The mutagenic effects of low level sub-acute inhalation exposure to benzene in CD-1 mice.

Benzene is a widely used chemical and common environmental contaminant. It is carcinogenic in man and animals and is genotoxic in mice, rats, and occupationally exposed humans at doses above one part per million. In order to evaluate the genotoxic effects of prolonged exposures to very low concentrations of benzene, we exposed CD-1 mice to benzene by inhalation for 22 h per day, seven days per week for six weeks at 40, 100 and 1000 parts per billion (ppb). Additional groups were exposed to purified air or were housed in standard plastic cages. The effects of in vivo exposure to benzene were evaluated by using an autoradiographic assay to determine the frequency of mutants which represent mutations at the hypoxanthine-guanine phosphoribosyl transferase (hprt) locus in spleen lymphocytes. At the end of the six weeks exposure period lymphocytes were recovered from the spleens of the mice and cryopreserved prior to assay. Mutant cells were selected on the basis of their ability to incorporate tritiated thymidine in the presence of 6-thioguanine. The weighted mean variant (mutant) frequencies (Vf) of female mice (three per group) were 7.2 x 10(-6) at 0 ppb; 29.2 x 10(-6) at 40 ppb; 62.5 x 10(-6) at 100 ppb and 25.0 x 10(-6) at 1000 ppb. The Vf of unexposed mice housed in standard cages was 13.2 x 10(-6). In male mice the same pattern of response was observed, but the increases in Vf in response to benzene were not as great. In both sexes of mice, the increases at 40 and 100 ppb were significantly greater than at 0 ppb (P less than 0.05). The increase in Vf with exposure to 100 ppb and the decline at 1000 ppb parallel the results observed for chromosome damage in spleen lymphocytes from the same animals (Au et al., Mutation Res., 260 (1991) 219-224). These results indicate that sub-chronic exposure to benzene at levels below the current Occupational Safety and Health Administration Permitted Exposure Limit may induce gene mutations in lymphocytes in mice.     

Quantitative analysis of radiation-induced changes in sperm morphology

When developing spermatogenic cells are exposed to radiation, chemical carcinogens or mutagens, the transformation in the morphology of the mature sperm can be used to determine the severity of the exposure. In this study five groups of mice with three mice per group received testicular doses of X irradiation at dosage levels ranging from 0 rad to 120 rad. A random sample of 100 mature sperm per mouse was analyzed five weeks later for the quantitative morphologic transformation as a function of dosage level. The cells were stained with gallocyanin chrome alum (GCA) so that only the DNA in the sperm head was visible. The ACUity quantitative microscopy system at Lawrence Livermore National Laboratory was used to scan the sperm at a sampling density of 16 points per linear micrometer and with 256 brightness levels per point. The contour of each cell was extracted using conventional thresholding techniques on the high-contrast images. For each contour a variety of shape features was then computed to characterize the morphology of that cell. Using the control group and the distribution of their shape features to establish the variability of a normal sperm population, the 95% limits on normal morphology were established. Using only four shape features, a doubling dose of approximately 39 rad was determined. That is, at 39 rad exposure the percentage of abnormal cells was twice that occurring in the control population. This compared to a doubling dose of approximately 70 rad obtained from a concurrent visual procedure.     

Therapeutic efficiency of spleen or bone marrow CFU in X-irradiated 89Sr marrow-ablated mice.

Experiments were carried out to compare the therapeutic efficiency (TE: number of CFU required to reduce the mortality from 100 to 50 per cent) of spleen or marrow (BM) stem cells (CFU) grafted into lethally irradiated mice (807 rad) which had been previously treated with 89Sr or splenectomized. It was found that during the reconstitution of the haemopoietic organs, the spleen does not provide more than 10 per cent of the functional cells necessary for survival. Besides, the BM-derived CFU growing in 89Sr marrow-ablated mice remain twice as efficient as the spleen-derived ones. Similarly, spleen-derived CFU transplanted into splenectomized mice are half as efficient as BM-derived ones. It may therefore be assumed that haemopoietic stem cells grafted into a foreign microenvironment retain their original kinetics of growth and differentiation during 7 to 10 days after their transplantation.     

