Langerhans cell number and morphology in mouse footpad epidermis after X irradiation

Effects of 250-kV X rays on epidermal Langerhans cells were studied in CBA/CaH mice. One group received 20 Gy to the feet, another 8 Gy to the whole body, and a third both the 8 Gy whole-body and a 12 Gy local dose to the feet. Mice from each group and controls were sacrificed at intervals from 1 to 64 days later. ATPase-positive cells in sheets of footpad epidermis were counted by light microscopy. The density of Langerhans cells in controls was 1515 +/- 36/mm2 (mean +/- SE; n = 34). By 3 days after irradiation they became rounded and less dendritic and numbers gradually reached a nadir by 10 days, at 18% of controls after 20 Gy and 57% of controls after 8 Gy. Some of the remainder exhibited bizarre morphology and ultrastructural abnormalities. After local irradiation of the feet Langerhans cell numbers recovered rapidly between 14 and 16 days, although their distribution was uneven until 30 days after irradiation. Repopulation was delayed after an 8 Gy whole-body dose by at least 3 weeks. These results demonstrate that high local doses of X rays substantially but transiently deplete the epidermal Langerhans cell population and support the hypothesis that functional hemopoietic tissue is required for extensive Langerhans cell replenishment.     

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。     

Reaction of a population of HeLa tumor cells in the stationary stage of growth to irradiation at different rates

The early stages of a repopulation process of HeLa cells under the action of irradiation 5 Gy dose and an influence of a preliminary 0.1 Gy dose irradiation at that process were investigated. As it was shown the fraction of cells with a great proliferation potential appeared in one day after lethal 5 Gy dose irradiation of the resting HeLa cells. If the other irradiation regime was used: 0.1 Gy dose plus 4.9 Gy dose in 3 min after the first action, the part of cells with a great proliferation potential became considerably less.     

Dose response of Langerhans cells in mouse footpad epidermis after X irradiation

Effects of a range (2-50 Gy) of single doses of 250 kV X rays on epidermal Langerhans cells in vivo were quantified in groups of CBA/CaH mice. Animals were sacrificed and compared with controls on the 10th day after local irradiation of their hind feet, when Langerhans cell numbers were at a minimum. ATPase-positive Langerhans cells in sheets of footpad epidermis were counted by light microscopy and cells with Birbeck granules were enumerated by electron microscopy. Both methods revealed a dose-dependent loss of Langerhans cells after ionizing radiation. Fifty percent of the ATPase-positive cells were lost after 14.4 +/- 1.3 Gy, and 50% of Birbeck granule-containing cells were lost after 17.9 +/- 4.2 Gy, suggesting that differentiated epidermal Langerhans cells are radioresistant. Loss of equivalent proportions of ATPase-positive and ultrastructurally identifiable cells after a range of doses indicates that X rays do not merely alter Langerhans cell surface markers but actually deplete the epidermal population of these cells.     

Evaluation of the expression of damage of the DNA-structural complex of rat thymocytes based on kinetics of alkaline lysis following irradiation

Rotary viscosimeters were used to study the postirradiation destruction of the DNA-structural complex (DSC) of rat thymocyte nuclei exhibited by a change in alkaline denaturation of DSC upon lysis. The S area, limited by the characteristic viscosity values obtained during alkaline lysis of thymocyte nuclei, was used as a characteristic of DSC. Immediately after irradiation the S area changed up to 81-84 per cent at 0.5-1.5 Gy and up to 56-44 per cent at 2-10 Gy. 6 to 24 h following irradiation a change in the profile of alkaline denaturation of DSC was a function of dose and dropped from 100 down to 11 per cent at doses of 0 to 10 Gy. After 2-3 days, the changes in S were also observed but they were not a strict function of dose and were the same with the values obtained immediately after irradiation.     

Induction of cytogenetic adaptive response in spermatogonia and spermatocytes by pre-exposure of mouse testis to low-dose (12)C(6+) ions

To investigate the effects of pre-exposure of mouse testis to low-dose (12)C(6+) ions on cytogenetics of spermatogonia and spermatocytes induced by subsequent high-dose irradiation, the testes of outbred Kun-Ming strain mice were irradiated with 0.05Gy of (12)C(6+) ions as the pre-exposure dose, and then irradiated with 2Gy as challenging dose at 4h after per-exposure. Poly(ADP-ribose) polymerase (PARPs) activity and PARP-1 protein expression were respectively measured by using the enzymatic and Western blot assays at 4h after irradiation; chromosomal aberrations in spermatogonia and spermatocytes were analyzed by the air-drying method at 8h after irradiation. The results showed that there was a significant increase in the frequency of chromosomal aberrations and significant reductions of PARP activity and PARP-1 expression level in the mouse testes irradiated with 2Gy of (12)C(6+) ions. However, pre-exposure of mouse testes to a low dose of (12)C(6+) ions significantly increased PARPs activity and PARP-1 expression and alleviated the harmful effects induced by a subsequent high-dose irradiation. PARP activity inhibitor 3-aminobenzamide (3-AB) treatment blocked the effects of PARP-1 on cytogenetic adaptive response induced by low-dose (12)C(6+) ion irradiation. The data suggest that pre-exposure of testes to a low dose of heavy ions can induce cytogenetic adaptive response to subsequent high-dose irradiation. The increase of PARP-1 protein induced by the low-dose ionizing irradiation may be involved in the mechanism of these observations.     

