Comparative Analysis of Behavioral Reactions and Morphological Changes in the Rat Brain After Exposure to Ionizing Radiation with Different Physical Characteristics.

The aim of this research was to study behavioral reactions and morphological changes in the brain of adult female Sprague Dawley rats after exposure to 170 MeV and 70 MeV protons and gamma radiation (60Co) at a dose of 1 Gy. The analysis of the behavioral reactions in the T-maze showed that exposure to ionizing radiation with different LETs led to an increase in number of repeated entries into the arms of the maze in the spontaneous alternation test. In the Open Field test a decrease in overall motor activity in the group of irradiated animals (70 MeV protons at the Bragg peak) was observed. A decrease in the number of standing positions was seen in all groups of irradiated animals. Morphological analysis showed the development of early amyloidosis, autolysis of the ependymal layer, an increase in the number of neurodegenerative changes in various structures of the brain, and the development of neuronal hypertrophy on the 30th day after irradiation in the cerebellum and hippocampal hilus. Exposure to protons at a dose of 1 Gy leads to the development of structural and functional disorders of the central nervous system of animals on the 30th day after irradiation. These data indicate a damage of short-term memory, a decrease in motor activity and exploratory behavior of animals. With an increase in LET, there is an increase in the number of amyloid plaques in the forebrain of rats, autolysis of the ependymal layer of the ventricles, and the development of dystrophic changes. Investigations of behavioral reactions and morphological changes in various parts of the brain of adult rats on the 30th day after influence of ionizing radiation with different physical characteristics at a dose of 1 Gy. Various negative patho-morphological and cognitive-behavioral changes observed.

DNA Damage in Splenocytes of Mice Exposed to Secondary Radiation Created by 650 MeV Protons Bombarding a Concrete Shielding Barrier.

The proportion of splenocytes with a high level of DNA double-strand breaks was determined in mice exposed to primary and secondary radiation created by bombarding of a concrete barrier (thickness 20, 40, and 80 cm) by 650 MeV protons. The proportion of splenocytes with a high level of DNA double-strand breaks was assessed by flow cytometric analysis of γH2AX+ and TUNEL+ cells. It is shown that concrete barrier can significantly reduce primary proton radiation; the severity of negative biological effects in mice irradiated in the center of the proton beam decreased with increasing the thickness of this barrier. However, the spectrum of secondary radiation changes significantly with increasing the barrier thickness from 20 to 80 cm and the distance from central axis of the beam from 0 to 20 cm, and the proportion of the neutron component increases, which also causes negative biological effects manifesting in a significant (p<0.05) increase in the percentage of splenocytes with a high level of DNA damage in mice irradiated at a distance of 20 cm from the center of the proton beam and receiving relatively low doses (0.10-0.17 Gy).

[DEMONSTRATION OF LIKELIHOOD OF THE NEGATIVE EFFECT OF PHYSICAL PROTECTION DURING TOTAL PROTON IRRADIATION OF MICE].

The experiments were performed with outbred CD-1 male mice (SPF category). Total irradiation at 1.0; 2.5 and 5.0 Gy by protons with the average energy of 170 MeV was conducted in a level medical beam of the phasotron at the Joint Institute of Nuclear Investigations. Targets were 2 points of in-depth dose distribution, i.e. beam entrance of the object, and modified Bragg peak. As a physical protector, the comb filter increases linear energy transfer (LET) of 170 MeV entrance protons from 0.49 keV/µm to 1.6 keV/µm and, according to the bone marrow test, doubles the biological effectiveness of protons when comparing radiation doses that cause 37% inhibition of blood cell formation in the bone marrow. Physical protection increases dose rate from 0.37 Gy/min for entrance protons to 0.8 Gy/min for moderated protons which more than in thrice reduces time of irradiation needed to reach an equal radiobiological effect.

[IMMEDIATE RADIOBIOLOGICAL EFFECTS IN MICE FOLLOWING γ-IRRADIATION BY LOW DOSES].

