[Assessment of Cyto- and Genotoxicity of Underground Waters from the Far Eastern Center on Radioactive Waste Treatment Site].

This study has been completed in the frames of activities on the environment assessment in the vicinity of the Far Eastern center (FEC) on radioactive waste treatment (a branch of Fokino, Sysoev Bay). Underground waters collected at the FEC technical site were surveyed both with instrumental techniques and bioassays. Concentrations of some chemicals (ranged to the third hazard category) in the samples collected are over the permitted limits. Activities of 137Cs and 90Sr in waters amount up to 3.8 and 16.2 Bq/l, correspondingly. The integral pollution index is over 1 in all the samples and could amount up to 165. The Allium-test application allows the detection of the sample points where underground waters have an enhanced mutagenic potential. Dependencies between biological effects and pollution levels are analyzed. The findings obtained could be used for the monitoring optimized and decision making on rehabilitation measures to decrease negative influence of the enterprise on the environment.

Drought response of mesophyll conductance in forest understory species--impacts on water-use efficiency and interactions with leaf water movement.

Regulation of stomatal (gs ) and mesophyll conductance (gm ) is an efficient means for optimizing the relationship between water loss and carbon uptake in plants. We assessed water-use efficiency (WUE)-based drought adaptation strategies with respect to mesophyll conductance of different functional plant groups of the forest understory. Moreover we aimed at assessing the mechanisms of and interactions between water and CO2 conductance in the mesophyll. The facts that an increase in WUE was observed only in the two species that increased gm in response to moderate drought, and that over all five species examined, changes in mesophyll conductance were significantly correlated with the drought-induced change in WUE, proves the importance of gm in optimizing resource use under water restriction. There was no clear correlation of mesophyll CO2 conductance and the tortuosity of water movement in the leaf across the five species in the control and drought treatments. This points either to different main pathways for CO2 and water in the mesophyll either to different regulation of a common pathway.

[Assessment of cyto- and genotoxicity of natural waters in the vicinity of radioactive waste storage facility using Allium-test].

Efficacy of bioassays of "aberrant cells frequency" and "proliferative activity" in root meristem of Allium cepa L. is studied in the present work for a cyto- and genotoxicity assessment of natural waters contaminated with 90Sr and heavy metals in the vicinity of the radioactive waste storage facility in Obninsk, Kaluga region. The Allium-test is shown to be applicable for the diagnostics of environmental media at their combined pollution with chemical and radioactive substances. The analysis of aberration spectrum shows an important role of chemical toxicants in the mutagenic potential of waters collected in the vicinity of the radioactive waste storage facility. Biological effects are not always possible to explain from the knowledge on water contamination levels, which shows limitations of physical-chemical monitoring in providing the adequate risk assessment for human and biota from multicomponent environmental impacts.

UV-B radiation impacts shoot tissue pigment composition in Allium fistulosum L. cultigens.

Plants from the Allium genus are valued worldwide for culinary flavor and medicinal attributes. In this study, 16 cultigens of bunching onion (Allium fistulosum L.) were grown in a glasshouse under filtered UV radiation (control) or supplemental UV-B radiation [7.0 µ mol·m(-2) · s(-2) (2.68 W · m(-2))] to determine impacts on growth, physiological parameters, and nutritional quality. Supplemental UV-B radiation influenced shoot tissue carotenoid concentrations in some, but not all, of the bunching onions. Xanthophyll carotenoid pigments lutein and ß -carotene and chlorophylls a and b in shoot tissues differed between UV-B radiation treatments and among cultigens. Cultigen "Pesoenyj" responded to supplemental UV-B radiation with increases in the ratio of zeaxanthin + antheraxanthin to zeaxanthin + antheraxanthin + violaxanthin, which may indicate a flux in the xanthophyll carotenoids towards deepoxydation, commonly found under high irradiance stress. Increases in carotenoid concentrations would be expected to increase crop nutritional values.

Changes in nitrate and nitrite concentrations over 24h for sweet basil and scallions.

Nitrate and nitrite concentrations were determined for sweet basil and scallions over 24h to determine if time of sampling or harvest impacts concentrations in raw vegetables. Also, nitrate and nitrite concentrations were determined separately for various edible parts of these plants. Basil had significant changes in nitrate and nitrite concentrations over a 24h period. Nitrate was correlated to changes in light intensity with a 3h lag time. The highest nitrate concentrations in basil (2777 ppm) occurred around 3h after the light intensity peaked and had low values (165-574 ppm) during the dark period. The scallion nitrate and nitrite concentrations were always low but nitrate showed a peak a few hours before sunrise. Nitrate and nitrite concentrations in some raw vegetables may be reduced by harvesting at the best time of day for each type of plant. Nitrate concentrations were different in the edible plant parts tested.

