Number Of Polychromatic Erythrocytes Research Articles

Biochemical and genotoxic effects of some pesticides on the Egyptian Toads, Sclerophrys regularis (Reuss, 1833)

The effect of the 1/10 the application rates of either copper sulfate, temephos, glyphosate, or bifenthrin on some biochemical and genotoxic markers of the Egyptian toads, Sclerophrys regularis, exposed for 96 hours under conditions similar to the real situation was investigated. The results illustrated that , brain acetylcholinesterase (AChE) was significantly inhibited following temephos treatment, while the enzyme activities markedly increased following either glyphosate or bifenthrin exposure. Liver glutathione-S-transferase (GST) activity was inhibited following either copper sulfate or bifenthrin, while the activity increased following glyphosate treatment. Also, the data illustrated that, The level of glutathione reduced (GSH) in liver and the activities of lactic dehydrogenase (LDH), transaminasaes (AST and ALT), and phosphatases (ACP and ALP) in liver and serum showed significant increases following all the treatments and bifenthrin was the potent one. Regarding the genotoxicity of the tested pesticides, the number of structural aberrations induced by the different treatments was enumerated in the present study. It was found that all the tested pesticides significantly increased either the frequency of abnormalities or the number of polychromatic erythrocytes (PCE) per 1000 cells (the frequency of micronucleus, MN) compared to the negative control, where bifenthrin and temephos were found to be more potent than cyclophosphamide (positive control) to induce these structural aberrations. According to the present results, the tested concentrations of the tested pesticides could lead to a collapse of cellular homeostasis and changes in the biochemical and genetic parameters in the Egyptian toads which can be considered as indicators of environmental pollution by pesticides.

Cytotoxic and Genotoxic Evaluation of the Aqueous and Hydroalcoholic Leaf and Bark Extracts of Crataegus oxyacantha in Murine Model.

Crataegus oxyacantha has been mainly used in traditional medicine for the treatment of cardiovascular diseases. However, its safety profile has not been fully established, since only the genotoxic effects of C. oxyacantha fruit have been described. Therefore, the objective of this work was evaluating the cytotoxicity and genotoxicity of the aqueous and hydroalcoholic leaf and bark extracts of C. oxyacantha by means of the micronucleus test in a murine model. Doses of 2000, 1000, and 500 mg/kg of both extracts were administered orally for 5 days in mice of the Balb-C strain. Peripheral blood smears were performed at 0, 24, 48, 72, and 96 h after each administration. The number of polychromatic erythrocytes (PCEs), micronucleated polychromatic erythrocytes (MNPCEs), and micronucleated erythrocytes (MNEs) was determined at the different sampling times. Our results showed that the leaf and bark of C. oxyacantha increase the number of MNEs at the 2000 mg/kg dose, and only the aqueous leaf extract decreases the number of PCEs at the same dose. Therefore, the aqueous and hydroalcoholic leaf and bark extracts of C. oxyacantha showed genotoxic effects, and only the aqueous leaf extract exhibited cytotoxic effects.

Fitting Binomial, Quasi – Binomial, Poisson and Quasi - Poisson Models in Analyzing In Vivo Micronucleus Assay Data

Micronucleus assay is an important analysis for testing the genotoxicity of a compound thus, a careful analysis should be done. The main focus of this study was to perform the recommendations of Hothorn and Gerhard (2009) on analyzing the number and proportion of micronucleus polychromatic erythrocytes for a given number of polychromatic erythrocytes. The compound TO1 was evaluated using the Binomial, Poisson, quasi-Binomial and quasi-Poisson models. Results showed that most doses of TO1 had no significant effect on the proportion of micronucleus. The result of quasi- Binomial and quasi-Poisson model revealed that most of the inferences agree with the results of the traditional models. However, some inferences can lead to a false positive decision if over-dispersion was not taken into account. The analysis also showed that TO1 was found to be a negative compound.

