Effects of γ-irradiation on antioxidant activity in soybean seeds

There are some reports that low doses of γ-irradiation could induce antioxidant activities in plant material, including soybean. Irradiation, required for the inactivation of some pathogens and induction of mutations, may have adverse effects on sensorial, nutritional and antioxidant qualities. The effects of different γ-irradiation doses (100–200 Gy) on antioxidant properties of soybean seeds was investigated. In this study, we report the results obtained by analysis of antioxidant enzyme activities, reduced glutathione, malonyldialdehyde (MDA) and hydroxyl (HO⁻) radical quantities, soluble protein content, and total antioxidant activity in irradiated soybean seeds. Antioxidant enzyme activities were affected due to high irradiation intensity. Significant changes of total antioxidant activity and MDA and HO.quantities were observed only under the highest irradiation dose, with a 15.7% reduction in total antioxidant activity, MDA quantity increase of 21.6%, and HO⁻ radical quantity increase of 79.3% compared to the non-irradiated control. The total soluble protein content increased slightly.     

Role of gamma irradiation in regulation of NO3 level in rocket (Eruca vesicaria subsp. sativa) plants

The changes in the growth indices and certain metabolic activities in response to different doses of γ-irradiation (0, 20, 50, 100, and 200 Gy) were studied, using rocket seedlings. The total yield, seed mass, and essential oil content increased significantly by 20 Gy dose of γ-irradiation as compared with the control samples. In addition, total sugars, total free amino acids, total soluble phenols, kinetin, GA₃, nitrate reductase activity, and total protein increased significantly at 20 Gy dose. Meanwhile, NO₃ content decreased significantly at 20 Gy dose. The nitrogen and potassium content increased at the same dose of gamma rays of 20 Gy. While the phosphorus content showed no significant effect at all the γ-irradiation doses used. The results obtained in this work revealed that the most effective dose was 20 Gy of gamma rays. It was suggested that pretreatment of rocket seeds before planting with 20 Gy dose of γ-irradiation, may oppose the harsh effect of NO₃ accumulation and increased the quality and quantity of rocket yield. Published in Russian in Fiziologiya Rastenii, 2006, Vol. 53, No. 2, pp. 215–219. The text was submitted by the author in English.     

Biotic and abiotic initiators for rishitin formation and accumulation in tomato

γ-Ray irradiation of pre-sowing seeds of tomato did not trigger the formation of the phytoalexin “rishitin” in either leaves or fruits of tomato plants through different growth seasons. Application of copper sulfate initiated rishitin formation in both leaves and fruits. Increasing of γ-ray dose was accompanied by decreasing rishitin accumulation in the presence of copper sulfate. Rishitin of tomato leaves was found to be reduced significantly, concomitant with increasing the disease incidence for late and early blight, andFusarium wilt disease, after applying γ-irradiation, in the case of biotic initiatorsPhytophthora infestans, Alternaria solani orFusarium oxysporum alone or together with the abiotic inducer copper sulfate. Shelf-extending γ-ray doses of 1.0, 1.5 and 2.0 kGy decreased rishitin amounts in tomato fruits treated with copper sulfate alone or infected withPhytophthora infestans. Also, the amount of formed rishitin was reduced by extending the storage period.     

Gamma irradiation induced apoptotic changes in the chromatin structure of human erythroleukemia K562 cells

Exponentially growing human erythroleukemia K562 cells were synchronized by centrifugal elutriation prior to and after Co⁶⁰ γ-irradiation (4 Gy). Forward scatter flow cytometry used for size analysis revealed the increase of an early apoptotic cell population ranging from lower (0.05 C-value) to higher DNA content (∼1 C) as the cells progressed through the S phase. The increase in cellular DNA content expressed in C-values correlated with apoptotic chromatin changes manifested as many small apoptotic bodies in early S phase and larger but less numerous disintegrated apoptotic bodies in late S phase. Most significant changes after exposure to γ-irradiation took place in early S phase resulting in an increase of nuclear size by more than 50%. Cell fractions containing irradiated cells showed enhanced growth arrest at 2.4 C-value, which was accompanied by apoptosis. Apoptotic cell cycle arrest near to the G₁/G₀ checkpoint and apoptotic changes indicate that the radiation resistance of K562 cells is related to the bypass of the early stage of the p53 apoptotic pathway. Apoptotic changes in chromatin structure induced by γ-irradiation indicate that these injury-specific changes can be identified and distinguished from chromatin changes induced by UV radiation or heavy metals.     