Outbred CD-1 mice females aged 4 to 4.5-months were investigated in 21-22 hours following total γ-irradiation at 10, 25, 50, 75, 100 and 200 mGy. Loss in bone marrow karyocytes, as well as spleen and thymus mass reductions were significant in the group of animals irradiated at 50 and 200 mGy and less dramatic in mice irradiated at 75 mGy. The orientative-trying behavior reaction (OTBR) in the open field tested in 19-20 hours after exposure to 10 and 25 mGy was reliably stronger than in the group of biological control; however, emotional status (ES) in the animals that received 10 mGy dropped significantly. Mice irradiated at 50 mGy were found to weaken the grip of their front limbs. Dose levels differing in opposite radiobiological effects on the parameters under study were established. Doses in the range from 10 to 25 mGy maximized OTBR and ES, while doses of 50, 100 and 200 mGy produced high reactions of the immune and hemopoietic organs.

[CYSTEAMINE-INDUCED MODIFICATION OF CYTOGENETIC DAMAGES TO THE CORNEAL EPITHELIUM OF MICE EXPOSED TO CORPUSCULAR RADIATION WITH VARYING LINEAR TRANSFER ENERGIES].

Cytogenetic damages to cells of the corneal epithelium were studied in mice exposed to protons (10, 25, 50 and 645 MeV), ions of boron, carbon and neon, and X-rays (180 keV) within the dose range from 25 to 750 cGy and injected with a radioprotector. Animals were subjected to a single exposure. The protective effect of ß-mercaptoethylamine was tested in the experiment. The radioprotector (0.2 ml) was introduced intraperitoneally 30 minutes before exposure in 350 mI/kg dose. Control animals received the same amount of sodium chloride solution. The animals were sacrificed by cervical dislocation in 24 and 72 hrs. after exposure. It was shown that cysteamine effectively protects in vivo corneal epithelium cells of mice exposed to electromagnetic radiation or protons in a broad energy spectrum (10 to 645 MeV), and to a broad range of radiation doses (25 to 750 cGy), as judged from levels of aberrant mitosis and mitotic activity. The radioprotector exhibited the highest effectiveness in animals exposed to the doses of 50 to 300 cGy. These findings prove that cysteamine may potentially be used for pharmacological protection from protons. The radioprotector failed to prevent chromosomal aberrations after exposure to heavy charged particles of boron, carbon and neon, which implies the need to design radioprotectors against this type of corpuscular radiation specifically.

[Cytogenetic damage to the corneal epithelium of mice due to the in vivo exposure to ionizing radiation with different levels of linear energy transfer].

Damages to corneal epithelium cells were studied in mice irradiated by protons with the energies of 10, 25, 50 and 645 MeV, 60Co γ-quanta and accelerated ions of boron, carbon and neon with the energies of 7.5; 2.5 and 6.0 MeV/nucleon, respectively. X-rays (180 keV) were used as a standard radiation. Animals were exposed to a single dose in the range from 25 to 760 cGy. The mitotic index and aberrant mitoses were counted in corneal preparations in 24 hrs after irradiation. No matter the type of radiation, the mitotic index had an inverse dose dependence, i.e. the higher the dose, the lower the mitotic index. Exposure to all types of radiation resulted in a sharp increase in the number of chromosomal aberrations in the corneal epithelium; frequency of aberrations was a function of dose and type of radiation. The number of chromosomal aberrations displayed a peculiar direct dose dependence irrespective of type of radiation; however, heavy ions of carbon and boron are the most damaging to the cytogenetic apparatus of epithelial cells. Protons at the Bragg peak and ensuing fall, and of 50 MeV also contribute to the production of chromosomal aberrations as compared with sparsely ionizing gamma- and X-rays and high-energy protons with low linear energy transfer. Coefficients of relative biological effectiveness were calculated based on the mitotic index and evidence of aberrant mitosis.

