Increased expression of telomerase RNA component is associated with increased cell proliferation in human astrocytomas.

Deregulation of telomerase, a ribonucleoprotein polymerase that compensates progressive loss of telomeric (TTAGGG)n repeats during DNA replication, has been suggested to facilitate tumorigenesis and cellular immortality by providing unlimited proliferation capacity for cancer cells. We investigated the relationship between tumor proliferation activity and in situ expression of the telomerase RNA component in 46 human grade I to IV astrocytomas. Heterogeneously distributed telomerase RNA expression was detected from all of the tumor samples as well as from normal human brain tissue. However, expression of telomerase RNA was significantly increased in highly malignant tumors (P = 0.024) and in tumors that showed increased proliferation activity determined by MIB-1 immunohistochemistry (P = 0.014). Interestingly, increased telomerase RNA levels were observed in a subgroup of grade II astrocytomas that showed significant increase in proliferation activity (P = 0.047), indicating that the telomerase RNA component is up-regulated already in early states of astrocytoma malignancy. Telomeric repeats amplification assays revealed telomerase activity in 4 of 6 glioblastomas and in 1 rapidly proliferating grade II astrocytoma. These results suggest that increased tumor proliferation activity triggers telomerase activation via mechanisms that involve increased production of the telomerase RNA component.     

Intermediate filament proteins in choroid plexus and ependyma and their tumors.

The intermediate filament protein types of normal choroid plexus and ependymal tissue and their putative tumors were investigated. In normal human choroid plexus tissue, but not in ependyma, keratin could be demonstrated immunohistochemically. By immunoblotting, keratins 8, 18, and 19 were found, but glial fibrillary acidic protein (GFAP) was absent. In mouse and rat, choroid plexus epithelium and ependymal lining cells were keratin-positive. In addition, many ependymal cells were vimentin-positive. Keratin was immunohistochemically found in three of four choroid plexus papillomas, two of two choroid plexus carcinomas, and the lining cells of three neuroepithelial cysts. GFAP-positive cells were present in some choroid plexus tumors. In contrast, none of the eight ependymomas contained keratin, but all were strongly positive for GFAP. The results show that choroid plexus lining cells and choroid plexus tumors have true epithelial characteristics in their cytoskeleton, in contrast to ependymomas, which do not show keratin positivity but show glial filaments, as would be seen in astrocytic tumors.     

Secretion of vascular endothelial growth factor/vascular permeability factor from human luteinized granulosa cells is human chorionic gonadotrophin dependent

Vascularization is a prominent event during corpus luteum formation, providing low density lipoproteins for steroid biosynthesis and enabling transport of secreted steroids. The process of vascularization is controlled by specific regulators. Vascular endothelial growth factor (VEGF), otherwise named vascular permeability factor (VPF), induces endothelial cell proliferation as well as angiogenesis in vivo and increases capillary permeability. Here we report the expression of VEGF/VPF mRNA by cultured human luteinized granulosa cells (GC) for at least 10 days. Without HCG VEGF/VPF expression declined after day 4 and by day 10 was reduced to approximately 30% of the value at day 4. However, after culture in the presence of 1 U/ml human chorionic gonadotrophin (HCG), expression of VEGF/VPF mRNA by GC was four times greater than control experiments by day 10, and increased 100% from day 4 to day 10. Simultaneously, HCG supplementation increased VEGF/VPF secretion by GC. Medium VEGF/VPF on day 3 was 13 pM without and 11 pM with HCG. Medium VEGF/VPF on day 10 was 6 pM without HCG and 29 pM with HCG. These results suggest that vascularization of the corpus luteum is induced by HCG-mediated effects of VEGF/VPF.      

Primary gastrointestinal stromal tumor of the esophagus in an HIV-positive patient

We describe a rare case of malignant gastrointestinal stromal tumor (GIST) of the esophagus presenting in an HIV-positive man. Not only did the tumor arise from an unusual anatomic site for GIST, namely, the esophagus, but it also had a predominant epithelioid cell morphology that is uncommon and preferentially associated with aggressive behavior. Exhaustive immunohistochemical studies showed strong reactivities to the classic GIST marker, CD34, and to the current more sensitive and more specific GIST marker, CD117/ c-kit protein. This immunophenotype corresponded to that of stromal tumors arising in the more common sites like stomach and small intestine as well as to that of a reported series of esophageal GISTs in the general population. Mutations of the c-kit protein was detected in the tumor, confirming previous observations. This further documents that esophageal GIST and the more common benign esophageal spindle cell lesions are pathologically distinct entities and despite its rarity, esophageal GIST should be recognized by pathologists and clinicians. The occurrence of this tumor in an HIV-positive patient is coincidental, and it resulted in an extremely unusual metastatic site that has not been reported for GISTs.     

Differential diagnosis of chordoma, chondroid, and ependymal tumors as aided by anti-intermediate filament antibodies.