Transcriptomic analysis of rat brain tissue following gamma knife surgery: Early and distinct bilateral effects in the un-irradiated striatum

Gamma knife surgery (GKS) is used for the treatment of various human brain disorders. However, the biological effects of gamma ray irradiation on both the target area, and the surrounding tissues are not well studied. The effects of gamma ray exposure to both targeted and untargeted regions were therefore evaluated by monitoring gene expression changes in the unilateral irradiated (60 Gy) and contralateral un-irradiated striata in the rat. Striata of irradiated and control brains were dissected 16 hours post-irradiation for analysis using a whole genome 44K DNA oligo microarray approach. The results revealed 230 induced and 144 repressed genes in the irradiated striatum and 432 induced and 239 repressed genes in the un-irradiated striatum. Out of these altered genes 39 of the induced and 16 of the reduced genes were common to both irradiated and un-irradiated tissue. Results of semiquantitative, confirmatory RT-PCR and western blot analyses suggested that γ-irradiation caused cellular damage, including oxidative stress, in the striata of both hemispheres of the brains of treated animals.     

Gamma-irradiation depletes endogenous germ cells and increases donor cell distribution in chimeric chickens

The production of chimeric birds is an important tool for the investigation of vertebrate development, the conservation of endangered birds, and the development of various biotechnological applications. This study examined whether gamma (γ)-irradiation depletes endogenous primordial germ cells and enhances the efficiency of somatic chimerism in chickens. An optimal irradiation protocol for stage X embryos was determined after irradiation at various doses (0, 100, 300, 500, 600, 700, and 2,000 rad). Exposure to 500 rad of γ-irradiation for 73 s significantly decreased the number of primordial germ cells (P < 0.0001). Somatic chimera hatchlings were then produced by transferring blastodermal cells from a Korean Oge into either an irradiated (at 500 rad) or intact stage X White Leghorn embryo. An analysis of feather color pattern and polymerase chain reaction-based species-specific amplification of various tissues of the hatchlings confirmed chimerism in most organs of the chick produced from the irradiated recipient; a lesser degree of chimerism was observed in the non-irradiated control recipient. In conclusion, the exposure of chick embryos to an optimized dose of γ-irradiation effectively depleted germ cells and yielded greater somatic chimerism than non-irradiated control embryos. This technique can be applied to interspecies reproduction or the production of transgenic birds.     

Effect of γ-radiation on some elements in certain organs of albino rats

Energy-dispersive X-ray fluorescence (EDXRF) was used to determine the concentrations of Ca, Cl, Fe, Ni, P, K, Se, S, and Zn in heart, lung, liver, spleen, and kidney of adult albino rats 2 mo after they were subject to a single gamma γ-radiation dose from⁶⁰Co at 5 gy. In female rats, K levels were significantly higher and the Ca levels significantly lower for the irradiated animals when compared to age-matched nonirradiated controls. Significant differences between irradiated and nonirradiated tissues were observed for other elements, although no sex-related differences could be found. Tissue damage and disturbances of biological functions were observed as a result of γ-irradiation.     

Changes in cellular proteins ofDeinococcus radiodurans followingγ-irradiation

  In order to examine radiation-induced proteins in an extremely radioresistant bacterium, Deinococcus radiodurans R ₁, changes in cellular proteins after γ-irradiation were analysed by two-dimensional gel electrophoresis and silver staining. Nine proteins (190, 120, 87, 60, 58, 52, 46, 41 and 41 kDa) were increased (or appeared) and more than 13 proteins diminished after γ-irradiation at 6 kGy. Increase of eight proteins (except for 190-kDa protein) was prevented when the cells were irradiated in the presence of chloramphenicol. Three proteins, 87, 60 and 46 kDa, continued to be synthesized during post-irradiation incubation, and the amounts of these proteins increased with higher doses in a range of 1 – 12 kGy. Changes in the amount of proteins after irradiation in the R  ₁ strain were compared with those in a moderately radioresistant mutant (rec 1) and in a highly radiosensitive mutant (rec30). These three proteins were increased in both R ₁ and rec 1, but not in rec 30, suggesting that they are characteristic for radioresistant strains. In addition, from the microsequence analysis, the 46-kDa protein was found to be homologous to the EF-Tu protein of Escherichia coli, whereas the remarkable homologous sequence to the N-terminal of the 60-kDa protein was not found among the known proteins. Received: 28 March 1995 / Accepted in revised form: 16 January 1996     

