Genotoxic activity of methyl mercury chloride and dimethyl mercury in human lymphocytes.

The genotoxicity of methyl mercury chloride (MMC, 0-25 x 10(-6) M) and dimethyl mercury (DMM, 0-434 x 10(-6) M) was evaluated by chromosome metaphase analysis in human lymphocytes treated in vitro for 24 h. Structural (CA) and numerical (AN) chromosomal aberrations were scored for the assessment of induced genotoxic effects, while the variation in mitotic index (MI) was considered a monitor for induced cellular toxicity. MMC induced CA and AN in a dose-related manner at doses exceeding 0.6 x 10(-6) M, and the proportion of cells with CA was constantly and significantly higher than that of cells with AN. DMM was able to induce both effects as well, although to a lesser extent than MMC, CA and AN being induced at doses exceeding 43.4 x 10(-6) M and 1.73 x 10(-6) M, respectively. MMC was 6-fold more effective in inducing CA than DMM at equivalent toxic doses. On the other hand, no significant difference was observed between the two compounds in inducing AN. Therefore MMC was much more clastogenic than DMM, whereas mitotic spindle disturbances appeared to be almost equally induced by both compounds.     

Genotoxic effects and induction of phytochelatins in the presence of cadmium in Vicia faba roots

This study investigates different effects in roots of Vicia faba (broad bean) after exposure to cadmium. Genotoxic effects were assessed by use of the well-known Vicia root tip micronucleus assay. Cytotoxic effects were evaluated by determining the mitotic index in root tip cells. Finally, molecular induction mechanisms were evaluated by measuring phytochelatins with HPLC. After hydroponical exposure of V. faba roots to a range of cadmium concentrations and during different exposure times, the results of this approach showed large variations, according to the endpoint measured: after 48 h of exposure, genotoxic effects were found between 7.5 x 10(-8) and 5 x 10(-7)M CdCl(2), and cytotoxic effects were observed between 2.5 x 10(-7) and 5 x 10(-7)M CdCl(2). Statistically significant phytochelatin (PC) concentrations were measured at >or=10(-6)M CdCl(2) for PC(2), and at >or=10(-5)M CdCl(2) for PC3 and PC4.     

Relative cataractogenic effects of X rays, fission-spectrum neutrons, and 56Fe particles: a comparison with mitotic effects

The eyes of Sprague-Dawley rats were irradiated with doses of 2.5-10 Gy 250-kVp X rays, 1.25-2.25 Gy fission-spectrum neutrons (approximately 0.85 MeV), or 0.1-2.0 Gy 600-MeV/A 56Fe particles. Lens opacifications were evaluated for 51-61 weeks following X and neutron irradiations and for 87 weeks following X and 56Fe-particle irradiations. Average stage of opacification was determined relative to time after irradiation, and the time required for 50% of the irradiated lenses to achieve various stages (T50) was determined as a function of radiation dose. Data from two experiments were combined in dose-effect curves as T50 experimental values taken as percentages of the respective T50 control values (T50-% control). Simple exponential curves best describe dose responsiveness for both high-LET radiations. For X rays, a shallow dose-effect relationship (shoulder) up to 4.5 Gy was followed at higher doses by a steeper exponential dose-effect relationship. As a consequence, RBE values for the high-LET radiations are dose dependent. Dose-effect curves for cataracts were compared to those for mitotic abnormalities observed when quiescent lens epithelial cells were stimulated mechanically to proliferate at various intervals after irradiation. Neutrons were about 1.6-1.8 times more effective than 56Fe particles for inducing both cataracts and mitotic abnormalities. For stage 1 and 2 cataracts, the X-ray Dq was 10-fold greater and the D0 was similar to those for mitotic abnormalities initially expressed after irradiation.     

Toxic and cytogenetic effects induced in Allium cepa with low concenrations of Cd and 232th

232Th (7.76 x 10(-7) M) and Cd (0.89 x 10(-8) M) in concentrations which do not exceed officially prescribed standards when entering with water do not increase the frequency of chromosome aberrations in comparison with the control. Such concentrations do not cause toxic effects in plants on the levels of tissues and of the whole organism but they do display their activity on the cell level damaging division spindle. Dependence "cadmium concentration-effect" is not linear for any type of cytogenetical damages. At the concentration of cadmium 0.89 x 10(-7) M its influence on formation of division spindle is weakened and the frequency of chromosome aberrations is reducing in comparison with the control and with the effects induced at lower concentrations of cadmium in solution (0.89 x 10(-8) M). Cadmium in high concentration (5.34 x 10(-5) M) causes significant toxic and mutagenic effects.     

