4,4'-Diaminodiphenylmethane: promoting effect on the development of thyroid tumors in rats treated with N-bis(2-hydroxypropyl)nitrosamine.

4,4'-Diaminodiphenylmethane (4,4'-methylenedianiline) (DDPM) promoted the development of thyroid tumors in rats treated with a subeffective dose of N-bis(2-hydroxypropyl)nitrosamine (2,2'-dihydroxy-N-nitrosodipropylamine) (DHPN) for thyroid tumorigenesis. Male inbred W rats were given a single ip injection of 280 mg DHPN/100 g body weight and fed diets with or without 1,000 ppm DDPM. Thyroid tumor incidences at the end of week 20 of the experiment were 90% (19/21) in rats given DHPN and then DDPM and 28% (6/21) in rats given DHPN alone. The incidence of thyroid cancers was 9.5% (2/21) in rats first given DHPN and then DDPM. Untreated rats and rats given DDPM alone had no thyroid tumors after 20 weeks. Incidences of kidney tumors were 38% (8/21) in rats given DHPN and then DDPM and 28% (6/21) in rats given DHPN alone. No tumors were found in the kidneys and lungs of rats given DDPM alone and in those of control rats. Treatment with DDPM alone slightly but not significantly decreased the serum concentrations of thyroxine and triiodothyronine; treatment with DHPN plus DDPM had no such effect.

Trisodium nitrilotriacetate monohydrate: promoting effects on the development of renal tubular cell tumors in rats treated with N-ethyl-N-hydroxyethylnitrosamine.

The effect of trisodium nitrilotriacetate monohydrate [N,N-bis(carboxymethyl)glycine trisodium salt] (Na3NTA X H2O) on development of renal tubular cell tumors induced with N-ethyl-N-hydroxyethylnitrosamine [CAS:13147-25-6; 2-(ethylnitrosamino)-ethanol] (EHEN) was studied. Six-week-old male inbred W rats were given a diet containing 1,000 ppm of EHEN for 2 weeks and then a diet containing a high (10,000 ppm) or low (500 ppm) concentration of Na3NTA X H2O for 30 weeks. The rats were killed during week 32. The higher concentration of Na3NTA X H2O enhanced the development of renal tubular cell tumors and increased the number and size of tumors in rats treated with EHEN, but the lower concentration of Na3NTA X H2O did not. The incidence of renal tubular cell tumors in week 32 was 33% in rats treated with 1,000 ppm EHEN for 2 weeks, 100% in rats treated with 1,000 ppm EHEN for 2 weeks plus high Na3NTA X H2O diet for 30 weeks, and 39% in rats treated with 1,000 ppm EHEN for 2 weeks and then given low Na3NTA X H2O diet for 30 weeks. Numbers of atypical cell foci per kidney area (No./cm2) were 17.0 +/- 7.6 in rats treated with EHEN and high Na3NTA X H2O, 7.3 +/- 2.2 in rats treated with EHEN and low Na3NTA X H2O, 3.7 +/- 1.4 in rats treated with EHEN alone, and 1.0 +/- 2.4 in rats treated with high Na3NTA X H2O diet alone. Atypical cell foci retained a tubular pattern and consisted of basophilic cells with a large nucleus or clear cells with a small nucleus.

DL-Serine: promoting activity on renal tumorigenesis by N-ethyl-N-hydroxyethylnitrosamine in rats.

Subcutaneous injection of DL-serine increased the number and size of renal tubular cell tumors in male W rats treated with 500 or 1,000 ppm N-ethyl-N-hydroxyethylnitrosamine [(EHEN) CAS: 13147-25-6, 2-(ethylnitrosamino)ethanol]. At the end of the 32-week experiment, the incidences of renal tumors were 95% in rats treated with 1,000 ppm EHEN for 2 weeks and then given three sc injections of DL-serine every 2 weeks, 33% in rats treated with 1,000 ppm EHEN for 2 weeks, and 28% in rats treated with 500 ppm EHEN for 2 weeks and then given three sc injections of DL-serine every 2 weeks. No renal tumors were found in rats treated with 500 ppm EHEN alone or given three sc injections of DL-serine alone every 2 weeks.

Histopathological and biochemical analyses of transplantable renal adenocarcinoma in rats induced by N-ethyl-N-hydroxyethylnitrosamine.