Effects of γ-ray treatment onCannabis saliva pollen viability

The viability and thein vitro germination capability of hemp pollen (cv. Carmagnola) were studied. Viability tests were based on the microscopic observation of the fluorescence of loaded fluorescein diacetate (FDA), while, for germinability tests, five different media were tested. The effects of irradiation with γ-rays on pollen viability and germination and on seed set were also studied, at three different irradiation doses (20, 60 and 100krad). The results show that in one of the media tested, about 85–90% of the pollen grains are viable and able to germinate in control samples, and that while viability measured by FDA test is not affected by increased γ-ray doses, the pollenin vitro germinability drops to about one-half of the controls at the maximum γ-ray dose employed, 100krad. Seed set of hemp plants pollinated with the irradiated pollen dropped to less than 1% of that of plants pollinated by untreated pollen for the higher dose used. The different media suitable forin vitro germination of hemp pollen, and the observed lack of correspondence between viability and germination capacity tests are discussed.     

Effects of pulsed electric fields on DNA of human lymphocytes

The effects of pulsed electric fields of low frequency (50 Hz) on DNA of human lymphocytes were investigated. The influence of additional external factors, such as hydrogen peroxide (H₂O₂) and γ-irradiation, as well as the repair efficiency in these lymphocytes, was also evaluated. The comet assay, a very sensitive and rapid method for detecting DNA damage at the single cells level was the method used. A significant amount of damage was observed after exposure to the electric fields, compared to the controls. After 2 h incubation at 37^°C, a proportion of damage was repaired. H₂O₂ and γ-irradiation increased the damage to lymphocytes exposed to pulsed electric fields according to the dose used, while the amount of the repair was proportional to the damage.     

Systematic review of experimental studies on the relative biological effectiveness of tritium

Tritium (³H) is a radioactive isotope of hydrogen. A number of factors combine to create a good deal of interest in tritium doses, both to workers and to members of the public. Tritium is ubiquitous in environmental and biological systems and is very mobile due to its occurrence as water. In this study we systematically review experimental data relating to tritium exposure with a view to assessing its low dose limiting relative biological effectiveness (RBE_max). Interpretation of published experimental studies is complicated by the fact that the reference radiations varied, and doses and dose rates were frequently much higher than those normally received by humans. The four available animal carcinogenicity studies gave RBE values of about 2.5 with chronically-delivered γ-ray reference, and about 1.2 with chronically-delivered X-ray reference. However, because of problems associated with the design and interpretation of the experiments, we do not consider that these RBE values should be taken to apply to the induction of cancer at low doses (i.e. they should not be interpreted as RBE_max). Combining the six studies with chronic γ-ray reference, with adequate quantitative data that examined endpoints apart from cell survival and related endpoints, yields an aggregate RBE estimate of 2.19 (95% CI 2.04, 2.33); the analogous combined RBE estimate using the three studies with chronic X-ray reference groups is 1.17 (95% CI 0.96, 1.39). Again, problems with the design, in particular the range of doses used in some of these studies, mean that these RBE values should also probably not be interpreted as RBE_max.     

Pre-sowing gamma irradiation of cotton seeds from a point source of Cs137 and its effect on emergence,growth and yield

Summary The effect of pre-sowing -irradiation of cotton seeds from a point source of Cs¹³⁷ on emergence, growth, flowering, and yield was studied. High radiation doses caused reduction of emergence, inhibition of growth retardation of flowering, and reduction in number of flowers and bolls produced by the plant. The yield was greatly reduced under high dosages and approached that of the control plants under low dosages. -rays caused a marked degree of sterility.No significant stimulating effect on growth and development was detected when the dry cotton seeds were subjected to -irradiation doses of 1.17 to 26.92 kr.     

