Synthetic activity of rat blood lymphocytes under acute and continuous gamma-irradiation – fluorescent microspectral study

The effects of different doses of acute and continuous gamma-irradiation on the synthetic activity of rat blood lymphocytes stained with acridine orange were studied by fluorescent microspectrometry. Male rats were exposed to acute gamma-irradiation with doses of 7.5, 4 and 3 Gy, or to continuous irradiation with dose rates of 14.4, 2.1, 1.1 and 0.43 cGy/day, respectively. The changes of the synthetic activity of blood lymphocytes occurred in three main stages after acute gamma-irradiation and in four stages under continuous irradiation. The stages reflect the processes of depression and activation of the immune system under irradiation. Essential differences between the acute and continuous effects were observed in the first stage. After acute gamma-irradiation, the synthetic activity decreased sharply, indicating the predominant contribution of the damaging effect of irradiation, whereas under continuous irradiation, as a result of the stimulatory effect of low-dose irradiation, the synthetic activity increased during the first stage. Received: 11 May 1998 / Accepted in revised form: 10 January 1999     

The individual prognosis of the gravity and of the outcome of acute radiation disease based on immunological indexes

The information significance of the immunological indexes for the prognosis of gravity of course and of outcome of an acute radiation disease for the people after the exposure of ionizing radiation in clinically significant doses is studied. The value of indexes of the C-reactive protein contents, of the complement contents and of the titer of haemagglutinins in serum of a blood of 147 patients damaged at Chernobyl NPP accident as a result of external radiation gamma-exposure in combination with internal irradiation from the incorporation in an organism predominantly beta-emitting radionuclides were compared to the weight of acute radiation disease and its outcome (survival or loss). Was determined, that indexes of the contents of C-reactive protein in a peripheral blood during primary reactions on the irradiation (1-2 day after irradiation) and in latent period of disease (3-9 day after irradiation), and also titer of a complement on 3-9 day after irradiation can serve a source of information for the prognosis of probable gravity of a radiation injury and its outcome at irradiation of the man in clinically significant doses.     

转基因MSC对放射损伤胸腺组织修复的研究

目的：探讨转基因MSC能否修复放射损伤的胸腺。方法：雌性BALB/C小鼠120只,随机分为4组：对照组30只,予以假照射;照射组30只,60Coγ射线胸腺单次照射,剂量为9Gy;照射＋MSC组30只,9Gy照射后2小时输注转基因MSC;对照＋MSC组30只,假照射后2小时输注转基因MSC。照射后1、7、14、21、28天检测各组小鼠血像、胸腺指数及胸腺病理。结果：照射＋MSC组血像和胸腺指数在各个时间点均优于照射组,胸腺病理显示组织结构恢复明显加快。结论：转基因MSC能归巢至放射损伤胸腺并能促进胸腺的修复,为干细胞的临床应用提供了实验依据。     

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。     

Effect of gamma-radiation on contents of lipid peroxidation products in animal blood

The dynamics of malonic dialdehyde (MD) dependence in the blood serum of Vistar line male rats on the intensity of irradiation after a total single irradiation with 60Co gamma-quantums in 1.0 and 5.0 Gy doses (during 30 days); 9.0 Gy (during 15 days) were studied. The MD contents values both in the norm and in 0.5, 1, 2, 4, 6, 8, 10, 15, 20 and 30 days after irradiation are given. The experimental data demonstrated the different sensitivity of the final LPO parameters to the doses of 1.0, 5.0, 9.0 Gy radiation depending on the dose power, the changes of these parameters being kept for a long time after irradiation.     

Quantitative changes in the arterial blood gases of mice following localized irradiation of the lungs

The arterial pH and partial pressures of oxygen (PaO2) and carbon dioxide (PaCO2) were evaluated in LAF 1 mice 15 and 38 weeks after localized irradiation of the animals' thoraxes. Graded radiation doses of 900 to 1200 rad were administered. These doses resulted in 0 to 100% lethality by 26 weeks (180 days) after irradiation. At 15 weeks after treatment mice receiving radiation doses which would subsequently result in lethality (by 180 days) exhibited significant reductions in their PaO2 and elevations in their PaCO2 values, respectively. However, there was no clear dose-response relationship between blood gas values and radiation dose, which may reflect the animals' ability to compensate for their poor blood gas exchange by an increased breathing frequency. At 38 weeks after irradiation the blood gas values were abnormal in mice from groups which had normal blood gas values at Week 15 (and no fatalities by Week 26) but in which animal deaths had occurred between Weeks 26 and 38. These data therefore indicated (i) that abnormal blood gas values occurred in the mice prior to fatalities resulting from the acute radiation pneumonitis syndrome and (ii) that mice surviving the initial radiation pneumonitis phase could still succumb to progressive pulmonary toxicity which was reflected by the increasing levels of animal lethality and altered blood gas tensions at the later times.     

