Micronuclei In Bone Marrow Cells Research Articles

High body temperature induces micronuclei in mouse bone marrow

The mouse micronucleus test was conducted to investigate the effect of high body temperature on micronucleus induction. Groups of 10 male ddY mice were exposed to 30°C for 1, 3 or 6 h, 37°C for 0.5, 1, 2, 3 or 4 h, and 40°C for 1 or 2 h. Bone marrow cells were sampled 24 h after heat exposure. Exposure of mice to 37°C for 3 or 4 h and 40°C for 1 or 2 h raised body temperature to approximately 40.5°C and produced statistically significant increases in micronucleated polychromatic erythrocyte frequencies (8.1±4.5, 6.0±2.1, 5.3±3.3, 7.5±2.9‰, respectively; control frequencies, 2.0±1.1‰). In addition, about 25% of the induced micronuclei were relatively large (diameter of micronucleus ≥1/4 diameter of cytoplasm). These results suggest that body temperatures of 39.5°C or higher for more than 30 min induce micronuclei in bone marrow cells, and one possible mechanism is disturbance of the mitotic apparatus.

Mechanistic studies on genotoxicity and carcinogenicity of salicylazosulfapyridine an anti-inflammatory medicine

Salicylazosulfapyridine (SASP), which has been in clinical use for over 50 years, was reported by the National Toxicology Program to increase rat (F344 strain) urinary bladder and mouse (B6C3F1 hybrid) liver tumours under ad libitum (AL) feeding conditions, while under a feed restriction (FR) regimen, these tumours were not increased. The present investigations were undertaken to assess the implications of these results for the safety of SASP in humans. SASP and its 2 major metabolites, 5-aminosalicylic acid (ASA) and sulfapyridine (SP) were tested for in vivo induction of micronuclei in mouse bone marrow cells with or without prefolic treatment and for in vivo formation of DNA adducts in rat and mouse liver and urinary bladder. None exhibited mutagenicity or DNA reactivity. SASP and SP have induced sister chromatid exchanges and micronuclei (MN) in cultured human lymphocytes in the absence of liver activation enzymes and in B6C3F1 mice (but not in rats) MN in bone marrow and peripheral RBC. Treatment with folate reduces the frequency of MN. Perhaps the short (28 days) RBC lifespan in mouse underlies the sensitivity of this species. Thus, SASP without folate supplementation is an aneuploidogen. In a 2-year study in AL fed SASP-treated (high dose 337.5 mg/kg) rats, urinary pH was increased and urinary specific gravity was reduced at 60 weeks. At the end, this SASP group showed urothelial hyperplasia and papillomas in the urinary bladders of male rats primarily. In the FR high dose SASP group, the hyperplasia was reduced from 82% to 14%. At the end of 2 years, the incidence of multi-organ leukemia was reduced in both AL and FR high dose SASP groups. Thus, SASP caused intraluminal bladder changes in the rat (especially males) consisting of chronic urothelial stimulation, concretions, hyperplasia which resulted in neoplasia. In the mouse, because of species differences in liver ratios (mouse > rat) and, increasing (3 times higher) liver perfusion in the mouse, compared to the rat, there was hepatocellular toxicity and resulting preneoplasia and neoplasia within 2 years. These findings occurred in all AL SASP groups (flat curve without dose response); but were reduced under FR conditions. In this species, the multiorgan lymphoma incidence was reduced in both AL and FR high dose SASP groups. Thus, SASP and its major metabolites are not genotoxic. Folate deficiency associated with SASP administration is probably responsible for aneuploidy in lymphocytes and erythrocytes. SASP does not induce neoplasia directly in either livers, urinary bladders or other organs. Accordingly, SASP is judged to pose no carcinogenic risk to humans.

Micronucleus induction by diuron in mouse bone marrow

Diuron, a widely used substituted urea herbicide, induced the formation of micronuclei in bone marrow cells of Swiss mice. A single i.p. dose of 340 mg/kg b.w. diuron which is maximum tolerated dose (MTD) increased significantly the number of micronuclei at 30 h and 48 h time period. The dose of 170 mg/kg b.w. also induced the micronuclei formation in the above time period. However, a dose of 85 mg/kg b.w. was ineffective at the time periods studied. No induction of micronuclei was observed at 72 h time period after all the doses of diuron studied as compared to the solvent control. The diuron-induced frequency of micronucleated erythrocytes was independent of the sex of the test animals.

