Recovery of hematopoiesis in mice following a continuous irradiation with a dose rate of 0.957 Gy/d and a total accumulated dose of 19.14 Gy. I. Bone marrow, spleen and thymus gland

This paper deals with the evaluation of histological changes in the bone marrow, spleen and thymus of mice after continuous irradiation with a dose rate of 0.957 Gy/day and a total accumulated dose of 19.14 Gy. Erythropoiesis in the spleen could be recovered quickly, significantly exceeding the spleen erythropoiesis of the controls on the seventh post-irradiation day. Myelopoiesis in the bone marrow could be recovered until the 21st day and erythropoiesis until the 28th day after the end of irradiation. Lymphopoiesis in the thymus could be recovered on the 28th day approximately and in the spleen roughly on the 60th day after the end of irradiation.     

Changes in mouse erythropoiesis during continuous irradiation with a daily dose of 957 mGy

Quantitative and qualitative changes in the blood producing organs and in the peripheral blood of mice are evaluated in this paper. The animals were irradiated for 42 days continually with a daily dosage of 957 mGy. Until the 7th day of irradiation a significant diminution of the cellularity of the bone-marrow and of the cellularity as well as the mass of the spleen could be observed. After the 14th day or irradiation a temporary stabilization of the cell number in the bone-marrow could be found until the 28th day, after that time there was a moderately strong decrease. The cellularity and the mass of the spleen increased temporarily until the 28th day of irradiation because of erythropoiesis and myelopoiesis increasing from 20% to 50%. The most significant changes in the peripheral blood could be observed in not granulated cells as a kind of sudden and permanent decrease. The diminution of the granulocyte and reticulocyte number proceeded somewhat more slowly, temporarily revealing an increasing tendency on the 28th day of irradiation. The erythrocyte numbers as well as the haematocrit and haemoglobin values decreased continually beginning from the 7th day of irradiation until the death of the test animals.     

Late changes in peripheral blood and spleen in mice after the effect of single whole body irradiation with the dose of 8 Gy.

Late haematopoietic changes were followed within 12-16 months after single whole body irradiation with the dose of 8 Gy. Permanently increased frequency of occurrence of cytopathologically altered leukocytes was observed in peripheral blood in irradiated animals as well as certain disorder of erythropoietic system. At the end of period followed the splenomegalia accompanied with marked macroscopic and microscopic changes was occurred. With simultaneous increase of occurrence of erythroblasts in peripheral blood the changes above are considered to be a symptom of erythroleukaemia.     

Rescue of lethally irradiated mice from hematopoietic death by pre-exposure to 0.5 Gy X rays without recovery from peripheral blood cell depletion and its modification by OK432.

Exposing mice to 0.5 Gy X rays 2 weeks before lethal irradiation has been reported to induce marked radioresistance and to rescue them from hematopoietic death. Here we examined effects of the 0.5-Gy pre-exposure on hematological changes in C57BL mice that were lethally irradiated with 6.5 Gy X rays. Approximately 77% of pre-exposed mice survived 30 days after this irradiation, whereas 80% of mice that did not receive this pre-exposure died by day 20. However, regardless of the pre-exposure, peripheral blood cell counts decreased markedly by day 3 and reached a nadir at day 20. CFU-S in femur and CFU-GM in spleen had started to recover at day 10 and 14, respectively, but recovery of functional peripheral blood cells occurred later. The effect of pre-exposure on survival was altered by OK432, a bioresponse modifier; the effect depended on the timing of its administration. OK432 given 2 days before 0.5 Gy enhanced the protective effect of pre-exposure, resulting in the survival of 97% of the mice. In contrast, injection of OK432 1 day before or 2 days after pre-exposure led to 100% mortality. Thus the survival-promoting effect of 0.5 Gy could be altered by OK432. The OK432-induced changes in the survival of mice could not be attributed solely to hematological changes, as shown by blood cell counts and progenitor cell contents. These results suggest that radioresistance induced by pre-exposure to 0.5 Gy X rays is not stable, but rather varies with the physiological conditions, and can be modulated by factors such as OK432.     

