Gamma-irradiated onions as a biological indicator of radiation dose.

Post-irradiation identification and dose estimation are required to assess the radiation-induced effects on living things in any nuclear emergency. In this study, radiation-induced morphological/cytological changes i.e., number of root formation and its length, shooting length, reduction in mitotic index, micronuclei formation and chromosomal aberrations in the root tip cells of gamma-irradiated onions at lower doses (50-2000 cGy) are reported. The capabilities of this biological species to store the radiation-induced information are also studied.     

低能N离子注入对蚕豆根尖细胞的诱变效应

为了研究低能N离子束的细胞遗传学效应, 以不同剂量的N离子束对蚕豆种子的种胚进行辐照, 观察分析根尖细胞的微核率、 有丝分裂指数和染色体畸变效应。 研究发现, 离子束的注入抑制了根尖细胞的有丝分裂, 干扰了正常的有丝分裂过程, 引发了染色体的结构、 行为和数目畸变; 随着离子注入剂量的增加, 微核率增加、 有丝分裂指数降低、 染色体畸变率增加。 In order to study the cytogenetic effects of low energy nitrogen ion irradiation, broad bean seed embryo was irradiated by different doses of nitrogen ions. Micronucleus rate, mitotic index and chromosome aberration in root tip cells were analyzed. The results showed that the injection of ions inhibited mitosis of root tip cells, interfered the normal process of mitosis, caused aberrations of chromosome structure, behavior and number. The frequency of micronucleus and chromosomal aberrations increased with the increasing radiation dosage, while mitotic index decreased.     

The influence of the electrostatic stress on cell proliferation in plants

The putative mutagen effect of the direct current discharge—often occurring in the natural environment of cultivated and spontaneous plant populations—was studied. The electric exposure was carried out on freshly germinated seeds, meristematic tissue samples being further investigated by cytogenetic means. The pharmaceutical plant species Echinacea purpurea was chosen due to its recent re-consideration as preventive and curative treatment for the human immune system. The cellular proliferation and the chromosomal aberrations in the somatic cells of the plantlet roots have been analyzed. The atmospheric electric exposure was simulated using a homemade device, various numbers of electric pulses—from 5 to 40—with amplitudes of 15 kV being tested. Slight stimulation of the mitotic index was noticed as well as various types of chromosomal aberrations (e.g., retard chromosomes, microuclei and chromosome bridges), which suggested that the electrostatic stress could cause genetic changes in the exposed plants during their early ontogenetic stages.     

Effect of aspirin on chromosome aberration and DNA damage induced by X-rays in mice

In order to reveal the anticlastogenic potency of aspirin, we evaluated the suppressive ability of aspirin on chromosome aberrations induced by X-ray. Aspirin at doses of 0.5, 5 and 50 mg/kg was administrated intraperitoneally or orally at 0.5 h after or before the X-ray irradiation. The anticlastogenic activity of aspirin on chromosome aberrations induced by X-ray was determined in the mouse micronucleus test and alkaline single cell gel electrophoresis (SCG) assay in vivo. The frequency by polychromatic erythrocytes with micronuclei (MNPCEs) was decreased by about 19–61% at 0.5 h after and about 23–62% at 0.5 h before the X-ray irradiation. DNA damage by X-ray was significantly decreased by oral administration of aspirin at 0.5 h after or before the X-ray irradiation for the SCG assay. We consider aspirin can be used as preventive agents against exposure of X-ray.     

