Radioprotective effect of chloropyllin, protoporphyrin-IX and bilirubin compared with amifostine® in Drosophila melanogaster.

The identification of substances that prevent or minimize the detrimental effects of ionizing radiation is an essential undertaking. The aim of this paper was to evaluate and compare the radioprotective potential of chlorophyllin, protoporphyrin and bilirubin, with amifostine®, an US Food & Drug Administration approved radioprotector Using the somatic mutation and recombination assay in the Drosophila melanogaster wing, it was found that pretreatment (1-9 h) with any of the porphyrins or amifostine® alone, did not affect the larva-adult viability or the basal frequency of mutation. However, they were associated with significant reductions in frequency of somatic mutation and recombination compared with the gamma-irradiated (20 Gy) control as follows: bilirubin (69.3 %)> chlorophyllin (40.0 %)> protoporphyrin (39.0 %)> amifostine® (19.7 %). Bilirubin also caused a 16 % increase in larva-adult viability with 3 h of pretreatment respect to percentage induced in 20 Gy control group. Whilst amifostine® was associated with lower genetic damage after pre-treatment of 1 and 3 h, this did not attain significance. These findings suggest that the tested porphyrins may have some potential as radioprotectant agents.

Different radiation dose rate as radioprotection and the cross effect with chromium using in vivo somatic cells of Drosophila.

Ionizing radiation plays a key role in the adaptation of an individual organism to environmental pollution, at the same time, it has biological effects that depend on radiation intensity or dose rate (DR). Although the effect of DR has been studied in vitro, the phenomenon known as the inverse effect of DR, which indicates as it decreases that the induction of damage is greater, has not been widely studied in vivo. The present study is aimed to test 0.5 and 1 Gy in somatic cells of the wing of D. melanogaster, administered at 5.4 or 34.3 Gy/h and from 0.037 to 0.3 mM of CrO3 as conditioning treatment. No changes were found in larva-to-adult viability. A protective as well as a cross effect of pre-exposure to different DR and CrO3 concentrations against genetic damage induced by 20 Gy or 1 mM CrO3 was evident.

Cytotoxic and genotoxic actions of Casiopeina III-Ea (Cas III-Ea) in somatic and germ cells of Drosophila melanogaster.

Casiopeinas® are a group of newly synthesized drugs designed to treat cancer. These copper (Cu) complexes exhibit cytostatic, cytotoxic, genotoxic, and antineoplastic activities through different mechanisms of action. To evaluate the influence of these compounds, some in vivo studies were performed using predominantly somatic cells. The aim of the present study was to examine the cytotoxic and genotoxic actions of Casiopeina III-Ea (Cas III-Ea) in somatic as well as germ cells of Drosophila melanogaster. For cytotoxicity, the productivity and some morphometric parameters were measured and genotoxicity was assessed by means of the somatic mutation and recombination test assay in the wing. For this purpose, second-instar larvae of the Canton-S strain were treated with different concentrations of Cas III-Ea. The emerged adults were weighed, the area of the wings determined, and the number of trichomes of the region C' counted. The productivity of treated males was measured by a brood method to monitor the influence of Cas III-Ea on spermatozoa, meiotic stage cells, and spermatogonia. For genotoxicity, mwh + /+ flr3 larvae 48 hr age were chronically treated within the same concentration range. Results indicated that Cas III-Ea at all concentrations tested significantly increased the productivity per couple in Brood III (spermatids) while at 1 mM a marked elevation was noted in the three broods tested. In contrast, the weight and size of individuals as well as the size and number of cells in the wing were decreased significantly. Data suggest that Cas III-Ea is a weak genotoxic but selective mutagen. Failure to obtain a dose-related genotoxic response suggests that one of the preferred mechanisms of action of Cas III-Ea is to induce apoptosis.

Study on the relationship of genotoxic and oxidative potential of a new mixed chelate copper antitumoral drug, Casiopeina II-gly (Cas II-gly) in Drosophila melanogaster.

The present study evaluates the superoxide dismutase (SOD) and catalase (CAT) activities in a wild strain of Drosophila melanogaster and the genotoxic potential induced by Cas II-gly (a new antineoplastic drug) using the somatic mutation and recombination test. Larvae 48h old were treated with Cas II-gly in a range of 0-1.5mM and aliquot were taken every 24h to have individuals treated for 24, 48, 72h and adulthood as well. A dose-dependent toxicity and a significant increase in SOD and CAT activities were found after a 24 and 48h treatment with 0.5-1.5mM concentrations. The comparison of the effect in enzymes with mutation indicated a positive correlation with increased genetic damage, after 24 and 48h of exposure for all concentrations tested. The addition of the genetic damage induced in each exposure time showed a significant effect, but only the small single spots had a concentration-related increase.

A study of the inhibition/promotion effects of sodium-copper chlorophyllin (SCC)-mediated mutagenesis in somatic cells of Drosophila.