Selenium prevents the growth stimulatory effects of cadmium on human prostatic epithelium

Cadmium has been implicated in the increase in prostate cancer incidence in men exposed to high levels. A decrease in zinc and a concomitant increase in cadmium levels in the human prostate has been shown. The role and mechanism of cadmium action in prostate carcinogenesis is not clear. Selenium, on the other hand, has been shown to inhibit carcinogenesis in several animal systems. Results show that cadmium stimulates the growth of human prostatic epithelium in vitro, between 10(-9) M and 10(-7) M concentrations. Selenium, at concentrations between 10(-12) M and 10(-7) M shows no growth stimulatory or inhibitory effects on these cells. However, when present at 10(-8) M level, selenium inhibits the growth stimulation induced by cadmium. These results suggest that selenium may be useful in counteracting the effects of cadmium in the human prostate and offer possibilities for investigations on the protective effects of selenium in cadmium-related carcinogenesis in man.     

Antimutagenic action of cobaltous chloride on radiation-induced mutations in cultured Chinese hamster cells

The effects of cobaltous chloride on 8-azaguanine (8AG)-resistant mutations induced by gamma-rays or ultraviolet (UV) light in cultured Chinese hamster V79 cells were examined. Cobaltous chloride alone had no significant effects on survival and mutations of V79 cells at concentrations less than 1 x 10(-5) M. Cobaltous chloride at a concentration of 3 x 10(-6) M had a marked effect in reducing 8AG-resistant mutations induced by gamma-rays of 2-6 Gy, when cells were incubated for 6-7 days in the presence of cobaltous chloride after gamma-ray irradiation (posttreatment). The pretreatment of cells with cobaltous chloride for 6 days before gamma-ray irradiation reduced 8AG-resistant mutations induced by gamma-rays. Pre- or post-treatment with cobaltous chloride had no such effect on UV-induced mutations, however. The difference in responsiveness to cobaltous chloride between bacterial and mammalian cell systems is discussed.     

Induction of sister-chromatid exchanges by DNA-damaging agents and 12-O-tetradecanoyl-phorbol-13-acetate (TPA) in synchronous Chinese hamster ovary (CHO) cells

Synchronous CHO cells were obtained by mitotic selection; synchrony was maintained up to the 5th cell cycle. The mitotic cells were seeded into T-25 flasks or P-60 plastic petri dishes, and cultured for 1 h at 37 degrees C, then the cells were treated by X-ray, UV light, and mitomycin C. The cells were then cultured for 2 cell cycles with TPA and BrdUrd and sister-chromatid exchanges (SCE) analyzed by the FPG method. Following X-irradiation, the frequency of induced SCE increased linearly with dose reaching a maximum of 19.8 times the control frequency after 200 rad. With higher doses, the SCE frequency declined. In the presence of TPA, SCE frequencies were 1.8 times control levels for all X-ray doses studied (0-800 rad), the frequency seen in non-irradiated cultures treated with TPA. The induced SCE frequency also increased linearly following treatment with UVL and mitomycin C, reaching levels higher than 1.8 times controls with doses exceeding 2.5 J/m2 UVL or mitomycin C (30 min). In the presence of TPA, the SCE frequencies increased to 1.8 times controls following low UVL and mitomycin C doses, but were not influenced by TPA in the higher dose range (above 2.5 J/m2 or 10(-10) M mitomycin C. Most of the SCE were induced by X-rays during the first S phase after treatment. Following higher UVL doses (5 J/m2), however, the SCE frequency remained elevated (1.5 times controls) for 4 cell cycles after exposure.     

Stimulation by N6,O2'-dibutyrul andenosine 3',5'-cyclic monophosphate of RNA and DNA synthesis and of cell proliferation of rat hepatocytes in primary tissue culture.