Chromosome aberration yields induced in human lymphocytes by 15 MeV electrons given at a conventional dose-rate and in microsecond pulses.

Yields of unstable chromosome aberration were analysed in human lymphocytes after in vitro exposure to 15 MeV electrons. Two dose-effect curves were prepared. In one, doses of 44 to 742 rad were given at 100 rad/min, and in the other doses of 53 to 764 rad were each delivered in single microsecond pulses. No significant difference could be found between the two sets of data when analysed in terms of the quadratic model of aberration production. Good agreement was observed with other dose-response studies in this laboratory, in which human lymphocytes were exposed to 250 kVp X-rays and 60CO gamma-rays at conventional rates of 100 and 50 rad/min, respectively. Comparison with the results of a low-LET dose-rate experiment shows that the yield of dicentric aberrations remains constant overa wide range, i.e. 25 to 6X 10(9), 100 to 1-5 X 10(10), and 150 to 3 X 10(10) rad/min, respectively, for doses of 100, 250 and 500 rad. Radiochemical consumption of oxygen occuring in the lymphocytes during the single microsecond exposures may amount to less than 5 per cent of the total oxygen present in the blood samples, immediately before irradiation. The data also indicate that the ultra-high dose-rates currently available are insufficient to overcome the therapeutic problem of hypoxic radioresistant tumour cells.     

The effect of low exposure-rate gamma irradiation on T and B lymphocyte function in the mouse

The effect of chronic irradiation on T and B cell numbers and function was studied in mice. Cobalt 60 gamma radiation at 6 R/hour reduced the numbers of anti-SRBC PFC in the spleen, with minimal levels recorded after total exposures of 1000-2000 R. Recovery was incomplete after 1000 R, reaching only 40-50 per cent of normal in four months and remaining at that level for the animal's lifetime. The long-term deficiency in PFC formation was not due to a quantitative lack of T or B cells since normal cell numbers were observed in the spleen 60-144 days after 1000 R. Adoptive transfer studies with combinations of bone marrow and thymus cells, or of splenic T and B cells, from normal and irradiated mice, revealed functional defects in both cell compartments during the first two months. Normal and near normal function of T and B cells occurred 100 days postirradiation, a time when the splenic in vivo response was still only 50 per cent of the controls. The latter observation suggests that the microenvironment of the chronically irradiated spleen alters factors regulating T and B cell interactions in response to a T-dependent antigen.     

The in vitro radiosensitivity of hemopoietic stem cells from control and preirradiated infant mice

Summary The radiosensitivity of hemopoietic stem cells isolated from infant mice (6 or 9 days of life), of infant preirradiated mice (exposed to 126 rad on day 6 and assayed at day 9 of life) and of adult C57/B1 mice was assayed on the basis of their capacity to form spleen colonies and to incorporate iododeoxyuridine after transplantation into heavily irradiated hosts. Stem cells of infant non-irradiated mice have a D₀ of 115 rad compared to 72 rad for adult mice whereas the D₀ of preirradiated infant mice has diminished to 80 rad. No significant difference in D₀ was seen between spleen and bone marrow cells or between total cells and cells not sensitive to³H-thymidine. It is postulated that this sensitization of stem cells caused by a preirradiation is responsible for the greater mortality of infant mice after fractionated exposure compared to a single one.     