Adaptive response in embryogenesis: V. Existence of two efficient dose-rate ranges for 0.3 Gy of priming irradiation to adapt mouse fetuses

The adaptive response is an important phenomenon in radiobiology. A study of the conditions essential for the induction of an adaptive response is of critical importance to understanding the novel biological defense mechanisms against the hazardous effects of radiation. In our previous studies, the specific dose and timing of radiation for induction of an adaptive response were studied in ICR mouse fetuses. We found that exposure of the fetuses on embryonic day 11 to a priming dose of 0.3 Gy significantly suppressed prenatal death and malformation induced by a challenging dose of radiation on embryonic day 12. Since a significant dose-rate effect has been observed in a variety of radiobiological phenomena, the effect of dose rate on the effectiveness of induction of an adaptive response by a priming dose of 0.3 Gy administered to fetuses on embryonic day 11 was investigated over the range from 0.06 to 5.0 Gy/min. The occurrence of apoptosis in limb buds, incidences of prenatal death and digital defects, and postnatal mortality induced by a challenging dose of 3.5 Gy given at 1.8 Gy/min to the fetuses on embryonic day 12 were the biological end points examined. Unexpectedly, effective induction of an adaptive response was observed within two dose-rate ranges for the same dose of priming radiation, from 0.18 to 0.98 Gy/ min and from 3.5 to 4.6 Gy/min, for reduction of the detrimental effect induced by a challenging dose of 3.5 Gy. In contrast, when the priming irradiation was delivered at a dose rate outside these two ranges, no protective effect was observed, and at some dose rates elevation of detrimental effects was observed. In general, neither a normal nor a reverse dose- rate effect was found in the dose-rate range tested. These results clearly indicated that the dose rate at which the priming irradiation was delivered played a crucial role in the induction of an adaptive response. This paper provides the first evidence for the existence of two dose-rate ranges for the same dose of priming radiation to successfully induce an adaptive response in mouse fetuses.     

Low dose of the gamma acute radiation syndrome (1.5 Gy) does not significantly alter either cognitive behavior or dopaminergic and serotoninergic metabolism.

The aim of this study was to evaluate the early-delayed effects of a low dose of the gamma acute radiation syndrome (1.5 Gy) on memory and on dopaminergic and serotoninergic metabolism in Swiss albino CD1 mice, of various ages (6, 10 and 20 weeks). At different times after irradiation (from 24 hr to three months), the mice were trained in a single-trial passive avoidance task and tested for retention either 24 hr or 5 days later. Their performance was compared to that of mice that were sham-irradiated. At the end of the behavioral test (days 3, 9, 30 and 93), the concentrations of dopamine (DA) and serotonin (5HT) and their metabolites were determined in hippocampus, anterior cortex and striatum of mice irradiated at the age of six weeks. No significant behavioral effect was observed whichever the age of the animals or the delay of observation. On the contrary at the moderate dose of 4.5 Gy we observed a significant memory deficit 9 days after the exposure. Considering the neurochemical study, in the striatum or in the frontal cortex, no significant modification was observed whichever the delay or the molecule. In the hippocampus slight modifications were noted: an increase (+144%, p = 0.002) in DA level on day 3 after exposure, and a decrease (-27%, p = 0.028) of 5HT level on day 30 post-irradiation. These modifications were only transient and not associated to modifications of the catabolites. This study demonstrates that total-body exposure to gamma radiation at low dose seems to induce only slight effects on the central nervous system.     

The cytokinesis-block micronucleus assay as a biological dosimeter in spleen and peripheral blood lymphocytes of the mouse following acute whole-body irradiation

To evaluate the application of the cytokinesis-block (CB) micronucleus (MN) assay as a biological dosimeter following in vivo exposure to ionising radiation we determined the micronucleus frequency in spleen and peripheral blood lymphocytes of the mouse, serially, for 14 days following acute whole-body irradiation. The baseline MN frequency of spleen lymphocytes (7.86 +/- 0.68, mean +/- 1 SD) was significantly (p less than 0.001) elevated when compared to that for peripheral blood lymphocytes (4.10 +/- 0.53). Immediately after irradiation there was a substantial dose-related increase in MN, but the MN frequencies in spleen lymphocytes (120.2 +/- 9.4 for 1 Gy; 409.5 +/- 38.4 for 2 Gy) were significantly (p less than 0.009) elevated compared to those in peripheral blood lymphocytes (78.0 +/- 7.0 for 1 Gy; 200.2 +/- 10.9 for 2 Gy). During the 14 days after irradiation, the MN frequency in spleen lymphocytes declined gradually to approximately half of the value observed immediately after irradiation. By contrast the MN frequency in peripheral blood lymphocytes increased during the week after irradiation, but ultimately MN frequencies in blood and spleen became approximately the same by day 14. Study of isolated murine lymphocytes irradiated in vitro showed that the number of MN generated by a given dose of radiation was approximately 2-3 times greater than the number generated by in vivo irradiation. These results suggest that measurement of MN in vivo after irradiation can be used as an in vivo dosimeter. However, precise dosimetry is probably affected by factors such as kinetic changes in different lymphocyte populations and possibly by in vivo factors which influence sensitivity of cells to radiation.     

