Vera Lucia Pannain

Histopathological stages of nonalcoholic fatty liver disease in type 2 diabetes: prevalences and correlated factors.

Background/Aims: Nonalcoholic fatty liver disease (NAFLD) is highly prevalent in type 2 diabetes mellitus (T2DM). However, data regarding the prevalence and correlates of its histopathological stages are scarce. The aim was to investigate the prevalence and correlates of the more severe histopathological features of NAFLD, nonalcoholic steatohepatitis (NASH) and advanced fibrosis, in T2DM.

Molecular Testing for Lipomatous Tumors: Critical Analysis and Test Recommendations Based on the Analysis of 405 Extremity-based Tumors

Ancillary molecular testing has been advocated for diagnostic accuracy in the differentiation of lipomas from atypical lipomatous tumors/well-differentiated liposarcomas (ALT/WDL); however, the implications and specific indications for use are not well-established in the current literature. Herein, we extend previous findings by quantitatively evaluating the impact of molecular testing of lipomatous neoplasms in our routine clinical practice, how it modifies the historical perspective of their clinical course, and the effect of distinct surgical procedures in modulating the risk of local recurrence for these tumors after molecular classification. On the basis of these analyses, we suggest a specific set of basic recommendations for complementary molecular assessment in the diagnosis of lipomatous tumors. Four hundred and five lipomatous neoplasms located in the trunk and extremities were analyzed histologically and for the presence of 12q13-15 amplification on paraffin-embedded tissues by assessing MDM2/CPM amplification. Survival analyses were calculated with Kaplan-Meier and compared with the log-rank. Multivariate analysis was evaluated by the Cox regression method. The 405 tumors were histologically classified as ordinary lipoma (n=324), intramuscular lipoma (n=29), and ALT/WDL (n=52). The level of agreement between the histologic diagnosis and the molecular diagnosis was high (96%) but pathologists showed a tendency to overestimate cytologic atypia and the diagnosis of ALT/WDL (precision, 79%; accuracy, 88%). Molecular assessment led to a major diagnostic reclassification in 18 tumors (4%). Eleven of the tumors histologically classified as ALT/WDL were reclassified as ordinary lipoma (n=5) and intramuscular lipoma (n=6); none of which recurred. Seven ordinary lipomas were reclassified as ALT/WDL, 6 of which were larger than 15 cm and deeply located; 2 recurred locally. After molecular data, the 5-year local recurrence rates for ordinary lipoma, intramuscular lipoma, and ALT/WDL were 1%, 12%, and 44%, respectively. Multivariate analyses after molecular assessment showed tumor type and type of resection to be associated with the risk of local recurrence. Complementary molecular testing refines the histologic classification of lipomatous tumors and better estimates the impact of surgical procedures on the risk of local recurrence. Pathologists tend to overestimate the degree of cytologic atypia and the indiscriminate use of molecular testing should be avoided, especially for extremity-based tumors. Molecular testing should be considered for “relapsing lipomas,” tumors with questionable cytologic atypia (even if widely excised), or for large lipomatous tumors (>15 cm) without diagnostic cytologic atypia.

Microvascular density of regenerative nodule to small hepatocellular carcinoma by automated analysis using CD105 and CD34 immunoexpression

BackgroundAngiogenesis is a proliferative process resulting in the development of new blood vessels from existing endothelial cells and is considered crucial for tumor growth and metastasis. Tumor angiogenesis can be quantified by microvascular density (MVD), which is evaluated in highly vascularized tumor areas (hot spots) by immunohistochemical assays using CD34 and CD31 pan-endothelial antibodies. More recently, CD105 has been successfully used for some tumor types because it could discriminate neovascularization. The expression of CD34 and CD105 in hepatocellular carcinomas (HCC) and hepatic precancerous lesions has been reported—although the results for CD105 are controversial—but to the best our knowledge, CD105 has not been previously investigated in dysplastic nodules (DN). We investigated and compared MVD-CD34 and MVD-CD105 immunoexpression in tissues containing different stages of hepatocarcinogenesis, including DN.MethodsA total of 31 regenerative nodules (RN), 26 DN and 25 small HCC from explants were used for immunohistochemical tests with CD34 and CD105 antibodies. Antibody expression was quantified by computerized image analysis measurement of MVD, areas containing highly positive endothelial cells within the nodules.ResultsThe median MVD for CD34 was higher in HCC than in DN and RN (p < 0.01), and was higher in DN compared with RN (p = 0.033). In contrast, MVD with CD105 was higher in RN, and the difference was significant in RN and DN compared with HCC (p = 0.019 and p = 0.012, respectively). When MVD with CD34 and CD105 were compared within a single group, there was a significant predominance of CD105 in RN and DN (p < 0.01). In addition, MVD-C34 in HCC predominated compared with MVD-CD105, but the difference was not statistically significant (p = 0.128).ConclusionsThis study identified a close relationship between CD105 and liver cirrhosis, and that CD34 antibody is a good endothelial marker for hepatic carcinogenesis. There was no difference between the use of CD105 and CD34 antibodies in preneoplastic lesions.