Sodium-copper chlorophyllin (SCC), a copper-porphyrin complex, has been shown to act as an inhibitor as well as a promoter of DNA-damage induction by a variety of mutagens in several test systems. In order to investigate the basis of this dual effect, experiments were carried out to compare the influence of pretreatment with intact SCC and that of its constituents, the metal-free protoporphyrin (PP-IX) and copper as CuCl(2). The wing-spot test was employed to monitor mutational events in somatic cells of Drosophila melanogaster. Heterozygous mwh+/+flr(3) larvae were treated for 24h with SCC, PP-IX, CuCl(2) or sucrose. Following this treatment, one group of larvae were immediately allowed to feed on instant medium containing 0.5mM N-nitroso-N-ethylurea (ENU) dissolved in phosphate buffer to reach pH 6. The remaining larvae received treatment with ENU with a delay of 1, 2 or 3days (DTD). Results revealed an (a) overall inhibitory effect for 0-DTD and 1-DTD after pretreatment with SCC, (b) only in 0-DTD after PP-IX, and (c) in all DTDs after treatment with CuCl(2). These results provide evidence that the copper ion plays a central role in the antimutagenic effect of SCC, and for a sustained period of time. Pretreatment with SCC and PP-IX produced a promoter effect at 2-DTD and 3-DTD. The results could be explained as an effect of the accumulation of metal-free porphyrin following the dissociation of the copper-porphyrin complex (SCC), the copper-ion reaching proteins to form complexes and participated in anabolic pathways.

Evidence that low concentrations of chlorophyllin (CHLN) increase the genetic damage induced by gamma rays in somatic cells of Drosophila.

It was first demonstrated in Salmonella that higher and lower concentrations of chlorophyllin (CHLN) may have effects in opposite directions, higher doses inhibiting and lower doses promoting the mutagenic activity of certain tobacco-related nitrosamines. Previous work of our group demonstrated that CHLN may have both a promoter and an inhibitory effect on mutagenesis in Drosophila. The present paper reviews the evidence obtained in our laboratory using gamma rays as the mutagenic agent, that higher and lower pretreatment concentrations of CHLN are associated with inhibitory and promoting effects, respectively, as in Salmonella. Employing the wing spot test, 48h larvae were pretreated with various concentrations of CHLN from 0 to 69 mM and then treated with 10 Gy gamma rays. With the highest concentration of CHLN, an approximate 54% reduction in mutagenesis was observed. At 35 mM a remnant of this inhibitory effect was found in that a significant decrease was limited to the twin spot category. Evidence of promotion was first seen at 4.3mM CHLN, an effect which persisted for the remaining five lower concentrations, the most pronounced evidence of promotion being found at the four lowest concentrations, 0.03-1.1 mM CHLN. It should be noted that no evidence of genotoxicity was found for CHLN alone, an observation consistent with the several reports in the literature. The results are taken as strong evidence that pretreatment with low concentrations of CHLN promotes DNA damage induced by gamma rays in somatic cells of Drosophila.

A long term study of the desiccation resistance of Drosophila melanogaster and D. simulans from the Mexican nuclear power plant "Laguna Verde".

The generation of electricity through the use of radioactive material at the nuclear power plant is inevitably associated with the production of wastes, some of which have potential impact on the biosphere. The objective of the present investigation was to provide information for evaluating the presumed impact of the Mexican Nuclear Power Plant "Laguna Verde" on the natural populations. Two sibling species that live in the immediate vicinity, Drosophila melanogaster and D. simulans have been studied for several traits in a long term study. The present study describes results for the desiccation resistance (DR) trait obtained during the period from 1995 to 2002. Flies were collected at two sites, one near the reactors and another farther away. The data obtained confirmed that D. melanogaster had higher DR values than D. simulans at both sites. The analysis of the results obtained from both species of the site closer to the reactor indicated that the values of the DR in the operational stage did not change, compared with those in the preoperational stage previously analyzed. Therefore, the significant differences found between the monitored sites did not seem to be associated to the operation of the reactors.

Ten-year viability study on natural populations of Drosophila sibling species from Laguna Verde, Veracruz, México.

The generation of electricity in nuclear power plants produces an increase in background radiation that could possibly have some impact on the organisms that live in that area. In order to identify and quantify any such possible effect, the natural populations of two sibling species, D. melanogaster and D. simulans, that live in the immediate vicinity of the first Mexican Nuclear Power Plant were analyzed for a period of 10 years. Collections of flies were made at two sites, one close to and one farther from the power plant, during the pre and operational stages of the reactor, and their egg-to-adult viability was analyzed. The data obtained indicate that in both sites, the egg-to-adult viability was generally higher in D. melanogaster than in D. simulans. Further, a relationship was found between egg-to-adult viability and the season of the year (warm-wet or cool-dry season). Some differences were found between the two sites. It may be concluded that there is no negative impact on the Drosophila populations studied.