Induction of micronuclei in wild-type and p53(+/-) transgenic mice by inhaled bromodichloromethane

Bromodichloromethane (BDCM) is commonly present in trace amounts in drinking water as a disinfection by-product. BDCM has been shown to be carcinogenic in mice and rats when given by gavage at relatively high doses. Genotoxic activity as well as induced regenerative cell proliferation may contribute to the carcinogenic potential of BDCM. The purpose of the current studies was to evaluate the ability of BDCM to induce micronuclei (MN) in bone marrow and blood of wild-type and p53(+/-) mice on the C57BL/6 and FVB/N genetic backgrounds using the inhalation route of exposure. Toxicity studies were being conducted in this laboratory with inhaled BDCM to select doses for longer-term cancer bioassays using wild-type and p53(+/-) transgenic mice on different genetic backgrounds. Bone marrow samples from these experiments were evaluated for the induction of MN after 1 and 3 weeks of exposure. Accumulation of MN in the peripheral blood was also evaluated at the 13-week time point of a cancer study with the p53(+/-) mice. For the 1-week time point, male C57BL/6 wild-type and p53(+/-) mice and FVB/N wild-type and p53(+/-) mice were exposed daily for 6h per day for 7 consecutive days to atmospheric BDCM concentrations of 0, 1, 10, 30, 100, or 150 ppm. In a second experiment, mice were exposed daily for 6h per day for 3 weeks to atmospheric BDCM concentrations of 0, 0.5, 1, 3, 10, or 30 ppm. Resulting levels of polychromatic erythrocytes (PCE) containing MN were assessed in the bone marrow. For all of the 1- and 3-week exposure groups, the only statistically significant increase in the percentage of bone marrow PCE cells containing MN was in the 1-week 100 ppm BDCM exposure group in the FVB/N wild-type mice (control 0.26% versus exposed 1.16%). C57BL/6 p53(+/-) mice and FVB/N p53(+/-) mice were exposed daily for 6 h per day for 13 weeks to atmospheric BDCM concentrations of 0, 0.5, 3, 10, or 15 ppm. MN were quantified in samples of peripheral blood. Statistically significant increases in the percentage of peripheral blood NCE cells containing MN were seen at the highest BDCM exposure group of 15 ppm in both the C57BL/6 p53(+/-) strain (control 0.36% versus exposed 0.67%) and the FVB/N p53(+/-) strain (control 0.36% versus exposed 0.86%). These data indicate weak induction of MN by BDCM, but only at high atmospheric concentrations relative to normal environmental exposures and with extended periods of exposure. Although comparisons are difficult because responses were negative or marginal, the p53 genotype or the genetic background did not appear to substantially alter susceptibility to the genotoxic effects of BDCM.     

The fate of cells with chromosome aberrations after total-body irradiation and bone marrow transplantation

Cytogenetic studies were done on bone marrow cells and peripheral lymphocytes of four patients (three with acute nonlymphocytic leukemia, one with aplastic anemia) at various intervals up to 861 days after total-body X irradiation (TBI) at doses between 4.5 and 10 Gy (450-1000 rad) followed by syngeneic or allogeneic bone marrow transplantation. Whereas no radiation-induced aberrations could be found in the bone marrow, apart from a transient finding in the patient with the lowest radiation dose, aberrant metaphases were seen in the peripheral lymphocytes of three patients in the range from 2.5 to 46% even at 861 days after the exposure. There were no demonstrable aberrations related to TBI in the only patient developing graft-versus-host disease. The dicentric yield as determined in the aberrant metaphases with 46 centromeres ranged between 3.4 +/- 1.3 and 4.9 +/- 0.4. In one patient it was demonstrated by BUdR-labeling that after 10 Gy (1000 rad) TBI the surviving and heavily damaged lymphocytes can go into cell cycle and reach at least the third mitosis. The percentage of aberrant cells diminished by about 25% at each mitotic division.     

Respiratory infection of mice with Histoplasma capsulatum

Summary Mice were infected by exposure to varying doses ofHistoplasma capsulatum yeast cells in the Henderson aerosol apparatus.Ten per cent of the animals were infected by inhalation of only 5 yeast cells. In the larger dosages used, 5, 400 to 24, 200 cells per animal, 90 to 100% infections were obtained. The disease spread rapidly from the lungs to the liver and spleen, with the liver generally containing the largest number of yeast cells. With small numbers of cells inhaled, mice were more highly positive at 4 weeks after exposure. In larger doses, per cent positive was highest at 2–4 weeks and mice sacrificed at later periods were less likely to yield as many positive animals.Presented in part at the annual meeting of the American Society for Microbiology, 1962.Work reported in this paper was supported in part by Grant E 1992 of the National Institutes of Health, U.S. Public Health Service.     

