Attila Patonai

Claudins and tricellulin in fibrolamellar hepatocellular carcinoma

Fibrolamellar hepatocellular carcinoma is a subtype of hepatocellular carcinoma occurring in non-cirrhotic liver at a younger age. The tumor expresses both hepatocellular and cholangiocellular markers. Previously, our group described overexpression of tight junction protein claudin 4 in cholangiocellular carcinoma in contrast to hepatocellular carcinoma. In the present study, tight junction protein expressions were studied to possibly clarify bipotential lineage of fibrolamellar hepatocellular carcinoma. Eleven fibrolamellar hepatocellular carcinomas were compared with seven “conventional” hepatocellular carcinomas, seven cholangiocellular carcinomas, and five normal liver samples. By immunohistochemistry, all fibrolamellar hepatocellular carcinomas were positive for HepPar1 and cytokeratins 7, 8, and 18, but negative for cytokeratin 19. Glypican-3 gave weak staining in two cases. Expression of claudin 1 was lower, while that of claudin 2 was higher in fibrolamellar hepatocellular carcinomas than in other tumors. Claudins 3, 4, and 7 were not detectable in fibrolamellar hepatocellular carcinomas as in the majority of “conventional” hepatocellular carcinomas, contrary to high expression observed in cholangiocellular carcinomas. Focal or diffuse claudin 5 expression was detected in nine of 11 fibrolamellar hepatocellular carcinomas contrary to other tumors. Tricellulin was significantly downregulated in all tumors compared with normal liver. Our findings showed claudins to exhibit specific expression patterns in fibrolamellar hepatocellular carcinomas not observed in other primary liver tumors, with unique claudin 5 expression and pattern features similar to common hepatocellular carcinoma, but different from cholangiocellular carcinoma. This is the first report describing the loss of tricellulin expression in human hepatic tumors.

Molecular Characteristics of Fibrolamellar Hepatocellular Carcinoma

Fibrolamellar hepatocellular carcinoma (FLC) occurs in non-cirrhotic liver and the etiopathogenesis is still obscure. Both hepatocellular and cholangiocellular markers are expressed in the tumor, however, molecular alterations and altered pathways playing role in the tumor pathogenesis are not clearly identified. The purpose of the present study was to compare the expression level of EGFR, syndecan-1 and ß-catenin in FLC, conventional hepatocellular carcinoma (cHCC) and cholangiocellular carcinoma (CCC) and to investigate the possibility of mutation both in EGFR and K-RAS. Eight FLCs were compared with 7 cHCCs, 7 CCCs and 5 normal liver samples. Cytokeratins 7, 8, 18, 19, HepPar1 (HSA), EGFR, syndecan-1 (CD138) and ß-catenin were detected by immunohistochemistry. In addition EGFR, ß-catenin and syndecan-1 were evaluated by digital morphometry and K-RAS, EGFR mutations in FLC cases using paraffin-embedded samples. All FLCs were positive for HepPar1 (HSA) and cytokeratins 7, 8, 18, but negative for cytokeratin 19 by immunohistochemistry. EGFR was significantly overexpressed in all three tumor types, being highest in FLCs (p = 0,0001). EGFR, K-RAS mutation analyses revealed no mutations in exons studied in FLCs. Our findings proved that expression of EGFR is higher in FLC than in other types of primary malignant hepatic tumors and no K-RAS mutation can be detected, so FLC is a good candidate for anti-EGFR treatment.

Experimental results of using autologous rectus fascia sheath for venous patch grafts in dogs

Autologous vascular patch grafts developed from the internal rectus sheath were implanted onto the bilateral common iliac vein and jugular vein of 4 experimental beagle dogs. During the development and implanting of the grafts no technical difficulties or perioperative complications were encountered. The follow-up lasted 6 months and 3 months in the case of the common iliac vein grafts and the jugular grafts, respectively. In the postoperative period, the morphological and functional characteristics of the implanted venous sections were examined by Doppler ultrasonography and CT angiography. Normal patency was detected, and none of these check-ups showed obturation or stenosis. The histological survey showed no mesothelial cell layer, but the insides of the grafts showed total restructuring and were covered by a normal endothelial layer. No difference could be detected between samples harvested 3 and 6 months after implanting. The immunohistochemical examinations using anti-claudin-5 and anti-CD31 antibodies confirmed the preliminary results of the histological examinations that the luminal surfaces of the implanted grafts developed a differentiated monolayer endothelium which was free of degenerative and inflammatory signs. The control examinations show the suitability of the internal rectus sheath as a venous wall donor.

Visceral Aluminum Deposition In Chronic Renal Insufficiency. (Light Microscopy and X-ray Microanalysis).

Aluminum is a common element in our environment, but has been proved to be toxic, mainly in chronic renal insufficiency. Most cases of ALU intoxication occur during hemodialysis due to treatment of aluminum-containing drugs. In the present case, we describe visceral manifestations of aluminum deposition in a middle aged, multidialysed, male patient. Light and polarization microscopy examinations and X-ray microanalysis revealed amorph, extracellular aluminum deposits in various parenchymal organs causing failure of heart, lung and kidney functions. There were no anamnestic data concerning aluminumcontaining drugs or occupational exposure.

Morphological evaluation of experimental autologous rectus fascia sheath vascular grafts used for arterial replacement in a dog model

Although experimental autologous patch or tubular conduit vascular grafts made from the internal rectus fascia sheath (IRFS) have been reported in the literature, thorough morphological evaluation and verification of the histological arterialisation of such grafts are lacking. Four purpose-bred Beagle dogs were utilised to create eight arterial internal rectus fascia sheath (ARFS) grafts implanted between bisected ends of the external iliac arteries. Four out of the eight ARFS grafts were patent after three months. Haematoxylin-eosin and Azan staining verified that the grafts gained a vessel-like layered structure with the presence of large amounts of collagen fibres. Although the inner surface of the intact IRFS was originally covered with claudin-5-negative and pancytokeratin-positive mesothelial cells in control samples, the internal cells of the ARFS grafts became claudin-5 positive and pancytokeratin negative like in intact arteries. Spindle-shaped cells of the wall of ARFS grafts were α-smooth muscle actin (α-SMA) positive just like the smooth muscle cells of intact arteries, but α-SMA immunoreactivity was negative in the intact IRFS. According to these findings, the fibroblast cells of the ARFS graft have changed into myofibroblast cells. The study has proved that ARFS grafts may be used as an alternative in arterial replacement, since the graft becomes morphologically and functionally similar to the host vessel via arterialisation.