On the response of a cell population to low-dose irradiation

The cell composition of a population of human blood lymphocytes was studied after irradiation at doses of 5 cGy, 1.0 Gy and 5 cGy + 1.0 Gy and the use of a cytokinesis block. The frequencies of uni-, bi- and multinucleate lymphocytes with and without micronuclei (MN) were taken into account. By the standard criterion the frequency of binucleate lymphocytes with MN among binucleate lymphocytes--the donors were characterized as follows: in with reduction of radiosensitivity after irradiation with 5 cGy + 1.0 Gy as compared to the values of radiosensitivity after irradiation with 1.0 Gy only (an adaptive response, AR); in with no change of radiosensitivity after exposure to these doses (no AR); and with an increased ofradiosensitivity after exposure to these doses (syndrome of increased radiosensitivity, IRS). It was found that upon exposure to 1.0 Gy and 5 cGy + 1.0 Gy in some donors with AR, without AR and with IRS the total numbers of damaged cells in the population and the number of binucleate cells with MN were equal. This result calls in question the involvement of the repair mechanism in the alteration of radiosensitivity of lymphocytes in these donors. It was also observed that in the same donors a simultaneous increase (or a decrease in the case of IRS) of the portion of undamaged binucleate cells in the population took place. Our results demonstrate the existence of a new, populational, mechanism involved in the alteration of radiosensitivity after exposure to the adaptive and challenge doses.     

The antiradiation properties of betamide under conditions of sublethal and lethal gamma irradiation]

Bone marrow cellularity in the femur, mass and cellularity of the spleen and small intestine, and nucleic acid concentration in the leukocyte mass of blood were investigated after the intraperitoneal administration of betamide (500 mg/kg) 15 min before gamma irradiation of mice with doses of 4, 7 and 9 Gy. The number of myelokaryocytes and splenocytes in the protected animals was shown to exceed considerably that in the controls on days 3 and 9 after irradiation with the three doses. With betamide injected on day 9 following irradiation the number of nucleated cells of the small intestine was larger and the nucleic acid concentration in leukocytes higher than the same indices in the irradiated control.     

The effect of high energy electron irradiation on blood-brain barrier permeability to haloperidol and stobadin in rats

Summary The heads of rats were irradiated by 4 MeV electrons in doses 90, 180, and 360 Gy. The observed times of deaths ranged 120–600, 60–420, and 150–370 min after 90, 180, and 360 Gy, respectively. A dose dependent decrease of the brain uptake index of haloperidol was observed 1 and 3 h post radiation. On the other hand an increased brain uptake index was found for stobadin after head irradiation with doses of 180 and 360 Gy. Regional cerebral blood flow, blood pressure, and heart rate were not significantly altered in the period following irradiation with 180 Gy. The observed changes in blood-brain barrier (BBB) permeability seem to be the result of the damaged function of morphological structures forming the BBB rather than altered regional blood flow.     

Vascular response to radiation injury in the rat lung.

Changes in relative left-to-right lung blood flow ratios were followed as an index of vascular radiation injury in left-hemithorax-irradiated Sprague-Dawley rats. Single doses of 11 to 21 Gy gamma radiation resulted in a dose-dependent decrease in relative blood flow to the irradiated lung from 3 to 5 weeks after exposure during the development of pneumonitis. Blood flow returned to near normal by 5 weeks after lower doses (11-13.5 Gy). After a single dose of 15 Gy the left-to-right blood flow ratio recovered to 75% of normal at 12 weeks and leveled off. Following 18 Gy irradiation a second period of reduced flow began 16 weeks after exposure. After 21 Gy irradiation flow to the irradiated side remained low for 1 year after exposure. Rats that received a single dose of 18 Gy to the left hemithorax were also treated with one or two of the following drugs: captopril, cyproheptadine, dexamethasone, diethylcarbamazine, penicillamine, or theophylline. Dexamethasone was most effective at preventing the decrease in blood flow to the irradiated lung when treatment was continued through the pneumonitis period and dose was not tapered until 8 weeks after radiation exposure. All other drugs and drug combinations were, for the most part, virtually ineffective after the pneumonitis period. There was a relatively poor correlation with earlier vascular permeability surface area product studies. This suggests that endothelial damage, as well as damage to other cell types, contributes to the development of post-irradiation fibrosis in the lung.     