Recovery response of dividing cells in the thymus of whole-body gamma-irradiated mice.

Mice were irradiated with different doses of gamma-rays 30 min after the administration of 32P-orthophosphate. The dose-response curves determined at 72 hours after exposure showed an inflection point in the total activity present in the DNA in thymus and spleen. In the low dose-range, the dose-response curves have D0 = 55 rad (n = 2-5) for thymus and DO = 95 rad (n = 2-5) for the spleen. Thirty minutes after the administration of 32P-orthophosphate, the dividing cells from thymus were partially synchronized by the administration of 80 mg per kg body-weight hydroxyurea. At different time-intervals, the mice were irradiated with 80 rad, and the total activity of DNA was determined at 72 hours after synchronization. A significant maximum of recovery was found at 5 hours (S phase) after the administration of hydroxyurea. In similar conditions, the dose-response curves corresponding to the G1, S and M phase of the division cycle were also determined. The synchronization of dividing cells induced by hydroxyurea failed in the spleen.     

Use of monoclonal antibodies to plague microbe antigens at the early stage of infection in white mice for enhancement of the late streptomycin and doxycycline treatment]

It was demonstrated that use for prophylaxy (after 5 h of infection) or for treatment (after 24 h after infection) of the monoclonal antibodies mixture to specific epitops of capsule antigen (fraction 1), lipopolysacharide, murine toxine can prevent development of plague pathogen at 100 of mice infected by approximately 1000 LD50 Yersinia pestis 231. 5-day course of prophylaxy by monoclonal antibodies provided survival of 50 per cent animals. Subsequent use of fraction 1 antigen for 5 days followed by treatment with streptomycin or doxycycline at 6-7-8-9-10 days after infection with Y. pestis 231 prevented infection manifestation at 80 per cent of animals, etiotropic therapy started at the same period was ineffective. When white mice were infected with Y. pestis 231 Fra-, with deleted ability to produce capsule antigen (fraction 1) 80% level of efficacy can be provided by subsequent administration of antibodies to fraction 1 combinated with lipopolysacharide, murine toxine and streptomycin. Use of monoclonal antibodies followed by doxycycline was ineffective.     

X-irradiation of equine peripheral blood lymphocytes stimulated with phytohaemagglutinin in vitro.

Small lymphocytes were isolated from the peripheral blood of horses and incubated at 37 degrees C in Eagle's medium supplemented with 20 per cent foetal calf serum. The addition of phytohaemagglutinin (PHA) to the cultures resulted in: increased RNA and protein synthesis; the enlargement of the small lymphocyte into a lymphoblast-like cell; the initiation of DNA synthesis, and cell division. When survival was measured 24 hours after X-irradiation by means of phase-contrast microscopy, the lymphoblast-like cell was much more radio-resistant (D0 = 250 rad) than the small lymphocyte (D0 = 20 rad). This increase in radioresistance, however, was not observed until 12-24 hours after PHA treatment. To investigate which of the changes occurring during the transformation of the small lymphocyte was responsible for the increased resistance to irradiation, the percentage of cells surviving irradiation was compared with the percentage of cells incorporating significant amounts of 3HTdR, 3H-UR, or 3H-leucine at the time of irradiation. For this comparison, a dose of 100 rad was used because 100 rad killed essentially all of the small lymphocytes, but less than 35 percent of the cells which had become radioresistant from the PHA treatment. The results indicated that the increase in radioresistance was not associated with DNA synthesis, but instead correlated with the increase in RNA and protein synthesis which the cells had attained at the time of irradiation.     

