Akira Nakagawara

Analysis of N-myc amplification in relation to disease stage and histologic types in human neuroblastomas

Both untreated and treated primary neuroblastomas from 52 patients were analyzed to determine the correlation between the amplification of N‐myc oncogene and various prognostic factors. Amplification of N‐myc was observed in eight of 28 untreated cases and in 12 of 24 treated cases. As a whole, 12 of 18 tumors (67%) in Stage IV had N‐myc amplification, but there were fewer cases in the unadvanced disease stage, as reported previously by others. Furthermore, the authors detected N‐myc amplification in three of nine tumors in Stage IV‐S, although the amplification was less than 50 copies. Analysis of progression‐free survival at 24 months revealed that amplification of N‐myc was associated with the worst prognosis (P < 0.001). In the untreated group, no amplification of N‐myc was detected in any of two ganglioneuromas and four ganglioneuroblastomas, whereas amplification of N‐myc was observed in all two round‐cell and six of 20 rosette fibrillary neuroblastomas. On the other hand, the authors detected amplification of N‐myc in three of eight less differentiated ganglioneuroblastomas in the treated group and observed the worst prognosis in these three patients. The total percentage of the cases from both untreated and treated groups suggest that amplification of N‐myc may occur more frequently in undifferentiated types of neuroblastomas than in less malignant types. In conclusion, the amplification of N‐myc in neuroblastomas was closely associated with the worst prognosis, which was suggested by both disease stage and histologic characteristics.

Generation of superoxide anions by leucocytes treated with cytochalasin E

Abstract Guinea pig polymorphonuclear leucocytes reduced cytochrome c when treated with cytochalasin E. The reduction was completely inhibited by superoxide dismutase and manganese ions, which indicates that superoxide anions are generated and released into the outside medium by the treatment. The reduction was inhibited by glycolytic inhibitors and cyclic AMP but not by cyclic GMP. The pattern is similar to the cyanide-insensitive respiration of leucocytes during phagocytosis. Nitroblue tetrazolium was also reduced by the leucocytes treated with the cytochalasin, which was inhibited by manganese ions, glycolytic inhibitors and cyclic AMP but was only partially inhibited by superoxide dismutase.

Malignant pheochromocytoma with ganglioneuroblastomatous elements in a patient with von recklinghausen's disease

A 14‐year‐old girl with numerous café‐au‐lait spots in her skin was hospitalized because of fever, weight loss, and a mass of the right upper quadrant of the abdomen. Despite intensive chemotherapy, she died 6 months after admission. The autopsy revealed a right adrenal tumor with metastases to liver, lungs, vertebrae, and lymph nodes. Histologically the tumor was a pheochromocytoma with small foci of ganglioneuroblastoma. The catecholamine contents of the tumor were markedly elevated, as confirmed by the catecholamine fluorescence technique. Electron microscopically, the tumor cells contained intracytoplasmic membrane‐bound chromaffin granules of varying sizes and shapes. This may be the first report of the concomitant occurrence of malignant catecholamine‐secreting pheochromocytoma with ganglioneuroblastomatous elements in a patient with von Recklinghausens disease.

Extraskeletal myxoid chondrosarcoma in young children

Two extraskeletal myxoid chondrosarcomas with a solid soft tissue mass occurred on the right upper arm of a 4‐year‐old boy and on the chest wall of a 1‐year‐old boy. Microscopically, both tumors were characterized by lobular configuration and were sparsely cellular with a background of myxoid matrix. The cells were small and round, and appeared undifferentiated, sometimes with a narrow eosinophilic cytoplasm. They grew in nests or strands and sometimes in a single file. They were strongly positive for S‐100 protein and vimentin. Ultrastructural features suggested that the cells had a poorly differentiated mesenchymal nature with chondrocytic differentiation. These are the sixth and seventh reported cases of extraskeletal myxoid chondrosarcoma occurring in children. There are definite differences between this tumor with immature features and the extraskeletal myxoid chondrosarcoma in adults. Problems of differential diagnoses from other small round cell sarcomas also are discussed.

N-myc oncogene and stage IV-S neuroblastoma : preliminary observations on ten cases

We studied the clinical significance of genomic amplification of N‐myc in Stage IV‐S neuroblastoma, with reference to spontaneous regression. Among 103 neuroblastomas in which N‐myc was measured, ten were Stage IV‐S (eight children were younger than and two were older than 1 year of age). The number of copies of N‐myc was 1 to 3 in five patients, four to ten in one patient, and more than ten in four patients, and the survivors of each group were four, one, and one (recurrent), respectively. Of 41 patients younger than 1 year of age, N‐myc amplification of more than three copies was found only in Stage IV‐S neuroblastoma. Cure with a tendency to regress spontaneously was seen in five of eight patients younger than 1 year of age. However, two patients older than 1 year of age classified as Stage IV‐S (one with N‐myc amplification) died of progressive disease. In two patients (1 and 3 months of age) with a huge hepatic involvement and in whom the tumor had an amplified N‐myc of more than ten copies, tumor regression occurred but there was a relapse to a progressive state later. The overexpression of N‐myc mRNA occurred in nine of ten stage IV‐S tumors and did not correlate with the prognosis. The vanillylmandelic acid (VMA) to homovanillic acid (HVA) ratio was low in tumors with an increased number of copies of N‐myc. Serum lactate dehydrogenase (LDH) levels were increased in Stage IV‐S patients with N‐myc amplification but not in those with regressing tumors and without N‐myc amplification. These data suggest that N‐myc amplification may affect the final outcome in the patient classified as Stage IV‐S, but tumor regression can occur early after birth and appears to be independent of N‐myc amplification.