Effect of the balanced inhibition of DNA and protein synthesis on the in vitro and in vivo proliferative activity of the lymphocytes]

Lymphocytes isolated from the rabbit peripheral blood were irradiated in vitro with 200, 400 and 1000 r doses and cultivated with phytohemagglutinin (PHA) doses and cultivated with phytohemagglutinin (PHA) at 37 degrees C during 48 hours. In several experiments cycloheximide, inhibiting protein synthesis, was added to the cells 60 minutes before and 30 minutes after irradiation. There was no apparent difference in the viability of irradiated cells with or without cycloheximide. The ability of lymphocytes of the popliteal lymph nodes for proliferation after PHA injection into one of the hind foot-pads of the irradiated mice was studied, as well. The injection of cycloheximide or puromycin into one of the hind foot-pads immediately after irradiation of the animals augmented the proliferation of lymphocytes in this extremity in comparison with contralateral one, 1.5-2 times. Cytosine arabinoside, inhibiting DNA synthesis, was not effective under these conditions.     

Sorption of bacteria by blood cells as an indicator of the reaction of animals to the action of different doses of gamma rays

In experiments with mice, guinea pigs, and dogs, a study was made of the sorptive capacity of peripheral blood cells after different gamma-radiation doses with reference to living Escherichia coli cells. The sorptive capacity of blood cells was inhibited in exposed animals the inhibition being maximum during the first 3 days following irradiation. Homologous immunoglobulin administered to mice 24 h before irradiation prevented the diminution of the sorptive capacity of cells and stimulated it during the first week following irradiation.     

Reliability of autoregulatory mechanisms of biosystems as affected by ionizing radiation

Reliability of biosystems in processes of adaptation and homeostasis after whole-body irradiation of rats with different doses was determined in terms of the stochastic process theory developed for the autoregulatory systems with random parameters. Fluctuations in the rate of spontaneous chemiluminescence of blood serum and accumulation of lipid peroxidation products by erythrocyte membranes of rat blood were taken as the initial material. The analysis of fluctuations of peculiar parameters permitted to predict the adaptability of the system and to make timely corrections.     

Mechanism of thymocyte death in exposure to ultrahigh doses of gamma radiation

A comparative study was made of the morphological and biochemical indices of rat thymus cells after gamma-irradiation with doses of 4-10 Gy (median), 20 Gy (high), and 200-400 Gy (superhigh). It was shown that 4 h after irradiation with superhigh doses the yield of polydeoxynucleotides (PDN) was twice as low as that observed after doses of 4-10 Gy. 24 h after irradiation the amount of the extracted PDN in thymocytes exposed to superhigh doses was markedly larger than that after 4 hours. After all doses applied chromatin degradation occurred at the internucleosome sites in a strict order, the activity of acid and alkaline nucleases being unchanged. A large number of cells have normal nuclear structure 4 h after irradiation (200-400 Gy), as was demonstrated by the electron microscopy data, while in 24 h no intact cells were virtually found in the thymus which correlated with the changes in the PDN yield. The mechanisms of the lymphoid cell death under the effect of different radiation doses are discussed.     

Radiation effects on expression of FC-receptor of alveolar macrophages of rat and their specific phagocytosis

BCG-activated alveolar macrophages (AM) of Wistar rats were irradiated with different doses of gamma-ray in vitro. The effects of radiation on the expression of their Fc-receptor and specific phagocytic activity were observed. AM, after irradiation with doses of 0, 100, 300 and 500 Gy, showed decreasing phagocytic activity to chicken red blood cells (CRBC) opsonized with anti-CRBC antibody with no change in phagocytic indices. The expression of Fc-receptor of AM was, however, increased.     