Variation in U.V. primary fluorescence-intensity of vital cells depending on 60Co gamma-radiation dose.

Using impulse-cytofluorophotometry in the ultra-violet spectral region the author has shown on vital, unstained Ehrlich ascites tumour cells that the primary fluorescence intensity of this tumour is on day 11 after transplantation 20 per cent higher than on day 8. Storage of the vital cells for 25 min at 20 degrees C has no effect on this result. When the cells are exposed to 60Co to gamma-radiation on day 6, a new stable fluorescence level is established afted 20 hours. Measurements of the primary fluorescence intensity depending on dose have shown a significant rise starting from 75 rad at 48 hours after irradiation. The fluorescence intensity rises by 42.5 per cent of the control value at 3000 rad, but only by 31.5 per cent on exposure to 4000 rad.     

Response of mouse intestine to 14 MeV neutrons.

At the Hamburg-Eppendorf Hospital neutron facilities the relative biological effectiveness (r.b.e.) of d,T-neutrons was determined with respect to survival of mouse intestinal crypts. (CBA/Rij x C57BL/Rij)F1 mice were irradiated to the whole body at different depths inside a tissue-equivalent phantom. Irradiations were carried out with a collimated neutron beam at about 6 rad/min given in single doses ranging from 450 to 1000 rad. For reference, gamma-rays from a 60Co therapy unit were used. The number of surviving intestinal crypts per circumference of the jejunum was determined 3 1/2 days after irradiation according to the method of Withers and Elkind. The number of surviving stem cells was calculated on the basis of Poisson statistics. The doses necessary to reduce survival to ten crypt stem cells per circumference amounted to 689 +/- 19 rad for neutrons and 1449 +/- 29 rad for 60Co gamma-rays. From these figures an r.b.e. of 2 . 1 +/- 0 . 1 is obtained. Measurements at different depths in the phantom did not show any variation of r.b.e. with depth along the axis of the neutron beam.     

Inhalation carcinogenesis of high-fired 241AmO2 in rats

Wistar rats were given a single inhalation exposure to high-fired 241AmO2 particles and examined over their life span. A total of 310 rats were used: 259 exposed to 241Am for life-span study, 30 exposed to 241Am for early metabolism study, and 21 unexposed life-span controls. The activity median aerodynamic diameter of the aerosols was 0.75-1.39 microns. About 55% of alveolarly deposited 241Am was cleared from the lung with a half-life of 0.5 days, 37% with a half-life of 7 days, and 8% with a half-life of 580 days. Group mean lung doses ranged from less than 5.7 rad up to 1500 rad. Significant early mortality due to radiation pneumonitis was seen only in the highest exposure group. The percentage of rats with lung tumors was 0% for controls (21 rats), 1% at lifetime lung doses less than 10 rad (139 rats), 7% at 10-50 rad (86 rats), 0% at 50-100 rad (9 rats), 60% at 100-500 rad (10 rats), and 7% at 500 rad (15 rats). Only one liver and one bone tumor were found in all exposed rats, both at lifetime tissue doses less than 10 rad. The fate and carcinogenicity of inhaled 241AmO2 in the lung of rats were similar to what has previously been described for inhaled 244CmO2.     

Mutagenicity studies with nitrofurans. I. Mutagenicity of nitrofurylacrylic acid for mammals

