Behavioral performance and microglial status in mice after moderate dose of proton irradiation.

Cognitive impairment is a remote effect of gamma radiation treatment of malignancies. The major part of the studies on the effect of proton irradiation (a promising alternative in the treatment of radio-resistant tumors and tumors located close to critical organs) on the cognitive abilities of laboratory animals and their relation to morphological changes in the brain is rather contradictory. The aim of this study was to investigate cognitive functions and the dynamics of changes in morphological parameters of hippocampal microglial cells after 7.5 Gy of proton irradiation. Two months after the cranial irradiation, 8- to 9-week-old male SHK mice were tested for total activity, spatial learning, as well as long- and short-term hippocampus-dependent memory. To estimate the morphological parameters of microglia, brain slices of control and irradiated animals each with different time after proton irradiation (24 h, 7 days, 1 month) were stained for microglial marker Iba-1. No changes in behavior or deficits in short-term and long-term hippocampus-dependent memory were found, but an impairment of episodic memory was observed. A change in the morphology of hippocampal microglial cells, which is characteristic of the transition of cells to an activated state, was detected. One day after proton exposure in the brain tissue, a slight decrease in cell density was observed, which was restored to the control level by the 30th day after treatment. The results obtained may be promising with regard to the future use of using high doses of protons per fraction in the irradiation of tumors.

Low-Intensity Femtosecond Radiation Activates the Natural Defenses of Mice in vivo.

The possibility of induction of cytogenetic damage in the bone marrow, changes in the cellularity of lymphoid organs and blood composition in mice irradiated with low-intensity femtosecond laser radiation at a power flux density of 5.1, 10.4, and 52 mJ/cm2 (0.5 mW for 5, 10, and 50 s) in vivo was shown. Using the radiation adaptive response test (0.1 Gy + 1.5 Gy), it was found that, when mice were exposed to femtosecond laser radiation in high doses, the body's natural defenses were activated in the same narrow range of energy flux density (2-16 mJ/cm2) as in the case of X-ray irradiation in a dose of 0.1 Gy (4 mJ/cm2). The data obtained suggest a similar mechanism of activation of the body's natural defense upon exposure to low doses of both ionizing and non-ionizing radiation.

Low dose of carbon ion irradiation induces early delayed cognitive impairments in mice.

People often encounter various sources of ionizing radiation, both in modern medicine and under various environmental conditions, such as space travel, nuclear power plants or in conditions of man-made disasters that may lead to long-term cognitive impairment. Whilst the effect of exposure to low and high doses of gamma and X-radiation on the central nervous system (CNS) has been well investigated, the consequences of protons and heavy ions irradiation are quite different and poorly understood. As for the assessment of long-term effects of carbon ions on cognitive abilities and neurodegeneration, very few data appeared in the literature. The main object of the research is to investigate the effects of accelerated carbon ions on the cognitive function. Experiments were performed on male SHK mice at an age of two months. Mice were irradiated with a dose of 0.7 Gy of accelerated carbon ions with an energy of 450 meV/n in spread-out Bragg peak (SOBP) on a U-70 particle accelerator (Protvino, Russia). Two months after the irradiation, mice were tested for total activity, spatial learning, as well as long- and short-term hippocampus-dependent memory. One month after the evaluation of cognitive activity, histological analysis of dorsal hippocampus was carried out to assess its morphological state and to reveal late neuronal degeneration. It was found that the mice irradiated with accelerated carbon ions develop an altered behavioral pattern characterized by anxiety and a shortage in hippocampal-dependent memory retention, but not in episodic memory. Nissl staining revealed a reduction in the number of cells in the dorsal hippocampus of irradiated mice, with the most pronounced reduction in cell density observed in the dentate gyrus (DG) hilus. Also, the length of the CA3 field of the dorsal hippocampus was significantly reduced, and the number of cells in it was moderately decreased. Experiments with the use of Fluoro-Jade B (FJB) staining revealed no FJB-positive regions in the dorsal hippocampus of irradiated and control animals 3 months after the irradiation. Thus, no morbid cells were detected in irradiated and control groups. The results obtained indicate that total irradiation with a low dose of carbon ions can produce a cognitive deficit in adult mice without evidence of neurodegenerative pathologic changes.

The Effect of Low and Medium Doses of Proton Pencil Scanning Beam on the Blood-Forming Organs during Total Irradiation of Mice.

The aim of this work was to study the effect of proton pencil beam scanning in the Bragg peak in the dose range of 0.1-1.5 Gy on the induction of cytogenetic damage in the bone marrow, reactive oxygen species (ROS) production in whole blood, and the state of lymphoid organs after total body irradiation of mice. Irradiation was carried out in the Prometeus proton synchrotron (Protvino) in the Bragg peak with proton energy at the output of 90-116 MeV. It was found that, under irradiation of mice in the range of low and medium doses of proton pencil beam scanning in the Bragg peak, the relative biological effectiveness (RBE) according to the criterion of cytogenetic changes was 1.15. In addition, it was found that the pathophysiological effect on the lymphoid organs and the production of ROS by blood cells were different as compared with the effect of X-rays.