Genotoxic and cytotoxic evaluation of Jatropha dioica Sessé ex Cerv. by the micronucleus test in mouse peripheral blood

Jatropha dioica Sessé ex Cerv. is a medicinal plant credited with low cytotoxicity in vitro. Thus, the objective of this work was to evaluate the possible genotoxic and cytotoxic effect in vivo of the J. dioica aqueous extract by means of micronucleus assay in mouse peripheral blood. Four different J. dioica aqueous extract dose-units were evaluated (30, 60, 100, and 300 mg/kg). The extract was administered orally to male Balb-C-strain mice every 24 h during 5 days. Blood samples were taken at 0, 24, 48, 72, 96, and 120 h from the mouse's tail and were performed in duplicate extensions. The number of Polychromatic Erythrocytes (PCE), Polychromatic Micronucleus Erythrocytes (PCEMN), and Micronucleus Erythrocytes (MNE) was determined at the different sampling times in the different study groups. Our results showed that the group that received 60 mg/kg of cyclophosphamide (positive control) presented a significant decrease in the PCE (p = 0.044) proportion and a significant increase in MNE (p = 0.032, p = 0.0001). The groups that received the different J. dioica aqueous extract doses did not present either a PCE decrease or an increase in PCEMN and MNE. J. dioica exerts neither a genotoxic nor a cytotoxic effect on mouse peripheral blood at high doses.

Neurobehavioral alterations and histopathological changes in brain and spinal cord of rats intoxicated with acrylamide

The aim of this project was to study the clinical manifestations, neurobehavioral, hematobiochemical, oxidative stress, genotoxicity, and histopathological changes during acrylamide toxicity in rats. A total of 30 adult male Wistar rats were divided in 5 equal groups and received 0, 10, 15, and 20 mg/kg body weight acrylamide as oral gavage, while group 5 was micronucleus (MN) control. Functional observational battery (FOB) parameters were studied at the 28th day of post treatment. Toxicological manifestations were evident in acrylamide-treated rats from 14th day onward. FOB revealed a significant change in central nervous system, neuromuscular, and autonomic domains. The hematological changes include significant decrease in concentration of hemoglobin, total erythrocyte count, packed cell volume, and mean corpuscular volume. The biochemical parameters aspartate aminotransferases, alkaline phosphatase, and albumin showed significant increase, while the levels of serum globulin and glucose were found to decrease significantly. The MN assay revealed the significant increase in frequencies of micronuclei and number of polychromatic erythrocytes. The oxidative stress parameters revealed no significant difference as compared to control rats. Histopathological changes observed in brain include neuronal degeneration, edema, and congestion, while spinal cord revealed demyelination in low-dose group and bilateral necrosis with malacia, liquefaction of white matter, and loss of myelin from gray matter in high-dose groups. The result indicates pathological alterations in brain and spinal cord and is responsible for neurobehavioral changes in rats. The FOB changes and histopathological alterations in spinal cord are in dose dependent to acrylamide intoxication. Various toxicological effects observed in experiment direct us to focus on a deep study and evaluate the possible causes pertaining to toxicity of this chemical. It would furnish the scientists with better options that would help them to search for a median path regarding the use of this chemical and take preventive measures to save the living beings from the hidden disasters of this chemical.

The Mitigating Effect ofCitrullus colocynthis(L.) Fruit Extract against Genotoxicity Induced by Cyclophosphamide in Mice Bone Marrow Cells

Possible genoprotective effect of Citrullus colocynthis (L.) (CCT) fruits extract against cyclophosphamide- (CP-)induced DNA damage in mice bone marrow cells was evaluated using micronucleus assay, as an index of induced chromosomal damage. Mice were preadministered with different doses of CCT via intraperitoneal injection for 7 consecutive days followed by injection with CP (70 mg/kg b.w.) 1 hr after the last injection of CCT. After 24 hr, mice were scarified to evaluate the frequency of micronucleated polychromatic erythrocytes (MnPCEs). In addition, the number of polychromatic erythrocytes (PCEs) among 1000 normochromatic erythrocytes (NCEs) per animal was recorded to evaluate bone marrow. Pretreatment with CCT significantly reduced the number of MnPCEs induced by CP in bone marrow cells (P < 0.0001). At 200 mg/kg, CCT had a maximum chemoprotective effect and reduced the number of MnPCEs by 6.37-fold and completely normalized the mitotic activity. CCT also led to marked proliferation and hypercellularity of immature myeloid elements after mice were treated with CP and mitigated the bone marrow suppression. Our study revealed that CCT has an antigenotoxic effect against CP-induced oxidative DNA damage in mice. Therefore, it could be used concomitantly as a supplement to protect people undergoing chemotherapy.