[MODIFICATION OF THE PROTON BEAM PHYSICAL PARAMETERS AND RADIOBIOLOGICAL CHARACTERISTICS BY ELEMENTS OF SPACECRAFT RADIATION PROTECTION].

The experiment was performed with outbred ICR (CD-1). female mice (SPF). The animals were irradiated by 171 MeV protons at a dose of 20 cGy. The spacecraft radiation protection elements used in the experiment were a construction of wet hygiene wipes called a "protective blind", and a glass plate imitating an ISS window. Physical obstacles on the path of 171 MeV protons increase their linear energy transfer leading to the absorbed dose elevation and strengthening of the radiobiological effect. In the experiment, two types of obstacles together raised the absorbed dose from 20 to 23.2 cGy. Chemically different materials (glass and water in the wipes) were found to exert unequal modifying effects on physical and biological parameters of the proton-irradiated mice. There was a distinct dose-dependent reduction of bone marrow cellularity within the dose range from 20 cGy to 23.2 cGy in 24 hours after exposure. No modifying effect of the radiation protection elements on spontaneous motor activity was discovered when compared with entrance protons. The group of animals protected by the glass plate exhibited normal orientative-trying reactions and weakened grip with the forelimbs. Rationalization of physical methods of spacecrew protection should be based as on knowledge in physical dosimetry (ionizing chambers, thermoluminescent, track detectors etc.), so the radiobiological criteria established in experiments with animals.

Dose distribution outside the target volume for 170-MeV proton beam.

Dose delivered outside the proton field during radiotherapy can potentially lead to secondary cancer development. Measurements with a 170-MeV proton beam were performed with passive detectors (track etched detectors and thermoluminescence dosemeters) in three different depths along the Bragg curve. The measurement showed an uneven decrease of the dose outside of the beam field with local enhancements. The major contribution to the delivered dose is due to high-energy protons with linear energy transfer (LET) up to 10 keV µm(-1). However, both measurement and preliminary Monte Carlo calculation also confirmed the presence of particles with higher LET.

[Mitigation of mice radiation damage after acute and prolonged γ-irradiation by a laser device].

Effects of 7 Gy 60Co γ-radiation (acute and prolonged exposure), and combined exposure to 650 nm laser and γ-radiation on survival, peripheral blood, karyocyte count and mitotic index of bone marrow cells were studied in young C57BL/6 mice. All mice died following acute γ-irradiation at the dose rate of 1.14 Gy/min for 5 days or combined exposure for 11 days. Thirty percent survival from prolonged exposure to the dose rate of 0.027 Gy/min was observed after 19-day γ- and 38-day combined irradiation. Peripheral blood parameters did not differ significantly after acute and prolonged exposure; however, hyperchromemia was observed in mice after 24 hours of acute γ-irradiation. The count of mitoses per 1000 nucleus-containing BM cells evidenced that BM was virtually collapsed after 72 hours since the acute γ-exposure. It was demonstrated that laser can manage protection from a broad range of ionizing radiation doses and mitigate the adverse effects of equally acute and prolonged radiation exposure.

[Radiobiological effects of total mice irradiation with Bragg's peak protons].

Outbred CD-1 female mice were irradiated in a proton beam (171 MeV, 5 Gy) on the phasotron at the Joint Institute of Nuclear Research (Dubna, Russia). Radiation was delivered in two points of the depth dose distribution: at the beam entry and on Bragg's peak. Technical requirements for studying the effects of Bragg's peak protons on organism of experimental animals were specified. It was recognized that protons with high linear energy transfer (mean LET = 1.6 keV/microm) cause a more severe damaging effect to the hemopoietic system and cytogenetic apparatus in bone marrow cells as compared with entry protons and 60Co gamma-quanta. It was shown that recovery of the main hemopoietic organs and immunity as well as elimination of chromosomal aberrations take more time following irradiation with Bragg's peak protons but not protons with the energy of 171 MeV.