Ultrastructure of cells of rat lymphatic organs during continuous irradiation

Changes in the ultrastructures of lymphatic organs of rats observed from the 2nd to 22nd day following continuous irradiation with gamma rays at a daily dose of 115 mGy (exposition: 12 R/day) are described. The maximum of destructive changes in lymphocytes was observed on the 11th to the 14th day of irradiation. A gradual balance between dystrophic and regenerative processes was achieved on the 18th day. In this connection to correlation could be determined between the ultrastructural changes described and the fluctuations of lymphocyte numbers in the lymphatic organs and peripheral blood during continuous irradiation.     

Insulin function in rats at early terms after 4 Gy whole-body irradiation

In the present study we made an attempt to estimate changes of insulin function at early terms after external irradiation of rats. Experimental conditions: male albino rats were studied 7; 14; 21; 28 days after the external whole-body gamma-irradiation (137Cs; 4 Gy). For this purpose the kinetics of 125I-insulin disappearance from blood plasma was investigated. Simultaneously dynamics of insulin blood concentration was studied in practically full and fasting animals. On the basis of the data received the following basic pharmacokinetic parameters were designed according to the two-compartmental model: central and peripheral compartment volumes, transfer and elimination rates, turnover and metabolic clearance rates. No substantial changes in insulin clearance were found compared to controls in all the postirradiation terms investigated. Hence, the changes in the turnover rate of insulin are proportional to blood hormone concentration. The significant increase of concentration and turnover was observed only 7 days after irradiation in rats with free access to food. The data received suggest that the insulin function of a pancreas in an organism exposed to a 4 Gy dose is maintained at a level sufficient for ensuring adequate regulation of the glucose homeostasis and of the carbohydrate metabolism.     

The dose dependence of the development of compensatory-restorative body reactions to irradiation. The EPR method]

The study deals with the mechanism of organism's adaptive responses to the effect of radiation in widely ranging dose. Post-irradiation metabolic changes were evaluated in canine blood as well as in murine blood, spleen, bone marrow and liver using the EPR spectroscopy. It was shown that the dynamics of changes in transferrin and ceruloplasmin pools and ribonucleotide reductase activity were phase-dependent with the maxima at the 2nd, 6th and 10-12th days after irradiation. Such dynamics was observed at various irradiation doses applied. The data allow us to suggest that the nonspecific compensatory--adaptive reactions of organisms develop as the response to irradiation. The dose-response function of the reaction intensity was found to be linear. The shape of the dose-response curve indicates that the minimum response of organism depends on the dose linearly up to 3.2 Gy (for dogs) as well as the maximum one. However, in the case of low-dose irradiation (0.25 or 0.5 Gy) there were deviations of maximum responses from the linearity, i.e. the amplification of the amplitude of compensatory adaptive reactions. These effect were shown to be dependent upon initial individual characteristics of animal blood and to be related to the "depressed" or "activated" state of organism prior to irradiation. The ribonucleotide reductase activity was measured in bone marrow and spleen of animals by the EPR method. The nature of non-repairable DNA damage is discussed in view of the inactivation of ribonucleotide reductase.     