Cytogenetic analysis of mouse bone-marrow cells, the dominant lethal test in mice and the cytogenetic analysis of human peripheral lymphocytes in vitro were used to study the mutagenicity of 3-(5-nitro-2-furyl)acrylic acid (5-NFA) for mammals. The bone-marrow cytogenetic analysis was performed in female mice exposed to 5-NFA administered intraperitoneally in single doses of 15--120 mg/kg and in 5 repeated doses of 15 and 30 mg/kg, intragastrically in single doses of 30--240 mg/kg and 5 repeated doses of 30 and 60 mg/kg, and perorally for 12 weeks to 5-NFA concentration of 10, 100 and 1000 mg 5-FNA/1 in drinking water. The bone-marrow analysis was performed in this case after 12 days, 3, 4, 6, 8, 10 and 12 weeks exposure. No increase in chromosome damage attributable to dosing with 5-NFA occurred in any of these experiments. Experiments in which mice were exposed to 5-NFA in drinking water for 12 weeks and then treated with a single i.p. dose of 2 mg of the mutagen TEPA [trix-(1-aziridinyl)phosphine oxide] per kg revealed that, at a concentration of 1000 mg 5-NFA/1, the clastogenic activity of TEPA was reduced to that in untreated animals. The dominant lethal test was performed in male mice exposed to 5-NFA applied intraperitoneally in single doses of 40--120 mg/kg and in 5 repeated doses of 10--30 mg/kg, intragastrically in 5 repeated doses of 20--60 mg/kg, and perorally for 4 weeks in drinking water containing 5-NFA at concentrations of 10, 100, 316 and 1000 mg/l. No significant differences were detected between the exposed and control groups of animals. Experiments in which male mice were exposed to 5-NFA in drinking water and treated after the 4-week exposure to 5-NFA with 1 mg TEPA/kg revealed that a concentration of 1000 mg 5-NFA/1 reduced TEPA-induced dominant lethality to within control values. A reduction in male fertility was observed after the single or repeated 5-NFA doses, but no changes when 5-NFA was applied in drinking water. The cytogenetic analysis of human peripheral lymphocytes exposed in vitro for the last 24 h of culture to concentrations of 1--100 micrograms 5-NFA/Ml did not show any compound-related chromosomal changes. The results of dominant-lethal and bone-marrow cytogenetic studies in mice after consumption of drinking water containing 1000 mg of 5-NFA/1 for 12 weeks and dosed subsequently with TEPA suggests that 5-NFA has some antimutagenic activity. Because none of the studies reported revealed any compound-related genetic activity, the results suggest that 5-NFA is not a chromosome-breaking agent in mammals.     

Cytogenetic effects of microwave irradiation on male germ cells of the mouse

Hybrid male mice were exposed to 2.45 GHz microwaves for 30 min/day, 6 days a week for two consecutive weeks at power densities of 1.0, 100 or 400 W m-2, with sham-exposed controls. Rectal temperatures before and after exposure were measured on days 1, 6 and 12. Measurements made on day 1 were treated with caution because of heterogeneity in rectal temperatures taken before exposure between the groups of mice given different treatments. On days 6 and 12, rectal temperatures rose by approximately 1 degree C in mice sham exposed, or exposed to 1 W m-2 or 100 W m-2. Only in the group of mice exposed to 400 W m-2 was the mean rise in rectal temperature during exposure (about 3 degrees C) significantly increased above the sham value. In groups killed 2-3 days after treatment (mainly meiotic exposure) frequencies of chromosome aberrations in spermatocytes showed no significant heterogeneity although the highest frequency of 1.5 per cent was at the highest (400 W m-2) power density. Another group killed 30 days after 100 W m-2 exposures (spermatogonial sampling) showed no significant increase over controls in chromosome aberration frequency. There was a small but significant increase in sperm count with increasing power density in mice killed 12-13 days after exposure, but a non-significant one in those exposed as spermatogonia (killed 41 days later). Thus effects were markedly less severe than those reported previously by Manikowska-Czerska et al. (1985) with a very similar radiation regime and were probably caused by the temperature enhancement.     

