Establishment of Methylation-Specific PCR for the Mouse p53 Gene.

Methylation-specific PCR (MSP) of the mouse p53 gene has not yet been reported. We searched the CpG islands, sequenced the bisulfited DNA, and designed PCR primers for methylation and unmethylation sites. DNA from a young mouse produced a strong PCR product with the unmethylated primer and a weaker band with the methylated primer.

Effects of calorie restriction on the age-dependent accumulation of mutations in the small intestine of lacZ-transgenic mice.

To understand the effect of calorie restriction on genome maintenance systems, the age-dependent accumulation of mutations in animals maintained on high and low calorie diets was examined using lacZ-transgenic mice. Mice were fed a diet of 95 kcal/w or 65 kcal/w from 2 to 17 months of age. The mutation frequencies in the lacZ gene in epithelial tissues from the small intestine were examined at 12 and 17 months.

Dynamics of delayed p53 mutations in mice given whole-body irradiation at 8 weeks.

Purpose: Ionizing irradiation might induce delayed genotoxic effects in a p53-dependent manner. However, a few reports have shown a p53 mutation as a delayed effect of radiation. In this study, we investigated the p53 gene mutation by the translocation frequency in chromosome 11, loss of p53 alleles, p53 gene methylation, p53 nucleotide sequence, and p53 protein expression/phosphorylation in p53(+/+) and p53(+/-) mice after irradiation at a young age.

The late effects of radiation on lifespan, lymphocyte proliferation and p53 haplodeficiency in mice.

Purpose: We investigated the effect of irradiation on the lifespan of eight-week-old mice, the number of lymphocytes in bone marrow and the levels of p53 protein expression in the splenocytes.

Methods And Materials: Eight-week-old mice, wild-type p53 (p53(+/+)) and heterozygous p53 (p53(+/-)), were irradiated with 3 Gy. The cell numbers and cell cycle phases of bone marrow cells were determined by flow cytometry.

Radiation dose-rate effect on mutation induction in spleen and liver of gpt delta mice.

The effect of dose rate on radiation-induced mutations in two somatic tissues, the spleen and liver, was examined in transgenic gpt delta mice. These mice can be used for the detection of deletion-type mutations, and these are the major type of mutation induced by radiation. The dose rates examined were 920 mGy/min, 1 mGy/min and 12.

A comparison of the mutagenic and apoptotic effects of tritiated water and acute or chronic caesium-137 gamma exposure on spleen T lymphocytes on normal and p53-deficient mice.

Purpose: This study was carried out to compare the mutagenic effects on spleen T lymphocytes of mice exposed to tritiated water (HTO) and chronic or acute (137)Cs gamma irradiation.

Materials And Methods: p53 wild type (p53(+/+)) and p53 null type (p53(-/-)) mice were exposed to a total dose of 3 Gy of HTO, chronic (137)Cs and acute (137)Cs.

Results: In spontaneous T-cell receptor (TCR) variant fractions and fractions following exposure to HTO, chronic (137)Cs and acute (137)Cs, TCR variant fractions in p53(+/+) mice were 5.

Measurement of mutant frequency in T-cell receptor (TCR) gene by flow cytometry after X-irradiation on EL-4 mice lymphoma cells.

It is well known that somatic mutations are induced by ionizing irradiation. We have previously reported the measurement of mutant frequency (MF) on the T-cell receptor (TCR) gene in mouse T-lymphocytes after irradiation by flow cytometry. In this study, we developed an in vitro system using murine EL-4 lymphoma cells and observed frequency of cells defective in TCR gene expression after exposure to ionizing irradiation.

Prolonged increase in T-cell receptor (TCR) variant fractions of spleen T lymphocytes in pregnant mice after gamma irradiation.

To investigate the relationship between the radiation-induced increase of T-cell receptor (TCR) defective variant fractions and physiological status such as pregnancy, C57BL/ 6N mice were irradiated with 3 Gy of gamma rays at various days of gestation, just before and just after pregnancy. While the highest level of variant fractions in spleen T lymphocytes appeared at 9 days postirradiation and resolved fairly rapidly for nonpregnant mice, the increased variant fractions for pregnant mice irradiated at 16.5 days of gestation reached a plateau at 14 days postirradiation and remained at high levels until 28 days after irradiation.