Mutation induction by very low dose rate gamma rays in cultured mouse leukemia cells L5178Y

Induction of cell killing and mutation to 6-thioguanine resistance was studied in growing mouse leukemia cells in culture following gamma rays at dose rates of 30 Gy/h, 20 cGy/h, and 6.3 mGy/h, i.e., acute, low dose rate, and very low dose rate irradiation. A marked increase was observed in the cell survival with decreasing dose rate; no reduction in the surviving fraction was detected after irradiation at 6.3 mGy/h until a total dose of 4 Gy. Similarly, the induced mutation frequency decreased after low dose rate irradiation compared to acute irradiation. However, the frequency after irradiation at 6.3 mGy/h was unexpectedly high and remained at a level which was intermediate between acute and low dose rate irradiation. No appreciable changes were observed in the responses to acute gamma rays (in terms of cell killing and mutation induction) in the cells which had experienced very low dose rate irradiation.     

Lipid peroxidation of rat spleen and thymus lymphocytes treated with x-rays (0-5-1.0 Gy)

It is established, that low doses of X-ray irradiation have affected activation of lipid peroxidation (LPO) in immunocompetent cells of the spleen and thymus. The amount of malonic dialdehyde (MDA) in lymphocytes of spleen and thymocytes increases 2 times twenty-four hours after animals' irradiation by X-rays in a dose of 0.5 Gy; when a dose grows to 1.0 Gy, the MDA content in the spleen lymphocytes increases from the 1st to the 6th days and in thymocytes from the 1st to the 3d days reaching its maximum at the 3d day. MDA accumulation in the immunocompetent cells of irradiated animals varies depending on the method of lipid peroxidation initiation.     

Quantitative and qualitative changes in the peripheral blood cells of rats in chronic irradiation of the animals

A study was made of qualitative and quantitative changes in peripheral blood lymphocytes of rats subjected to continuous totalbody gamma-irradiation at dose--rates from 0.002 to 0.35 Gy/day and cumulative doses from 0.7 to 20.0 Gy. After termination of irradiation (on the 400th-500th day), the number of lymphocytes changed in an undulatory manner, i. e., increased at a dose--rate of 0.008 Gy/day, and decreased down to minimum control values at a dose--rate of 0.35 Gy/day, and the fraction of young macrocellular forms increased. The effect increased with increase in dose--rate and cumulative dose.     

Vascular response to fractionated irradiation in the rat lung.

The effects of fractionated hemithorax irradiation on normal lung tissue were examined by measuring changes in the vascular permeability surface area product (PS) and relative lung blood flow in Sprague-Dawley rats. The rats received five daily fractions per week of either 3.0 or 4.0 Gy for 4 weeks to the left lung. Between 3 and 5 weeks after the start of irradiation, the average PS was approximately 50% above normal for the group of rats that received 3.0 Gy/day and 200-300% above normal in the group of rats that received 4.0 Gy/day. Treatment with cyproheptadine, indomethacin, or theophylline had no effect, but treatment with dexamethasone significantly reduced PS to near normal levels. Left-to-right blood flow ratios in the group of rats that received 3.0 Gy/day decreased to 66% of normal levels by 4 weeks. In the group of rats that received 4.0 Gy/day, blood flow decreased to 46% of normal levels by 4 weeks. Treatment with dexamethasone maintained normal blood flow until the drug dose was reduced. These results agree with earlier studies using single-dose irradiation and indicate that the methods used to measure PS and blood flow are sensitive at low doses.     

Morphology of mononuclear cells in the peripheral blood of rats after single irradiation

The morphology of nucleated cells in the peripheral blood of rats after single dose irradiation with a dose of 5.5 Gy was followed during 28 days after irradiation. During profound agranulocytopenia and granulocytopenia the number of lymphocyte-like mononuclear cells was increased from the days 7-10 after irradiation and the number of monocyte-like mononuclear cells increased from day 14. The cell population discussed in the paper differed markedly from typical lymphocytes and monocytes in particular cytomorphologic parameters.     