Effects of zerumbone on cisplatin-induced clastogenesis in Sprague-Dawley rats bone marrow cells

Zerumbone (ZER) is derived from Zingiber zerumbet smith from the Zingiberaceae family. It has been shown to have anti-cancer and apoptosis-inducing properties against various human tumour cells. The aim of our study was to assess the effect of ZER on cisplatin-induced clastogenesis in Sprague-Dawley rat bone marrow polychromatic erythrocytes (PCEs) using micronucleus test (MN). Animals treated with two ZER doses for 4 consecutive days plus a single dose of cisplatin following treatment, presented a non-significant effects of ZER on cisplatin-induced clastogenesis. The results also indicated that ZER has no clastogenic effects on rat bone marrow polychromatic erythrocytes after 4 days treatment with 250 and 500 mg/kg body weight when compared to one dose cisplatin of 45 mg/kg body weight. On the other hand, significant decrease in the number of PCE was observed in all treatment groups, indicating cytotoxicity of ZER and cisplatin. Under the present experimental conditions, ZER could not prevent cisplatin-induced clastogenesis in rat.

Preventive role of aluminosilicate clay against induction of micronuclei and chromosome aberrations in bone-marrow cells of Balb/c mice treated with Zearalenone

Zearalenone (ZEN) is a potent estrogenic metabolite produced by some Fusarium species. No treatment has been successfully employed to remove ZEN contamination in foods. This study was conducted to evaluate the ability of hydrated sodium calcium aluminosilicate (HSCAS) to protect Balb/c mice against cytotoxicity and genotoxicity induced by ZEN. HSCAS was given via the oral route, either alone or simultaneously with a toxic intra-gastric dose of ZEN. The experimental approach comprised treatments of seven groups of mice. The first three groups received 400, 600 or 800 mg/kg bw of HSCAS. Two experimental groups received, respectively, ZEN alone (40 mg/kg bw, representing 8% of the LD 50) and ZEN in combination with HSCAS at 400 mg/kg bw. The two control groups received distilled water and olive oil, respectively. The positive control groups received colchicine (4 mg/kg bw) for the micronucleus assay and mitomycin C (1 mg/kg bw) for the chromosome aberration test. Forty-eight hours after treatment, the femur and tibia were dissected out and analyzed. The results show that ZEN was cytotoxic and genotoxic to Balb/c mice, as indicated by the increase in the frequencies of micronucleated polychromatic erythrocytes (PCEMN) and of chromosomal aberrations in bone-marrow cells. The simultaneous intra-gastric administration of HSCAS with ZEN resulted in a reduction in the number of PCEMN and a decrease of the chromosomal aberration frequency, and an increase in the number of polychromatic erythrocytes (PCE) in bone-marrow cells, compared with those in the group treated with ZEN alone. It could be concluded that HSCAS itself was safe and efficient in the prevention of the toxic effects of ZEN in the gastrointestinal tract.

The evaluation of micronucleus frequency by acridine orange fluorescent staining in peripheral blood of rats treated with lead acetate

The data concerning the mutagenic, clastogenic and carcinogenic properties of inorganic lead compounds have been conflicting. Here, we evaluated the frequency of micronuclei in the peripheral blood of female rats treated with three different lead acetate doses. Outbred female Wistar rats were treated by gavage once per week for 10 weeks with cumulative doses of 140, 250 and 500 mg/kg body weight (body wt) of lead acetate. Mitomycin C (MMC) 2 mg/kg body wt was used as a positive control. The aim of the present study was to investigate the possible cytotoxic and genotoxic effects of lead acetate on peripheral blood reticulocytes using the micronucleus test following chronic exposure. The results show the effects of lead acetate in peripheral blood reticulocytes. These effects are both cytotoxic and genotoxic because of a decrease in the number of polychromatic erythrocytes in the peripheral blood and an increase in frequency of micronucleated reticulocytes, respectively.