High body temperature induces micronuclei in mouse bone marrow cells

High body temperature induces micronuclei in mouse bone marrow cells

Effects of new quinolone antibacterial agents on mammalian chromosomes.

The clastogenic effects of several new quinolones (ciprofloxacin, enoxacin, levofloxacin, nalidixic acid, ofloxacin, pipemidic acid, and N1-cyclopropyl quinolones for drug candidate) were studied cytogenetically using Chinese hamster lung cells (CHL) and the mouse micronucleus test. Some N1-cyclopropyl quinolones strongly induced chromosomal aberration on CHL cells, and some, but not all, were also capable of inducing micronuclei in mouse bone marrow cells. Levofloxacin showed weak clastogenicity in CHL cells but did not induce either micronuclei in mouse bone marrow or unscheduled DNA synthesis (UDS) in rat hepatocytes when administered to intact live animals. The lack of concordance between in vitro and in vivo assays could reflect the differences in the tissue levels of the drugs and the in vitro conditions.

In-vivo genotoxicity of the alkaloid drug pilocarpine nitrate in bone marrow cells and male germ cells of mice

Pilocarpine nitrate, an alkaloid drug of plant origin induces spindle disfunction in bone marrow cells of mice. Further studies were carried out to investigate its mutagenic effects in somatic and germ cells of mice by assessing chromosome aberrations at mitotic metaphase and as micronuclei in bone marrow cells and sperm-shape abnormality in cauda epididymides. The dose and time yield effects of the drug were investigated. The statistically significant results that were obtained for both chromosomal aberrations and micronucleus test but not for the sperm-shape abnormality test, indicated the genotoxicity of this compound in somatic cells but not in germ cells.

Mutagenic activity of isoxazolylnapthoquinoneimines assayed by micronucleus bone marrow test

Studies were undertaken to evaluate the ability of various quinoneimines to induce micronuclei in bone marrow cells as a measure of their genotoxicity. Accoddingly, 2-hydroxy- N-(3,4-dimethyl-5-isoxazolyl)-1,4-napthoquinone-4-imine (I), its 2-acetyl derivative (II) and 2-[(5-methyl-3-isoxazolyl)amino]- N-(5-methyl-3-isoxazolyl-1,4-naphthoquinone-4-imine (III), as well as two of their precursors, 2-hydroxynaphthoquinone (NQ-2-OH) and 3,4-dimethyl-5-aminoisoxazole (DMAI) were given by intraperitoneal injection at 5, 50, 100 and 200 mg/Kg doses to S.J.L. Swiss mice with 24 h sampling time. Compounds I and II displayed highly significant differences at 50, 100 and 200 mg/kg doses (p < 0.01) and their mutagenic dose response curves correlated closely with an inverted U-shaped form whose interpretation is still the subject of controversy. NQ-2-OH only produced a significant increase in micronucleus frequency at 50 mg/kg, whereas no mutagenic activity was found for compound III and DMAI at the doses assayed. At 50 mg/kg the order of relative mutagenic potencies was I > II > NQ-2-OH. Mechanisms advanced to explain loss of drug activity at high doses include capture saturation, enzymatic induction during metabolism and participation of and independent defense system.

Ambient air pollutants in upper Silesia: partial chemical composition and biological activity.

The air monitoring system in Upper Silesia has provided abundant data on airborne pollutants. Air quality in this region is bad: a concentration of several gases, volatile compounds, metals, and complex mixtures of organic compounds carried by small particulate matter exceeds both daily and yearly admissible levels. About 250 individual polycyclic aromatic hydrocarbonds (PAHs) were identified in airborne pollutants, and hundreds of not identified compounds are seen on gas chromatographic profiles as minor peaks. Among PAHs are present compounds with known carcinogenic potency for humans. Seasonal variation with distinctly lower concentration of pollutants in summer than in winter was noticed. Fifteen PAHs including benzo[a]pyrene (B[a]P) determined by GC-MS method in 20 measuring points showed constant relative proportions. Thus B[a]P could be used as a representative compound for other PAHs. In urban areas, a core of Silesia values for B[a]P concentration ranged from 60 to 90 ng/m3 in winter to 5 to 20 micrograms/m in summer. Mutagenicity tested on Salmonella strains showed seasonal variation with distinctly higher values in winter. Environmentally exposed humans showed a higher level of PAH-DNA adducts in WBC than the control population from rural area. Total organic extract of small particulate matter exhibited both direct and indirect mutagenic activity, induced formation of micronuclei in bone marrow cells of BALB/c mice, induced chromosomal rearrangements, and increased sister chromatid exchange index.