Induction of Heat Stress Tolerance in Barley Seedlings by Pre-Sowing Seed Treatment with Glycinebetaine

Heat stress adversely affects plant growth and development, while glycinebetaine (GB) plays a protective role under stressful conditions. The objective of this study was to assess the optimum level of GB for use as a presowing seed treatment and the subsequent effect on the heat tolerance of barley (Hordeum vulgare L. cv. Haider-93) seedlings. Among a range of GB levels, the 20 mM concentration emerged as the most effective in enhancing seed germination, shoot fresh and dry weight and shoot water content under heat stress, and this level was selected for further studies. Time course changes revealed that the seedlings developing from 20 mM GB treated seeds had greater shoot dry weight, net photosynthetic rate (P_N), leaf water potential (ψ_w) and reduced relative membrane permeability (RMP), compared to no-GB treated plants under heat stress. Correlations between dry weight and high P_N (r = 0.881), low ψ_w (r = −0.938) and RMP (r = −0.860) of shoots suggested the involvement of GB in heat stress tolerance. Leakage of Ca²⁺ and NO₃⁻ was the greatest followed by K⁺ and PO₄³⁻ under no-GB seed treatment, and GB application under heat stress appreciably reduced the leakage of all these ions, particularly Ca²⁺, K⁺ and NO₃⁻. In conclusion GB absorbed by seeds, after translocation to the seedlings, enhanced their capacity to maintain greater water content, and higher seedling vigor by virtue of increased P_N, reduced RMP and leakage of important ions under heat stress. These results have implications for final field stand under the conditions where the ambient temperature is supra-optimal for barley growth.     

Estimation of relative biological effectiveness of tritium according to chromosome aberration frequency in human blood lymphocytes

The goal of this work was to determine the relative biological effectiveness (RBE) of tritium β-radiation according to the chromosome aberration frequency in the peripheral blood lymphocytes after in vitro and in vivo radiation exposures. The experimental RBE assessment of tritium β-radiation relative to ⁶⁰Co γ-radiation according to unstable chromosome aberration frequency in the peripheral blood lymphocytes under particular conditions is described. It has been demonstrated that tritium β-radiation is, in general, more effective in the dose range of up to 1 Gy, which is most pronounced at low doses. The RBE value of tritium β-radiation at minimum doses reached 2.2 and decreased at higher doses (1 Gy) to 1.25. The data on comparative analysis of the frequency of stable chromosome aberrations in the blood lymphocytes of professional nuclear workers (Sarov, Russia) after long-term chronic exposure to tritium β-radiation, as compared with γ-irradiation, are reported for the first time. The higher biological effectiveness of tritium β-radiation was demonstrated and was estimated as 2.5.     

Estimation of the levels of radiation-induced <Emphasis Type="Italic">P</Emphasis>-element transposition in <Emphasis Type="Italic">Drosophila melanogaster</Emphasis> experimental populations and laboratory strains

When experimental P + M populations were exposed to chronic γ-irradiation (0.31 mGy/h), the highest instability level of the singed-weak (sn _w ) locus was observed in F₃–F₁₀ with a subsequent decrease and stabilization of the mutation rate. The sn ^w mutation rate was within the range of spontaneous variation in conditions of P-M hybrid dysgenesis and irradiation of males of the Harwich laboratory strain with active P elements at 1.61 mGy/h. The instability of the sn ^w locus was significantly higher at lower dose rates (0.23 and 0.31 mGy/h), suggesting a nonlinear dose-effect relationship.     