[Dynamics of the induced chromosomal instability in welsh onion (Allium fistulosum L.): gamma irradiation of the seeds of different storage periods].

The chromosome aberrations in root meristem cells of welsh onion (Allium fistulosum L.) seeds after gamma-irradiation (5 and 10 Gy) of different-aged seeds (7, 19, 31, 43 and 55 months of storage) were studied. The irradiation dose of 5 Gy significantly increased the frequency of aberrant anaphases (FAA) for 31- and 43-months seeds; the dose of 10 Gy significantly increased the FAA in seeds of all age groups. The irradiation of young (7 months) seeds resulted in decreasing of the fraction of bridges to the control level of the old (55-months) seeds for the dose of 5 Gy and below the control level of the old seeds--for the dose of 10 Gy. Some peculiarities of cytogenetic parameters of genome instability and the germinating capacity of the seeds made it possible to suppose that the third year of storage is a critical period for the welsh onion seeds.

[Number of aberrations per cell as a parameter of chromosome instability. 2. Comparative analysis of the factors of different nature]. / Kil'kist' aberatsii na klitynu iak parametr khromosomnoï nestabil'nosti. 2. Porivnial'nyi analiz vplyvu faktoriv riznoï pryrody.

The average number of aberrations per aberrant cell was concluded to carry out information on chromosome instability peculiarities induced by different mutagens as it was shown in our previous work. The purpose of the current study was to present comparative analysis of intercellular distribution of number of aberrations and their theoretical approximations. Distribution of numbers of aberrations per cell in Allium cepa L. and Allium fistulosum L. root tip cells induced by different mutagenic factors (gamma-irradiation, thiotepa, formaldehyde and seed aging) have been studied. The results were approximated to theoretical Poisson, geometric and negative binomial distributions. The intercellular distribution of aberrations did not correspond to any of the used theoretical distributions when A. cepa seeds were gamma-irradiated. There was some, but not regular, accordance with theoretical distributions when chemical mutagens thiotepa in A. cepa and formaldehyde in A. fistulosum and seed aging in both species were evaluated. During seed aging frequency of aberrant cells increased more quickly in A. fistulosum in comparison with A. cepa.

[Chronic irradiation with low doses can be characterized by significant biological efficacy]. / Khronichne oprominennia v malykh dozakh mozhe kharakteryzuvatysia velykoiu biolohichnoiu efektyvnistiu.

Chromosomal aberrations (ChA) level was analyzed in the onion root meristem after the chronic irradiation with different dose capacities. It was shown that after the chronic irradiation with doses of 0.87 cGy, 2.61 cGy and 4.35 cGy the level of chromosomal aberrations depended on the dose capacity. Its value may also correspond to those which have been induced with accute irradiation. Biological efficacy of chronic irradiation may be from 20 to 1000 time folder in order to compare it with accute irradiation and this value depends on the irradiation regime.

[Genotoxicity and toxicity assay of water sampled from the underground nuclear explosion site in the north of the Perm region (Russia)]. / Otsenka geno- i tsitotoksichnosti khimicheskogo i radionuklidnogo zagriazneniia vodoemov v raione provedeniia podzemnogo iadernogo vzryva na severe Permskoi oblasti.

The results of our study revealed a local biologically relevant surface water contamination in the radionuclide anomaly in the north of Russia (Perm region) by means of Allium shoenoprasum L. the anaphase-telophase chromosome aberration assay. This radionuclide anomaly was formed in 1971 as a result of an underground nuclear explosion with soil excavation. Specific activities of main dose-forming radionuclides in all examined reservoirs are below intervention levels officially adopted in Russia for drinking water. We found that 90Sr significantly contribute to induction of cytogenetic disturbances. Our previous and described here data suggest that metal ions and radionuclides combined exposure on the various biota species (with the dose below permissible exposure limits for human) may cause substantial biological effects in part be due to synergic response. The findings described here indicated that development of a new concept of radiation protection for humans and biota should be based on the clear understanding of biological effects of low doses of radiation in chronic exposure to multi-pollutant mixtures.