Eosinophils involved in fulminant hepatic failure are associated with high interleukin-6 expression and absence of interleukin-5 in liver and peripheral blood

Background/Aims: Although eosinophils are considered to play an important role in the pathogenesis of various parasitic, allergic and autoimmune digestive diseases, their role in fulminant hepatic failure (FHF) is unknown. Our contribution was to identify and quantify eosinophils and cytokine levels [interleukin (IL)‐6, IL‐5 and macrophage inflammatory protein (MIP)‐1α] in liver parenchyma and peripheral blood from FHF patients at pre‐ and post‐transplantation steps.

Analysis of morphological variables and arterialization in the differential diagnosis of hepatic nodules in explanted cirrhotic livers.

BackgroundMany terminologies have been given to dysplastic hepatocellular nodules, which are preneoplastic lesions. In 1995, the International Working Party meeting established the nomenclature and morphological criteria for hepatocellular nodular lesions. Nevertheless, an unequivocal differential diagnosis is sometimes difficult, particularly among large regenerative nodules, dysplastic nodules and hepatocellular carcinoma. Angiogenesis is observed during hepatocarcinogenesis and the presence of the isolated arteries may help to discriminate these nodules. The relevance of the International Working Party histological variables and presence of the isolated arteries were analyzed with regard to the diagnosis of large regenerative nodules, low and high grade dysplastic nodules and hepatocellular carcinoma, in order to evaluate which have a real contribution in such diagnoses.MethodsOne hundred and seven nodular hepatocellular lesions over 5 mm (or smaller nodules with a different color) from explanted cirrhotic livers were analyzed and classified following the criteria of the International Working Party. Classifications were as follows: large regenerative nodules, low grade dysplastic nodules, high grade dysplastic nodules and hepatocellular carcinoma. The presence of isolated arteries (not related to the portal tracts or fibrosis) was verified for the nodules.ResultsAmong the 107 nodular lesions studied, 17 were classified as large regenerative nodules, 38 as low grade dysplastic nodules, 28 as high grade dysplastic nodules and 24 as hepatocellular carcinoma. The most relevant International Working Party variables in the differential diagnosis of the nodules were cellularity, trabeculae thickness, cytoplasmic staining, nuclear atypia, pseudoacinar pattern, portal tracts, nucleocytoplasmic ratio and mitosis. The isolated arteries, identified by hematoxylin and eosin staining, were important discriminating between two groups: low grade lesions (large regenerative nodules/low grade dysplastic nodules) and high grade lesions (high grade dysplastic nodules/hepatocellular carcinoma) (P < 0.001).ConclusionThe International Working Party criteria allow for the classification of the majority of hepatocellular nodules. However, other features such as cytoplasmatic tintorial affinity and pseudoacinar pattern may contribute to these diagnoses. The finding of isolated arteries in a nodular lesion should be investigated carefully, since the nodule could be a dysplastic lesion or hepatocellular carcinoma.

Vascular Immunohistochemical Markers: Contributions to Hepatocellular Nodule Diagnosis in Explanted Livers

INTRODUCTION The process of hepatic carcinogenesis involves a progression including large regenerative nodules, to dysplastic nodules, and finally to hepatocellular carcinoma. Angiogenesis is fundamental to the development of malignant tumors. Changes in sinusoidal capillarization and isolated arteries occur early in hepatic carcinogenesis. However, sometimes differentiation of hepatocellular nodules can be difficult for the general pathologist. The aim of this study was to evaluate angiogenesis by immunohistochemistry using CD34 and HHF35 antibodies for differential diagnosis of large regenerative nodules versus dysplastic nodules versus hepatocellular carcinoma using explanted cirrhotic livers. METHODS Seventy-nine nodules obtained from 29 explanted cirrhotic livers were classified according to the International Working Party as follows: 17 large regenerative, 23 low-grade dysplastic, 23 high-grade dysplastic, and 16 hepatocellular carcinomas. These nodules were submitted to immunohistochemistry with antibodies to CD34 and HHF35 to analyze sinusoidal capillarization and arterialization, respectively. RESULTS Semiquantitative analysis revealed that CD34 expression was >30% in dysplastic nodules and hepatocellular carcinoma; the staining in 93.8% of cases was diffuse, almost involving the entire sinusoidal lining in hepatocellular carcinoma. The number of isolated arteries was high in hepatocellular carcinoma (average, 4.369), which positively correlated with the other nodules (P < .005). CONCLUSION Quantification of sinusoidal capillarization and isolated arteries in hepatocellular nodules, as detected with CD34 and HHF35 antibodies, respectively provided an important tool to differentiate dysplastic nodules from hepatocellular carcinoma.

HBV-DNA levels in HBsAg-positive blood donors and its relationship with liver histology.

