 receptor alpha knockout mice is comparable to observations in patients with PBC. The correlation of AMA and anti-MMTV suggests the hypothesis that MMTV infection may trigger the production of AMA. Crown copyright (C) 2011 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.