[Mechanism of induction of cytogenetic damage in plant meristematic cells caused by combined effect of heavy natural radionuclides and heavy and alkaline metals]. / Zakonomernosti induktsii tsitogeneticheskikh narushenii v kletkakh meristem rastenii pri sochetannom deistvii tiazhelykh estestvennykh radionuklidov, tiazhelykh i shchelochnykh metallov.

The analysis of experimental data on the combined effect of 232Th and water from melted snow containing metal ions from industrally loaded territories on Tradescantia (clone 02) and on Allium shoenoprasum meristematic root tip cells treated with heavy natural radionuclides from natural water sources in the area of radioactive wastes storehouse is presented. It was demonstrated that plant meristematic cells response to heavy metals and heavy natural radionuclides, both from two-component model system and from natural water sources, have commom features.

[The number of aberrations in aberrant cells as a parameter of chromosomal instability. 1. Characterization of dose dependency]. / Kil'kist' aberatsii na aberantnu klitynu iak parametr khromosomnoï nestabil'nosti. 1. Kharakterystyka dozovykh zalezhnostei.

Analysis of chromosome instability (CI) is of great importance in view of pollution of the environment by genotoxic factors. Frequency of aberrant cells, spectrum of chromosome aberrations, damages of aberrant cell and distribution of aberrations in the cells are the most conventional parameters of CI. We have carried out the comparative analysis of the frequency of aberrant cells and the dynamics of aberrant cell damages induced by different mutagenic factors (alpha-irradiation from 241Am, gamma-irradiation from 60Co and tioTEPA) in Allium-test. This comparative analysis denotes that the studied parameters have different dynamics characterizing different mechanisms of CI in Allium cepa L.

In vitro bulb development in shallot (Allium cepa L. Aggregatum Group): effects of anti-gibberellins, sucrose and light.

Bulbing was studied in shallot plants cultured in vitro. Bulbing occurred under a 16 h photoperiod with fluorescent + incandescent light and 30-50 g 1(-1) sucrose in the culture medium. Exogenous gibberellin (10 microM GA3) inhibited leaf and root growth and bulbing. When added to the medium at a concentration of 10 microM, three inhibitors of gibberellin biosynthesis (ancymidol, flurprimidol and paclobutrazol) promoted bulb formation and the percentage of bulbing. When ancymidol was used in combination with GA3, it did not reverse the effect of GA3 applied alone. Under treatments with 30-70 g l(-1) sucrose, bulbing ratios greater than those found in control plants were achieved by addition of ancymidol, and bulb fresh weight was increased in the same way. Ancymidol caused a 66% decrease in sucrose content in leaf bases but greatly increased the glucose, fructose and fructan contents. The increase in fructan content by ancymidol could result from the three-fold rise in total [14C]sucrose uptake per plant from the culture medium associated with a marked increase in leaf base labelling at the expense of root labelling. The possible role of ancymidol is discussed and evidence supports a major regulatory role for gibberellins in bulbing.

[Cytogenetic effects of 241Am in the Allium test]. / Tsitogeneticheskie éffekty 241 Am v Allium-teste.

The purpose of this study was to elucidate the genome damage induced by 241Am-irradiation using different parameters of cytogenetic evaluation in Allium-test. The root tip cells test-system for the cytogenetic effects studying was used. 241AmCl3 of different concentrations was used (1.5 x 10(-9)-1.5 x 10(-7) g/l). Water solution-to-plant transfer factor for 241Am was found to be 0.18 +/- 0.04. The internal doses of 241Am accumulated during germination were 0.37-37.00 cGy. The impact of 241Am-irradiation was evaluated on the mitotic index (MI), the yield of aberrant anaphases (AA), the distribution of chromosome aberrations number in cells and the average level of lesion of aberrant cell (LAC). Probably all these parameters are differ in sensitivity to damage factor, but only some changes in MI was revealed. It is supposed, that the absence of any changes in the distribution of chromosome aberrations number in cells and the average level of LAC in 241Am-irradiated cells confirm the absence of significant 241Am-impact on chromosomes, as the alpha-irradiation should cause significant damages in chromosomes. Although solutions of 241Am were high-concentrated, the seedlings didn't accumulate high internal doses. It appears the distribution of 241Am is a significantly heterogeneous hence it is possible the absorbed doses in nuclei can't reach the level necessary for revealing of cytogenetic effects.

A fast comet assay variant for solid tissue cells. The assessment of DNA damage in higher plants.