Effect of mexidol on hemopoietic system in conditions of an emotional stress after exposure to ionizing radiation

The emotional stress after an irradiation can complicate the current of radiative defeats. At an emotional stress developing in early terms after an irradiation in low doses, are reduced adaptive and compensator capabilities of hemopoietic system. The emotional stress after a lethal dose irradiation inhibits the post-radiation recovery of haemopoiesis, aggravates the course of acute radiation disease and decreases the efficiency of the radioprotector--indralin. These disorders are especially pronounced under a prolonged and intensive stress. The use of the mexidol, having anxiolytic and antistress by activity, made it possible to arrest completely disturbances in the development of adaptive reactions to stress in the blood system and to normalize its compensatory potentialities in animals under conditions of combined influence of intensive long-term emotional stress and of low-dose irradiation. In the case of lethally irraiated animals, the treatment of stressed animals with mexidol favorably influenced the course of acute radiation disease, enhanced recovery processes in the blood system. Under these conditions, the use of mexidol completely removes the negative effect of emotional stress in indralin-protected animals. The pharmacological correction by mexidol from displays of an intensive emotional stress, developing after an irradiation in various doses, can be a part in system of medical measures.     

Structural changes in the DNA of the blood nucleotides and the development of late sequelae in irradiated rats

Fluorescent analysis of structural changes of nucleotide in the blood associated with AT fragments has been studied in rats, gamma-irradiated with doses of 2 and 4 Gy. It was demonstrated that the haematological, gerontological and cancerogenic late consequences were preceded by the structural genomic disturbances determined by 30 day after irradiation.     

Changes in the EPR spectra of the nitrosyl complexes of blood proteins in the low-intensity whole-body irradiation of mice

After NO adding to mice blood and isolated erythrocytes ESR signal of nitrozyl complex HbNO (g = 2.07, g = 1.98) and NO-induced MetNg (g = 6.0) were registered. It was shown that the intensity of ESR spectra of these complexes increased after radiation of mice with a dose of 0.06, 0.6 and 5.4 cGy. Low-dose irradiation (0.6 and 0.06 cGy) caused the change in the form of ESR spectra of HbNO (g = 2.07), which is indicative of the shift from T-structure to R-structure and of the preferred formation of R-conformations of oxyhemoglobin in blood. It was found that dependence of NO-induced MetHb signal on irradiation dose is bimodal that may be connected with nonlinear response of the cells to irradiation and retarded adaptive response after radiation with low doses.     

Blood Glutathione as an Index of Radiation-Induced Oxidative Stress in Mice and Humans

The effect of x-rays on GSH and GSSG levels in blood was studied in mice and humans. An HPLC method that we recently developed was applied to accurately determine GSSG levels in blood. The glutathione redox status (GSH/GSSG) decreases after irradiation. This effect is mainly due to an increase in GSSG levels. Mice received single fraction radiotherapy, at total doses of 1.0 to 7.0 Gy. Changes in GSSG in mouse blood can be detected 10 min after irradiation and last for 6 h within a range of 2.0–7.0 Gy. The highest levels of GSSG (20.1 ± 2.9 ), a 4.7-fold increase as compared with controls) in mouse blood are found 2 h after radiation exposure (5 Gy). Breast and lung cancer patients received fractionated radiotherapy at total doses of 50.0 or 60.0 Gy, respectively. GSH/GSSG also decreases in humans in a dose–response fashion. Two reasons may explain the radiation-induced increase in blood GSSG: (a) the reaction of GSH with radiation-induced free radicals resulting in the formation of thyl radicals that react to produce GSSG; and (b) an increase of GSSG release from different organs (e.g., the liver) into the blood. Our results indicate that the glutathione redox ratio in blood can be used as an index of radiation-induced oxidative stress. © 1997 Elsevier Science Inc.     

The cytogenetic effects of low doses of accelerated charged particles in human blood lymphocytes in vitro

The aim of the investigation was the study of cytogenetic effects in human blood lymphocytes of low doses of ionizing radiation in vitro. The analysis of unstable chromosome aberrations in human lymphocytes after irradiation by the accelerated ions 12C with the energy 500 MeV/nucleon and LET 10.7 keV/microm was carried out. Blood samples were irradiated on Nuclotron of the High Energy Laboratory of the Joint Institute for Nuclear Research. The doses of irradiation were in the range from 0.05 up to 1.0 Gy. Was shown that the frequency of unstable chromosome aberrations depends from the dose of ionizing radiation and can be described by linear function. At the doses 0.25-0.50 Gy the dose-independent curve was obtained for dicentrics and centric rings. The frequencies of dicentrics and centric rings as markers of the radiation action were slightly different for different donors that could be explained by different radiosensitivity. Using the calibration curve obtained earlier for gamma-rays coefficients of relative biological efficiency of accelerated 12C with the energy 500 MeV/nucleon were defined: they varied from 1.0 at the doses (0.5-1.0 Gy) up to 3.2 at the lower doses (0.05-0.25 Gy).