Biofortification of chromium in fenugreek seeds

BackgroundFenugreek and chromium are known to have anti-diabetic properties and this has been well demonstrated by multiple studies. Researches have been undertaken to study thebiofortification of chromium (Cr³⁺⁾ in fenugreek. Some of the researchers have studied the biofortification from the soil irrigated with tannery waste water or from soil amended by tannery-sludge, with a view of enhancing the anti-diabetic effect of fenugreek plants. The present research work was also undertaken to increase the chromium content of fenugreek seeds, but through direct treatment of chromium picolinate solution to fenugreek seeds.MethodsFenugreek seeds were procured and divided in four groups having three batches of 10 g seeds- treatment₁, treatment₂, control₁ and control₂. Control₁ group was kept raw and was given no treatment and control₂ group was germinated using double distilled water. Treatment₁ and treatment₂, on the other hand, were given treatment of 0.02 g and 0.04 g chromium picolinate solution, on first day and 0.01 g and 0.02 g chromium picolinate solution on second day, respectively. The germinated samples were then completely dried, powdered, digested with di-acid mixture and assayed using Inductively Coupled Plasma optical emission spectrometry method for chromium content. The treatment₁ sample was selected for further nutritional analysis along with control₁ and control₂ group to compare the nutritional composition of raw, germinated and chromium treated fenugreek seed flour. Fifteen sprouts from treatment₁ group (treatment_1A group) and fifteen sprouts from control₂ group (control₃ group) were sown in earthen pots for the analysis of chromium content in seeds of new plants.ResultsThe fenugreek seeds treated with two different concentrations of chromium picolinate viz. treatment₁ and treatment₂ group attained 55 and 80 times higher chromium content as compared to control₂ group, respectively. All the estimated minerals and bioactive compounds were significantly high (p ≤ 0.01) in germinated fenugreek seed flour and chromium treated fenugreek seed flour compared to raw fenugreek seed flour. Germinated fenugreek seed flour and chromium treated fenugreek seed flour were statistically comparable to each other in respect of all the parameters analysed. Hence, it was evident that enriching fenugreek seeds with chromium, did not affect the nutritional content of fenugreek seed by any mean. Also, there was no significant difference between the chromium content in seeds of control₃ group and T_1A group.ConclusionsTreatment of fenugreek seeds with chromium solution seems to be an efficient and safe method for increasing their chromium concentration as compared to application of chromium to the soil for biofortification with minimal to no chance of chromium accumulation and inheritance in next generation plants. However, there is a need of more research to see how reliable these observations would be when different chromium salts and/or varied chromium concentration are used.     

Comparative study on aging, UV treatment, and radiation on cataract formation

Four randomized groups of male mice Fl (C57Black/CBA) were investigated: (a) UV-irraidated (UV-A, 15 min daily over 10 months, 51 ± 7 W/m²); (b) γ-ray irradiated (2 Gy), single; (c) influence of combination of UV- and γ-ray treatment; (d) aging. The lens opacities were measured at the seventh and tenth month. An expert method based on a six-point scale was used for cataract measurement. At the seventh month, the median of lens opacities were Aging group = 0; UV-irradiated group = 2.5; γ-irradiated group = 4.75; γ- and UV-irradiated group = 6.0. The difference between all groups was significant (p < 0.004, Kruskall-Wallis ANOVA test). The Conover post hock test has shown a significant difference for all comparison pairs (p < 0.002) with the exception of the γ- and UV-irradiated group. At the tenth month, the lens opacities strongly increased: Aging group = 2.5; UV-irradiated group = 5.0; γ-irradiated group = 6.5; γ- and UV-irradiated group = 7.5 (median). The difference between groups was significant (p < 0.0001, Kruskall-Willis ANOVA test). The Conover post hock test has shown a significant difference for all comparison pairs (p < 0.003) with the exception of the UV-irradiated group and γ-irradiated group. No formation of specific lens opacities for any group was found. Morphology and protein composition were investigated at the tenth month. The results of a study of morphological changes show destructive and degenerative impairments of the capsule, epithelium cages, and lens fibers. However, no specific changes related to certain particular actions have been found. In addition, there were no specific changes in the protein composition of either water-soluble and water-insoluble fractions estimated by the differential gel electrophoresis technique. The data mean that aging, UV treatment, and γ-radiation causes similar lens changes. It was supposed that UV treatment and/or γ-radiation act as an aging factor on the lens.     