Evidence suggesting that chlorophyllin (CHLN) may act as an inhibitor or a promoter of genetic damage induced by chromium(VI) oxide (CrO3) in somatic cells of Drosophila.

In Drosophila, 48h-old larvae were pretreated for 24h with chlorophyllin (CHLN) or sucrose and then treated with chromium(VI) oxide (CrO(3)) immediately following completion of the pretreatment period (0-day delay) or delayed 1, 2 or 3 days. The effects were scored in the wing spot test. After delays of 0 and 1 day, clear evidence of a protective effect of CHLN was found. Contrarily, after delays of 2 and 3 days, the results showed a reversal, i.e. CHLN-related events appeared more frequently than those in the sucrose control suggesting a promoting effect. It would appear prudent that CHLN be tested in a variety of situations in any given organism before decisions are reached regarding its inhibitor/promoter effects.

Radioresistance of sibling Drosophila species from Laguna Verde, Veracruz, México.

PURPOSE: To identify and quantify if the generation of electricity by nuclear power plants produces an increase in background radiation that might affect the radioresistance of organisms that live in that area. MATERIALS AND METHODS: Natural populations of two sibling species of Drosophila, D. melanogaster and D. simulans, living in the immediate vicinity of the Mexican Nuclear Power Plant in Laguna Verde were studied for 10 years. Collections of flies were made at two sites, one close to and one further from two reactors, during both the pre-operational and operational stages of the reactors. The effect of exposure to various doses of gamma-rays on egg-to-adult survival of the flies was analysed. RESULTS: The data obtained indicate that in both sites, egg-to-adult survival was higher in D. melanogaster than in D. simulans. There was an increase in the egg-to-adult survival during the pre-operational period of one of the reactors and the possible causes are discussed. No differences were found between the two sites. CONCLUSIONS: The analysis indicates that the reactors do not have a negative impact on the Drosophila populations studied.

Evidence that chlorophyllin (CHLN) may behave as an inhibitor or a promoter of radiation-induced genetic damage in somatic cells of drosophila.

Irradiation of 96h old Drosophila following a 24h pretreatment with 5% chlorophyllin (CHLN) was delayed 0-4 days. The antimutagenic effect of CHLN in somatic cells monitored by the wing spot test persisted for 3 days after completion of the pretreatment and appeared to terminate at a time corresponding to the cessation of mitotic divisions of wing anlagen cells. Within the same population of cells, CHLN demonstrated both an inhibitory effect as measured in mwh single spot classes, and contrarily, a promoting effect in the class of mwh/flr twin spots and to an extent in the class of large flr spots. The reason for the contrasting effects of CHLN remains to be determined.

On the persistence of the radioprotective effect of chlorophyllin (CHLN) in somatic cells of Drosophila.

By delaying the time of gamma irradiation of 72 h larvae, pretreated at 48 h with 5% chlorophyllin (CHLN), it was established that the overall inhibiting effect of CHLN in somatic cells of Drosophila, as measured in the wing spot test, persists for about 4 days or until the time of cessation of the proliferation of wing anlagen. In the same population of cells, some spot classes gave evidence of an inhibitory effect whereas others did not arguing against the suggestion that the radioprotective effect of CHLN is a consequence of an induced delay in development, shrinking of the potential radiation target and lowering the probability of induced events. Other observations of interest are described.

On the recovery of single spots with the flr phenotype in the wing spot test in Drosophila.

Graf et al. (U. Graf, F.E. Würgler, A.J. Katz, H. Frei, H. Juon, C.B. Hall, P.G. Kale, Somatic mutation and recombination test in Drosophila, Environment Mutagen. 6 (1984) 153-188.) described the overall results of assays of a series of compounds in the Drosophila wing spot test as indicating that single mwh spots appeared most frequently, followed by less frequent twin spots with both mwh and flr cells and lastly the 'quite rare' single flr spots. Data are presented below demonstrating that some compounds behave in a manner consistent with the above description, whereas others do not in that the frequency of single flr spots is equal to or exceeds that of twin spots and cannot be described as occurring 'rarely'. It is suggested that (large) flr singles be used as a measure of mutations/deletions directly from treated transheterozygotes. An argument is presented questioning the use of treated mwh +/+ TM3 individuals as an assay of mutations/deletions at the mwh+ locus.

Evidence for an effect of exposure to low levels of alpha particle irradiation in larval cells of Drosophila as measured in the wing-spot test.

Larvae of Drosophila were exposed to a range of concentrations of alpha particles from 3 to 318 mRad, and genetic effects measured in the wing-spot test. The results were positive, and evidence of a linear relationship between exposure and response observed. The induction of chromosome breakage is suggested by the significantly higher frequency of twin spots in the treated series compared with controls.