Cytogenetic effects of lambda-cyhalothrin on Wistar rat bone marrow

In this study, the genotoxic and cytotoxic potential of lambda-cyhalothrin (LCT), a synthetic pyrethroid insecticide, was investigated in Wistar rat bone-marrow cells, using the structural chromosomal aberration (SCA) and micronucleus (MN) test systems. LCT was administrated to adult female albino rats as repeated i.p. doses of 6.12, 3.06, 0.8 mg/kg BW for 13 days at 48 h intervals. Mitomycin C (MMC) was used as a positive control (2 mg/kg BW). All the doses of LCT increased the number of structural chromosomal aberrations and the frequency of micronucleated erythrocytes, compared with the control group. It was also observed that LCT caused a significant decrease in the number of polychromatic erythrocytes. Our results demonstrate that LCT has a clastogenic/genotoxic potential as measured by the bone marrow SCA and MN tests in Wistar rats.

WR-2721: inhibitor of cisplatin-induced micronuclei.

The modulatory effect of S-2-/3-aminopropylamino/ethyl-phosphorothioic acid, (WR-2721, Amifostine) on induction of micronuclei by cis-diamminedichloroplatinum[II] (CDDP) was studied. The adult, male Swiss mice were treated with WR-2721, at a dose of 200 mg/kg or 400 mg/kg body weight, and/or with CDDP, at a dose 5 mg/kg or 10 mg/kg body weight. WR-2721 was given alone or 30 min before CDDP administration. The frequency of micronucleated polychromatic erythrocytes (MNPCEs) and also the number of polychromatic erythrocytes (PCEs) in the bone marrow and peripheral blood, at 24 h after the drug application, were determined. After administration of CDDP, the frequency of MNPCEs distinctly increased, and the number of PCEs decreased. As compared with the animals injected with CDDP only, in mice treated with WR-2721 before CDDP application, the number of MNPCEs was reduced and the frequency of PCEs was increased. However, WR-2721 given alone, without subsequent administration of cis-diamminedichloroplatinum[II], caused an increase in the number of micronucleated polychromatic erythrocytes and a decrease in the number of PCEs. The geno- and cyto-toxicity and chemoprotection were dependent on the doses of the agents WR-2721 and CDDP applied. In mice injected with CDDP and/or WR-2721, the patterns of changes in the frequency of MNPCEs and PCEs were similar in the bone marrow and peripheral blood, respectively. The protective effect of the aminothiol compound WR-2721 against induction of micronuclei and apoptotic cell death in the erythropoietic system by CDDP was shown.

Correction of an Error in Calculation in the Article "Frequency of Micronuclei in the Peripheral Blood and Bone Marrow of Cancer-Prone Mice Chronically Exposed to 2450 MHz Radiofrequency Radiation" (Radiat. Res. 147, 495-500, 1997)

Upon re-examination of the data in our manuscript entitled Frequency of Micronuclei in the Peripheral Blood and Bone Marrow of Cancer-Prone Mice Chronically Exposed to 2450 MHz Radiofrequency Radiation, which appeared in the April 1997 issue of Radiation Research (Vol. 147, pages 495-500), we realized that an error was made in the statistical analysis. In the last column in Tables I and II, the group mean micronuclei was based on 1000 polychromatic erythrocytes (PCEs), but the calculation of the standard deviation was based on 2000 PCEs. When the correction is made using the same number of PCEs, the two-tailed Student's t test indicated that the difference between the mice chronically exposed to radiofrequency (RF) radiation and the sham-exposed mice is statistically significant for both peripheral blood and bone marrow in Table I, and not for either tissue in Table II. We apologize for the miscalculation. Having corrected the error, we wish to point out that, although the difference was statistically significant within this (one) experiment, the biological relevance must be put in proper perspective. The increase in the frequency of micronuclei in the 62 mice exposed to RF radiation, compared to the frequency in the 58 shamexposed mice, was only 1 additional micronucleus per 200 PCEs, or an increase of 0.05%. This is a very small change in a large number of animals exposed to RF radiation ov r a very long period (18 months). Based on this evidence alone, it would be premature to conclude that the RF radiation exposure acted as a weak mutagen. Furthermore, the statistical increase in the frequency of micronuclei was not correlated with an outcome of carcinogenesis. Neither the latent period nor the total number of tumors was reported to be different between the exposed and sham-exposed mice (Frei et al., manuscript in preparation), the same animals for which the micronucleus frequencies were determined. As one final point, and as pointed out in the paper, the incidences of micronuclei in both the mice exposed to RF radiation and the sham-exposed mice were all within the range of the spontaneous incidence of micronuclei reported for mice in the literature.