Cytotoxic effect and induction of sister chromatid exchange in exponentially growing rat 9L gliosarcoma cells after brief exposure to BrdU

The magnitude of DNA modulation in rat 9L gliosarcoma cells after a brief exposure to bromodeoxyuridine (BrdU) was studied by assaying colony-forming efficiency (CFE) and the number of sister chromatid exchanges (SCEs) per metaphase. The CFE assay showed that a 1-hr exposure to BrdU, at concentrations ranging from 10 to 1000 microM, produced a maximum cell kill of 5%. After a 2-hr exposure to 20 microM BrdU, the surviving fraction was 0.99, and even at a BrdU concentration of 1000 microM, 77% of the 9L cells survived. Compared with control cultures, the relative number of SCEs per metaphase in treated cultures was increased after a 1-hr exposure to BrdU at concentrations of 100 microM or more and after a 2-hr exposure to concentrations of 20 microM or more; no increase was observed in cells treated for 30 min with BrdU at concentrations up to 1000 microM. When the treated cells were allowed to grow in BrdU-free growth medium, the number of SCEs per metaphase returned to the control level within 24 hr, even after exposure to BrdU at concentrations as high as 1000 microM. These results demonstrate that exposure to BrdU at concentrations of up to 1000 microM for 30 min, 100 microM for 1 hr, and 20 microM for 2 hr causes little modulation of DNA.     

Magnetic field affects thymidine kinase in vivo

Whole mice on normal or vitamin E deficient diet were immobilized by Nembutal anaesthesia and exposed to a stationary magnetic field of 1.4 tesla for up to 60 min. Thymidine kinase (TdR-K) was assayed in the high-speed supernatant of bone marrow cells which were collected into optimally adjusted nutrient medium of pH 7.3-7.4 containing 1350 mg NaHCO3 per litre and were then destroyed by sonication. In parallel, uptake of 125I-labelled 5-I-2'-deoxyuridine (125IUdR) into DNA of whole bone marrow cells, of various tissues and of the whole body was measured. The results indicate the following. The magnetic field exposure caused in bone marrow cells an increase of activity of TdR-K and of uptake of 125IUdR to about 130 per cent of control. The effect depended on immobilization of the mice in the field and on the presence of NaHCO3 in the nutrient medium used for cell collection. There was no field-induced change in body temperature. The effect on 125IUdR uptake was similar in isolated tissues and the whole body following intraperitoneal injection of the tracer. It increased to a maximum of about 135 per cent of control, during exposure times over 30 min. This effect is not explained as a result of a temporary change in the rate of cell proliferation. Vitamin E deficiency caused a depression of activity of TdR-K and of uptake of 125IUdR in bone marrow cells to about 75 per cent of control. This depression was similar to that observed after whole body gamma-irradiation with about 0.01 Gy (1 rad). The inhibitory effects of vitamin E deficiency on TdR-K were overcome by exposure to the magnetic field. Immediately after cessation of the magnetic field for 60 min, 125IUdR uptake was normal; normalization of uptake was delayed with exposure times shorter than 60 min. A 60 min exposure to the magnetic field had no long term effect on turnover of labelled cells in the mice. The data imply the non-specific control of thymidine kinase by charged molecular species and the modification of this control by the magnetic field.     

Modification of radiation-induced division delay by caffeine analogues and dibutyryl cyclic AMP

The mitotic selection procedure for cell cycle analysis was utilized to investigate the concentration-dependent modification of radiation-induced division delay in Chinese hamster ovary (CHO) cells by methyl xanthines (caffeine, theophylline, and theobromine) and by dibutyryl cyclic AMP. The methyl xanthines (concentrations from 0.5 to 1000 micrograms/ml) all reduced radiation-induced division delay with the effect being linear between approximately 100 and 1000 micrograms/ml. After doses of 100-300 rad, delay was reduced by 75, 94 or 83 per cent at 1000 micrograms/ml for each drug, respectively. However, the addition of dibutyryl cyclic AMP had an opposite effect: radiation-induced delay was increased by the concentration range of 0.3 to 300 micrograms/ml. These results indicate that in mammalian cells the control of cell cycle progression and the modification of radiation-induced division delay are not simply related to intracellular levels of cyclic AMP. Rather, there appear to be at least two competing mechanisms which are differentially affected by caffeine analogues or by direct addition of dibutyryl cyclic AMP. The direct effect of caffeine and the methyl xanthines on membrane calcium permeability is considered.