Contribution of secondary particles to the dose in 12C radiotherapy and other heavy ion beams.

The results of experimental studies performed in a radiotherapy (12)C ion beam with a nominal energy of 500 MeV/amu and in (16)O and (56)Fe ion beams with a nominal energy of 1 GeV/amu have been described. Linear energy transfer (LET) spectra have been established by means of an LET spectrometer based on a chemically etched track detector, and the measured results were also compared with theoretical calculations obtained using the program Stopping and Range of Ions in Matter (SRIM). It was observed that with increasing depth in a beam, the LET spectra are shifted towards higher values of LET; one can also observe an important widening of the spectra along the range, as well as an increasing amount of nuclear reaction products and/or of fragments in the spectra. The relative contribution of these secondary particles to the total absorbed dose was assessed.

Biological weighted effective dose in 205 MeV clinical proton beam.

The contribution of high linear energy transfer (L) charged particles to dosimetric and microdosimetric characteristics in a clinical proton beam was experimentally studied using an ionization chamber and track etched detectors. The particles mentioned are produced by proton nuclear interactions; at the Bragg peak region slowed down protons also contribute in the L region above several keV microm(-1). Due to these particles the biological weighted effective dose (BWED) of the beam changes with depth. The spectra of particles with L above 7 keV microm(-1) were established by means of track etched detectors, which permitted us to determine their contribution to dosimetric and microdosimetric characteristics of clinical proton beams. The studies were realized in the clinical proton beam of the JINR Dubna Phasotron, with a primary energy of 205 MeV. The relative contribution to the absorbed dose of the particles with L above 7 keV microm(-1) increases from several per cent at the beam entrance to several tens of per cent at the Bragg peak region. The relative biological weighted efficiency (RBWE) for radiotherapy has been calculated using a biological weighting function. It increases with depth from 1.02 at the beam entrance to about 1.25 at the Bragg peak region.

Passive spectrometry of linear energy transfer: development and use.

A linear energy transfer (LET) spectrometer based on the evaluation of particle track parameters in a chemically etched polyallyldiglycolcarbonate (PADC) track detector has been developed at our laboratory. It permits us to determine LET spectra between 10 and 700 keV microm(-1) in tissues. The LET spectra obtained permit us to calculate total dose and dose equivalent corresponding to particles with etchable tracks also. We have recently been able to verify the calibration curves used by using C, Mg, Ne, Si and Fe ion beams with different energies. The calibration curves obtained are presented and compared with those originally used, and a good correlation is found. The LET spectrometer with new calibration was used to analyse the radiation quality of the radiotherapy proton beam at the Joint Institute for Nuclear Research (JINR). The radiation quality was studied along the proton's range, particular attention being devoted to the region of the Bragg peak. It was found that the biologically weighted effective dose (BWE) reaches a value of about 1.25 at the Bragg peak region. At the beam entrance this value increases to about 1.02 due to secondary particles created through primary proton nuclear reactions in tissues.

Microdosimetric characteristics of the clinical proton beams at JINR, Dubna.

High-energy proton radiotherapy beams give rise to secondary heavy charged particles with elevated linear energy transfer (LET), which contribute to the dose in a patient. This contribution to the characteristics of radiotherapy proton beams was experimentally studied by means of a LET spectrometer based on a track detector. The spectrometer permits LET spectra to be established in the region above 10 keV.micron-1 in tissue. Sets of track detectors were exposed in the various depths of a phantom irradiated with protons of two energies, 150 and 205 MeV. It was observed that the contribution of particles with the values of LET mentioned increases with the depth, representing from about 2 (at the surface) up to few tens% close to Bragg peak region of the total dose. There, some of primary protons contribute also above 10 keV.micron-1. Using the 'biological weighted function' proposed, the clinical RBE was calculated, it could approach 1.3. This effect has to be taken into account during the clinical beam production and the radiotherapy.