An antigenicity study of calcipotriol (MC903)

Antigenicity of calcipotriol (MC903), an anti-psoriasic agent, was investigated in mice and guinea-pigs. 1. In mice, MC903 administered alone or with an adjuvant (Alum) did not result in the production of MC903-specific IgE antibody. 2. In guinea-pigs sensitized with MC903 alone or plus an adjuvant (FCA), systemic anaphylaxis was not induced by challenging with MC903. IgG1 antibody in MC903-sensitized guinea-pigs was not detected by the 4-hr PCA test. 3. Ovalbumin induced the production of ovalbumin-specific IgE antibody in mice, and ovalbumin-specific IgG1 antibody in guinea-pigs. Intense systemic anaphylaxis in the ovalbumin-sensitized guinea-pigs was induced by challenging with ovalbumin. 4. Results obtained in the present study suggest that MC903 does not induce the production of IgE antibody in mouse, and IgG1 antibody in guinea-pig, and in MC903-sensitized guinea-pig systemic anaphylaxis is not induced by challenging with MC903.

A mutagenicity study of calcipotriol (MC903)

Calcipotriol (MC903), an anti-psoriasic agent, was examined for mutagenicity in the reverse mutation test and the chromosomal aberration test in vitro, and the micronucleus test in Slc:ddY mice. 1. In the reverse mutation test using Salmonella typhimurium (TA100, TA1535, TA98, TA1537) and Escherichia coli (WP2uvrA), MC903 did not significantly increase revertant colonies in any of the test strains with or without metabolic system. 2. In the chromosomal aberration test with a Chinese hamster fibroblast cell line (CHL), MC903 did not significantly increase aberrant cells in the direct method or in the activation method. 3. In the micronucleus test with male mice, MC903 did not significantly increase in the number of polychromatic erythrocytes with micronuclei in the bone marrow. These results suggest that MC903 has no mutagenic as well as clastogenic effects under the present experimental condition.

Dominant-lethal mutations and micronucleus induction in male BALB/c, BDF1 and H mice by tobacco smoke

The mutagenicity of whole tobacco smoke (TS) was examined in male BALB/c, BDF1 and H mice using the dominant lethal and micronucleus tests in bone marrow polychromatic erythrocytes. Male mice (about 80 days old) from the three strains were treated with TS on 5 days/week for 8 weeks. The animals were divided into three groups for every strain: control and two experimental groups. Two doses - low (1h treatment/day) and high (2 h treatment/day) - were used. In the first case 8 exposures of 7.5 min each with 1-min intervals were given and to realize the high dose this was repeated 4 h later. It was found that TS induces significant dominant-lethal mutations in both experimental groups of BALB/c, BDF1 and H mice ( p < 0.001), but some strain differences existed. The data obtained suggest that in BALB/c and BDF1 mice TS induces dominant-lethal mutations mainly in spermatocytes, spermatogonia and gonial stem cells, while in H mice only spermatids and spermatocytes are affected. The bone marrow from each strain and each dose group was investigated for the presence of micronucleated polychromatic erythrocytes (PCE) at 5, 19, 38, 54, and 63 days after beginning of the treatment with TS, using the same exposure regimen. Exposure of male mice to TS caused an up to 2–3-fold increase in the number of PCE in the bone marrow of BALB/c and BDF1 mice in both dose groups. In H mice this effect is observed only on days 19 and 38 of sampling. No cumulative or dose-dependent effects were detected. Increased formation of micronuclei within PCE of femoral bone marrow after passive smoking was regarded as being due to the effect of TS.