Six chordomas, nine chondrosarcomas, and three myxopapillary ependymomas of the filum terminale were evaluated immunohistochemically for the expression of intermediate filament proteins by the use of monospecific antibodies against intermediate filament proteins of keratin, vimentin, and glial fibrillary acidic protein (GFAP) type. All chordomas were positive for keratin but negative for GFAP, whereas chondrosarcomas and ependymomas were negative for keratin. Chondrosarcomas showed strong vimentin positivity, whereas ependymomas were positive for GFAP. Chordomas showed desmosomelike junctions by electron microscopy, whereas chondrosarcomas of different types showed no junctions or only primitive ones. By electron microscopy chordomas often showed prominent intermediate filaments also associated with desmosomes, and poorly differentiated chondrosarcomas also showed prominent intermediate filaments. Keratin positivity of chordomas suggests their epithelial nature, while vimentin positivity of chondrosarcomas is in line with their mesenchymal derivation. The results also show that antibodies against different intermediate filament proteins can be applied as diagnostic aids in making the distinction between chordomas, chondroid tumors, and ependymal tumors.     

Fine mapping of a gene responsible for regulating dietary cholesterol absorption; founder effects underlie cases of phytosterolaemia in multiple communities

Sitosterolaemia (also known as phytosterolaemia, MIM 210250) is a rare recessive autosomal inherited disorder, characterised by the presence of tendon and tuberous xanthomas, accelerated atherosclerosis and premature coronary artery disease. The defective gene is hypothesised to play an important role in regulating dietary sterol absorption and biliary secretion, thus defining a molecular mechanism whereby this physiological process is carried out. The disease locus was localised previously to chromosome 2p21, in a 15 cM interval between microsatellite markers D2S1788 and D2S1352 (based upon 10 families, maximum lodscore 4.49). In this study, we have extended these studies to include 30 families assembled from around the world. A maximum multipoint lodscore of 11.49 was obtained for marker D2S2998. Homozygosity and haplotype sharing was identified in probands from non-consanguineous marriages from a number of families, strongly supporting the existence of a founder effect among various populations. Additionally, based upon both genealogies, as well as genotyping, two Amish/Mennonite families, that were previously thought not to be related, appear to indicate a founder effect in this population as well. Using both homozygosity mapping, as well as informative recombination events, the sitosterolaemia gene is located at a region defined by markers D2S2294 and Afm210xe9, a distance of less than 2 cM.     

Alpha-1-antitrypsin and lysozyme. Their limited significance in fibrohistiocytic tumors

A wide range of tumors were immunohistochemically analyzed for alpha-1-antitrypsin (AAT) and lysozyme in order to evaluate their specificity as histiocytic markers and their significance in the diagnostic and histogenetic evaluation of fibrohistiocytic tumors. Besides histiocytic lesions, AAT immunoreactivity was commonly found in different types of carcinomas and sarcomas, and strong immunoreactivity was found in carcinoid tumors, malignant melanomas, and schwannomas, which, however, had negative results for lysozyme. The AAT immunoreactivity could be abolished with the absorption of the antibody with purified AAT also in nonhistiocytic tumors. The neoplastic pleomorphic cells in malignant fibrous histiocytomas (MFHs) usually had strongly positive results for AAT, whereas only entrapped histocytes had positive results for lysozyme and for two monoclonal antibodies to histomonocytic cells. The results show that AAT has a relatively low specificity as a histiocytic marker, and one should be careful in concluding the histiocytic nature of tumors, such as MFHs, based on AAT immunostaining. It seems also questionable whether AAT can be used as a diagnostic marker for MFH. The reason for the widespread AAT immunoreactivity in various tumors may be that AAT is taken up from serum to various types of nonhistiocytic tumor cells.     

Widespread immunoreactivity for alpha-1-antichymotrypsin in different types of tumors

The presence of alpha-1-antichymotrypsin (AACT) immunoreactivity was investigated in a broad range of different tumors. Because optimal AACT immunoreactivity of histiocytes was achieved by use of a protease pretreatment of paraffin sections, all tumors were studied by using pepsin treatment of the sections. In such conditions, more than 80% of different carcinomas, sarcomas, melanomas, and lymphomas were positive for AACT. The results indicate that AACT immunoreactivity is not specific for histiocytic tumors and therefore cannot be used as a histiocytic marker and to substantiate the concept of histiocytic nature of tumors, such as malignant fibrous histiocytoma. The concept of AACT as a histiocytic marker may be based on the fact that histiocytes, for some reason, stain more intensively for AACT, and immunoreactivity can also be found in them at unoptimal immunostaining conditions.     

Fixation of femoral shaft osteotomy with an intramedullary composite rod: an experimental study on dogs with a two-year follow-up

A novel composite material with an ultra-high strength and a low elastic modulus called carbon fibre-reinforced liquid crystalline polymer (LCP/CF) has been developed. An experimental diaphyseal osteotomy of the proximal femur in fourteen Beagle dogs was fixed with an intramedullary LCP/CF rod of 4.5 mm in diameter and 80 mm in length. The radiological follow-up intervals were 1, 3, 6, 12, 24, 52, and 104 weeks. Five dogs were killed at 1 year and three dogs at 2 years for histological studies; six dogs were retained for longer follow-up. Radiographs showed an uncomplicated healing of the diaphyseal osteotomy with an external callus formation in all dogs in 12 weeks. Histological analysis revealed a benign host tissue response with few inflammatory cells. Both bone and fibrous tissue were seen at the LCP/CF-host tissue interface. The cross-sectional cortical area of the operated femur was slightly greater than that of the control femur in the 2-year follow-up. LCP/CF showed promising properties for high-load applications.