Combined Effect of Low-Intensity Helium-Neon Laser and X-Ray Radiation on in Vivo Cellular Response of the Whole Blood and Lymphoid Organs in Mice.

We studied the effect of exposure to helium-neon laser (dose range 0.16-50 mJ/cm2) on activation of natural protection reserve in mice using the adaptive response test. DNA comets method revealed a protective response manifested in DNA damage level in whole blood leukocytes of mice and in lymphoid organs by the thymus and spleen weight index; preexposure to laser did not induce the adaptive response. ROS level in the whole blood was assessed by the level of zymosan-induced luminol chemiluminescence. In mice subjected to adaptive laser irradiation in doses of 0.16-5 mJ/cm2 followed by X-ray irradiation in a dose of 1.5 Gy, the activation index calculated as the ratio of induced to spontaneous area of luminescence was by 1.4 times lower than that in non-irradiated animals, which attested to reduced ROSgeneration reserve capacity of neutrophils.

Induction of the Adaptive Response in Mice Exposed to He-Ne Laser and X-Ray Radiation.

We studied the dose-dependent induction of in vivo adaptive response in the bone marrow and blood of mice exposed to low-intensity radiation of He-Ne laser (633 nm) and X-ray radiation by the severity of cytogenetic injury and intensity of ROS production, respectively. Induction of the adaptive response in mice preexposed to He-Ne laser and X-ray radiation depended on the adaptive dose and the interval between the adaptive and main doses and correlated with changes in ROS generation. The adaptive response after exposure to low-intensity ionizing and non-ionizing radiation was observed in the same dose range, which attests to similar mechanisms of its induction.

[Induction of DNA damage in blood leucocytes and of cytogenetic injuries in bone marrow polychromatic erythrocytes in mice exposed to low-LET and high-LET radiation and in their progeny].

The present work was aimed at studying the molecular and cellular levels of the response of the hematopoietic system in mice and their progeny to the action of low-LET and high-LET radiation at different times after exposure. The damage to the genome at the molecular level was assessed by the comet assay in peripheral blood leucocytes, whereas at the cellular level it was estimated by means of the micronuclear test in the marrow cells, after exposure of mice to X-radiation of 1, 3 and 5 Gy and to a high-LET low-intensity radiation at thedoses of 0.14 and 0.35 Gy, as well as to a combined effect of these types of radiation. When accessing the level of the DNA damage to individual cells by the comet assay, we also used, apart from a commonly accepted parameter %TDNA, additional characteristics: the proportions of leucocytes with an intact and highly fragmented DNA. Using these parameters, we detected the changes characterizing the dynamics of the leukocyte population in mouse blood at different times after the action of X-ray and high-LET radiation. It was found that: (1) the DNA damage increases with the dose of high-LET radiation; (2) the level of damage in the progeny of the animals exposed to high-LET radiation does not differ from that in unirradiated animals both at the molecular and cytogenetic levels; and (3) a decrease in the radiosensitivity of the progeny of the mice exposed to high-LET radiation at a dose of 0.35 Gy makes itself evident only at the molecular level, which may point to the possible transgeneration transmission of genomic lesions.

[Influence of low-dose-rate red and near-infrared radiations on the level of reactive oxygen species, the genetic apparatus and the tumor growth in mice in vivo].

The effect of low-dose-rate red and near-infrared radiations from the matrix of light emitted diode (650 nm and 850 nm) and a He-Ne laser (633 nm) on activation of the reserve of a natural defense system in the mice exposed to radiation in vivo was studied by the level of reactive oxygen species (ROS) production in blood cells, the induction of cytogenetic adaptive response in bone marrow cells, thymus and spleen, and the rate of Ehrlich ascites carcinoma growth in a solid form. As a positive control animals were irradiated with X-rays by the scheme of the radiation-induced adaptive response (0.1 Gy + 1.5 Gy). The levels of ROS production was assessed in whole blood by luminol-dependent chemiluminescence, of cytogenetic damage--by the "micronucleus test" in the bone marrow, the weight of the thymus and spleen--by index of organ, and the rate of tumor growth--according to its size for 30 days after inoculation. Adaptogenic and anticarcinogenic effects of studied radiations were revealed. The values of these effects were not different from those in animals pre-irradiated with the X-rays. The relationship between the level of ROS production and adaptive response induction in the mice under the influence of non-ionizing radiation was first ascertained. The experimental data obtained may indicate a similar mechanism of induction of protective responses to ionizing and non-ionizing radiations in mice in vivo.