The effects of nano-TiO<Subscript>2</Subscript> on seed germination,development and mitosis of root tip cells of <Emphasis Type="Italic">Vicia narbonensis</Emphasis> L. and <Emphasis Type="Italic">Zea mays</Emphasis> L

This study aimed to provide new information about phyto-toxicology of nano-TiO₂ on plant systems. To contribute to the evaluation of the potential harmful effects of the nanoparticles on monocots and dicots we considered their effects on seed germination and root elongation applying a concentration range from 0.2 to 4.0‰ in the plants Zea mays L. and Vicia narbonensis L. Moreover, we achieved a genotoxicity study at cytological level in root meristems by means of traditional cytogenetic approach, to evidence possible alterations in mitotic activity, chromosomal aberrations, and micronuclei release. From these analyses it comes out that nano-TiO₂ particles, after short-term exposure and under our experimental conditions, delayed germination progression for the first 24 h in both materials. Root elongation was affected only after treatment with the higher nano-TiO₂ concentration. Further significant effects were detected showing mitotic index reduction and concentration-dependent increase in the aberration emergence that evidenced a nano-TiO₂-induced genotoxic effect for both species.     

The incidence of mitotic abnormalities in cryopreserved eight-cell early and compacted mouse embryos

The effects of slow freezing-rapid thawing on viability and chromosomal complement of eight-cell early and eight-cell compacted mouse embryos were investigated. The abnormalities connected with damage to the mitotic apparatus, and with chromosome damage were investigated in cryopreserved embryos by cytological analysis. The embryos were preserved using 1M glycerol as cryoprotectant and a slow cooling regime to -30 degrees C before transfer to liquid nitrogen. The proportion of mitotic abnormalities in compacted embryos was significantly higher (13.1%) than in early embryos (5.9%) and unfrozen control embryos (3.9%) in these studies performed after thawing. This was reflected after cryopreservation by a reduced viability of the embryos as judged by culture to the hatching blastocyst stage--compacted 8-cell embryos (71.8+/-4.7%) versus controls (78.4+/-5.9%) and cryopreserved early-stage 8-cell embryos (86.1+/-4.0%)--p<0.05 in each case. However, aneuploidy rates were low in all groups in both fresh and cryopreserved embryos (around 3%).     

Cytogenetic changes induced by aqueous ferrofluids in agricultural plants

In this paper, the authors present their results regarding the cellular division rate and the percentage of chromosomal aberrations in the root meristematic cells of agricultural plants when cultivated in the presence of different concentrations of aqueous ferrofluid, ranging between 10 and 250 μL/L. The agricultural species (Zea mays) with a major role in the life of people was chosen for the experimental project. The water-based ferrofluid was prepared following the chemical co-precipitation method, using tetramethylammonium hydroxide as magnetite core stabilizer. Microscopic investigations (cytogenetic tests) resulted in the evaluation of the mitotic and chromosomal aberration index. They appeared to increase following ferrofluid addition.     

Development of cataract and corneal opacity in mice due to radon exposure

This work investigates the radiation damage on the eye of albino mice exposed to effective radon doses ranging from 20.92 to 83.68 mSv. These doses were taken over 2–8 weeks using a radon chamber constructed by the National Institute for Standard (Egypt). The guidance on the quality assurance program for radon measurements was followed. Therefore, the estimated doses received by the laboratory animals meet the requirements of national standard. The refractive index(RI) and protein concentration were measured for soluble proteins of both corneas and lenses. In addition, the sodium dodecyle sulfate polyacrylamide gel electrophoresis (SDSPAGE) technique was used. The results show increasing of the RI of both cornea and lens proteins and decreasing in total protein concentration of exposed animals. These results were accompanied with changes in the SDSPAGE profile for both cornea and lens. Therefore, radon exposure produces substantial hazards to the cornea and lens of experimental animals and has a crucial role in the development of cataract and corneal opacity.     

Activation of mononuclear bone marrow cells treated in vitro with a complex homeopathic medication

Canova is a Brazilian homeopathic medication with immunomodulatory properties, recommended for patients where the immune system is depressed. Previous studies demonstrated that Canova induces up-regulation in numbers of leukocytes. The bone marrow microenvironment is composed of growth factors, stromal cells, extracellular matrix and progenitor cells that differentiate into mature blood cells. We now report the effect of in vitro administration of the medication on the mononuclear differentiation of the bone marrow cell. Swiss mice femurs were dissected cleaned and the cells of the marrow were flushed. The cells were plated, treated or not, incubated for different times and processed for light, transmission and scanning electron, and confocal microscopy analysis. Bone marrow cells showed an enhanced proliferation in vitro in response to Canova medication and Canova plus M-CSF and an increase was also observed in the numbers of the cell niches and ring-shaped nuclei cells. Confocal and transmission and scanning electron microscopy showed the stages of monocyte maturation, with resting and activated cells. With Canova treatment there was a marked increase in cell size, which is mainly attributable to the augmented cytoplasm, an increase in the number of mitochondria, expansion of the RER and an enlarged Golgi. The response to Canova treatment indicates that it influences mononuclear differentiation and activation of bone marrow progenitor and stromal cells.     

