Hiromichi Mannoji

Glial fibrillary acidic protein in medulloblastoma

SummaryTwenty-four cases of classical medulloblastoma and one case of desmoplastic medulloblastoma were examined for glial fibrillary acidic protein (GFAP) using the immunoperoxidase method to assess astrocytic differentiation. In 16 cases of classical medullablastoma GFAP-positive cells were present in variable numbers.These cells were classified as three different types according to size and shape. The type 1 cell was morphologically identical to the ordinary tumor cell, with a hyperchromatic nucleus and a scanty cytoplasm. The type 2 cell had a fairly rich cytoplasm with short cytoplasmic processes. The type 3 cell was characterized by a relatively large nucleus with sparse chromatin and well-developed cytoplasmic processes, and was considered a reactive astrocyte. The type 1 and some of the type 2 cells seemed to be neoplastic, displaying astrocytic differentiation. The remaining type 2 cells may have been reactive astrocytes.In one case of desmoplastic medulloblastoma, the majority of GFAP-positive cells were arranged in “islands”, and had delicate fibrillated processes. GFAP-positive cells were also observed outside these “islands”, though they were less numerous. Most of them were regarded as type 3 cells, but some were type 2. This may be interpreted as meaning that the glial character of the tumor was expressed more within than outside these “islands”.

Interferon effects on multiplication, cytoplasmic protein and GFAP content, and morphology in human glioma cells.

Beta-type human fibroblast interferon inhibited the multiplication of human glioma cells in a dose-dependent manner. The inhibitory effects were thought to be due to suppression of the cells entering the S-phase. After interferon treatment with 5 \sX 103 IU/ml for ten days, the mean cell volume, the soluble protein, and the glial fibrillary acidic protein increased to 970%, 190%, and 380% that of the controls, respectively. The interferon-treated cells changed shape and resembled mature astrocytes. It is presumed that beta-type human fibroblast interferon inhibits DNA synthesis of cultured glial cells and subsequently induces specific differentiated proteins and morphologic alteration.

Intraventricular cavernous malformation associated with medullary venous malformation.

We reported a case of cavernous malformation (CM) associated with medullary venous malformation in the same area. The CM was located in the trigone of the lateral ventricle in the dominant hemisphere and drained into the longitudinal caudate vein of Schlesinger via deep medullary veins. By a transsylvian transventricular approach, the CM was totally removed with successful preservation of the medullary venous malformation. This approach is available for trigonal lesions, especially in cases with enlarged inferior horn. We stress that CM removal can be conducted with preservation of the adjacent medullary venous malformation.

Cilostazol Improves Outcome after Subarachnoid Hemorrhage: A Preliminary Report

Background: Cerebral vasospasm (VS) is the most common cause of morbidity and mortality after aneurysmal subarachnoid hemorrhage (SAH). Reversal of VS by intra-arterial infusion of cyclic adenosine monophosphate (cAMP)-elevating agents has been reported; however, the preventive role in the development of VS is not fully understood. This study is designed to evaluate the possible efficacy of using cilostazol, a selective inhibitor of phosphodiesterase type 3 and a cAMP-elevating agent, in patients with SAH. Methods: In this prospective randomized study, we enrolled 100 SAH patients who met the following criteria: neck clipping within 72 h after onset, Hunt and Hess (HH) score ≤4, modified Rankin scale (mRS) score ≤2 prior to ictus, and no serious cardiovascular complications. Patients were divided into control and cilostazol groups; we focused on the effects of cilostazol on the decrease in the incidence of symptomatic VS, cerebral infarction, and the mRS score at discharge. Result: Patients’ age, male/female ratio, mRS score prior to ictus, HH grade, Fisher group, site of the aneurysm, drugs prescribed during the observation period, and length of hospital stay were not different between the groups. Cilostazol did not significantly decrease the incidence of symptomatic VS (37.3% in the control vs. 22.4% in the cilostazol group, p = 0.183) and cerebral infarction (27.5% in control vs. 10.2% in the cilostazol, p = 0.091). However, mRS score was significantly improved at discharge (2.6 in controls vs. 1.5 in the cilostazol group, p = 0.041). Patients’ age being ≤65 years (OR = 8.47, 95% CI = 2.45–29.32, p = 0.0007), Fisher group ≤3 (OR = 4.64, 95% CI = 1.00–21.45, p = 0.049), HH grade ≤2 (OR = 4.31, 95% CI = 1.27–14.59, p = 0.019), no hydrocephalus (OR = 8.55, 95% CI = 1.72–19.23, p = 0.0046), and cilostazol use (OR = 5.52, 95% CI = 1.61–18.90, p = 0.0065) were independent predictors of good outcomes (mRS score ≤2). Conclusion: Cilostazol may improve outcomes after SAH, but further double-blind, placebo-controlled studies are required for a definitive conclusion.

Establishment and Maintenance of a Human Glioma Transplanted Serially to Hereditary Asplenic-Athymic (Lasat) Mice

A tumor line in asplenic-athymic (lasat) mice designated KNS-GL-2 was established from a recurrent anaplastic astrocytoma of the cerebrum in a 15-year-old boy (NS761072). The tumor line increased in terms of the rate of tumor take and growth with serial passage, forming a nodular appearance and massive intratumoral hemorrhage at the third generation. Microscopically, tumors developed in lasat mice wee similar to the original tumor and consisted of cells with round and oval nuclei and a clear cytoplasm, which was positive for S-100 and GFA proteins by the immunoperoxidase method. Large clusters of tumor cells with dark scanty cytoplasm seen at the third generation lost their astrocytic appearance and were negative for both S-100 and GFA proteins. This was considered to be evidence for dedifferentiation or malignant transformation. There was a scattering of tumor cells with clear cytoplasm in these clusters, with a positive reaction to S-100 protein. It was possible that tumor cells dedifferentiated during serial passage but still retained the potential to differentiate into astrocytes.

Alternate culture and animal passage of human glioma

SummaryThe method of the alternate culture and animal passage was introduced in the study of human glioma. For animal passage the hereditary asplenic-athymic (lasat) mice were used as a carrier. Because the lasat mice have practically no cellular and only little humoral immunity, the rate of tumor take was expected to be raised, and successful results were obtained. Ultrastructural and immunohistochemical (GFA protein) studies were also done. The overgrowth of stromal elements in reculture of tumors in lasat mice was less vigorous than in athymic nude mice.After four passages through lasat mice, an established human glioma cell line, KNS-42, showed numerous well differentiated cell nests, and this alternate culture and animal passage suggested to enhance the differentiation and growth capacity. After three passages through lasat mice, the tumor line, KNS-42-L, produced tumors also in athymic nude mice and their histological features were essentially the same as those in lasat mice, and hence the lasat mice could be saved.