Keratin 19 protein expression is an independent predictor of survival in human hepatocellular carcinoma.

AIM: We aimed to assess the clinicopathological relevance and prognostic significance of expression of the hepatic progenitor cell markers keratin 19 (K19), epithelial cell adhesion molecule (EpCAM) and CD117 (c-KIT) in a White series of hepatocellular carcinoma (HCC). METHODS: We evaluated the immunohistochemical expression of K19, EpCAM and CD117 in 89 surgical specimens of HCC from Greek patients (mean age 66.7±11.3 years, male 75.2%) followed up for 39.6±25.3 months. RESULTS: K19, EpCAM and CD117 expression was detected in tumour cells of 10.11, 15.38 and 3.7% HCCs, respectively. Female sex was correlated with EpCAM immunohistochemical expression (P=0.035), whereas no other significant relationship with clinicopathological parameters was observed. K19 positivity tended to be correlated with microvascular invasion (P=0.054). In univariate analysis, K19 positivity and microvascular invasion were found to be associated with decreased recurrence-free survival (P<0.001 and P=0.004, respectively) and overall survival (P=0.002 and P=0.029, respectively). EpCAM and CD117 positivity was not correlated with patient survival. In multivariate analysis, K19 positivity emerged as an independent predictor of recurrence-free survival (odds ratio=7.84, 95% confidence interval=2.658-22.912, P<0.001) and overall survival (odds ratio=3.845, 95% confidence interval=1.401-10.549, P=0.009). CONCLUSION: Our study confirms the prognostic significance of K19 expression in Caucasian patients with HCCs, providing further evidence that it may be used to stratify HCC according to tumour aggressiveness.

Liver regeneration: immunohistochemical study of intrinsic hepatic innervation after partial hepatectomy in rats.

BACKGROUND: We examined the intrinsic hepatic innervation after partial hepatectomy (PH) in rats and the presence and pattern of neural sprouting in regenerating liver. METHODS: Male Wistar rats (age 9-13 weeks-w, weight 204-356 g), were submitted to two-thirds PH. Rats were sacrificed at postoperative days (d) 1, 3, 5, 7, at 2 and 4 w, and at 3 and 6 months (m) (6-7 animals/group, control group n = 4). Immunohistochemistry for the pan-neural marker protein gene product 9.5 (PGP9.5) and growth-associated protein 43 (GAP-43), a marker of regenerating nerve axons, was performed on tissue sections from the R1 lobe of the regenerating liver. Portal tracts (PTs) with immunoreactive fibers were counted in each section and computer-assisted morphometric analysis (Image Pro Plus) was used to measure nerve fiber density (number of immuno-positive nerve fibers/mm2 (40x)). RESULTS: Immunoreactivity for PGP9.5 was positive in all groups. The number of PGP9.5 (+) nerve fibers decreased from 0.32 +/- 0.12 (control group) to 0.18 +/- 0.09 (1d post-PH group), and gradually increased reaching pre-PH levels at 6 m (0.3 +/- 0.01). In contrast, immunoreactivity for GAP-43 was observed at 5d post-PH, and GAP-43 (+) PTs percentage increased thereafter with a peak at 3 m post-PH. GAP-43 (+) nerve fiber density increased gradually from 5d (0.05 +/- 0.06) with a peak at 3 m post-PH (0.21 +/- 0.027). At 6 m post-PH, immunoreactivity for GAP-43 was not detectable. CONCLUSIONS: Following PH in rats: 1) nerve fiber density in portal tracts decreases temporarily, and 2) neural sprouting in the regenerating liver lobes starts at 5d, reaches peak levels at 3 m and disappears at 6 m post-PH, indicating that the increase in hepatic mass after PH provides an adequate stimulus for the sprouting process.

Ontogeny of intrinsic innervation in the human thymus and spleen.

The ontogeny of the innervation of human lymphoid organs has not been studied in detail. Our aim was to assess the nature and distribution of parenchymal nerves in human fetal thymus and spleen. We used the peroxidase immunohistochemical technique with antibodies specific to neuron-specific enolase (NSE), neurofilaments (NF), PGP9.5, S100 protein, and tyrosine hydroxylase (TH) and evaluated our results with image analysis. In human fetal thymus, NSE-, NF-, S100-, PGP9.5-, and TH-positive nerves were identified associated with large blood vessels from 18 gestational weeks (gw) onwards, increasing in density during development. Their branches penetrated the septal areas at 20 gw, reaching the cortex and the corticomedullary junction between 20 and 23 gw. Few nerve fibers were seen in the medulla in close association with Hassall's corpuscles. In human fetal spleen, NSE-, NF-, S100-, PGP9.5-, and TH-positive nerve fibers were localized in the connective tissue surrounding the splenic artery at 18 gw. Perivascular NSE-, NF-, S100-, PGP9.5-, and TH-positive nerve fibers were seen extending into the white pulp, mainly in association with the central artery and its branches, increasing in density during gestation. Scattered NSE-, NF-, S100-, PGP9.5-, and TH-positive nerve fibers and endings were localized in the red pulp from 18 gw onward. The predominant perivascular distribution of most parenchymal nerves implies that thymic and splenic innervation may play an important functional role during intrauterine life.