Mutagenic potential of acute exposure to organophosphorus and organochlorine compounds

The cytogenetic and cytotoxic effects of the pesticides methyl parathion, Bayleton and Hinosan were evaluated in mammalian test systems. The frequency of chromosome aberrations and micronuclei in bone marrow cells and the arginase enzyme profile in the liver tend to show the genotoxicity of organophosphorus and organochlorine pesticides in a single-exposure response study. Methyl parathion was the most hazardous among the three, showing definite pathology in the livers of treated rats.

Differentiation of micronuclei in mouse bone marrow cells: a comparison between CREST staining and fluorescent in situ hybridization with centromeric and telomeric DNA probes.

Immunofluorescent staining (CREST) of kinetochore proteins and in situ hybridization (FISH) with centromeric DNA probes are able to distinguish between micronuclei (MN) containing lagging chromosomes or acentric fragments. Different frequencies of signal-positive MN induced by mitomycin C (MMC) were obtained by Miller et al. (Mutagenesis, 6, 297-302, 1991) between CREST labelling and FISH with the mouse major-gamma-satellite DNA probe (major probe). Both modes of identifying the presence of an entire chromosome in a MN are theoretically prone to misclassification. Breaks induced in pericentric heterochromatin can produce fragment-containing MN with a major signal. Alternatively, alterations of kinetochore proteins can produce CREST-negative MN containing lagging chromosomes. To improve the reliability of MN differentiation two additional DNA probes, the mouse minor satellite DNA probe (minor probe) and the telomere repeat (5'-TTAGGG-3')7, have been used and double labelling has been employed with minor/major and minor/telomere probes. At 1 mg/kg of MMC the labelling frequencies of MN with CREST and the minor probe were identical (18.5 and 19%, respectively) and the major probe showed a higher labelling rate (30.5%). Using double-labelling the difference between minor and major probe responses was confirmed (17 and 31.5%, respectively). At 5 mg/kg of MMC, CREST labelling gave the lowest (6%), the minor probe gave intermediate (10 and 11.5% after single- and double-labelling, respectively) and the major probe gave the highest signal frequencies (16.5 and 15% after single- and double-labelling, respectively). The CREST and the minor signal frequencies were not significantly different at either dose of MMC whereas the minor and the major signal frequencies were significantly different.(ABSTRACT TRUNCATED AT 250 WORDS)

Decrease in deoxyribonucleotide triphosphate pools and induction of alkaline-labile sites in mouse bone marrow cells by multiple treatments with methotrexate

Methotrexate (MTX), an inhibitor of dihydrofolate reductase (DHFR), slightly induced micronuclei in bone marrow and peripheral blood cells, and this induction was enhanced by multiple treatments with the drug. Furthermore, we have suggested that the multiple-dose effect on the induction of micronuclei by MTX might be explained by intracellular accumulation of the drug, resulting in an enhancement of DHFR inhibition. An imbalance or decrease in the deoxyribonucleotide (dNTP) pool would be generated by this enzyme inhibition. Therefore, we attempted to determine the level of the dNTP pool in mouse bone marrow cells. The levels of three dNTPs (dTTP, dATP, dGTP) as determined by HPLC were only 1/10–1/40 of the levels previously found in mammalian cell lines, but dCTP levels could not be determined precisely because they approached the limits of detectability. The levels of dTTP, dATP and dGTP in mouse bone marrow cells 3 h after four injections of MTX (4 mg/kg/day) decreased to 21.2%, 47.0% and 38.1%, respectively, of those in the control group. The level of dTTP 3 h after four injections of 100 mg/kg of the drug decreased to almost 0%. The results of alkaline elution assays suggested that alkaline-labile sites were generated in mouse bone marrow cells 6 h after four injections of MTX (4 mg/kg). These findings suggest that the multiple-dose effects of MTX on micronucleus induction in mouse bone marrow cells may be explained by the decrease in the dNTP pool and subsequent generation of alkaline-labile sites (possibly apurinic/apyrimidinic sites).

