[Analysis of transmission of Burkholderia cepacia isolates in an intrahospital by randomly amplified polymorphic DNA-PCR method].

Strains of Burkholderia cepacia isolated in our hospital from November 1995 to September 1996 were classified with randomly amplified polymorphic DNA-PCR (RAPD-PCR) and conventional biochemical tests (ID test.NF-18, API20NE, and Neg Combo 4J kit), and intrahospital isolates of B. cepacia were analysed. During the period 28 strains from inpatients and 2 from medical apparatus were isolated. Twenty four of 28 (85.7%) were from sputum. In 1996 from January to February, 20 strains were detected from 8 inpatients, and two strains were from the nebulizers at the Trauma and Critical Care Center (TCC). With typing of B. cepacia by conventional methods no epidemiological relations among isolates were found. However, DNA patterns of original isolates from the nebulizers at TCC by RAPD-PCR were identical with those of isolates in sputa from patients in other wards who had stayed at TCC, indicating that TCC was an initial source of transmission and the strain was transmitted with the patients to the wards. These results suggest that RAPD-PCR method might be a useful tool to analyse an epidemiological survey for intrahospital transmission of isolate.

Interspecies comparison of c-myc gene in human and rat glioma cell lines.

Interspecies difference in expression of the c-myc gene between two human and three rat glioma cell lines was studied with use of a human c-myc probe. The c-myc deoxyribonucleic acid (DNA) fragments detected at higher stringency in Southern blotting, showed a difference in size and gene copy number between human and rat glioma cells. The c-myc transcript was detected at both higher and lower stringencies in Northern blotting in human glioma cells, whereas it was demonstrated only at lower stringency in rat glioma cells, and the c-myc transcript was seen in cytoplasms of both glioma cells by in situ hybridization. The c-myc protein, if examined with anti-human c-myc protein monoclonal antibody, was observed as two separate components in Western blotting and localized immunocytochemically in nuclei in human glioma cells, whereas it was detected as three separate forms in Western blotting and shown in both nuclei and cytoplasm in rat glioma cells. The above discrepancy in manifestation of c-myc DNA fragments, transcript and protein could be due to the difference in nucleotide sequence of c-myc gene between human and rat glioma cells.

Amplification and expression of a multidrug resistance gene in human glioma cell lines.

Two human glioma cell lines were examined for multidrug resistance (MDR). A vincristine (VCR)-resistant glioma cell line showed a cross resistance to Adriamycin (doxorubicin, ADR) and etoposide (VP-16) to varying extents, suggesting the presence of MDR; the resistance to VCR was considerably decreased by calcium entry blockers. On the other hand, another VCR-sensitive glioma cell line exhibited no cross resistance to ADR or VP-16. Double minute chromosomes and homogeneously staining regions as well as clonal aberrations of chromosome 7 were not observed in cytogenetic studies of multidrug-resistant and multidrug-sensitive glioma cell lines. In Northern and Southern blot analyses, MDR gene 1 (MDR1) messenger ribonucleic acid (mRNA) was shown to be overexpressed without any amplification of the MDR1 gene in multidrug-resistant glioma cell lines as compared to multidrug-sensitive glioma cell lines. It would be reasonable to suggest that amplification of the MDR1 gene may not be a sine qua non for acquisition of MDR and that the MDR1 mRNA level may be correlated with the extent of MDR.

Lectin histochemistry of human gliomas.

Formalin-fixed and paraffin-embedded specimens of 24 human gliomas were examined histochemically with five lectins; concanavalin A (Con A), wheat germ agglutinin (WGA), Ricinus communis agglutinin 1 (RCA-1), peanut agglutinin (PNA), and Ulex europaeus agglutinin 1 (UEA-1). Although the staining intensity with lectins was variable, tumor cells in five astrocytomas, three oligodendrogliomas, six ependymomas, and one gliosarcoma, were generally positive for Con A, WGA, and RCA-1, and negative for PNA and UEA-1, whereas those in nine glioblastomas were usually positive for Con A and WGA and negative for RCA-1 and PNA as well as UEA-1. The malignancy in neoplastic astrocytes was correlated with the decrease in binding with lectins, especially RCA-1. Blood vessels, particularly the endothelial layers, in all gliomas were stained intensely with all lectins used. Macrophages showed two staining features with lectins; stippled and granular. The former macrophages were positive for Con A, WGA, RCA-1, and PNA, and negative for UEA-1, whereas the latter macrophages were positive for all lectins used. Thus, the staining characteristics with lectins of macrophages were different from those of any glioma cells and very useful for identification of macrophages in gliomas.