Introduction and Objectives The clinical meaning of viremia, especially at low levels, in chronic hepatitis B virus (HBV)-infected patients remains unknown. The objective of the present study was to determine serum HBV-DNA levels and its relationship with liver histology in HBsAg-positive blood donors. Methods A cross-sectional study was conducted on 78 blood donors, with alanine aminotransferase (ALT) and HBeAg evaluation and quantitative determination of HBV-DNA by polymerase chain reaction (Amplicor, HBV Monitor, Roche; lower limit of sensitivity 1,000 copies/mL). Liver biopsy was obtained from all patients with detectable viremia irrespective of ALT and HBV-DNA levels. Results Among 78 blood donors, serum HBV-DNA was detected in 47 (60%) patients; 39 (83%) were males; mean age 37.6±10.4 years; 31 (66%) were HBeAg-negative, and ALT was elevated in 26 (55%). The median of HBV-DNA levels was 24,000 copies/mL and 31 (40%) subjects had no detectable serum HBV-DNA. Although the histologic lesions were mild in the majority of patients, HBV-DNA levels were significantly higher in patients with chronic hepatitis or cirrhosis when compared with patients without histologic liver disease (25,260,000 vs. 9480 copies/mL; P<0.001). There was a significant correlation between HBV-DNA levels and necroinflammatory score (r=0.59) and fibrosis (r=0.50); however, in the subset of HBeAg-negative patients with HBV-DNA levels below 30,000 copies/mL, 25% presented histologic disease related to HBV. Conclusions Most HBsAg-positive blood donors show low viral load. There is a significant association between viral replication and liver damage; however, low HBV-DNA levels do not exclude the presence of histologic disease.

RASSF1A and DOK1 Promoter Methylation Levels in Hepatocellular Carcinoma, Cirrhotic and Non-Cirrhotic Liver, and Correlation with Liver Cancer in Brazilian Patients

Hepatocellular carcinoma (HCC) is the second most common cause of cancer mortality worldwide. Most cases of HCC are associated with cirrhosis related to chronic hepatitis B virus or hepatitis C virus infections. Hypermethylation of promoter regions is the main epigenetic mechanism of gene silencing and has been involved in HCC development. The aim of this study was to determine whether aberrant methylation of RASSF1A and DOK1 gene promoters is associated with the progression of liver disease in Brazilian patients. Methylation levels were measured by pyrosequencing in 41 (20 HCC, 9 cirrhotic, and 12 non-cirrhotic) liver tissue samples. Mean rates of methylation in RASSF1A and DOK1 were 16.2% and 12.0% in non-cirrhotic, 26.1% and 19.6% in cirrhotic, and 59.1% and 56.0% in HCC tissues, respectively, showing a gradual increase according to the progression of the disease, with significantly higher levels in tumor tissues. In addition, hypermethylation of RASSF1A and DOK1 was found in the vast majority (88%) of the HCC cases. Interestingly, DOK1 methylation levels in HCC samples were significantly higher in the group of younger (<40 years) patients, and higher in moderately differentiated than in poorly differentiated tumors (p < 0.05). Our results reinforce the hypothesis that hypermethylation of RASSF1A and DOK1 contributes to hepatocarcinogenesis and is associated to clinicopathological characteristics. RASSF1A and DOK1 promoter hypermethylation may be a valuable biomarker for early diagnosis of HCC and a potential molecular target for epigenetic-based therapy.

Fusion of HMGA1 to the LPP/TPRG1 intergenic region in a lipoma identified by mapping paraffin-embedded tissues

Ordinary lipoma frequently harbors rearrangement of HMGA2. LPP is the most common partner gene to HMGA2, but has not been seen fused to HMGA1. We report the fusion of HMGA1 to the intergenic region between LPP and TPRG1 in a lipoma. Conventional cytogenetic analysis of an abdominal-wall lipoma diagnosed in a 60-year-old woman showed a t(3;6)(q27;p21). Molecular cytogenetic mapping of available paraffin-embedded tissues revealed the fusion of HMGA1 to a 139-kb genomic region between the LPP and TPRG1 loci. No rearrangement of HMGA2 was found. The biological function of this novel fusion could be similar to the role of HMGA2-LPP in tumorigenesis.

Comparative study of the different degrees of risk of gastrointestinal stromal tumor

OBJECTIVE To evaluate the applicability of the main categories of risk and morphological factors in the prognosis of gastrointestinal stromal tumors. METHODS we retrospectively studied fifty-four cases of GIST, assessing the main prognostic factors of this neoplasis: risk levels, topography, size, mitotic index, necrosis, histological subtype and immunophenotype. We also verified their association and the reduction of overall survival. RESULTS Univariate analysis showed that tumors with mitoses number greater than 5 per 50CGA (high-power fields), the presence of necrosis and a high risk for both the systems proposed by Fletcher and Miettinen had a significant association with reduced survival (p = 0.00001, 0.0056, 0.03 and 0.009, respectively). The remaining analyzed factors (size, histological subtype, topography and immunophenotype) had no such association. Multivariate analysis (Jacard index) showed that the Miettinen degree of risk was the one that best correlated with prognosis. CONCLUSION the risk criteria of Fletcher and Miettinen are important in assessing the prognosis of patients with gastrointestinal stromal tumors, especially the latter, which adds to the mitotic index and the presence of tumor necrosis.