Transplantable renal adenocarcinoma can be readily induced in Wistar strain male rats by initiation with N-ethyl-N-hydroxy-ethylnitrosamine followed by promotion with beta-cyclodextrin. The transplantability rates of the tumors by s.c. inoculation in newborn rats were 33 and 50%, respectively, for tumors of the first and second passages, and 100% for both third and fourth passages. The transplantability rates were affected by route of inoculation; rates of 50 and 100% were observed for s.c. and i.p. inoculations, respectively. The growth rate of tumors induced by i.p. inoculation was 3-fold higher than that induced by s.c. injection. Macroscopically, most of the tumors grew in the s.c. tissue of inoculation sites. However, invasive growth of tumors in spleen, liver, stomach, peritoneum, and intestine were seen in 50% of the animals inoculated i.p.; metastatic cancers to lung were seen in 16%. Histologically, the tumors were well-differentiated adenocarcinomas composed of uniform cells resembling kidney tubular cells and appeared to be derived from normal kidney tissues. A 5-fold decrease in gamma-glutamyl transferase activity in tumor tissues was found as compared with that of nontumorous kidney tissues. Electrophoretic analysis of cellular proteins in polyacrylamide gels revealed that tumor tissues exhibited five new polypeptides with molecular weights of 81,000, 64,000, 59,000, 50,000, and 36,000 which were either lacking or undetectable in the nontumourous area and control kidney. In addition, protein banding patterns of transplantable renal tumor appeared to be more heterogeneous than those of primary kidney tumor.

Chrymutasins: novel-aglycone antitumor antibiotics from a mutant of Streptomyces chartreusis. I. Taxonomy, mutation, fermentation, isolation and biological activities.

Three novel antibiotics, named chrymutasins A, B and C, were isolated from the fermentation products of a mutant strain obtained by NTG (N-methyl-N'-nitro-N-nitrosoguanidine) treatment. The mutant strain produced the chrymutasins, which differed in the aglycone moiety from chartreusin, and related compounds. The production of these compounds needed a characteristically long fermentation period. The antitumor activity of chrymutasin A is better in vivo than that of chartreusin, the cytotoxic activity against cell lines in vitro is equivalent, and the antimicrobial spectrum is narrower.

Chrymutasins: novel-aglycone antitumor antibiotics from a mutant of Streptomyces chartreusis. II. Characterization and structural elucidation.

Chrymutasins A, B and C are glycosidic antibiotics produced by a mutant of the chartreusin producer-organism Streptomyces chartreusis. We report here the structure elucidation of these compounds. The sugar moieties involved were determined by comparison with the related chartreusins. The structure of the aglycone, the same in all three compounds, was elucidated by NMR, incorporation studies of labeled compounds and synthesis of derivatives. The chrymutasin aglycone differs from that of chartreusin by a single carbon and an amino group.

Novel antitumor antibiotics, saptomycins. I. Taxonomy of the producing organism, fermentation, HPLC analysis and biological activities.

Streptomyces sp. HP530 was found to produce novel antitumor antibiotics, saptomycins, closely related to the pluramycin-group and was further found to mutate frequently. The natural mutant produced several new saptomycins as determined by HPLC analyses. We isolated saptomycins A, B, C1, C2 and F from the parent strain and saptomycins D, E, G and H from the mutant. The saptomycins showed antimicrobial activities and potent antitumor activities against human or murine tumor cell lines in vitro and against Meth A fibrosarcoma in vivo. In particular, saptomycin D was most effective component in vivo of all saptomycins.

Novel antitumor antibiotics, saptomycins. II. Isolation, physico-chemical properties and structure elucidation.

A complex of novel antitumor antibiotics related to the pluramycin-group was isolated from the fermentation of actinomycete, named Streptomyces sp. HP530. The producing strain mutated frequently. The products isolated from the parent strain were designated saptomycins A, B, C1, C2 and F, while those of the mutant were named saptomycins D, E, G and H. These structures were elucidated by several NMR spectral analyses and other spectroscopic experiments.

Novel cytocidal compounds, oxopropalines from Streptomyces sp. G324 producing lavendamycin. I. Taxonomy of the producing organism, fermentation, isolation and biological activities.

In the course of our investigation aimed at the discovery of novel antitumor antibiotics from microorganisms, Streptomyces sp. G324 was found to produce the antitumor antibiotic, lavendamycin, and also, to yield the novel beta-carboline compounds, oxopropalines. We isolated five compounds as oxopropalines A, B, D, E and G. Oxopropalines B, D and G showed cytocidal activities against human or murine tumor cell lines in vitro.

Novel cytocidal compounds, oxopropalines from Streptomyces sp. G324 producing lavendamycin. II. Physico-chemical properties and structure elucidations.

Novel cytocidal compounds designated oxopropalines A, B, D, E and G were isolated from the fermentation of an actinomycete named Streptomyces sp. G324, a strain that also produced an antitumor antibiotic, lavendamycin. All these compounds possessed a beta-carboline chromophore. The structures of the oxopropalines were elucidated by several NMR spectral analyses and other spectroscopic experiments.

Glycolytic enzyme Pgk1 is strongly expressed in the developing tooth germ of the mouse lower first molar.

This study examined detailed in situ expression patterns and possible functional roles of phosphoglycerate kinase 1 (Pgk1) gene in the developing tooth germ of the mouse lower first molar. The strong expression of Pgk1 mRNA was seen in the odontogenic epithelial cells and surrounding mesenchymal cells of the tooth germ from embryonic day 10.5 (E10.5) to E18.0. Western blotting analysis demonstrated that Pgk1 protein formed 84-kDa protein complex in these embryonic organs. The results of immunoprecipitation-western blotting also suggested this complex to be formed with glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Moreover, the immunofluorescence expression of those proteins was shown to overlap each other in the tooth germ at E15.0. A strong immunofluorescence expression of both Pgk1 and GAPDH also corresponded to the in situ expression of those mRNAs. These results suggested that Pgk1 plays some functional roles in the development of tooth germ and other embryonic organs by forming protein complex with GAPDH.