Effect of infrared and X-ray radiation on thymus cells and the rate of growth of Ehrlich carcinoma.

We studied the effect of infrared light with a wavelength of 850 nm and modulated frequency of 101 Hz and X-ray radiation on the induction of cross-adaptive and radiation responses in the thymus and on the rate of tumor growth in mice in vivo. Preliminary exposure to infrared and X-ray radiation was shown to result in recovery in thymus weight after irradiation in a dose of 1.5 Gy and also inhibited the growth rate of Ehrlich carcinoma. These data attest to common mechanisms of the adaptive response induced by infrared and X-ray radiation in mice. Infrared light can be used as an adaptogen to adapt the animals to adverse factors.

Delayed effects of chronic low-dose high linear energy transfer (LET) radiation on mice in vivo.

In the present work, the delayed effects of chronic high linear energy transfer (LET) radiation in polychromatic erythrocytes (PCEs) of mice bone marrow were investigated in vivo. Irradiation of the two-month-old SHK white mongrel random-bred male mice was performed in the radiation field behind the concrete shield of the accelerator of 70 GeV protons to accumulate doses of 0.005-0.16 Gy. The dependence of the biological response on dose, adaptive response (AR) and genomic instability (GI) in F(1) and F(2) generations from males irradiated with doses of 0.005 and 0.16 Gy and from males exposed to combined action of immunomodulator-bendazol hydrochloride (BH) and of 0.16 Gy irradiation, were examined using the micronucleus formation test. The data demonstrated that irradiation of mice with these doses lead to an increase in the level of cytogenetic damage and induces no AR. With analysis of the bone marrow radiosensitivity to 1.5 Gy of X rays and the capacity to AR it was found that the chronic high-LET irradiation of parents induced the GI at least two generations. The combined exposure to BH and the dose of 0.16 Gy induces no AR in F(0) generation but induces AR in F(1) and F(2) offspring.

[Study of the genetic instability in generations of mice irradiated of a low-dose rate of high-LET radiation].

In present work, we investigated the genetic instability in mice of F1, of F2 and of F3 generations born from males irradiated by a low-dose rate of high-LET radiation that simulates the spectral and component composition of radiation fields formed in the conditions of high-altitude flights in vivo in polychromatic erythrocytes of bone marrow using the micronucleus test. Two-month-old males of SHK white mongrel mice were used. Irradiation was performed for 24 h a day in the radiation field behind the concrete shield of the U-70 accelerator of 70 GeV protons (Serpukhov) to accumulate doses of 11.5, of 21.5 and of 31.5 cGy (1 cGy/day). The experiments demonstrated that in mice of F1 generation born from males irradiated with doses of 11.5, 21.5 and of 31.5 cGy, an increase in sensitivity to additional irradiation with a dose of 1.5 Gy of gamma-radiation and the absence of adaptive response compared with the descendants of unirradiated males occur. In contrast to F1 generation genetic instability in mice of the F2 and F3 generations was revealed only by the absence of adaptive response. These data indicate a genetic instability in F1, F2 and F3 generations born from irradiated males.

[Peculiar effects of low doses of high let radiation on mice].

Experiments with exposure of mice to low doses of chronic high-LET radiation were carried out in the radiation field behind the concrete wall of the Serpukhov accelerators of protons with the energy of 70 GeV. The goal was to study dose dependence, radiation adaptive response (AR), and genetic instability. Mice (SHK strain) were irradiated continuously 15, 24 and 31 days which corresponded to the doses of 11.5, 21.5 and 31.5 Gy. Cytogenetic damages were determined using the micronuclear test in marrow polychromatophil erythrocytes. It was shown that all the experimental doses aggravated the cytogenetic damage; however, no AR induction in marrow cells was observed. Males of the F1 generation born from the males irradiated at 11.5 Gy had same level of spontaneous cytogenetic damage as males born from non-irradiated parents. Yet, they displayed an exaggerated sensitivity to additional exposure to 1.5 Gy and no AR induction by the standard gamma-protocol which is indicative of genetic instability.

[The effect of low-dose rate radiation simulating the high-altitude flight conditions on mice in vivo].

In present work, we investigated the peculiarities of the effect of a low-dose rate high-LET radiation that simulates the spectral and component composition of the radiation field formed in the atmosphere at a height of 10 km on mice in vivo. The dose dependence and adaptive response were examined. Irradiation of mice was performed for 24 h a day in the radiation field behind the concrete shield of the Serpukhov accelerator of 70 GeV protons for the time (15-31 days) necessary to accumulate the required doses. The experiments demonstrated that irradiation of mice in vivo in the dose range of 11.5-31.5 cGy leads to an increase in cytogenetic damage to bone marrow cells and induces no adaptive response in bone marrow cells.