Hepatoblastoma Producing Both Alpha-Fetoprotein and Human Chorionic Gonadotropin Clinicopathologic Analysis of Four Cases and a Review of the Literature

A clinicopathologic study was done of four cases of hepatoblastoma with precocious puberty, together with an analysis of 21 such cases reported in the literature. All four patients were males, the age ranging from 11 to 35 months. Three of the four patients died within 12 months after operation, but the fourth is living. Histologically, all patients had a hepatoblastoma with the coexistence of both fetal and embryonal type cells, although predominantly fetal in two and predominantly embryonal in the other two. Tumor giant cells were rarely encountered in all these cases. Both alpha‐fetoprotein (AFP) and human chorionic gonadotropin (hCG) in the serum or urine increased in our four cases and in five other cases reported in the literature. Though the serum AFP level paralleled the severity of clinical symptoms, the serum or urine hCG did not necessarily correspond to the clinical course. It is likely that these poorly prognostic virilizing hepatoblastomas secrete two different tumor markers, AFP and hCG, from different cells, and that these functioning tumor cells may not always exist concurrently in the recurrent or metastatic tumor. Cancer 56: 1636‐1642, 1985.

Patterns of destruction of mouse neuroblastoma cells by extracellular hydrogen peroxide formed by 6-hydroxydopamine and ascorbate

SummaryThe patterns of the cytolytic effects of 6-hydroxydopamine (6-OHDA), with/without ascorbate, on C-1300 and three other cloned mouse neuroblastoma cell lines (N1E-115, NS-20, N-18) were studied in vitro. The sensitivity to 6-OHDA differed and the three cloned cell lines were more sensitive than the wild type C-1300 cell line. Ascorbate synergistically potentiated the cytolytic effect of 6-OHDA to all four cell lines. The 6-OHDA cytotoxicity was eliminated by the addition of exogenous catalase but not by addition of other oxygen free radical scavengers, thereby suggesting that the hydrogen peroxide formed might influence the cells, extracellularly. In addition, the critical time for tumor cell lysis was the first 60 min of the reaction. The cytotoxicity induced by the unmasked cyclophosphamide, 4-hydroperoxycyclophosphamide, was synergistically enhanced in the presence of a nontoxic concentration of 6-OHDA and ascorbate. These data suggest that reactive oxygen intermediates may prove to be a good tool for destroying neuroblastoma cells.

Role of cytoskeletal elements in cytochalasin E-induced superoxide production by human polymorphonuclear leukocytes

The release of superoxide anions from human polymorphonuclear leukocytes induced by cytochalasin E was greatly enhanced by the pretreatment of the cells either with deuterium oxide or with concanavalin A. Colchicine, vinblastine and cyclic AMP inhibited the release. Cytochalasins A and B also suppressed the superoxide release. These observations suggest the involvement of microfilament-microtubule system in the production and release of superoxide anions induced by cytochalasin E.

Combined effects of vitamin E (alpha-tocopherol) and cisplatin on the growth of murine neuroblastoma in vivo

Combined effects of vitamin E (alpha-tocopherol) and cisplatin on the growth of two murine neuroblastomas (C1300, NS-20) was investigated in vivo. Five groups of mice were prepared; group 1 were fed the control diet, group 2 were fed a vitamin E-deficient diet, group 3 were fed a vitamin E-supplemented diet, group 4 were fed the control diet and plus vitamin E solution given intraperitoneally during the treatment (solvent i.p. group), and group 5 were given vitamin E in the same manner (20 mg/kg/day; vitamin E i.p. group). Cisplatin (6 mg/kg) was injected intraperitoneally into the mice of each group during the treatment. In case of the C1300 neuroblastoma, the antitumor activity of cisplatin was most enhanced in the mice receiving vitamin E i.p., and the intra-tumor vitamin E and platinum levels were significantly higher in this group than in the other groups (P less than 0.01, and P less than 0.05 respectively). In contrast, in animals transplanted with the NS-20 murine neuroblastoma, which proved to be a cisplatin-tolerant tumor in separate experiments, no combined effect of those drugs was observed, although the intra-tumor level of platinum was elevated. The possibility was that vitamin E increases the influx of cisplatin into the tumor cells and acts after incorporation of cisplatin through the plasma membrane. Vitamin E did not accentuate the cisplatin-induced renal impairment in vitamin E-loaded groups. Those results suggested that vitamin E should be considered as a co-agent of cisplatin for the treatment of neuroblastoma.

Identification of a carrier mother of a female patient with chronic granulomatous disease.

5. Havlikova D, Vychytil O, and Jelinek J: The syndrome of congenital pancreatic insufficiency; chronic respiratory disease and chronic liver damage, Acta Paediatr Scand 56:676, 1967. 6. Townes PL: Proteolytic and lipolytic deficiency of the exocrine pancreas, J PEDIATR 72:221, 1969. 7. Morris MD, and Fisher DA: Trypsinoge n deficiency disease, Am J Dis Child 114:203, 1967. 8. Grand RJ, Rosen SW, and di Sant Agnese PA, et al: Unusual case of XXY Klinefelters syndrome with 10. pancreatic insufficiency, hypothyroidism, deafness, chronic lung disease, dwarfism and microcephaly, Am J Med 4U478, 1966. Donowitz M, and Gryboski JD: Pancreatic insufficiency and the congenital rubella syndrome, J PEDIAT~ 87:241, 1975. McKusick VA, Eldridge R, and Hostetler JA, et al: Dwarfism in the Amish II. Cartilage hair hypoplasia, Bull Hopkins Hosp 116:285, 1965.