Congenital patent urachus in an adult: a case report.

A rare case of the congenital patent urachus in an adult is reported. The patient, a 42-year-old man, was admitted with the chief complaint of macroscopic hematuria. During the cystoscopy perfusion water discharged from the umbilicus. Diagnostic imaging revealed a tubular fistula between the bladder and umbilicus and the patient underwent urachal resection and partial cystectomy. Microscopically the lumen was lined by transitional, columnar and squamous epithelium and there was no evidence of malignancy. To date 11 cases of congenital patent urachus in an adult have been reported in Japan. From a review of the Japanese literature, the common chief complaint is urine discharge from the umbilicus accompanying cystitis and/or urachal inflammation. Urachal resection is performed in all cases. We also clarified the epithelial histology of the patent urachus in this patient.

Effect of varying the duration of exposure to phenobarbital on its enhancement of N-bis(2-hydroxypropyl)nitrosamine-induced thyroid tumorigenesis in male Wistar rats.

Male Wistar rats were injected s.c. with a single dose of 280 mg N-bis(2-hydroxypropyl)nitrosamine (DHPN) per 100 g body weight and the effect of subsequent exposure to 500 p.p.m. phenobarbitone (PB) for 6, 12 and 19 weeks on development of thyroid tumors was studied. Exposure to PB for 12 or 19 weeks enhanced the development of thyroid tumors in the rats. The incidences of thyroid tumors in DHPN-treated rats at the end of the experiment for 20 weeks were 87%, 83%, 42% and 37%, respectively, in rats given PB for 19 weeks, 12 weeks, and 6 weeks, and in those given no PB. The incidence of thyroid cancers was 12% in rats given DHPN and then PB for 19 weeks. The total numbers of thyroid tumors were 241, 125, 65 and 23, respectively, in DHPN-treated rats given PB for 19 weeks, 12 weeks, and 6 weeks, and in those given no PB. Thus, the total numbers of thyroid tumors were correlated with the period of PB treatment.

Effect of propylthiouracil on the thyroid tumorigenesis induced by N-bis(2-hydroxypropyl)nitrosamine in rats.

The effect of 0.15% propylthiouracil (PTU) on thyroid tumorigenesis was studied in male Wistar rats given a single i.p. injection of 280 mg of N-bis(-2-hydroxypropyl)nitrosamine (DHPN) per 100 g body weight. The mean weights of the thyroid of rats treated with DHPN followed by PTU and with PTU alone were significantly higher than those of rats treated with DHPN only and control rats. The incidences of follicular adenoma at the end of week 20 of the experiment were 100% (21/21) in rats treated with DHPN followed by PTU, and 19% (4/21) in rats given DHPN alone. Papillary adenoma was observed in one rat treated with DHPN followed by PTU. The incidence of follicular carcinomas with invasive growth into the capsule, adipose tissues or blood vessels was 52% (11/21) in rats given DHPN and then PTU. No papillary carcinomas or solid tumors were found in any rats. Rats given PTU alone and untreated rats had no thyroid tumors. The serum concentration of T4 in rats treated with PTU alone was significantly lower than that in the control group. The serum concentration of T4 in rats treated with DHPN followed by PTU was slightly, but not significantly, lower than that in control rats. The serum concentrations of T3 in rats treated with DHPN followed by PTU, DHPN alone and PTU alone were also slightly, but not significantly, lower than that in controls.

Promoting effect of phenobarbital on N-bis (2-hydroxypropyl)nitrosamine thyroid tumorigenesis in rats. Effect of varying duration of exposure to phenobarbital.

Male Wistar rats were injected subcutaneously with a single dose of 280 mg N-bis(2-hydroxypropyl)nitrosamine (DHPN) per 100 g body weight and then given a diet containing 500 mg/kg phenobarbital (PB) for 6, 12 and 19 weeks. Exposure to PB for 12 or 19 weeks enhanced the development of thyroid tumours in rats treated with DHPN. The incidences of thyroid tumours at the end of the experiment were 87% in rats given DHPN and then PB for 19 weeks, 83% in rats given DHPN and then PB for 12 weeks, 42% in rats given DHPN and then PB for 6 weeks, and 37% in rats given DHPN. The incidence of thyroid cancer was 12% in rats given DHPN and then PB for 19 weeks. The total numbers of thyroid tumours were: 241 in rats given DHPN and then PB for 19 weeks, 125 in rats given DHPN and then PB for 12 weeks, 65 in rats given DHPN and then PB for 6 weeks, and 23 in rats given DHPN alone. The total numbers of thyroid tumours were proportional to the period of exposure to PB.