Tatsuo Murakami

A NEW HUMAN PLEOMORPHIC HEPATOCELLULAR CARCINOMA CELL LINE, KYN‐2

A new human hepatocellular carcinoma (HCC) cell line, KYN‐2, has been established from a surgical specimen obtained from a 52‐year‐old Japanese male HCC patient. The originally resected HCC was classified as pleomorphic HCC corresponding to Edmondson‐Steiners grade III with a thick trabecular to solid arrangement. The cell line has been maintained for 17 months through 35 passages. Morphologically, the KYN‐2 cells have retained the characteristics of the original HCC, being pleomorphic and composed of various types such as cells with relatively small, polygonal, eosinophilic cytoplasm and oval‐shaped nuclei with a marked tendency to pile up, flat cells with abundant clear cytoplasm and oval‐shaped nuclei, and many multinucleated giant cells, proliferating in a pavement‐like cell arrangement. Some junctional complexes and a number of microvilli are evident between the cells by electron microscopy. Functionally, these cells were found to secrete albumin, α,‐acid glycoprotein, α1‐antitrypsin, ceruloplasmin, transferrin, complement C, fibrinogen, fibronectin, prothrombin, retinol‐binding protein (serum type), α‐fetoprotein (AFP), carcinoembryonic antigen (CEA), ferritin and β2‐microglobulin in chemically defined medium (CDM). The secretion of AFP and CEA is apparently dependent upon culture medium and passage. The doubling time of cells growing in serum‐containing medium at the 14 th passage was 84 h, and those of cells in serum‐containing medium, HB101 (serum‐free medium) and CDM at late passage were 28,68, and 42 h, respectively. Chromosome analysis revealed that the chromosome number ranged from 56 to 69 without a mode, and the presence of marker chromosomes. HB virus DNA sequence was not detected by hybridization analysis. The tumorigenicity of KYN‐2 cells was identified by development of tumors in nude mice after subcutaneous injection of the cells; the tumors showed an appearance basically similar to that of the original HCC. Thus, these findings suggest that the KYN‐2 cell line is available as a new human HCC cell line and should be useful for various studies on HCC. ACTA PATHOL JPN 38: 953‐966, 1988.

Argininemia: Report of a New Case and Mechanisms of Orotic Aciduria and Hyperammonemia

Arqininemia is a rare inborn error of ureagenesis due to a deficiency of arginase activity (Terheggen et al., 1969; Terheggen, et al., 1970a; Terheggen et al., 1970b; Terheggen et al., 1975; Cederbaum et al., 1976; Snyderman et al., 1977; Cederbaum et al., 1977; Michels and Beaudet, 1978; Snyderman et al., 1979). Despite the apparent defect in enzyme activity, hyperammonemia is only intermittently observed in this disease, unlike other enzymopathies of the urea cycle. Increase in orotic acid excretion is also another biochemical characteristic of this disease (Snyderman et al., 1977; Bachmann and Colombo, 1980). However, the mechanisms of hy-perammonemia and orotic aciduria have not been thoroughly explained. The purpose of this communication is to describe clinical features at 2 hours of testing. Concentration of arginine was elevated also in cerebrospinal fluid (1.91 mg/dl, normal range; 0.15–0.55 mg/dl), orotic aciduria.

HETEROTRANSPLANTATION OF AN ALPHA‐FETOPROTEIN‐PRODUCING HUMAN GALLBLADDER CARCINOMA INTO NUDE MICE

An alpha‐fetoprotein (AFP)‐producing human gallbladder carcinoma showing direct invasion into the liver was transplanted into BALB/c‐nu/nu nude mice. Although patient serum levels of AFP and carcinoembryonic antigen (CEA) were within normal limits, they were elevated to 1,040 ng/ml and 22.1 ng/ml, respectively, after cholecystectomy. Prominent liver metastasis was demonstrated by diagnostic imaging techniques shortly after the operation. Pathologically, the resected tumor consisted of papillotubular adenocarcinoma and the part which had Invaded the liver showed a solid growth pattern with no papillo‐tubular structure. The transplanted tumor showed both papillo‐tubular and solid growth patterns, in which positive reactions for AFP, CEA, ferritin (FER), carbohydrate antigen 19‐9 (CA 19‐9), albumin (ALB) and fibrinogen (FIB) were confirmed by the avidin‐biotin‐peroxidase complex method. Serum levels of AFP, CEA, CA 19‐9, β2‐microglobulin (BMG) and FER were elevated in the nude mice bearing tumor transplants. Twenty‐five percent of the serum AFP from nude mice with tumor transplants bound with concanavalin A (Con A), suggesting that the tumor was of gastrointestinal rather than hepatic origin.

Argininemia: Report of a Case

A case report of argininemia is described. A female neonate born to nonconsanguineous and healthy parents after uncomplicated pregnancy and delivery was admitted to the hospital because of convulsion. She began vomiting after feeding on the 18th day of life. Sb.e developed fever complicated by clinic convulsion and opisthotonic posture on the 23rd day of life. Convulsive episodes have recurred intermittently and her neurological developrnent has been arrested since her early infancy. At 4 years of age, she was admitted to hospital because of pneumonia. Physical examination on admission revealed a markedly retarded girl with microcephaly, spastic tetraplegia and intermittent convulsion. Hyperammonemia (960 pg/dl, normal range: 70120 pg/dl) and a marked increase in orotic acid in urine (6,900 pg/mg creatinine, normal range: 5-10 pg/mg creatinine) were found on the 13th day of pneumonia. Serum arginine was slightly elevated (3.1 mg/dl) and ornithine and citrulline were low but within the normal ranges at this time. After protein feeding was reinstituted, serum arginine rose to a maximum value of 19.4 mg/dl, whereas plasma ammonia and orotic acid excretion declined to normal or slightly elevated values. Blood urea nitrogen remained low (1.4-5.7 mg/dl) during this period. Arginase activity of the patient’s