The effects of DBcAMP in doses from 1.5 x 10(-8) to 1.5 x 10(-3) M on the compartmental apparent surface area (ASA) and (5(-3H)uridine radioactivity concentration (URC), (methyl-3H)thymidine labelling index per 1 hour ([Me-3H]Tdr LI/h) and per cent mitotic index (MI%) and colchicine metaphase index (CMI%) of young rat differentiated hepatocytes in primary tissue culture were investigated by morphometric and radioautographie methods. In these cells DBcAMP was found to elicit: (1) progressive increments in the ASA of nucleoli, karyoplasm and cytoplasm; (2) peak increases in nucleolar URC at 1.5 x 10(-8) and 10(-5) M, but a slight decrease at 1.5 x 10(-3) M; (3) singificant increments in karyoplasmic and total nuclear URC at all doses, except at 1.5 x 10(-6) and 10(-4) M, when such parameters remained at control levels; (4) steady and progressive increases in cytoplasmic and total cell URC values; (5) marked increments in (Me-3H)Tdr LI/h, MI% and CMI% up to the dose of 1.5 x 10(-4) M, but at 1.5 x 10(-3) M these parameters were found to be either much less enhanced or to approach closely to control values. cAMP in doses from 1.5 x 10(-8) to 10(-4) M also markedly incremented the in vitro hepatocyte CMI%, while having a lesser stimulatory effect at 1.5 x 10(-3)M. Finally of the various possible metabolites of DBcAMP administered at 1.5 x 10(-8) M to liver cultures, N6- and O2'-MBcAMP and, again, cAMP significantly increased the CMI%, of cultured hepatocytes, whereas 5'-AMP, adenosine and allantoin had no significant effect and Na-butyrate slightly decreased it. The present observations strengthen the hypothesis that cAMP and its butyrated derivatives, by possibly amplifying the template activity of the liver chromatin, accelerate the flow of differentiated primary young rat hepatocytes into the various stages of the mitotic cell cycle.     

Clastogenic effects of zinc chloride on human peripheral blood leucocytes in vitro

The clastogenic effects of three different concentrations of zinc chloride on human peripheral blood leucocytes were studied in vitro. The highest concentration (1.5 x 10(-3) M) was lethal after 48 and 72 h of culture and no blast cells were formed. The two lower concentrations (3.0 x 10(-4) M and 3.0 x 10(-5) M) significantly reduced the frequency of cell division, induced chromatid breaks and damaged cells in frequencies significantly higher than in control experiments maintained in sodium chloride and in distilled water.     

Effects of helio-neon laser radiation upon cellular cycle in a plant model

The scope of this study was to investigate possible relationships between He-Neon laser radiation and mitotic and phase indices in meristematic cells of Allium cepa L. bulbs. Our results indicate that mitotic index increased after irradiation depending this modification on the time exposure and the potency of the He-Neon beam. Phase indices were also modified: frequency of prophase increased, while inter- meta- and anaphase decreased: telophases remain unchanged. These variations were significative only when the preparations were irradiated a) with 5 mW for 10 min. or more, b) with 10 mW or c) when the preparations were processed 60 min. after irradiation. These findings could not be attributed to thermal changes. Modifications in RNA or protein synthesis could be responsible.     

Hyperaccumulation of cadmium by roots, bulbs and shoots of garlic (Allium sativum L.)

The effects of cadmium chloride concentration on root, bulb and shoot growth of garlic (Allium sativum L.), and the uptake and accumulation of Cd2+ by garlic roots, bulbs and shoots were investigated. The range of cadmium chloride (CdCl2 x 2.5H2O) concentrations was 10(-6) - 10(-2) M. Cadmium stimulated root length at lower concentrations (10(-6) - 10(-5) M) significantly (P < 0.005) during the entire treatment period. The seedlings exposed to 10(-3) - 10(-2) M Cd exhibited substantial growth reduction (P < 0.005), but did not develop chlorosis. Garlic has considerable ability to remove Cd from solutions and accumulate it. The Cd content in roots of garlic increased with increasing solution concentration of Cd2+. The roots in plants exposed to 10(-2) M Cd accumulated a large amount of Cd. approximately 1,826 times the control. The Cd contents in roots of plants treated with 10(-3), 10(-4), 10(-5) and 10(-6) M Cd were approximately 114, 59, 24 and 4 times the control, respectively. However, the plants transported only a small amount of Cd to their bulbs and shoots and concentrations in these tissues were low.     