Effect of gamma-radiation on ultraviolet fluorescence of rat lymphocytes

Changes in the level of the proper UV-fluorescence ( UVF ) of rat blood lymphocytes are considered as a function of radiation dose and time after gamma-irradiation of animals. 24 h following irradiation with a dose of 0.5 Gy the UVF intensity decreased; with doses of 1-8 Gy, the level of cytofluorescence exceeded the control values. The effect of fluorescence at this time was not related to changes in a cell diameter whereas at later times following irradiation part of the effect was related to the growth of cell dimensions.     

γ射线对人淋巴细胞cx43和ANLN基因转录表达的影响

目的研究γ射线对人外周血淋巴细胞cx43和ANLN基因转录表达的影响。方法对数生长期的淋巴细胞,分别给予1、2、3、4、5、6 Gy的^60Coγ射线照射,照射后12h,以及2Gy照射后4、8、12、24、36、48、72h,分别提取总RNA,反转录成cDNA。利用实时荧光定量PCR技术,检测各组cx43和ANLN基因表达改变。结果人外周血淋巴细胞cx43 mRNA表达水平在2Gy照射后4、8、12h明显增高,分别为对照组（未照射组）的6．74、9．06、7．22倍（P〈0．05）;24～72h,其表达水平与对照组相比没有明显变化。1、2、3、4、5、6Gy剂量照射后12h,cx43 mRNA表达水平显著增高（P〈0．05）。ANLN mRoNA表达水平在2Gy不同时间点及1～5Gy照射后12h,表达降低（P〈0．05）,6Gy照射后12h其表达开始升高,为对照组的6．08倍（P〈0．05）。结论γ射线照射2Gy不同时间点及不同剂量照射后12h,cx43基因表达上调,ANLN基因表达下调。1～3Gy剂量照射后12h,cx43 mRNA表达在此范围内有时间和剂量的依赖性。cx43可能会发展为核事故受照射人员的分子生物学剂量标记物。     

Effects of single gamma-irradiation on the activity of ornithine decarboxylase in the thymus and pulmonary tissue]

In studying the dose (0.1-6 Gy) and time (2 h to 180 days) dependence of ornithine decarboxylase activity, it was found that deviations from the control were more pronounced in the thymus than in the pulmonary tissue. The radiation effect was a function of dose and time after irradiation. A nonmonotonous type of the dose-response curve was observed 7 days after irradiation: the radiation effect with a low dose (0.1 Gy) was opposite to that with sublethal doses (1-6 Gy).     

Catecholamine inhibition of Ca2+-induced insulin secretion from electrically permeabilised islets of Langerhans

Noradrenaline (1-10 microM) inhibited Ca2+-induced insulin secretion from electrically permeabilised islets of Langerhans with an efficacy similar to that for inhibition of glucose-induced insulin secretion from intact islets. The inhibition of insulin secretion from permeabilised islets was blocked by the alpha 2-adrenoreceptor antagonist, yohimbine. Adenosine 3',5'-cyclic monophosphate (cAMP) did not relieve the noradrenaline inhibition of Ca2+-induced secretion from the permeabilised islets, although noradrenaline did not affect the secretory responses to cAMP at substimulatory (50 nM) concentrations of Ca2+. These results suggest that catecholamines do not inhibit insulin secretion solely by reducing B-cell adenylate cyclase activity, and imply that one site of action of noradrenaline is at a late stage in the secretory process.     

Irradiation to ensure quarantine security for Cryptophlebia spp. (Lepidoptera: Tortricidae) in sapindaceous fruits from Hawaii