The relative biological effectiveness of a high energy neutron beam for micronuclei induction in T-lymphocytes of different individuals

In assessing the radiation risk of personnel exposed to cosmic radiation fields as it pertains to radiological damage during travel in civilian aircrafts, it is particularly important to know the relative biological effectiveness (RBE) for high energy neutrons. It has been the subject of numerous investigations in recent years using different neutron energies and cytogenetic examinations. Variations in the radiosensitivity of white blood cells for different individuals are likely to influence the estimate of the relative biological effectiveness for high energy neutrons. This as such observations have been noted in the response of different cancer cell lines with varying inherent sensitivities. In this work the radiosensitivities of T-lymphocytes of different individuals to the p(66)/Be neutron beam at iThemba LABS were measured using micronuclei formations and compared to that noted following exposure to ⁶⁰Co γ-rays. The principle objective of this investigation was to establish if a relationship between neutron RBE and variation in biological response to ⁶⁰Co γ-rays for lymphocytes from different individuals could be determined. Peripheral blood samples were collected from four healthy donors and isolated lymphocytes were exposed to different doses of ⁶⁰Co γ-rays (1–5 Gy) and p(66)/Be neutrons (0.5–2.5 Gy). One sample per donor was not exposed to radiation and served as a control. Lymphocytes were stimulated using PHA and cultured to induce micronuclei in cytokinesis-blocked cells. Micronuclei yields were numerated using fluorescent microscopy. Radiosensitivities and RBE values were calculated from the fitted parameters describing the micronuclei frequency dose response data. Dissimilar dose response curves for different donors were observed reflecting varying inherent sensitivities to both neutron and gamma radiation. A clear reduction in the dose limiting RBE_M is noted for donors with lymphocytes more sensitive to γ-rays (p = 0.032, R² = 0.94). Unlike observations made with different cancer cell lines exposed to the same clinical neutron beam, the variations in neutron RBE observed in T-lymphocytes of different individuals is related to the cellular radioresistance to photons.     

X射线诱导的HeLa细胞旁效应中ROS和NO关系研究

活性氧(ROS)和一氧化氮（NO)是辐射诱导的旁效应信号通路中的两个重要信号分子。 实验研究了这两种信号分子在HeLa细胞旁效应信号通路中的关系。 通过微核实验, 发现X射线辐照过的HeLa细胞及其旁观者细胞微核形成明显增加, 而二甲亚砜（DMSO）预处理显著抑制了微核形成。 另外还发现, 接受条件培养基的旁观者细胞的增殖速率增加, 而DMSO预处理产生条件培养基的受辐照细胞则使旁观者细胞的增殖速率降低。 以上的结果从不同角度证实了HeLa细胞存在X射线诱导的旁效应, 且其可以被DMSO预处理所抑制。Western blotting和DAF FM DA荧光探针检测分别显示出辐照后细胞的诱导型一氧化氮合酶（iNOS）和NO水平均升高, 而DMSO预处理则降低其水平。 因此, 可以推测X射线诱导的HeLa旁效应当中ROS是NO的上游信号。 Accumulating evidence indicates that irradiated cells can release signals which induce a series of biological responses in non exposed cells. This is known as irradiation induced bystander effects. Both reactive oxygen species(ROS) and nitric oxide(NO) play important roles in bystander effects. In this study, we determined the relationship of ROS and NO in the signaling pathway of bystander effects. HeLa cells were treated with or without dimethye sulfoxide(DMSO) before X ray irradiation, and micronuclei formation as well as cell proliferation rate was detected in both irradiated and bystander cells. In addition, we also detected inducible nitric oxide synthase(iNOS) expression and NO level in irradiated cells using Western blotting and DAF FM DA fluorescent probe, respectively. Our results showed that micronuclei were induced in irradiated and bystander cells while DMSO treatment significantly suppressed the formation of micronuclei in both of them. We also found that when cells were irradiated their proliferation rate was suppressed while DMSO treatment eliminated this inhibition effect. In contrast, the cells received conditioned medium from irradiated cells proliferated more quickly than the cells received medium from non irradiated cells while DMSO treatment reduced the difference. Finally, we found that irradiated cells had higher level of iNOS and NO compared to non irradiated controls, whereas DMSO treatment decreased their levels. These results suggest that ROS is the upstream signal of NO in X ray induced bystander effects in HeLa cells.     