Interaction between radiation and cadmium or mercury in mouse embryos during organogenesis

The response of mouse embryos to different concentrations of cadmium or mercury with or without low LET radiation was measured in terms of gross morphological anomalies and cellular changes. Single doses of the heavy metals (2 mg/kg body weight) were injected i.p. on day 8 of gestation 30 min before whole-body irradiation. Combined exposures to CdCl2 and X-rays led to a significant reduction in the rate of exencephaly compared with the high frequency after cadmium alone. The hypothesis that metallothionein, a sulphur-rich and metal-binding protein, may be responsible for the antagonism observed could not be confirmed. Mercuric chloride alone induced a low rate of exencephaly and the data on combined treatment suggest additivity with 0.5 and 1.0 Gy X-rays. Regarding cellular criteria, cell death in the eye anlage on day 9 of gestation was significantly suppressed after 0.5 Gy and especially after CdCl2 plus 0.5 Gy compared to cadmium alone. It is assumed that the reduction of cell lethality is correlated with the low occurrence of exencephaly. Concerning the proliferation of neuroblasts, cadmium stimulated the mitotic activity whereas X-rays depressed the proliferation capacity. After the combined treatment a distinct antimitotic effect was established.     

Effect of polyamines on the cellular radiation reaction

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.     

Toxic effects of mercury(II), cadmium(II) and lead(II) porphyrins on Trypanosoma brucei brucei growth

The effects of free mercury(II), cadmium(II) and lead(II) ions and their metalloporphyrin-derivatives on Trypanosoma brucei brucei growth in culture were studied. All experiments were conducted in the dark. IC(50) values on growth obtained in 24-h time-course experiments were 1.5 x 10(-7), 2.4 x 10(-6), 4.4 x 10(-6) and 2.6 x 10(-5) M for mercury(II) porphyrin, cadmium(II) porphyrin, lead(II) porphyrin and free base porphyrin, respectively. While the IC50 values for Hg2+, Cd2+ and Pb2+ were 3.6 x 10(-6), 1.5 x 10(-5) and 1.6 x 10(-5) M, respectively. These results clearly indicate that the toxicity of the metalloporphyrin complexes of mercury(II), cadmium(II) and lead(II) to T. b. brucei parasites was much higher compared to their free metal ions and free base porphyrin at low concentrations. It was also observed after 8 h incubation that the metalloporphyrins were effective in inhibiting the division of the parasites at concentrations >1.25 x 10(-7) M for mercury(II) porphyrin, concentrations >1.2 x 10(-6) M for cadmium(II) and lead(II) porphyrins and at concentrations >3.6 x 10(-6) M for Hg2+ ion. These observations were not detected in samples treated with the free metal ions and the free base porphyrin at the same concentrations. Interestingly, trypanosomes treated with metalloporphyrin complexes displayed different morphological features from those cells treated with free base porphyrin or metal ions. The chemotherapeutic potential of the metalloporphyrins of H2TMPyP for treatment of African trypanosomiasis is discussed.     

Interactive effects of organic and inorganic selenium with cadmium and mercury on spermatozoal oxygen consumption and motility in vitro

The effects of cadmium (Cd), mercury (Hg), and three different chemical forms of selenium (Se) (selenite, selenocystine, and selenomethionine) on ram spermatozoal motility and oxygen consumption in vitro were studied over a 4-mo period. Concentrations of 10(-6) to 10(-2) M Cd and Hg were injurious to spermatozoa as indicated by depressed motility and reduced oxygen uptake. Equimolar concentrations of Se as selenite, selenocystine, or selenomethionine counteracted the toxicity of Cd and Hg at low concentrations (10(-5) and 10(-6) M) but not at higher concentrations (10(-4) to 10(-2) M). Gel filtration (Sephadex G-75) of seminal plasma and solubilized sperm prepared from semen incubated with Cd or Hg with or without the Se compounds revealed that Cd or Hg eluted with the void volume proteins in all treatments. Incubation of ram spermatozoa with any of the three chemical forms of Se ranging from 10(-6) to 2.5 X 10(-5) M significantly improved sperm motility and oxygen consumption.     

The G2 checkpoint activated by DNA damage does not prevent genome instability in plant cells

Root growth, G2 length, and the frequency of aberrant mitoses and apoptotic nuclei were recorded after a single X-ray irradiation, ranging from 2.5 to 40 Gy, in Allium cepa L. root meristematic cells. After 72 h of recovery, root growth was reduced in a dose-dependent manner from 10 to 40 Gy, but not at 2.5 or 5 Gy doses. Flow cytometry plus TUNEL (TdT-mediated dUTP nick end labeling) showed that activation of apoptosis occurred only after 20 and 40 Gy of X-rays. Nevertheless, irrespective of the radiation dose, conventional flow cytometry showed that cells accumulated in G2 (4C DNA content). Simultaneously, the mitotic index fell, though a mitotic wave appeared later. Cell accumulation in G2 was transient and partially reversed by caffeine, thus it was checkpoint-dependent. Strikingly, the additional G2 time provided by this checkpoint was never long enough to complete DNA repair. Then, in all cases, some G2 cells with still-unrepaired DNA underwent checkpoint adaptation, i.e., they entered into the late mitotic wave with chromatid breaks. These cells and those produced by the breakage of chromosomal bridges in anaphase will reach the G1 of the next cell cycle unrepaired, ensuring the appearance of genome instability.     

