Kunyu Harada

Clinical evaluation of 33 patients with histologically verified germinoma

We evaluated 33 patients with histologically verified germinoma between 1978 and 1992. The patients consisted of 23 men and 10 women between 6 and 32 years of age. All patients underwent either biopsy, partial resection, or total resection. All patients received radiotherapy, and all had a complete remission. Recurrence occurred in three patients. Two recurrences were outside of the irradiated area. Though there were four deaths, none were caused by germinoma. Three resulted from the inadequate hormonal replacement. Accurate staging of patients with germinoma using magnetic resonance imaging (MRI), including whole spinal MRI, and development of a radiochemotherapy or chemotherapy method that preserves the hypothalamic-pituitary axis function are needed.

Telomerase activity and the expression of telomerase components in pituitary adenoma with malignant transformation

BACKGROUND Telomerase activity responsible for cellular immortality may participate the development of human cancers. Telomerase is a multisubunit ribonucleoprotein composed of at least three components: hTERT, hTERC, and TEP1. This is the first report showing telomerase activity and telomerase component expression in pituitary adenoma with histological malignant transformation. DESCRIPTION A 16-year-old male with a prolactin-producing pituitary adenoma with metastasis is presented. The patient underwent three partial resections of an intra- and suprasellar lesion over a 2-year period and received focal irradiation. Eight years after the first admission, a metastatic lesion to the subarachnoid space around the medulla oblongata was detected and the lesion was resected as the fourth operation. Furthermore, the suprasellar lesion showed regrowth and partial resection was performed as the fifth operation. The last two specimens were diagnosed as pituitary carcinoma. Radiotherapy with gamma knife was performed for the residual suprasellar lesion and a new lesion in the left temporal lobe after the fifth operation. Telomerase activity was examined by TRAP/TRAP-HPA methods, qualitatively and quantitatively. Telomere length was examined by Southern blot analysis, and the expression of telomerase components (hTERT, hTERC, and TEP1) was examined by reverse transcriptase-polymerase chain reaction (RT-PCR). The MIB1 index, telomerase activity, and hTERT expression increased according to histologic malignancy chronologically in this patient. None of the specimens showed immunoreactivity for p53, EGFR, or bc12. No telomerase activity was detected in pituitary adenomas without malignant transformation, other benign brain tumors, or normal brain tissues. CONCLUSION We report a patient with pituitary adenoma transforming to carcinoma. The tumor cells acquired immortality and revealed malignant transformation during the course of the disease, that was proved by an increase of telomerase activity and hTERT expression.

Growth Inhibition of Human Glioma Cells by Transfection-induced P21 and its Effects on Telomerase Activity

The aim of this study is to investigate the effect of the p21 gene transfection on the growth of cultured human glioma cell lines, and analyze the telomerase activity, and detection of telomerase components in p21 transfectant.The p21 gene was transfected into human glioma cell lines, U251MG and T98G with our novel liposome. The cell growth was assessed by counting the number of trypan blue-excluding cells in a hemocytometer and flow cytometry analysis. The expression of P21 protein and its mRNA were examined by Western and Northern blot analysis. The telomerase activity was assayed by TRAP (telomerase repeat amplification protocol)/TRAP–HPA (hybridization protection assay) method qualitatively and quantitatively. The length of telomere was measured by Southern blot analysis. The expression of telomerase components (hTERT, hTERC and TEP1) were examined by RT-PCR (reverse transcriptase-polymerase chain reaction).The p21 transfectant demonstrated the expression of P21 protein and its mRNA. The p21 transfection of human glioma cells results in growth inhibition and G0/G1 arrest. The p21 transfectant revealed a decrease of telomerase activity and hTERT expression as compared with control cells.These results suggest that p21 transfection induces G0/G1 arrest in human glioma cells which associates with the reduction in the telomerase activity and hTERT expression.

Telomerase activity in central nervous system malignant lymphoma

Primary central nervous system malignant lymphoma (PCNSL) has been regarded as a rare neoplasm. Recently, however, its incidence has been rapidly increasing. Despite active clinical trials, its clinical and biologic features remain unknown and there has been no effective treatment or prognostic factor. The current study attempted to elucidate telomerase activity and expression of the telomere‐related RNA of PCNSL as novel prognostic factors.

Dermoid cyst in the posterior fossa accompanied by Klippel-Feil syndrome.

Abstract  Objects: Dermoid cysts accompanied by Klippel-Feil syndrome are uncommon, and the coincidence of these two rare diseases suggests an interesting idea about the pathogenesis of a dermoid cyst, which the authors consider with reference to an actual case. A posterior fossa dermoid cyst with dermal sinus and Klippel-Feil syndrome are reported in the same patient. A 23-year-old woman was admitted to hospital with progressive headaches. Methods: Cervical radiography demonstrated C4–5 vertebral fusion, and magnetic resonance imaging revealed a large cystic mass in the midline of the posterior fossa. The cystic lesion was totally removed along with the accompanying dermal sinus. It was histopathologically diagnosed as a dermoid cyst. Conclusion: The rare coincidence of a dermoid cyst and Klippel-Feil syndrome suggests the possibility that a disturbance in the mesoderm before the fourth week of gestation might play an important part in the causation of dermoid cyst.