Contribution of histiocytic cells to sarcomatous development of the gliosarcoma. An immunohistochemical study.

The expression of glial fibrillary acidic protein, fibronectin (FN), factor VIII-related antigen (FVIII/RAG), and of three monohistiocytic markers, lysozyme, alpha-1-antitrypsin and alpha-1-antichymotrypsin was examined in five gliosarcomas (GS) by peroxidase-antiperoxidase immunostaining of formalin-fixed and paraffin-embedded specimens, and compared with vascular changes in 16 glioblastomas (GB). In contrast to GB, endothelial proliferations of GS were sheathed by sarcomatous tissue (perivascular sarcoma), which was contiguous with fibrosarcomatous areas. Cells with conspicuous intracytoplasmic FN content (FN+ cells) were seen in the vascular stroma of GB and dominated in the sarcomatous parts of GS. Most FN+ cells of GS were of varying size and shape and clearly neoplastic. Monohistiocytic markers were demonstrable in small infiltrating mononuclear cells as well as in many sarcomatous cells including FN+ cells. FVIII/RAG was restricted to lumen-lining endothelium and was not found in sarcomatous cells. These results suggest that a major part of sarcoma in GS is less likely to develop from proliferated endothelial cells than from histiocytic cells in the perivascular spaces of GB. By FN mediation, histiocytic cells might also guide and promote sarcomatous proliferations of other mesenchymal cells, leading to fibrosarcomatous development. Prominent monstrous giant cells of one GS seemed to be degenerating glioma cells.

Fibrous xanthoma in the gasserian ganglion: case report.

Occurrence of fibrous xanthoma has been reported increasingly in the skull and the central nervous system, but is extremely rare in the gasserian ganglion. We report on the clinical presentation, radiological appearance, surgical treatment, and histological makeup of a fibrous xanthoma arising from the left gasserian ganglion.

Development of stroma in malignant lymphomas of the brain compared with epidural lymphomas. An immunohistochemical study.

The relation of lymphoma cells to gliomesenchymal stroma within nervous tissue was studied by peroxidase-antiperoxidase immunostaining of formalin-fixed and paraffin-embedded surgical specimens for fibronectin (FN), factor VIII-related antigen and glial fibrillary acidic protein in 17 malignant non-Hodgkin lymphomas of the brain. For comparison, 9 non-Hodgkin lymphomas, 6 Hodgkin lymphomas, and 19 plasmacytomas of the spinal or cranial epidural spaces were studied with the same methods. Lymphoma cells were consistently negative for all markers. All lymphomas of the brain showed conspicuous concentric perivascular circles of immunoreactivity for FN in parts infiltrating brain tissue. Such structures are considered to derive from splitting of basal laminae of preexisting brain vessels; they were not seen in tumors of the epidural space. Cells with conspicuous FN content were found in brain as well as in epidural lymphomas. A monohistiocytic origin of those cells was confirmed by presence of monohistiocytic markers lysozyme and alpha-1-anti-chymotrypsin. Thus, additional immunostaining for FN seems to be useful for detecting monohistiocytes/macrophages in brain tumors.

Effect of selective inhibitor of thromboxane A2 synthetase on cerebral vasospasm after early surgery.