不同剂量的X-射线全身照射对小鼠健康状况及涎腺的影响

目的:通过直线加速器全身照射昆明小鼠建立辐射损伤模型,探索不同放射剂量对小鼠健康状况及涎腺功能和结构的影响。方法:选取八种不同剂量对昆明小鼠行体外全身照射,于照射后一个月内观察小鼠生长情况、体重变化;照射后一周、一个月检测各组小鼠血象的变化;测定放射半数致死剂量;照射后两个月,测定各组小鼠的唾液流量及唾液淀粉酶含量,并对下颌下腺组织切片行HE染色。结果:13Gy和15Gy照射组小鼠的体重逐渐下降,一周后死亡,其余组小鼠体重最终呈增加趋势。X-射线全身照射的半数致死量为10Gy。照射后一周,照射组小鼠的白细胞数目明显降低,与对照组比较有明显统计学差异(P0.01);在其他血象方面,除了7Gy组外,其他照射组与对照组比较也均有统计学差异(P0.05)。照射一个月后,各照射组小鼠的血象均恢复正常。照射后两个月,9Gy组和11Gy组小鼠的唾液流量及唾液淀粉酶含量均显著低于0Gy组,且11Gy组较9Gy组亦明显降低,差异均有统计学意义(P0.05)。随照射剂量的增加,小鼠的下颌下腺腺泡细胞数目逐步减少,结构排列紊乱,组织损伤逐渐加重。结论:X-射线全身照射引起小鼠健康状况受损,免疫功能减低,损伤程度与放射线强度呈剂量依赖性,小鼠半数致死量为10Gy,该剂量适合建立全身放射损伤模型。     

Early changes in intracellular ATP concentration after 5 Gy irradiation by routine regimen and regimen modified by low doses (0.1+4.9 Gy)]

Comparative studies were made of immediate (within the first minutes) changes in intracellular ATP concentration after irradiation by different regimens. ATP concentration in cells at a stationary phase of growth increased immediately after irradiation with a dose of 5 Gy, reaching its maximum within 30-40 min. Irradiation with the same total dose, by the regimen of 0.1 + 4.9 Gy at a 3-minute interval between the doses, did not cause alterations in the ATP content in cells. It is concluded that the absence of the increase in the parameter under study (ATP) after irradiation by the latter regimen indicates that preirradiation at a dose of 0.1 Gy inhibits the adequate development of an early response to irradiation with a dose of 4.9 Gy.     

Combined action of virus injection and local tumor irradiation on tumor growth in mice

The dynamics of SCCVII transplantable tumor growth in C3H/H mice was determined after local tumor irradiation and/or virus (NDV LaSota) i.p. injection. The virus applied alone significantly suppressed tumor growth, particularly until the 19th day after tumor transplantation. Local irradiation with 30 Gy resulted in tumor disappearance followed with its regrowth about 15 days later. However, if the virus was injected after the irradiation, there was no tumor growth until the end of the 31 day observation period. It should be noted that virus application prior to local irradiation did not have any additional influence on tumor growth. Thus, the pronounced efficacy of virus applied after tumor irradiation deserves attention. It is possible that the virus injected after irradiation induced a chain of cytokine production joining the action of tumor destruction induced by irradiation. This should be further studied in clarifying the approaches to combined tumor therapy with possible cell-free vaccine production.     

Lipids of neocortex after X-ray irradiation of the rat head at a dose of 200 Gy

X-ray irradiation at a dose of 200 Gy with local exposure of the rat head induced the change of the lipid content in the neocortex tissue. The amount of phosphatidylinositol was decreased, the amount of free fatty acids, diglycerols, sphingomyelin was increased, and the amount of cholesterol had a growth trend in 2 h after X-ray exposition. The results testify in favor of participation of phosphatidylinositol- and sphingomyelin-relating signal systems and cholesterol in early stages of the cerebral radiation syndrome. We suggest that the change of the lipid content in early periods after the effect of a super-high dose of X-ray irradiation indicates the lipid dependence in the elimination of motion damages and the restoration of the functions of nerve cells. Effects on the lipid metabolism in the nerve tissue are promising for correcting the cerebral radiation syndrome.     

Morphological changes in spleen, bone marrow and blood of rats after continuous gamma irradiation and indomethacin treatment.

A possibility of modifying the radiation damage of haematopoiesis by indomethacin administered during continuous irradiation was followed. Indomethacin was given in drinking water (average dose of 740.7 micrograms/kg per day) within 7-days continuous irradiation with gamma rays (a daily dose rate of 2.055 Gy, total accumulated dose of 14.385 Gy). Under the given conditions no marked protective effect of indomethacin on haematopoiesis was found with the exception of mild stimulation of myelopoiesis and an increased release of developmentally younger cells of myeloid and erythroid series to the peripheral blood as well.