Combined antitumor effects of TNF and G-CSF on a human medulloblastoma xenograft line

SummaryThe antitumor effects of TNF and G-CSF on a xenograft line of human medulloblastoma were examined. (Method): 1) A human medulloblastoma xenograft line was transplanted into nude mice. Tumor bearing nude mice were divided into the following eight groups: untreated controls (C); those receiving a subcutaneous injection of G-CSF for one week (G1); for four weeks (G2); those receiving an intratumoral injection of TNF for four weeks (Tit); an intravenous injection of TNF (Tiv); those receiving a combination of G1 and Tit (G1 + Tit); a combination of G2 and Tit (G2 + Tit); and a combination of G2 and Tiv (G2 + Tiv). The relative tumor weight in each group was calculated and any antitumor effects were examined by calculating a tumor growth inhibition ratio. 2) Tumor bearing nude mice were divided into the following two groups: those receiving a subcutaneous injection of G-CSF and an intravenous injection of TNF (G + T); and only an intravenous injection of TNF (T). We evaluated the pathological findings from the tumors at 0 h, 0.5 h, 1 h, 3 h, 6 h, 12 h, 24 h and 48 h after the TNF injection. Routine H.E. staining and immunostaining using antigranulocyte and antimacrophage antibodies were performed. (Results): 1) The tumor growth inhibition ratio was 0.112, 0.190, 0.287, 0.451, 0.347, 0.635, and 0.622 at G1, G2, Tit, Tiv, G1 + Tit, G2 + Tit, G2 + Tiv group. A combined antitumor effect was clearly seen in the G2 + Tit and the G2 + Tiv groups. 2) The tumor was fragmented by the infiltration of many inflammatory cells 24 hours after TNF injection. Many more macrophages were observed in the tumors of G + T mice than in the T mice. Granulocytes were observed only in the tumors of the G + T mice.

New approach for the treatment of medulloblastoma by transfection with glial fibrillary acidic protein gene

Glial fibrillary acidic protein (GFAP) is one of the intermediate filaments found in mature normal astrocytes and differentiated glioma cells. It seems to be able to stabilize the cytoskeleton of the astrocyte and may play a role in maintaining astrocyte cell shape, in association with other cytoskeletal components such as microfilaments and microtubles. However, its tissue-specificity remains unclear. To clarify the effect of GFAP expression in brain tumour cells, transfer of the GFAP gene into the human medulloblastoma cell line, DAOY-1 (which does not express GFAP) was carried out using liposomes. Upon transfection, we observed the alterations in the characteristics of GFAP transfected cells. Cell growth, morphology and sensitivity to anticancer drugs were compared between GFAP gene transfected DAOY-1 and control DAOY-1 cells. Growth inhibition and increase of sensitivity to anticancer drugs were observed with GFAP expression in GFAP gene-transfected DAOY-1 cells. However, no morphological changes were noted.

Cavernous sinus meningioma presenting as orbital apex syndrome. Diagnostic methods of dynamic MRI, spoiled GRASS (SPGR) image.

Orbital apex syndrome is a symptomatologic complex. In this paper, the usefulness of several clinical diagnostic MR methods in preparing for surgery is discussed. These include dynamic MRI and the spoiled GRASS (SPGR) image for a cavernous sinus meningioma presenting with orbital apex syndrome. A 53 year old man, who had right cavernous sinus tumor presenting with orbital apex syndrome, was examined by several new MRI techniques. The tumor was partially removed and fibroblastic meningioma was confirmed pathologically.

Cytokine Gene Therapy of Malignant Glioma by Means of DNA/Liposomes

During the past two decades, there have been major advances in our understanding of tumor-associated genes that control cell growth and differentiation and in techniques using a variety of viral and nonviral vectors for delivering genes into mammalian cells. These developments have provided us with opportunities of gene therapy for cancer patients, and there is a rapidly growing number of cancer gene therapies worldwide. In this chapter, molecular abnormalities in the evolution of malignant glioma and recent basic experimental studies of gene therapy are reviewed. Our approach to gene therapy for brain tumors is selective in vivo gene transfer by means of immunoliposomes entrapping a plasmid vector. We have demonstrated in our experimental studies that a remarkable antitumor effect could be obtained by intratumoral injection of the liposome-entrapped interferon-β gene or the tumor necrosis factor-α gene.

Incidental Meningiomas in Autopsy Cases in Computed Tomography and Magnetic Resonance Imaging Era

Between January 1980 and June 1994, autopsy was performed on 1979 cases at the Division of Neuropathology, Montefiore Medical Center. Intracranial meningioma was detected incidentally in 31 of these autopsies. These 31 cases were analyzed clinically and pathologically: the male-to-female ratio was about 1:2, and the average age at death was 76.8 years. A positive correlation was noted between tumor diameter and age. On histological study, the percentage of the psammomatous type of tumor was higher than that previously reported for clinical cases. When the proliferative activity was examined using antiproliferating cell nuclear antigen (anti-PCNA) antibody and silver-stained nucleolar organizer regions (Ag-NORs), the proliferative activity appeared lower in the psammomatous type than in any other histological type. These results suggest that clinical cases of meningioma incidentally detected in elderly persons have a low proliferative activity and persist in the individual for long periods of time.