Protective effect of sulfoethylglucan against hexavalent chromium

The effect of pretreatment with sulfoethylglucan (SEG) on the frequency of micronuclei and the liver alkaline phosphatase activity induced by potassium bichromate (Cr(VI)) in mice was evaluated. Simultaneous application of SEG and Cr(VI) decreased the frequency of micronuclei in bone marrow cells ( P < 0.01) and the level of liver alkaline phosphatase activity in comparison to the Cr(VI) group. Pretreatment with SEG 24 h prior to the first Cr(VI) application resulted in a more pronounced decrease in the Cr(VI)-induced frequency of micronuclei. The mechanisms of the protective effects of sulfoethylglucan could be explained either by the formation of Cr ion complexes with sulfoethyl groups of glucan or by the scavenging ability of SEG to trap hydroxyl radicals.

Evaluation of the genotoxic potential of zinc pyrithione in the Salmonella mutagenicity (Ames) assay, CHO/HGPRT gene mutation assay and mouse micronucleus assay.

The mutagenic potential of zinc pyrithione (Znpt) was evaluated in vitro in the Salmonella/mammalian microsome plate incorporation mutagenicity (Ames) assay and the CHO/HGPRT gene mutation assay. The clastogenic potential of Znpt was evaluated in vivo using the mouse micronucleus test. Znpt was negative in the Ames test in five tester strains in the presence and absence of rat liver microsomal enzymes when assayed at concentrations ranging between 10 and 333 micrograms per plate and between 0.03 and 33 micrograms per plate, respectively. Znpt also produced negative results in the CHO/HGPRT assay. No significant increases in mutant frequencies were seen in the presence and absence of rat liver microsomal enzymes. In each case, the highest concentrations reduced cellular viability by 83% and 85%, respectively. Znpt also did not induce increased frequencies of micronuclei in mouse bone marrow cells when tested at the maximally tolerated dose (MTD) (44 mg kg-1). These data support the conclusion that Znpt lacks genotoxic activity under the conditions of these tests.

The size of micronuclei in mouse bone marrow cells induced by rotenone

The size of micronuclei in mouse bone marrow cells induced by rotenone

Induction of micronuclei in mice bone marrow cells by home made aguardientes collected in southern Chile and their incidence in gastric cancer.

Induction of micronuclei in mice bone marrow cells by home made aguardientes collected in southern Chile and their incidence in gastric cancer.

Genotoxic potential of the organochlorine insecticide lindane (γ-BHC): an in vivo study in chicks

The purpose of this work was to evaluate the genotoxic potential of lindane (γ-isomer of benzene hexachloride (BHC)) in chicken in vivo test: the bone marrow chromosome aberration and micronucleus tests. With the highest dose (100 mg/kg) a significant enhancement of chromosome aberrations was noticed after 24 and 48 h and with the second highest dose (75 mg/kg) after 24 h. A significant increase in the incidence of micronuclei in bone marrow cells was induced by all three doses (100, 75 and 50 mg/kg) given either intraperitoneally or orally while in peripheral erythrocytes only the two higher intraperitoneal doses (100 and 75 mg/kg) gave significant increases. On the basis of these results, lindane may be considered genotoxic in this test system and it is suggested that the chick in vivo system may be used as an alternative to a mammalian system for screeining environmental chemicals for genotoxicity.

Induction of kinetochore positive and negative micronuclei in mouse bone marrow cells by salicylazosulfapyridine and sulfapyridine

Salicylazosulfapyridine (SASP) and its major metabolite sulfapyridine (SP) have been shown to induce chromosomal damage in vivo. Both chemicals were tested in the micronucleus (MN)/kinetochore (KC) staining test to gain insight into the question of whether chromosomal breakage, aneuploidy-inducing events, or both were important to the observed production of MN in bone marrow cells of mice. In this test, both SASP and SP were shown to be strong inducers of kinetochore positive (KC +) MN. Although small increases in kinetochore negative (KC −) MN were also observed in SP treated mice, as well as in mice receiving the highest dose of SASP tested, the results suggest that both chemicals induce predominantly aneuploidogenic type damage.