The authors report the results of inhibition of thromboxane A2 synthetase in 49 consecutive patients with subarachnoid hemorrhage (SAH). These unselected Grade I to IV patients all had a ruptured aneurysm of the anterior circle of Willis, and were operated on within 72 hours after SAH. Twenty-seven patients were treated postoperatively by an intravenous infusion of sodium (E)-3- [4-(3-pyridylmethyl)-phenyl] -2- propenoate (OKY-1581), a selective inhibitor of thromboxane A2 synthetase, at 5 micrograms/kg/min for 10 to 14 days, and the remaining 22 patients did not receive this drug. Both groups of patients had similar age distribution and preoperative neurological conditions. A suggestive but statistically insignificant improvement was found in postoperative angiographic vasospasm, ischemic symptoms, and overall outcome in the group receiving OKY-1581. The incidence of low-density areas on the postoperative computerized tomography scans was significantly decreased in patients treated with OKY-1581 infusion.

Neoplastic cerebral aneurysm from lung cancer. Case report.

A 56-year-old man with lung cancer developed a massive intracerebral hematoma resulting from rupture of a small neoplastic aneurysm in a cortical branch of the left posterior temporal artery. Histologically, undifferentiated squamous cells formed an embolus in the aneurysm and destroyed the aneurysm wall. No tumor mass was evident around the aneurysm.

Immunohistochemical study of fibronectin in hemangioblastomas and hemangiopericytomas.

Eight hemangioblastomas and two hemangiopericytomas were studied using indirect immunoperoxidase stains for fibronectin (FN) and glial fibrillary acidic protein (GFAP) in formalin-fixed, paraffin-embedded surgical specimens. Stromal cells in hemangioblastomas were GFAP-negative and showed variable FN expression, while GFAP-positive cells were FN-negative, thus suggesting that the stromal cells are not derived from astrocytes. Hemangiopericytoma cells were poorly to intermediately FN-positive. The origin of stromal cells is discussed in the light of their fine structure and the immunohistochemical stains with other cell markers.

Immunohistochemical study of fibronectin in human glioma and meningioma.

The presence of fibronectin (FN) and glial fibrillary acidic protein (GFAP) in two astrocytomas, 17 glioblastomas, and five meningiomas was studied by indirect immunoperoxidase staining of formalin-fixed and paraffin-embedded surgical specimens. Angiogenesis in tumor was scored by the microscopic angiogenesis grading system (MAGS), and plasma FN levels were measured by single radial immunodiffusion. In astrocytomas and glioblastomas, GFAP-positive tumor cells had no FN expression and FN was confined to proliferating vessel walls and the leptomeninges, showing a mutually exclusive FN and GFAP expression. GFAP-positive tumor cells were occasionally surrounded by a network of FN-positive matrix produced by cells derived from the leptomeninges or blood vessels. In meningiomas, FN expression was found in vessel walls and meningioma cells including whorl formations and psammoma bodies. In general, deep immunoperoxidase staining for FN was shown in the endothelial cells and the psammoma bodies. Plasma FN levels were correlated significantly not to the degree of leptomeningeal proliferation but to the MAGS scores in gliomas.

Translational mobility of concanavalin A receptors in normal and neoplastic glial cells.

The dynamics of cell-associated Concanavalin A (Con A) in astrocytes of the newborn rat (RNA), the rat glioma (AC), and the human glioblastoma (GB) were studied in vitro by fluorescence and electron microscopy. Con A receptors on the cell surface were seen usually as a continuous thin layer, and Con A accumulations in fluorescence microscopy were actually Con A receptors on complicatedly infolded cell membrane and collection of Con A pinosomes. No capping occurred in the three types of glial cells. The translational movement of Con A receptors on the cell surface was rapid in the AC, slow in the RNA, and intermediate in the GB, and partly associated with Con A internalization. Con A pinosomes were more numerous in the RNA compared with those in the AC and the GB. Colchicine accelerated the translational mobility of surface Con A receptors more markedly in the AC and the GB than in the RNA. The translational movement Con A receptors, when treated with cytochalasin B, was retarded in the RNA and the GB and rather accelerated in the AC. Con A pinosomes were decreased in the three types of glial cells by treatment with colchicine or cytochalasin B.