Studies were undertaken to determine whether irradiation treatment at 250 Gy, an accepted treatment for disinfestation of fruit flies in spindaceous fruits from Hawaii, would also disinfest fruit of two species of Cryptophlebia. Cryptophlebia illepida (Butler) was determined to be more tolerant of irradiation than Cryptophlebia ombrodelta (Lower); therefore, C. illepida was the focus for detailed tests. Using the criterion of success in developing to the adult stage, the pattern of tolerance to irradiation in C. illepida was generally eggs < early instars < late instars < pupae. The most tolerant stage potentially occurring in harvested fruits was late (fourth and fifth) instars. Development to adult was reduced slightly in late instars receiving an irradiation dose of 62.5 Gy, whereas development to adult was dramatically reduced in late instars receiving irradiation doses > or = 125 Gy. No C. illepida larvae receiving an irradiation dose > or = 125 Gy emerged as adults and produced viable eggs, indicating sterility can be achieved at doses well below 250 Gy. In large scale tests, when 11,256 late instars were irradiated with a target dose of 250 Gy, 951 pupated (8.4%) and none eclosed as adults. Within the pupal stage, tolerance increased with age; 7- to 8-d-old pupae (the oldest pupae tested) treated with an irradiation dose of 125 Gy produced viable offspring, whereas those treated with a dose of 250 Gy produced no viable offspring. Irradiation of adults with a target dose of 250 Gy before pairing and mating resulted in no viable eggs. Irradiation of actively ovipositing adult females resulted in no subsequent viable eggs. Therefore, the irradiation quarantine treatment of a minimum absorbed dose of 250 Gy approved for Hawaii's fruits will effectively disinfest fruits of any Cryptophlebia in addition to fruit flies.     

Characterization of altered absorptive and secretory functions in the rat colon after abdominal irradiation: comparison with the effects of total-body irradiation.

The aim of this work was to determine the alterations in the absorptive and secretory functions of the rat colon after abdominal irradiation and to compare the effects of abdominal and whole-body irradiation. Rats received an abdominal irradiation with 8 to 12 Gy and were studied at 1, 4 and 7 days after exposure. Water and electrolyte absorption was measured in vivo by insertion of an agarose cylinder into the colons of anesthetized rats. In vitro measurements of potential difference, short-circuit current and tissue conductance were performed in Ussing chambers under basal and agonist-stimulated conditions. Most of the changes appeared at 4 days after abdominal irradiation. At this time, a decrease in water and electrolyte absorption in the colon was observed for radiation doses > or = 9 Gy. The response to secretagogues (VIP, 5-HT and forskolin) was attenuated after 10 and 12 Gy. Epithelial integrity, estimated by potential difference and tissue conductance, was altered from 1 to 7 days after 12 Gy abdominal irradiation. These results show that the function of the colon was affected by abdominal irradiation. Comparison with earlier results for total-body irradiation demonstrated a difference of 2 Gy in the radiation dose needed to induce changes in the function of the colon.     

不同剂量x射线对大鼠精子CRISP2mRNA表达水平的影响

目的：观察不同剂量x射线对大鼠精子CRISP2mRNA表达水平的影响,探讨其在电离辐射所致大鼠精子功能改变中的作用。方法：用吸收剂量为1、2、4、和6Gy的x射线分别照射活体SD大鼠的外生殖系统1…4812、24h后,用PCR技术检测精子CRISP2基因mRNA表达水平;用光学显微镜观察精予活力。以未照射组为对照。结果：4、6GyX射线照射不同时间（1、4、8、12、24h时）后大鼠精子的CRISP2mRNA相对表达量均较对照组显著下降（P．〈0．05）,其中6Gb,照射24小时后相对表达量最低（P〈0．01）,而4Gy照射组与6Gy照射组相比较差异无统计学意义（P〉0．05）;2Gyx射线照射8h后CRISP2mRNA相对表达量下降有统计学意义（P〈0．05）;2GyX射线照射1、4h后及1GyX射线照射不同时间（1、4、8、12、24la）后大鼠精子的CRISP2mRNA相对表达量较对照组下降,但差异无统计学意义（P〉O．05）。1、2GyX射线照射不同时间（1、4、8、12、24小时）及4GyX射线照射（1、4、8h）后,精子活力与正常对照组相比无明显改变（P〉0．05）;4GyX射线照射12、24h后大鼠精子活力显著低于正常对照;6GyX射线照射不同时间（1、4、8、12、24h）后,精子活力明显低于对照组（P〈0．05）。结论：不同剂量X射线照射不同时间可导致SD大鼠精子活力下降,这可能与其下调CRISP2基因的mRNA表达水平有关。     

Bimodal changes in germinability of pea seeds under the influence of low doses of gamma-radiation

