Subclassification of hepatocellular adenomas: practical considerations in the implementation of the Bordeaux criteria.

Hepatocellular adenomas are benign liver lesions with a risk of rupture and malignant transformation. Various molecular subgroups have been identified which appear to have characteristic morphological and immunohistochemical features. We examined the morphology and immunohistochemical profile of a series of 121 HCA from 97 patients to identify the HCA subtypes present and determine the number at risk for malignant transformation according to the World Health Organization (WHO) criteria for hepatocellular adenomas. There were 34 HNF1α inactivated HCA (28%), 61 inflammatory HCA (50%), 15 ß-catenin activated HCA (12%) and 11 unclassified adenomas (9%). This proportion of cases was similar to that seen in other series utilising molecular classification. The morphological features of the adenomas were suggestive but not definite indicators of the subtypes present. Morphological features that showed overlap between the subtypes included steatosis within the lesion, a ductular reaction and focal atypia, so that immunohistochemical typing was required for accurate classification. In conclusion, immunohistochemistry is a clinically useful surrogate for identifying underlying molecular changes in the HCA subtypes.

Low-Dose Lipopolysaccharide Causes Biliary Injury by Blood Biliary Barrier Impairment in a Rat Hepatic Ischemia/Reperfusion Model.

This study explored whether bacterial endotoxins, in the form of lipopolysaccharides (LPS), could have an injurious effect on the biliary tract in conjunction with ischemia. A total of 64 rats were randomly assigned to 4 groups: sham operation (sham group), 1 mg/kg LPS intraperitoneal (LPS group), hepatic ischemia/reperfusion (IR; IR group), and IR combined with LPS (IR+LPS group). Following 1 or 6 hours of reperfusion, serum liver tests, bile duct histology, immunofluorescence microscopy (zonula occludens-1 [ZO-1]), bile composition (bile salts, phospholipids, lactate dehydrogenase), hepatic gene expression (bile salt transporters and inflammatory mediators), as well as serum and biliary cytokine concentrations were quantified and compared between the study groups. In addition, the integrity of the blood biliary barrier (BBB) was assayed in vivo using horseradish peroxidase (HRP). LPS administration induced severe small bile duct injury following 6 hours of reperfusion. Furthermore, total bile salts and bilirubin concentrations in serum were increased in the LPS groups compared with sham controls (LPS, + 3.3-fold and +1.9-fold; IR+LPS, + 3.8-fold and +1.7-fold, respectively). The BBB was impaired in the LPS groups as evidenced by elevated levels of HRP in bile (+4.9-fold), and decreased expression of claudin 1 (-6.7-fold) and claudin 3 (-3.6-fold). LPS was found to be a potent inducer of small bile duct injury following hepatic ischemia and 6 hours of reperfusion. This injury was associated with increased permeability of the BBB and impaired hepatic bile salt clearance. Liver Transplantation 23 194-206 2017 AASLD.

Inconsistent hepatic antifibrotic effects with the iron chelator deferasirox.

BACKGROUND AND AIM: Development of effective antifibrotic treatments that can be translated to clinical practice is an important challenge in contemporary hepatology. A recent report on ß-thalassemia patients demonstrated that deferasirox treatment reversed or stabilized liver fibrosis independent of its iron-chelating properties. In this study, we investigated deferasirox in cell and animal models to better understand its potential antifibrotic effects. METHODS: The LX-2 stellate cell line was treated with 5 µM or 50 µM deferasirox (Exjade, Novartis Pharmaceuticals Australia, North Ryde, NSW, Australia) for up to 120 h. Three-week-old multidrug resistance 2 null (Mdr2(-/-) ) mice received oral deferasirox or vehicle for 4 weeks (30 mg/kg/day). Cells and liver tissue were collected for assessment of fibrosis and fibrogenic gene expression. RESULTS: In LX-2 cells treated with 50 µM deferasirox for 12 h, α1(I)procollagen expression was decreased by 25%, with maximal reductions (10-fold) seen following 24-120 h of treatment. Similarly, α-smooth muscle actin (αSMA) expression was significantly lower. Alterations in matrix remodeling genes, specifically decreased expression of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2, were observed. There was no significant difference in hepatic hydroxyproline content in Mdr2(-/-) mice following deferasirox administration (vehicle: 395 ± 27 µg/g vs deferasirox: 421 ± 33 µg/g). Similarly, no changes in the expression of fibrogenic genes were observed. CONCLUSION: Despite reductions in α1(I)procollagen and αSMA expression and alterations in matrix degradation genes in LX-2 cells, deferasirox did not exhibit antifibrotic activity in Mdr2(-/-) mice. Given the positive outcomes seen in human trials, it may be appropriate to study deferasirox in other animal models of fibrosis and/or for a longer duration of therapy.