Evaluation of the genotoxic potential of sorbic acid and potassium sorbate

The genotoxic potential of sorbic acid and potassium sorbate was investigated in vivo and in vitro. Oral administration of sorbic acid (up to 5000 mg/kg body weight) did not induce sister chromatid exchanges or the formation of micronuclei in bone marrow cells of mice. Intraperitoneal treatment of rats with 400–1200 mg potassium sorbate/kg body weight did not alter the elution profile of DNA from isolated liver cells in the in vivo alkaline elution assay. Sorbic acid did not induce DNA repair in cultured human A549 cells in the unscheduled DNA synthesis (UDS) assay. In vitro incubation of the cells with 1–1000 μg potassium sorbate/ml, in the absence or presence of rat liver homogenate, did not result in the formation of DNA single-strand breaks in the alkaline elution assay. These results demonstrate that sorbic acid and its potassium salt are not genotoxic in vivo or in vitro. In contrast to sorbic acid and potassium sorbate, sodium sorbate is very sensitive to oxidative degradation; the main oxidation product was identified to be 4,5-oxohexenoate, which was mutagenic in the Ames test.

Data selection and treatment of chemicals tested for genotoxicity and carcinogenicity.

A database containing qualitative and quantitative results of experimental studies in the fields of genotoxicity and carcinogenicity has been developed. By analyzing results of the studies performed by the U.S. National Toxicology Program, or by a similar program developed in Japan, or reported in the scientific literature, as well performed by private organizations, information has been collected relating to 3389 chemicals, identified by their CAS number. The studies considered for the database include three genotoxicity/mutagenicity short-term test (STTs), namely, two in vitro (Salmonella, gene mutation assay, and mammalian cells/human lymphocytes chromosome aberration assay) and one in vivo, the rodent bone marrow micronucleus assay. To investigate the possible predictive value of these STT assays for carcinogenicity, the results of animal long-term bioassays have also been collected. We have re-evaluated all the genotoxicity studies and the majority of those cases studied in different laboratories with contrasting results has been resolved; a small proportion of questionable cases is, however, still present in the database. In total, 2898 (85.5%) of the chemicals have been tested in the Salmonella assay; 1399 (41.3%) have been tested in the in vitro chromosome aberration assay; 319 (9.4%) have been tested in the in vivo rodent bone marrow cell micronucleus assay; 716 (21.2%) of the chemicals have been tested in the in vivo animal long-term bioassay. For 1118 chemicals tested in the Salmonella assay, 30,650 quantitative studies have been included in the database, thus allowing a possible classification of mutagenic chemicals according to their mutagenic potency.(ABSTRACT TRUNCATED AT 250 WORDS)

Genotoxicity testing of extracts of a Swedish moist oral snuff

The present study was designed to investigate the potential genotoxicity of aqueous and methylene chloride extracts of Swedish moist oral snuff. The test systems were selected to provide optimal data for the prediction of carcinogenicity in rodents and included assays for the induction of mutation in bacteria, sister-chromatid exchanges (SCE) in human lymphocytes, of chromosome aberrations and gene mutations in V79 Chinese hamster cells and of micronuclei in mouse bone marrow cells. In addition, the methylene chloride extract was tested for the induction of sex-linked recessive lethal mutations in Drosophila melanogaster. The aqueous extract of ‘Snus’ induced SCE in human lymphocytes and chromosome aberrations in V79 cells, the latter effect being observed both with and without metabolic activation. No induction of point mutations was detected with the Ames test or in V79 cells and the micronucleus test in mice was negative. It was demonstrated that the induction of chromosome aberrations without metabolic activation may be due to a high salt concentration, indicating that the clastogenic agent(s) in this extract required metabolic activation. The methylene chloride extract showed genotoxicity in the Ames test, the SCE test and the chromosome aberration test, whereas no induction of gene mutations in V79 cells was observed. Once again, the results suggested that metabolism is required for genotoxicity. The methylene chloride extract did not cause induction of micronuclei in mice or of sex-linked recessive lethal mutations in Drosophila melanogaster. These combined data on genotoxicity were analyzed using various models for the prediction of carcinogenicity. In a sequential testing model, the probabilities that the aqueous and methylene chloride extracts of ‘Snus’ are carcinogenic due to a genotoxic mechanism were both predicted to be low. Using carcinogenicity prediction by battery selection (CPBS), the probabilities of the methylene chloride and aqueous extracts being correctly identified as non-carcinogens are 71 and 77%, respectively. Up to date, the CPBS approach has been validated primarily for individual compounds, so some caution should at present be exercised in interpreting the results using this method. Based on these results, the carcinogenic potential of Swedish ‘Snus’ should be considered to be low, a conclusion in agreement with the low incidence of oral cancer in Sweden compared to other countries.