TP53 and TP53-related genes associated with protection from apoptosis in the radioadaptive response.

We investigated the effect of administering priming low-dose radiation prior to high-dose radiation on the level of apoptosis and on the expression of TP53 and TP53-related genes in mouse splenocytes. The percentage of apoptotic cells was significantly lower in TP53(+/+) mice receiving priming radiation 2 to 168 h before the high-dose irradiation, compared to TP53(+/+) mice exposed to 2 Gy alone. In contrast, TP53(+/-) mice exhibited a reduced level of apoptosis only when priming was performed for 2 or 4 h prior to the high-dose irradiation.

The delayed manifestation of T-cell receptor (TCR) variants in X-irradiated mice depends on Trp53 status.

The influence of Trp53 on the radiation-induced elevation of T-cell receptor (TCR) variant fractions was examined in splenic T lymphocytes of Trp53-proficient and -deficient mice. Wild-type Trp53+/+, heterozygous Trp53+/- and null Trp53-/- mice were exposed to 3 Gy of X rays at 8 weeks of age. The fraction of TCR-defective variants was measured at various times after irradiation.

Ionizing radiation enhances the expression of the nonsteroidal anti-inflammatory drug-activated gene (NAG1) by increasing the expression of TP53 in human colon cancer cells.

The induction of apoptosis in cells of human colon cancer cell lines after gamma irradiation was investigated to determine whether apoptosis was mediated by TP53 and the subsequent expression of its downstream target, the NSAID-activated gene (NAG1). HCT116 (TP53(+/+)), HCT15 (TP53 mutant) and TP53 null HCT116 (TP53(-/-)) cells were irradiated with gamma rays, and apoptosis was measured at various times after irradiation. In HCT116 TP53(+/+) cells, apoptosis was increased after irradiation; the increase was dependent on the time after treatment and the dose of gamma rays.

Radioadaptive response for protection against radiation-induced teratogenesis.

To clarify the characteristics of the radioadaptive response in mice, we compared the incidence of radiation-induced malformations in ICR mice. Pregnant ICR mice were exposed to a priming dose of 2 cGy (667 muGy/min) on day 9.5 of gestation and to a challenging dose of 2 Gy (1.

Trabecular bone mass and bone formation are preserved after limb immobilisation in p53 null mice.

Objectives: To determine whether disruption of the p53 gene leads to preservation of trabecular bone volume (BV) after limb immobilisation.

Materials And Methods: Tibias of immobilised hind limbs of p53 gene knockout (p53(-/-)) and wild-type (p53(+/+)) mice were compared. Right knee joints of 8 week old mice were immobilised in full extension for 7 days.

Effect of extended exposure to low-dose radiation on autoimmune diseases of immunologically suppressed MRL/MpTn-gld/gld mice.

The purpose of this paper is to analyze the relationship between alterations of splenic T-cell subpopulations and the amelioration of autoimmune diseases of MRL/MpTn-gld/gld mice (MRL/gld mice) after extended exposure to low-dose radiation. After the onset of disease, 4-month-old MRL/gld mice were exposed to doses of 0.05, 0.

Role of p53 gene in apoptotic repair of genotoxic tissue damage in mice.

When DNA is damaged by exposure to a small amount of radiation, it is repaired efficiently by innate mechanisms. However, if cellular damage is more extensive, DNA repair cannot be adequately completed. To clarify the role of the p53 gene in apoptotic tissue repair, the incidence of in-vivo radiation-induced somatic mutation was evaluated by measuring the T cell receptor (TCR) gene expression in p53(+/+) and p53(-/-) mice.

Micronuclei induced by low dose rate irradiation in early spermatids of p53 null and wild mice.

To obtain evidence of the dose-rate effect on induction of micronuclei in early spermatids, we observed frequencies in wild-type p53(+/+), heterozygous p53(+/-) and null p53(-/-) mice 14 days after gamma rays irradiation at a high (1,020 mGy/min) or a low (1.2 mGy/min) dose-rate. A dose- and dose-rate-related increase in micronuclei was seen in early spermatids with no difference between the different p53 status.

Apoptosis and p53 expression in chondrocytes relate to degeneration in articular cartilage of immobilized knee joints.