~(12)C~(6+)高能重离子辐照大葱损伤及其分子生物学效应

利用30,90,180Gy3种剂量的12C6+重离子束辐照大葱种子,研究其在细胞水平和农艺性状的诱变效应并进行RAPD分析。通过与M1代的研究结果比较后表明:经过不同剂量12C6+重离子照射后能有效地诱导大葱细胞形成微核和染色体畸变,这种诱变效应,在M2代仍然有所表现。M1代大葱结果期的株高、白长、花序直径和种子产量随辐照剂量增加产生明显差别,其中30Gy辐照组增幅最大。大葱总水溶性蛋白质和维生素C的含量在30Gy组中积累最多,在90Gy组有明显下降。与M1代一致,M2代中大葱染色体微核率及RAPD分析所得的DNA多态性比率仍然与辐照剂量呈正相关,但比率整体下降;说明高能量重离子辐照造成的DNA变异在M2代被修复和淘汰。     

Characterisation of cryopreserved cells freshly isolated from human bone marrow

Osteoblast progenitor cells (OBPCs) isolated from bone marrow have the ability to differentiate into osteoblasts and thus potential therapeutic use to tissue-engineer bone. In order for OBPCs to be available for clinical use a means of storing viable cells is necessary. The aim of this study was to determine whether a simple method of cryopreservation had an effect on osteogenic differentiation or growth of OBPCs isolated from fresh human bone marrow. Stro-1 was used to identify the isolated OBPCs. The osteoblastic potential of the marrow cells was confirmed as culture with osteogenic supplements (OS) significantly increased osteoblastic protein production (alkaline phosphatase (ALP), osteopontin and osteocalcin) compared with standard conditions (P less than 0.05). Ten further marrow aspirates were harvested; each was halved for either cryopreservation or control culture. Primary cultures from both populations formed colonies with recognised OBPC morphology. OS stimulated both cryopreserved and control populations to produce significantly more osteoblastic proteins (P less than 0.05) and there was no significant difference between the increase in osteogenic proteins when cultured with OS (P great than 0.2). The proliferation rate after 5 days in culture was not significantly affected by cryopreservation (P greater than 0.05). It has been suggested that OBPCs are immuno-privileged; so allogenic cells could be implanted into patients for tissue engineering bone without causing a hypersensitivity reaction. Our study demonstrates a method of storage, which allows OBPCs to be available for use without affecting osteoblastic potential or viability.     

激光扫描共聚焦显微技术对水稻不育系花粉减数分裂的特性研究

以不育系新稻97200A为材料,利用激光扫描共聚焦显微镜对其花粉母细胞减数分裂及成熟花粉育性进行观察研究,此外还调查了不育系自交结实率和人工授粉异交结实率。结果表明：不育系异常减数分裂占15.8%,包括拖拽染色体、落后染色体、三极纺锤体、染色体桥、微核和子染色体分离不同步等。成熟花粉育性检验典败花粉占6.15%,圆败花粉占10.26%,染败花粉占80.77%。绝大部分的花粉败育可能是由于减数分裂过程中出现的染色体异常配对、异常分裂、染色体断裂及细胞质分布不均匀等使得花粉粒发育不能正常进行而导致的。     

