A novel galacto-oligosaccharide (GOS) manufactured by a two-step enzyme reaction of lactose was examined in a comet assay for its potential to induce DNA damage in vivo by estimating the DNA fragmentation level in the cellular nuclei of the glandular stomach, colon, and peripheral blood. GOS was orally administered at doses of 0 (vehicle alone), 500, 1000, and 2000 mg/kg/day to five male Crl: CD(Sprague Dawley) rats per group three times (48, 24, and 3 h before the animals were terminated). The specimens were prepared in accordance with the standard protocol (version 14.2) of the “International Validation of the In Vivo Rodent Alkaline Comet Assay for the Detection of Genotoxic Carcinogens” organized by the Japanese Center for the Validation of Alternative Methods. No significant differences in the percentage of DNA in the tail were observed between the GOS-treated groups and vehicle controls in any of the organs evaluated. Additionally, no GOS-related clinical signs or effects on body weight were seen. Based on these results, the comet assay of GOS in the glandular stomach, colon, and peripheral blood using rats was judged negative. Therefore, it is concluded that GOS did not induce DNA damage in vivo under the conditions employed in this study.

A series of safety tests were undertaken on a novel galacto-oligosaccharide (GOS) produced from lactose by a two-step enzymatic process involving Sporobolomyces singularis and Kluyveromyces lactis. Bacterial reverse mutation and chromosomal aberration tests, with or without metabolic activation, were performed. These tests showed no mutagenesis in the Ames assay or in Escherichia coli WP2uvrA, and no chromosomal aberrations in cultured fibroblast cells from Chinese hamster lungs (CHL/IU). Micronuclei were not induced in the reticulocytes of mouse peripheral blood following oral administration of GOS. In a 90-day repeated oral dose toxicity study in rats, GOS was administered at 0, 500, 1000 and 2000 mg/kg to male and female Sprague-Dawley rats. There were no GOS-related changes in clinical signs, body weight, water intake, feed intake, urinalysis, ophthalmology, haematology, blood chemistry, organ weights, gross pathology or histopathology in any of the treatment groups compared to the control group. The no observed adverse effect level (NOAEL) of GOS was at least 2000 mg/kg/day in both males and females.

The in vivo micronucleus test using mouse colonic epithelial cells was evaluated as the 11th collaborative study organized by the Collaborative Study Group on the micronucleus test (CSGMT) with three model chemicals that were known to induce chromosome damage in mouse colonic cells. Five laboratories participated in this validation study. All three model chemicals, i.e. 1,2-dimethylhydrazine dihydrochloride (1,2-DMH), N-methyl-N-nitrosourea (MNU), and mitomycin C (MMC), induced micronucleated colonic epithelial cells in a 4-day exposue protocol in all participating laboratories. We confirmed that the present single cell suspension method could be used to detect the model chemicals as micronucleus inducers in mouse colonic epithelial cells. Advantages of this method are that experiments are easy to perform and that intact cells can be analyzed. The present study suggested that the colon micronucleus assay proposed here is useful for mechanistic studies of colon carcinogenesis.

患者は72歳, 女性.下腹部不快感と便秘を主訴として来院した.腹部理学的所見にて臍右側に手拳大の腫瘤を触知した.腹部超音波検査にて臍右側にpseudo-kidney signを認めた.注腸X線検査では横行結腸に内腔のほとんどを占める陰影欠損を認め, 大腸内視鏡検査では白色の隆起性病変を認めた.生検にて良性潰瘍組織であったが腫瘍性病変と診断し, 横行結腸切除術を施行した.組織検査ではAntoni A型良性神経鞘腫と診断された.大腸神経鞘腫は比較的少なく, とくに横行結腸に発生した症例は稀であり, 報告した.

Recent reports indicate that 7% of pregnant mothers in North America use cannabis. This is concerning given that in utero exposure to Δ9-tetrahydrocannabinol (Δ9-THC), the main psychoactive component in cannabis, causes fetal growth restriction and may alter replication and survival of pancreatic β-cells in the offspring. Accordingly, we hypothesized that maternal exposure to Δ9-THC during pregnancy would impair postnatal glucometabolic health of offspring. To test this hypothesis, pregnant Wistar rats were treated with daily intraperitoneal injections of either 3 mg/kg Δ9-THC or vehicle from gestational day 6 to birth. Offspring were subsequently challenged with glucose and insulin at 5 months of age to assess glucose tolerance and peripheral muscle insulin sensitivity. Female offspring exposed to Δ9-THC in utero were glucose intolerant, associated with blunted insulin response in muscle and increased serum insulin concentration 15 min after glucose challenge. Additionally, pancreata from male and female offspring were harvested at postnatal day 21 and 5 months of age for assessment of endocrine pancreas morphometry by immunostaining. This analysis revealed that gestational exposure to Δ9-THC reduced the density of islets in female, but not male, offspring at postnatal day 21 and 5 months, culminating in reduced β-cell mass at 5 months. These results demonstrate that fetal exposure to Δ9-THC causes female-specific impairments in glucose homeostasis, raising concern regarding the metabolic health of offspring, particularly females, exposed to cannabis in utero.