Objective: We have reported that articular cartilage showed early stage degeneration at 7 and 14 days after immobilization, moderate degeneration at 28 days, and severe degeneration at 42 days in rabbits. To test whether apoptosis occurs in association with p53 expression in chondrocytes during the process of articular cartilage degeneration, we investigated the degree of cartilage degeneration, the frequency of apoptotic cells, and the levels of p53 mRNA in rabbits and mice after knee immobilization.

Methods: Right knees of male Japanese white rabbits were immobilized in full extension with fiberglass casts for up to 42 days.

Effects of a 4.7 T static magnetic field on fetal development in ICR mice.

In order to determine the effects of a 4.7 T static magnetic field (SMF) on fetal development in mice, we evaluated fetal teratogenesis and endochondral ossification following exposure in utero. Pregnant ICR mice were exposed to a 4.

Disruption of the p53 gene results in preserved trabecular bone mass and bone formation after mechanical unloading.

We tested the hypothesis that mechanical unloading facilitates signaling of p53, an important modulator of cell cycling and apoptosis, in bone marrow cells and thereby reduces trabecular bone volume (BV). We performed histomorphometric analyses and bone marrow cell cultures in tail-suspended (TS) p53 null (p53-/-) and wild-type (p53+/+) mice. Eight-week-old male mice were assigned to four groups after 1-week acclimatization: p53+/+ + ground control (GC), p53+/+ + TS, p53-/- + GC, and p53-/- + TS.

Sequential changes in transforming growth factor (TGF)-beta1 concentration in synovial fluid and mRNA expression of TGF-beta1 receptors in chondrocytes after immobilization of rabbit knees.

We have previously reported that a combination of transforming growth factor (TGF)-beta1 and basic fibroblast growth factor (bFGF) synergistically increases the proliferation of chondrocytes obtained from knee joint immobilized for 7-14 days in male Japanese white rabbits. In the present study, we performed experiments with chondrocytes and syn ovial fluid obtained from rabbit knees immobilized for 0-42 days, to clarify the sequential changes in TGF-beta1 and bFGF concentrations in synovial fluid and the mRNA expression of TGF-beta1 receptor type I (RI) and II (RII) in chondrocytes after immobilization. The combination of TGF-beta1 and bFGF had a synergistic effect on the proliferation of chondrocytes obtained from knee joints immobilized for 7-14 days.

Threshold effect for teratogenic risk of radiation depends on dose-rate and p53-dependent apoptosis.

Purpose: To obtain evidence that the p53 gene is indispensable for reduction of high teratogenic risk of radiation at a high dose-rate to zero risk by lowering the dose-rate.

Materials And Methods: Wild-type p53(+/+), heterozygous p53(+/-) and null p53(-/-) mice were exposed to gamma-rays at high or low dose-rates during days 9.5-10.

Radiotherapy after hyperbaric oxygenation improves radioresponse in experimental tumor models.

We examined the effect of radiotherapy after hyperbaric oxygen (HBO) breathing in experimental tumors using a tumor growth delay assay. Tumor models used were SCCVII (radiobiological hypoxic fraction: approximately 10%) and 9L tumors (containing less hypoxic cells) subcutaneously transplanted into C3H/He mice and Fisher 344 rats, respectively. Irradiation using X-rays was locally administered to the tumors immediately after decompression.

Concentrations and consequent radiation doses of uranium-238, thorium-232 and potassium-40 contained in the front glass of CRTs.

Concentrations of 238U, 232Th, and 40K contained in the front glass of CRTs (cathode ray tube) of three television sets are measured by gamma-ray spectrometry on the assumption of radioactive equilibrium in uranium and thorium series. The concentrations of 238U and 232Th are also measured by neutron activation analysis. The obtained results by the two different methods agree well within experimental uncertainty, showing that the radioactive equilibrium is found to be established in the present samples, and that the concentrations of 238U and 232Th in the glass can be measured by the gamma-ray spectrometry as well as by the neutron activation analysis.

[Accurate measurement of doses in the sample chamber of a 137Cs gamma-ray irradiation apparatus].

Dose rates in the sample chamber of a 137Cs gamma-ray irradiation apparatus (Nordion International Inc., Gammacell 40 Exactor) were accurately measured using an ionization chamber dosimeter. The dose rates obtained were 5.