Pleiotropic morphological and abiotic stress resistance phenotypes of the hyper-abscisic acid producing Abo™ mutant in the periwinkleCatharanthus roseus

The pleiotropic properties of aabo abo (Abo^™) γ-ray induced mutant ofCatharanthus roseuscv. Nirmal, selected among the M₂ generation seeds for ability to germinate at 45°C, are described. The mutant produced seeds possessing tricotyledonous embryos, unlike the typically dicotyledonous embryos present in the wild type Abo⁺ seeds. In comparison to Abo⁺ adults, the mutant plants had short stature and lanceolate leaves. The vascular bundles in the leaves and stem were poorly developed. Leaf surfaces were highly trichomatous, epidermal, cortex and mesophyll cells were small sized and a large majority of stomata were closed. Besides high temperature, the mutant was salinity and water-stress tolerant. The abscisic acid (ABA) content in the leaves was about 500-fold higher. The genetic lesionabo responsible for the above pleiotropy was recessive and inherited in Mendelian fashion. The seedlings and adult plants of the mutant accumulated higher proline than Abo⁺ plants. The phenotypes ofabo abo mutants permitted the conclusions that (i) the mutant synthesizes ABA constitutively, (ii) both ABA-dependent and ABA independent pathways for proline and betaine accumulation are functional in the mutant, and (iii) cell division, elongation and differentiation processes in embryo and adult plant stages are affected in the mutant.     

Nitration of γ-tocopherol in plant tissues

Nitration of γ-tocopherol has been suggested to be an important mechanism for the regulation and detoxification of reactive nitrogen oxide species in animal tissues. To investigate whether this reaction does also occur in plants, reversed phase high-performance liquid chromatography (HPLC) and mass spectrometry (LC-MS) were used for analysis of 5-nitro-γ-tocopherol (5-NγT) in leaves and seeds. 5-nitro-γ-tocopherol (5-NγT) could be detected in an in vitro system where it was most likely generated by the reaction of γ-tocopherol with a nitric oxide radical. In vivo 5-NγT was identified in leaves of the Arabidopsis mutant line (vte4), which has insertion in the gene encoding γ-tocopherol methyltransferase and consequently lacks α-tocopherol and accumulates high levels of γ-tocopherol. Quantification of NO_x in leaves revealed that the vte4 mutant in comparison to wild type and the mutant vte1, which does not contain any tocopherol, has a reduced NO_x concentration. The level of 5-NγT in leaves of the vte4 mutant was shown to depend on the developmental stage and on the duration of light exposure. 5-NγT was also detectable in germinating seeds of Brassica napus, Nicotiana tabacum and Arabidopsis thaliana. These seeds have in common high γ-tocopherol contents. The rate of germination at two days after imbibition inversely correlated with the γ-tocopherol content of the seeds. The result suggests that γ-tocopherol or its respective derivative, 5-NγT, may prolong early development by reducing the level of NO_x.     

Effect of γ-radiation on the ratio of [18F]2-fluoro-2-deoxy-D-glucose to glucose utilization in human glioblastoma cellsin vitro

The glucose consumption in tumoursin vivo as reflected by uptake of [¹⁸F]2-fluoro-2-deoxy-D-glucose (¹⁸FDG) using positron emission tomography (PET) is currently under investigation as a measure of tumour response to radiotherapy. The calculation of cerebral metabolic rate of glucose from¹⁸FDG-PET data requires a proportionality factor referred to as the lumped constant. In the presentin vitro study, the utilizations of¹⁸FDG and glucose have been measured in a human glioblastoma cell line (86HG-39) as a function of γ-radiation dose with various post-irradiation times and of different fractionation modes. The ratio of utilization of¹⁸FDG to that of glucose (R_F/G), assumed to correspond to the lumped constant, was observed to increase 12 and 24 h after single fraction γ-exposure by factors ranging from 1.2 to 1.5 compared with the non-irradiated controls. It decreased after multiple fraction γ-exposure (4 × 2 Gy) by a factor of 0.7 compared with the single fraction schedule (1 × 8 Gy). The results suggest that the affinities of glucose transporters or hexokiriase enzyme or both for¹⁸FDG and glucose could be influenced by γ-irradiation in this tumour cell linein vitro. Apparent changes of the glucose consumption determined with PET in human tumours following radiotherapy may, therefore, not be solely due to changes in cellular metabolism or cell number but may also be due to changes in R _F/G .