Radiation response of cell organelles

The cellular responses to various form of radiation, including ionizing- and UV-irradiation or exposure to electromagnetic fields is manifested as irreversible and reversible structural and functional changes to cells and cell organelles. Moreover, beside the morphological signs related to cell death, there are several reversible alterations in the structure of different cell organelles. The radiation-induced changes in the supramolecular organization of the membranes, including plasma membrane, and different cell organelle membranes, play a significant role in the development of acute radiation injury. These signs of radiation-induced reversible perturbation biological membranes reflect changes in the organization and/or composition of the glycocalix, modified activity and/or distribution of different membrane domains, including enzymes and binding sites. The observed changes of the cell surface micromorphology and the alteration of intercellular connections are closely related to the reorganization of the cytoskeletal elements in the irradiated cells. The mitochondria, endoplasmic reticulum, Golgi-complex, the lysosomal system have long been considered to be direct intracellular targets of irradiation. The listed morphological alterations of nuclear chromatin (e.g. changes of fine structure, altered number of nucleolar organizing regions and micronuclei, development of chromosome aberrations) may originate from the radiation-induced damage to the supramolecular organization of DNA and/or nucleus specific proteins. These endpoints of radiation effects resulted as direct consequence(s) of absorbed radiation energy, and indirectly altered intra-, intercellular communication or modified signal transduction. Some complementary data suggest that all these effects are not strictly specific to radiation and may be best considered as general stress responses, similar to those observed after application of various injurious agents and treatments to cells. Moreover, they may be equally responsible for direct degradation of supramolecular component of cells, altered signal transduction, or changes in the amount or ratio of any extracellular mediators upon irradiation. Nevertheless, qualitative and/or quantitative evaluation of any changes of chromosomes by different techniques (morphological analysis of metaphase chromosomes, fluorescent in situ hybridization, development of micronuclei etc.) are useful biological indicators as well as "biological dosimeters" of radiation injury. It is suggested, that some modern methods such as immunohistochemical detection of different proteins, specific markers of cell organelles and cytoskeleton, inspection of distribution of cell surface charged sites and different membrane domains and application of tracer substances may all be included into protocols for evaluation of cell alterations induced by different types and intensities of radiation.     

Atomic force microscope tracking observation of Chinese hamster ovary cell mitosis

CHO cells possess easily identifiable karyotypes, and CHO cell chromosomes are large and few in number, making these cells ideal for mutational and drug toxicity studies and suitable for investigations of animal chromosome structure. Here, we used atomic force microscopy (AFM) in the tapping mode for detailed visualizations of Chinese hamster ovary (CHO) cell chromosomes during various mitotic phases, including typical prophase, prometaphase, metaphase, anaphase and telophase. Based on our detailed observations, we were able to divide metaphase and anaphase into sub-phases: metaphase I, II and III, and anaphase I and II. Furthermore, we used the AFM error-signal mode to visualize chromosomal ultrastructures and cytokinesis. While these visualizations were all successful, we found that the image quality was affected by cellular debris, contamination. Collectively, our results show that the AFM technique has great potential for the detailed study of chromosomes and chromosomal ultrastructures during all phases of the cell cycle, but that careful standards of sample preparation must be maintained.     

The effect of boundary conditions on guided wave propagation in two-dimensional models of healing bone

Guided wave propagation has recently drawn significant interest in the ultrasonic characterization of bone. In this work, we present a two-dimensional computational study of ultrasound propagation in healing bones aiming at monitoring the fracture healing process. In particular, we address the effect of fluid loading boundary conditions on the characteristics of guided wave propagation, using both time and time-frequency (t-f) signal analysis techniques, for three study cases. In the first case, the bone was assumed immersed in blood which occupied the semi-infinite spaces of the upper and lower surfaces of the plate. In the second case, the bone model was assumed to have the upper surface loaded by a 2mm thick layer of blood and the lower surface loaded by a semi-infinite fluid with properties close to those of bone marrow. The third case, involves a three-layer model in which the upper surface of the plate was again loaded by a layer of blood, whereas the lower surface was loaded by a 2mm layer of a fluid which simulated bone marrow. The callus tissue was modeled as an inhomogeneous material and fracture healing was simulated as a three-stage process. The results clearly indicate that the application of realistic boundary conditions has a significant effect on the dispersion of guided waves when compared to simplified models in which the bone's surfaces are assumed free.     