Lack of efficacy of mTOR inhibitors and ACE pathway inhibitors as antifibrotic agents in evolving and established fibrosis in Mdr2⁻/⁻ mice.

BACKGROUND & AIMS: Mammalian target of rapamycin and angiotensin-converting enzyme inhibition has been shown to have antifibrotic activity in models of liver fibrosis. The aim of our study was to determine the efficacy of rapamycin, everolimus, irbesartan and captopril, alone and in combination, as antifibrotic agents in the Mdr2(-/-) model of cholestasis both in early injury and established disease. METHODS: Mdr2(-/-) mice were treated for 4 weeks with vehicle, rapamycin (1 mg/kg) or everolimus (5 mg/kg) every second day or with captopril (30 mg/kg/day), irbesartan (10 mg/kg/day) or vehicle. Further groups of 3-week-old Mdr2(-/-) mice were treated with rapamycin and irbesartan in combination (1 mg/kg/day and 10 mg/kg/day) or with rapamycin (2 mg/kg/day) for 4 weeks. Liver injury and fibrosis were compared between treated and untreated animals. RESULTS: There were no significant improvements in liver injury, histology, hepatic hydroxyproline or profibrogenic gene expression following treatment with rapamycin, everolimus, captopril or irbesartan at any time point studied. Likewise, there were no improvements in liver histology or profibrogenic gene expression following combination therapy or high-dose rapamycin treatment. CONCLUSIONS: The antifibrotic effects of rapamycin, everolimus, captopril and irbesartan seen in other models of fibrosis were not replicated in the Mdr2(-/-) model in this study. This highlights the clear need to test specific antifibrotic agents in a number of different animal models. We believe this animal model is ideal to study usefulness of antifibrotic agents in cholestatic liver disease because of the similarity in genetics and hepatic histopathology to human cholestatic liver disease.

Alpha B-crystallin, a new independent marker for poor prognosis in head and neck cancer.

OBJECTIVES: Gene expression profiling has provided many insights into tumor progression but translation to clinical practice has been limited. We have previously identified a list of potential markers by the differences of expression profiling of seven matched head and neck cancer (HNSCC) tumors with autologous normal oral mucosa (NOM). Alpha B-crystallin (CRYAB) was in the top 5% of genes identified with statistically significant differences in expression between tumor and NOM at the mRNA level. The objective was to confirm this in routine paraffin sections at the protein level. STUDY DESIGN: The level of alpha B-crystallin was determined in tumors of 62 HNSCC patients whose prognosis was known for 5 years. METHODS: Immunohistochemical detection of alpha B-crystallin expression was performed on HNSCC paraffin sections. RESULTS: Univariate survival analysis identified lack of alpha B-crystallin staining as an independent prognostic marker for disease-free interval (P < 0.001) and overall survival (P < 0.002) of HNSCC patients over the 5-year observation period. Notably, all 13 patients (100%), including 5 patients with nodal disease whose tumors lacked alpha B-crystallin had no recurrences (P < 0.001). Nineteen of 27 node-negative patients stained positive for alpha B-crystallin and seven of the 19 (36.8%) had recurrences. CONCLUSION: Presence or absence of expression of alpha B-crystallin was a powerful marker for prognosis in this series of patients.