The single-cell gel electrophoresis or comet assay, under high alkaline conditions, detects low levels of DNA damage. In it, broken DNA migrates from the nucleus to the anode providing images similar to comets. To adapt this assay to solid tissue cells, nuclei were directly obtained from Allium cepa L. roots. The surface of each single fresh sharply cut meristem was exposed to a small drop of 50 mM Sörensen buffer at pH 6.8, placed on a regular agarose-coated slide. By immediately adding low melting point agarose at 30 degrees C, nuclei resulted embedded in agarose. A final layer of this agarose ended the preparative steps. Conventionaly prepared leukocytes were used as a control. The treatment with detergent (lysis step of the conventional assay) proved to be unnecessary for the nude nuclei. A 20 min-long electrophoresis (at 0.65 V. cm-1, 230 mA and 10 degrees C) was more sensitive than a 10 min-long one for detecting the differential response of plant nuclei to 2 and 4 Gy of gamma-irradiation. A short fixation in methanol transformed the preparations into semi-permanent ones, without altering their later DNA staining by ethidium bromide. The use of instantaneously isolated nuclei simplifies and expands the use of this technique to any eukaryotic cell from solid tissues.

Light inactivation of Arabidopsis photomorphogenic repressor COP1 involves a cell-specific regulation of its nucleocytoplasmic partitioning.

Arabidopsis COP1 acts as a repressor of photomorphogenesis in darkness, and light stimuli abrogate this suppressive action. COP1, when fused to beta-glucuronidase (GUS), is enriched in the nucleus in darkness, but not in the light, in hypocotyl cells of Arabidopsis seedlings and epidermal cells of onion bulbs. In Arabidopsis hypocotyl cells, the nuclear GUS-COP1 level changes in response to dark-light transitions and quantitatively correlates with the extent of repression of photomorphogenic development. In root cells, GUS-COP1 is constitutively nuclear, consistent with an established role of COP1 in suppressing root chloroplast development in both light and darkness. We conclude that COP1 acts inside the nucleus to suppress photomorphogenesis and that light inactivation of COP1 involves a cell type-specific control of its nucleocytoplasmic partitioning.

Variation of RBE between p(75) + Be and d(50) + Be neutrons determined for chromosome aberrations in Allium cepa.

The RBE of p(75) + Be neutrons relative to d(50) + Be neutrons has been determined for chromosome aberrations induced in Allium cepa (onion) roots. Two biological criteria were selected: the average number of aberrations (mainly fragments) per cell in anaphase and telophase, and the percentage of aberration-free cells. The influence of sampling time (3 to 7 h incubation) between irradiation and fixation was investigated systematically. This factor did not significantly influence the results. The RBE values of p(75) + Be neutrons compared to those of d(50) + Be neutrons were 0.85 (0.79-0.91) and 0.87 (0.80-0.95) for the first and the second criteria, respectively. In previous experiments for the same beams, we found an RBE of 0.90 (0.86-0.94) for survival of V79 cells (D0 ratio), 0.96 (0.93-0.99) for the intestinal crypt cell system, and 0.83 (0.70-0.96) for Vicia faba growth delay.

[Effect of polyamines on the cellular radiation reaction]. / Vliianie poliaminov na radiatsionnuiu reaktsiiu kletok.

The influence of polyamines (e.g. putrescine, spermine and spermidine) on the survival rate of HeLa cells and the mitotic index of A. cepa meristem cells, as well as a change in a radiation response of cells under the effect of polyamines have been investigated. Putrescine was shown to produce the lowest cytotoxic effect on mammalian cells, whereas the cytotoxic effect on plant cells was either insignificant or absent at all. One-hour incubation of HeLa cells with putrescine of 5 x 10(-4)-5 x 10(-5) M prior to or after irradiation with a dose of 6 Gy increased the survived cell fraction. Spermine of 10(-3) M increased considerably the mitotic index of the exposed meristem as compared to irradiated meristem untreated with spermine. The role of polyamines in the formation of radiation damage to a cell is discussed.

UV-mediated toxic bioactivity of harmine in the meristematic cells of Allium cepa.

The photodynamic effect of harmine and UV light was studied by measuring the number of sister-chromatid exchanges and micronuclei induced by this treatment in Allium cepa meristematic cells. A significant increase in the frequencies of both cytogenetic events was observed when proliferating cells were treated for 4 h with harmine followed by irradiation with UV light for 3 min.