Time-course of aberrations and their distribution: impact of LET and track structure

The biological response to high linear energy transfer (LET) radiation differs considerably from that to low LET radiation and this has been attributed to differences in the spatial energy deposition of both radiation qualities. In the case of X-rays the energy is deposited uniformly within the cell nucleus and produces damages in a purely stochastic manner. In contrast, for particles the energy is deposited inhomogeneously along the ion trajectory and the local dose decays with the square radial distance from the center of the track. This nonuniformity affects the yield and the distribution of aberrations among cells. Moreover, after high LET exposure a relationship between the aberration yield and cell cycle delay was observed. In this study, we present a detailed analysis of the distribution of aberrations in human lymphocytes reaching mitosis at early and later times after low and high LET exposure. Aberration data were fit to stochastic distributions demonstrating that the delay is related to the number of particle traversals per cell nucleus. To further elucidate this relationship, we introduce a Monte Carlo phenomenological model which incorporates the number of particle hits per nucleus. This value was derived by fitting theoretical distributions to the experimental data. Additionally, the probability that a cell traversed by a particle reaches mitosis at a given time was calculated. The analysis of biological data and numerical simulations clearly show the impact of the track structure on the formation of chromosome aberrations and their distribution among cells.     

Analysis of MIR-18 results for physical and biological dosimetry: radiation shielding effectiveness in LEO.

We compare models of radiation transport and biological response to physical and biological dosimetry results from astronauts on the Mir space station. Transport models are shown to be in good agreement with physical measurements and indicate that the ratio of equivalent dose from the Galactic Cosmic Rays (GCR) to protons is about 3/2:1 and that this ratio will increase for exposures to internal organs. Two biological response models are used to compare to the Mir biodosimetry for chromosome aberration in lymphocyte cells; a track-structure model and the linear-quadratic model with linear energy transfer (LET) dependent weighting coefficients. These models are fit to in vitro data for aberration formation in human lymphocytes by photons and charged particles. Both models are found to be in reasonable agreement with data for aberrations in lymphocytes of Mir crew members: however there are differences between the use of LET dependent weighting factors and track structure models for assigning radiation quality factors. The major difference in the models is the increased effectiveness predicted by the track model for low charge and energy ions with LET near 10 keV/micrometers. The results of our calculations indicate that aluminum shielding, although providing important mitigation of the effects of trapped radiation, provides no protective effect from the galactic cosmic rays (GCR) in low-earth orbit (LEO) using either equivalent dose or the number of chromosome aberrations as a measure until about 100 g/cm 2 of material is used.     

Effects of recombinant human erythropoietin on the haemopoietic bone marrow monitored by magnetic resonance spectroscopy in patients with end-stage renal disease

Volume selective magnetic resonance (MR) proton spectroscopy was used to investigate changes in the haemopoietic bone marrow in patients with end-stage renal disease undergoing treatment with recombinant human erythropoietin (rHuEPO). Significant changes could be detected in the spectra 14 days after the beginning of treatment, before any response was seen in the haemoglobin concentration of peripheral blood. The spectral changes indicate an alteration in cellular composition of haemopoietic bone marrow with an increase in the amount of haemopoietic active tissue. One patient showed a major change in the spectrum four days after treatment began, indicating that MR spectroscopy may detect early changes in the cellular composition of the bone marrow. This noninvasive method may be useful in evaluating treatment effects of recombinant human haemopoietic growth factors in the bone marrow, as well as investigating bone marrow response from different modes of rHuEPO administration.