Blue Light Inhibits Mitosis in Tissue Culture Cells

Irradiation of the mitotic (prophase and prometaphase) tissue culture PK (pig kidney embryo) cells using mercury arc lamp and band-pass filters postponed or inhibited anaphase onset. The biological responses observed after irradiation were: (i) normal cell division, (ii) delay in metaphase and then normal anaphase and incomplete cytokinesis, (iii) exit into interphase without separation of chromosomes, (iv) complete mitotic blockage. Cell sensitivity to the light at wavelengths from 423 and 488 nm was nearly the same; to the near UV light (wavelength 360 nm) it was 5–10 times more; to the green light (wavelength >500 nm) it was at least 10 times less. To elucidate the possible mechanism of the action of blue light we measured cell adsorption and examined cell autofluorescence. Autofluorescence of cytoplasmic granules was exited at wavelengths of 450–490 nm, but not at >500 nm. In mitotic cells fluorescent granules accumulated around the spindle. We suppose blue light irradiation induces formation of the free radicals and/or peroxide, and thus perturb the checkpoint system responsible for anaphase onset.     

Cytogenetic studies on the effect of chronic gamma irradiation onVicia faba

Vicia faba seeds (cv. Giza 1) were planted in the Inshas gamma radiation field where they were chronically irradiated during the whole life of the plant. The percentage of the induced abnormal P.M.Cs, as well as the frequency of abnormal P.M.Cs in the different meiotic stages were proportional with the given doses. The main types of chromosome aberrations were anaphase and telophase bridges, fragmentation and lagging chromosomes. The nearest plants to the source showed an inhibition of shoot growth, flower and seed sterility and irregular branching. At the dosage levels used irradiation had no effect on pollen fertility. Seeds of the 1st filial generation were used for both mitotic and meiotic studies. The percentage of the mitotic abnormalities was proportional with the doses. The most dominant type of anomaly was the presence of micronuclei in the different stages of mitosis and in the resting cells. Irradiation affected also other types of anomaliese.g. lagging chromosomes, fragments, bridges...etc. Meiosis, and pollen fertility (2nd generation) were normal.     

The toxic effects of cadmium on cell division and chromosomal morphology of Hordeum vulgare

The effects of cadmium at different concentrations (0.5–20 ppm) on root growth, cell division and chromosomal morphology of Hordeum vulgare were studied. The rate of root growth and mitotic index decreased progressively with increasing cadmium concentration and treatment duration. Different concentrations of cadmium could cause mitotic irregularities comprising c-mitoses, anaphase bridges, breaks, stickiness, lagging and vagrant chromosomes and micronuclei. The intensity of the toxic effects is basically dependent on the cadmium concentration and duration of treatment.     

Comparative investigation of structural and gene somatic mutations in workers of nuclear chemical plants. II. Frequency of lymphocytes mutant in T-cell receptor loci

Frequency of lymphocytes mutant at T-cell receptor (TCR) loci was defined in 42 workers of nuclear chemical plants. In 11 persons mainly exposed to external radiation the mean frequency of TCR-mutant lymphocytes was statistically significant by higher compared with control group of unexposed donors: 9.1 x 10(-4) vs 3.5 x 10(-4) correspondently (p < 0.01). Frequency of TCR-mutant lymphocytes did not correlate neither the frequency of structural mutations non doses of external exposure. In group of workers exposed to combined external and internal radiation (n = 31) the average frequency of TCR-mutant lymphocytes was higher compared with control level: 8.9 x 10(-4) vs 3.5 x 10(-4) correspondently (p < 0.01). Correlations between the frequency of TCR-mutant cells and Pu content in organism (r = 0.5; p = 0.005) and between the frequency of chromosome aberration of unstable and stable types (r = 0.5; p = 0.002 and r = 0.6; p = 0.036, correspondently) were set. Comparison of results of analysis of structural and gene mutations allows us to supose that in case of external exposure the observed disturbances can result from genome instability in remote period after irradiation. In case of combined exposure the genetic changes were possibly caused by the constant action of alpha-radiation from Pu containing in the body.