We investigated the effect on cell death of reactive oxygen species induced by water-soluble cationic metalloporphyrins with superoxide dismutase (SOD) activity. The SOD activity of 5,10,15,20-tetrakis(4-N-methylpyridyl)]porphine (MPy4P) containing Fe, Mn or Cu was measured using a cytochrome c assay by the xanthine/xanthine oxidase system and stopped-flow kinetic analysis. Cell viability of four cell lines treated with metalloporphyrins, mitomycin c (MMC), or cisplatin was estimated by a trypan blue exclusion assay. FeMPy4P with a high SOD activity showed a significant cytotoxicity compared with MMC and cisplatin, while CuMPy4P without SOD activity exhibited no cytotoxicity. However, MnMPy4P showing an SOD activity as high as that of FeMPy4P did not indicate cytotoxicity. These findings suggest that FeMPy4P as SOD mimic converts intracellular O2⋅− to H2O2 and that it rapidly reacts with H2O2 to form ⋅OH, causing DNA damage and inducing cell death. On the other hand, MnMPy4P did not participate in the Fenton reaction, so that DNA damage in the cells treated with MnMPy4P was not observed. In addition, the cytotoxicity by the metalloporphyrin was inversely correlated with the SOD activity of the cells and the selective damage at cellular and DNA levels was confirmed. We believe that for an anticancer drug with antioxidant ability O2⋅− is useful as a target molecule to induce selective cell death between cancer and normal cells and that metalloporphyrins showing SOD activity and Fenton-like reaction are a new class of anticancer agents.

Flow cytometry is a powerful tool to determine ploidy level and is commonly applied to measure DNA content and analyze the cell cycle. Here, we analyzed ploidy among the progeny derived from a population of self-pollinated triploid Senno (Lychnis senno Siebold et Zucc.). Each plant within the progeny was estimated to be an aneuploid showing a different ploidy level. For a large population, identification of chromosome number by chromosome observation is time-consuming. Therefore, a simple procedure to categorize different ploidy levels and chromosome number is required to facilitate breeding efforts. We applied flow cytometry to estimate chromosome number using internal standards for reference; these estimates were compared with the results obtained following chromosome observation. Leaf samples from progeny of triploid Senno were measured by flow cytometry in combination with an internal standard to evaluate DNA content accurately. By using appropriate internal standards, chromosome number was effectively estimated. When the parent triploid plant (strain MS, 2n = 3x = 36) was used as the internal standard, diploid (2n = 24) and different aneuploids (2n = 25, 44, 45, 55) were discriminated. For measurement of presumed triploids, strain No. 27 (2n = 25) was used as another internal standard. As a result, triploid (2n = 3x = 36) and different aneuploids (2n = 33, 35) were detected. The exact number of chromosomes as per chromosome observation corresponded well with the number estimated through flow cytometry. This procedure will facilitate ready, accurate, and reliable estimation of chromosome number using an adequate reference sample for internal standard. We discuss the possibility to estimate chromosome number by flow cytometry and the applicability of the procedure for determining the ploidy level as well as aneuploids in large populations derived from parental triploid Senno plants.

THP-adriamycin was administered in a dose of 5 to 20 mg, 3 times a week, and the results showed 4 cases of CR, 5 cases of PR, 3 cases of MR and 2 cases of NC with the accumulated total dose of less than 100 mg. Histological effects were slight in a total dose of less than 50 mg, while the effects appeared in a total dose of 60 to 70 mg and further effects were obtained after about 1 week of the termination of the therapy. Leukopenia occurred in 6 of 14 cases. The side effects of THP-adriamycin were milder than those of ADM.

Primary effects and side-effects of a new anthracycline antineoplastic agent, THP-adriamycin (THP-ADM), were examined in 20 patients with malignant head and neck tumors. According to a classification by tumor sites, there were 6 cases of oropharynx, 6 of nose and sinuses, 3 of tongue, 2 of floor of mouth, 2 of neck and 1 of external auditory meatus. According to the tissue classification, there were 11 cases of squamous cell ca., 6 cases of malignant lymphoma, each one case of anaplastic ca., carcino sarcoma and adeno cystic ca. There were 16 previously untreated cases and 4 pretreated cases. Twelve cases received THP-ADM by intraarterial injection and 8 cases by the same dose. 10 to 20 mg/body 3 times a week, in a total of 40 to 100 mg. 2 CR, 5 PR, 3 MR and 4 NC in 14 cases with carcinoma, and 2 CR, 3 PR and 1 MR in 6 cases with malignant lymphoma were obtained. Among 12 cases receiving intraarterial injection, 3 CR and 5 PR were obtained. The decrease of WBC counts below 3000/mm3 after THP-ADM administration was observed in 9 cases. Side effect of THP-ADM appeared to be less severe than that of Adriamycin.

Combination therapy of 5-FU, vitamin A and radiation (FAR therapy) was given to 50 patients with squamous cell carcinoma of the head and neck. The primary effect was observed in 22 patients who received FAR therapy as radical therapy. Among them, only 4 patients had recurrence or metastasis, and death was none. FAR therapy was given to 4 patients as pre-operative therapy and to 2 patients as post-operative therapy. However, the number of cases was not enough for us to draw any conclusion. In the group of 22 patients consisting of patients with recurrence or metastasis and patients who refused operation or were unresectable, no primary effect was observed only in 4 patients. Among a total of 50 patients, only 4 patients did not show a primary response to FAR therapy, 19 had relapse, 19 died (6 cases of them died from other causes), and 31 are still alive. There were no patients having severe side effects. From these results, FAR therapy seemed to be worth trying in treatment of head and neck cancer.