Novel markers for poor prognosis in head and neck cancer.

Head and neck cancer (HNSCC) is one of the most distressing human cancers, causing pain and affecting the basic survival functions of breathing and swallowing. Mortality rates have not changed despite recent advances in radiotherapy and surgical treatment. We have compared the expression of over 13,000 unique genes in 7 cases of matched HNSCC and normal oral mucosa. Of the 1,260 genes that showed statistically significant differences in expression between normal and tumor tissue at the mRNA level, the three top ranking of the top 5% were selected for further analysis by immunohistochemistry on paraffin sections, along with the tumor suppressor genes p16 and p53, in a total of 62 patients including 55 for whom >4-year clinical data was available. Using univariate and multivariate survival analysis, we identified SPARC/osteonectin as a powerful independent prognostic marker for short disease-free interval (DFI) (p < 0.002) and poor overall survival (OS) (p = 0.018) of HNSCC patients. In combination with other ECM proteins found in our analysis, PAI-1 and uPA, the association with DFI and OS became even more significant (p < 0.001). Our study represents the first instance of SPARC as an independent prognostic marker in HNSCC.

Changing patterns of knowledge, reported behaviour and sexually transmitted infections in a South African gold mining community.

BACKGROUND: In 1998, a major HIV intervention project was started in a mining community in Carletonville, South Africa. This included community-based peer education, condom distribution, syndromic management of sexually transmitted infections (STI), and presumptive STI treatment for sex workers. OBJECTIVES: To investigate changes in sexual behaviour and the prevalence of STI before and 2 years after the start of the HIV prevention programme. METHODS: Cross-sectional surveys were carried out in 1998 and 2000 among mine workers, sex workers and adults in the community. Demographic and behavioural factors were recorded and participants were tested for syphilis, gonorrhoea and chlamydial infection and, at the start of the intervention, for HIV. RESULTS: In 1998, the prevalence of HIV among men and women in the general population, mine workers, and sex workers, was 20%, 37%, 29% and 69%, respectively. In 2000, syphilis, gonorrhoea and chlamydial infection had increased among mine workers; chlamydial infection had increased among men and women, and syphilis had increased among women. There was evidence of positive behaviour change but this was not substantial or universal. Knowledge of HIV/AIDS and awareness of the epidemic were high but condom use remained low. CONCLUSION: There was little evidence of significant behaviour change and the prevalence of curable STI increased. The prevention programme had had less impact than expected. Reasons for the reduced impact, and the lessons for future intervention projects are discussed. There is a need for further monitoring of the HIV epidemic especially as its impact increases.

Locating research informants in a multi-ethnic community: ethnic identities, social networks and recruitment methods.

OBJECTIVE: Recruitment of informants can 'make or break' social research projects, yet this has received little research attention. Drawing on our recent qualitative research into health and social capital in a multi-ethnic neighbourhood in South England, this paper presents a detailed analysis of the complexities encountered in recruiting research informants who described themselves as African-Caribbean, Pakistani-Kashmiri and white English. METHODS: Three methods of recruitment were used: (1) advertisements and articles in local media, (2) institutional contacts through local voluntary organisations and (3) interpersonal contacts, referrals and snowballing. We compare and contrast the experiences of ethnically matched interviewers who conducted research amongst the three aforementioned ethnic groups. These experiences were recorded by means of lengthy interviewer 'debriefing questionnaires' that focused on factors that had served to help or hinder them in finding research participants. These questionnaires formed the basis of a discussion workshop in which the interviewers and researchers sought to identify the factors impacting on the recruitment process. RESULTS: Our findings suggest that local advertisements and media contact worked best for recruiting members of the white English community in our South English town. Interpersonal contacts were crucial in recruiting Pakistani-Kashmiri informants. Institutional contacts were the most useful way of accessing African-Caribbean individuals. CONCLUSION: We conclude that local ethnic identities and social networks produce qualitatively different responses to recruitment attempts in different communities. Such differences necessitate the employment of a range of recruitment methodologies and detailed formative research in a target community before commencing recruitment.