An analytical model for the phase behavior of cholesteryl esters in intracellular inclusions.

The cholesteryl ester-rich, intracellular inclusions that characterize atherosclerotic plaque are capable of existing in a metastable, relatively fluid state for long periods of time. We have developed an analytical model which explains this metastability, and other aspects of the phase behavior, of physiologically relevant, phospholipid-stabilized dispersions of cholesteryl ester mixtures. The model, based on classical nucleation theory, incorporates temperature, time and lipid composition as independent variables. Differential scanning calorimetry was used to elucidate the model. The dispersions consisted of cholesteryl palmitate and an ester containing a long-chain, unsaturated or polyunsaturated, fatty acid. When a dispersion of approx. 1-microns droplets is melted, then cooled, crystallization is preceded by the formation of small crystalline nuclei (homogeneous nucleation). Nucleation is energetically unfavorable until (typically) well below the melting point. sigma, the tension between the surface of the crystal nucleus and surrounding fluid, is a measure of the difficulty in forming nuclei. This parameter was found to increase with the content of unsaturated ester. sigma was found to increase with increasing triacylglycerol content, and to decrease upon addition of free cholesterol.

The adaptive response modifies latency for radiation-induced myeloid leukemia in CBA/H mice.

We have investigated the effect of the adaptive response on acute myeloid leukemia (AML) induced in CBA/Harwell mice by a chronic radiation exposure. Groups of mice irradiated with a total dose of 1. 0 Gy at two different chronic dose rates (0.5, 0.004 Gy/h) had similar frequencies of AML. Compared to control animals that did not develop AML, irradiation at either of these dose rates did not change the longevity of the mice that did not die of leukemia. The survival rates of irradiated mice that did develop leukemia in the two groups were not different from each other, indicating that the dose rates produced similar responses and therefore were both chronic exposures. We then tested the ability of a chronic 10-cGy (0. 5 Gy/h) exposure to ionizing radiation, mild hyperthermia (40.5 degrees C whole-body, 60 min) or treatment with interleukin-1 (1500 U i.p.) to induce an adaptive response and modify the frequency or latency of AML which resulted from a subsequent (24 h later) 1.0-Gy (0.5 Gy/h) chronic radiation exposure. The frequency of radiation-induced leukemia was not changed in mice given any of the three adapting treatments 24 h prior to the chronic 1.0-Gy dose that induced leukemia. However, the latent period for development of AML was significantly increased by both the prior low radiation dose and mild hyperthermia treatment. Injection of interleukin-1, in contrast, may have reduced the latent period. Similar to the single 1.0-Gy chronic exposure alone, none of the adapting treatments prior to that exposure influenced the survival of animals that did not develop AML. These results indicate that an earlier exposure to a small adapting dose of radiation or to a mild heat stress can influence secondary steps in radiation-induced carcinogenesis.

A method for hyperthermic treatment of mouse skin.

The Sencar mouse skin system is a recognized model for tumour initiation, promotion and progression. The current interest in the effect of hyperthermia on this multi-stage tumorigenesis model prompted the need for a technique to accurately heat a section of dorsal skin of a large number of mice for 30 min per heat treatment. In the technique described, experimental groups of 25 female Sencar mice were treated at 7-8 weeks of age under general methoxyflurane anaesthesia. Treatment consisted of the application of initiating and/or promoting agents with or without hyperthermia. For hyperthermic skin treatments, each group of mice was placed onto a platform in a water bath so that the dorsal skin of the mice was in contact with 44 degrees C temperature controlled water.

A simple transport system for radiation treatment of specific pathogen-free mice in lifetime studies.

To prevent the introduction of pathogens, specific pathogen-free (SPF) facilities generally have a "once out, never back" policy with respect to animals and materials. In a lifetime study of the long-term effects of ionizing radiation exposure in mice, large numbers of SPF mice needed to be transported from clean-animal barrier labs to a multiuser conventional building for radiation treatment and then back into the animal facility. The conventional building is known to harbor wild mice as well as insects, spiders, and mites, and this situation might potentiate the transfer of wild mouse pathogens to laboratory animals. Introduction of pathogens into the mouse population would jeopardize the entire study, but the radiation treatments were an essential component of the study. These considerations prompted development of a system for transporting individual animals out of and back into the facility without exposure to pathogens. The system consists of reusable transport/treatment vessels and transport protocols designed to minimize the potential for pathogen exposure.

Skin tumor promotion by Vitamin E in mice: amplification by ionizing radiation and Vitamin C.

We have shown previously that Vitamin E acts as a tumor promoter in 7,12-dimethylbenz(a)anthracene (DMBA) initiated mouse skin. We now show that high concentrations (80 micromol) of Vitamin E are required for promotion, and that 10-fold lower concentrations do not promote tumor formation. The same high concentration of the water-soluble anti-oxidant Vitamin C did not act as a tumor promoter, but did amplify the promoting effect of high, but not low, concentrations of Vitamin E. Oxidizing free radicals generated by beta-radiation exposure of the skin at the time of Vitamin E treatment also enhanced promotion by high (but not low) concentrations of Vitamin E. The results are consistent with a process whereby tumor promotion by the lipid-soluble Vitamin E occurs as a result of alpha-tocopherol acting as a free radical scavenger, with the formation and subsequent transfer of the alpha-tocopherol free radical center to the surrounding lipids, resulting in lipid oxidations.