Effect of Sevidol on the Induction of Micronuclei in the Bone-Marrow Erythrocytes of Mice

SUMMARYSevidol, an insecticidal mixture comprising carbaryl and lindane, was studied for genotoxicity evaluation in the bone-marrow erythrocytes of Swiss albino male mice. EMS was taken as the reference mutagen. Sevidol was dissolved in DMSO and three dose levels (1/10 LD50, 1/2 LD50, a dose close to LD50) were studied. Sevidol was found to increase the number of micronuclei significantly in the polychromatic erythrocytes at the three doses tested. A significant decrease in the number of polychromatic erythrocytes was also observed at the highest dose tested, indicating the toxic nature of the compound.

Micronucleus test in bone marrow of mice treated with 1-nitropropane, 2-nitropropane and cisplatin

Micronucleus tests were carried out in bone marrow of mice treated with 1-nitropropane, 2-nitropropane and cisplatin. For 1-nitropropane and 2-nitropropane the results were negative. With cisplatin a dose-dependent increase in the number of polychromatic erythrocytes with micronuclei was observed. The lowest positive dose was 0.1 mg/kg ( P < 0.001, Mann-Whitney-Wilcoxon test). The hepatocarcinogen 2-nitropropane showed clastogenic activity in human lymphocytes in vitro in the presence of S9 (Bauchinger et al., 1987). The negative results in bone marrow suggest that short-lived genotoxic metabolites may be formed in the liver but do not reach the bone marrow.

Relative distribution of mature erythrocytes, polychromatic erythrocytes (PE) and micronucleated PE on mouse bone-marrow smears: control observations

Maps are presented showing the distribution of normal erythrocytes (NE), polychromatic erythrocytes (PE) and micronucleated PE (MPE) on 3 mouse bone-marrow smears. Adjacent areas of each slide were assessed along their whole length, and it was found that both the incidence of MPE and the ratio of PE: NE varied between different regions of the slide. These two variables were essentially independent of each other. The maps give the impression of clusters of islands of MPE on the slide, but these become less significant when the number of erythrocytes in those areas is taken into account. The present data confirm our previous conclusion that the resolving power of the bone-marrow micronucleus assay is related to the number of PE assessed for MPE per slide. Our data also illustrate that in instances where small departures from control values are observed, clarification/confirmation of the result can be obtained by extending the assessment of slides for MPE. It is proposed that the present recognition of the inherent sensitivity contained within this assay necessitates careful examination of the criteria for a positive response. The selection of PE suitable for assessment is discussed, and our recent 24-h oral corn oil control data are analysed.

The effect of benzene in the micronucleus test

The potential cytogenetic effect of benzene was studied by means of the micronucleus test in developing erythrocytes of male and female mice at dosages ranging from 0.0625 to 2.0 ml/kg/day. Groups of mice were given two daily oral doses and sacrificed 6, 18, 24 or 48 h, or 5, 9 or 16 days after the last dose. The doses were given 24 h apart. Bone marrow from the mice was processed for visualization of micronuclei in polychromatic erythrocytes. The results showed a significant ( P < 0.05) increases in the numbers of polychromatic erythrocytes with micronuclei from mice sacrificed 6 h after being given dosages of 0.25 ml/kg/day or more of benzene. Similarly, significant ( P < 0.05) increases in polychromatic erythrocytes with micronuclei were seen in mice given 0.125 ml/kg/day or more of benzene and sacrificed 18 or 24 h after the second dose. However, in a second experiment negative results were obtained at 24 h from mice at 0.0625, 0.125 and 0.25 ml/kg. One group of mice (0.25 ml/kg/day and killed at 48 h) showed significant increases in the numbers of polychromatic erythrocytes with micronuclei. Mice killed 5 days after treatment with 2 doses of 0.125 ml/kg or more of benzene showed significant numbers of micronuclei in polychromatic erythrocytes. The groups of mice at 0.5, 1.0 or 2.0 ml/kg/day and killed 9 or 16 days after the second dose of benzene showed values that were comparable to the corresponding negative controls. In all cases there were sufficient numbers of cells to analyze. Although benzene is a known hematopoietic poison and bone-marrow depressant, there were sufficient numbers of cells to evaluate in the micronucleus test. Thus, these experiments uphold the usefulness of the micronucleus test as a screening procedure in cytogenetics.