Pea seeds (cv. 'Nemchinovskii-85', harvest of 2002, 80%-germination percentage) were exposed to gamma-radiation with doses ranging from 19 cGy to 100 Gy. One week after the irradiation with doses of 19 cGy and 3 Gy. the germination percentage decreased to 58 and 45%, respectively; at doses of 7 and 10 Gy it was 73 and 70% respectively. At greater doses (25, 50, and 100 Gy), germination percentage decreased in proportion. Anomalous changes in seed germination percentage (as a function of irradiation dose) were caused by the redistribution of irradiated seeds between fractions I and II. The measurements of room temperature phosphorescence in air-dry seeds and the phosphorescence of endogenous porphyrines of imbibing seeds showen that the germination decrease after the irradiation with low doses (19 cGy and 3 Gy) was caused by the increase in the number of weak seeds of fraction II, which had high rates of water uptake and suffered from hypoxia under seed coat. Some of these seeds suffocated from hypoxia, and other seeds produced seedlings with morphological defects (such seeds were considered incapable of germination). During storage of seeds irradiated at doses 19 cGy-10 Gy, the recovery of germination percentage (after its initial decrease) was caused by the decrease in seed number in fraction II. The subsequent germination decrease was caused by seed death. The higher was the irradiation dose, the faster were changes in germination percentage during storage of irradiated seeds. Bimodal changes in pea seed germination with the increase of y-irradiation dose has apparently the same origin as the changes in seed germination during accelerated ageing.     

Beta-cell function in isolated human pancreatic islets in long-term tissue culture

Human pancreatic islets were isolated by collagenase treatment of pancreatic tissue obtained from 27 individuals aged 12 to 69 years. The islets were maintained free floating in tissue culture medium RPMI 1640 supplemented with calf or human serum. In two cases the insulin production was followed up to nearly two years. The insulin production rate of the individual islet preparations varied between 0.2 and 8 ng per islet per day. No significant correlation with donor age or sex was found. The glucose concentration in the medium influenced the insulin release in a dose dependent manner. The acute response of the cultured islets to glucose was evaluated both by batch incubation and by perifusion. Both in the acute and the chronic experiments maximal insulin release was found at 10 mM glucose. In conclusion, these experiments indicate that viable islets of Langerhans can be obtained from adult human pancreatic tissue and that their beta-cell function can be maintained for up to two years. The variation in insulin production rate could not be ascribed to age or sex and may reflect both physiological and methodological factors.     

Uptake of carbon monoxide by C3H mice following X irradiation of lung only or total-body irradiation with 60Co

Carbon monoxide uptake (Vco) and ventilation rate (VR) of C3H mice were determined at 14 weeks following either X irradiation of lungs only or total-body irradiation with 60Co at different dose rates. Following localized X irradiation of lung at 97 cGy/min there was a reduction in Vco, which was inversely related to radiation dose, with a small reduction below control levels being detected at 7 Gy, the lowest dose tested. An increase in VR could be detected only at doses of 11 Gy, or more. Another group of animals received 11.5 Gy total-body irradiation at either 26.2 or 4.85 cGy/min followed by transplantation with syngeneic bone marrow. Following total-body irradiation, Vco was significantly reduced by about 37% at the higher dose rate and 23% at the lower dose rate. In contrast, a trend toward elevated VR was detected only at the higher dose rate. The results indicate that Vco is a sensitive indicator of radiation-induced lung injury and that under the experimental conditions used Vco is a more sensitive indicator of radiation-induced lung injury in C3H mice than VR.     

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.     

氧离子辐射对体外人精子自化学发光,运动性,顶体反应和存活率的影响

Effects of 16O+6 ion irradiation with different doses on human sperm spontaneous chemiluminescence (SCL), motility, acrosome reaction (AR) and viability were examined. Spermatozoa were irradiated with 0, 0.25, 0.5, 1, 2, 4, 8, 16, 32, or 64 Gy 16O+6 ion beam at the energy of 3.17 MeV/u. After irradiation, samples were analyzed by SCL measurement at 1, 2 and 3 h of incubation; motility was determined by the transmembrane migration method within 2 h of incubation; the percentage of AR and viability was evaluated by the triple-stain technique at 3.5 h of incubation. The results showed: sperm SCL was significantly increased with irradiation doses and the lowest effective dose was 0.5 Gy; compared with controls, the transmembrane migration ratio of spermatozoa progressively elevated with irradiation doses at 0.5, 1, and 2 Gy; the percentage of sperm AR markedly increased in 0.5-4 Gy irradiation and the optimal dose was 2 Gy, and then significant decreased with further increase of irradiation doses; the viability had no significant change within 0.25-8 Gy, but was progressively decreased at 16, 32 and 64 Gy. These data suggested that heavy ion at low doses increased motility and AR, whereas had deleterious effects at higher doses, which are associated with free radical reactions induced by heavy ion irradiation.     

Jejunal crypt stem-cell survival after fractionated gamma-irradiation performed at different dose rates

Jejunal crypt survival after fractionated total body irradiation of C3H mice given at dose rates between 1.2 and 0.08 Gy/min was studied and the results analysed according to the linear quadratic model. Whereas alpha was independent of dose rate beta decreased with dose rate to approach zero at about 0.01 Gy/min. During the period of recovery, sublethal damage from doses given at high dose rate interact with low dose rate irradiation given immediately after, and increases its effectiveness.     

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.