Mouse Bone Marrow Erythrocytes Research Articles

Genetic and prenatal developmental evaluation of anthraquinone

Anthraquinone is a recently identified contaminant present in teas globally, and its potential teratogenic and genotoxic impacts have yet to be fully comprehended. Hence, this study's objective was to determine anthraquinone's genotoxicity using various studies such as the Ames test, Mammalian erythrocyte micronucleus test, and in-vitro mammalian chromosome aberration study. Additionally, the study assessed its effects on maternal gestational toxicity and the fetus's teratogenicity through prenatal developmental toxicity research in rats. Results indicated that anthraquinone did not manifest mutagenic effects on Salmonella typhimurium histidine-deficient, did not cause chromosomal aberrations in Chinese hamster ovary cell subclone CHO-K1, and did not exhibit a genotoxic effect on mouse bone marrow erythrocytes. However, in the prenatal developmental toxicity study, administering anthraquinone orally to pregnant rats from day 5 to day 19 of gestation resulted in decreased body weight and food consumption of pregnant rats, along with a higher number of visceral malformations in the fetuses in the highest dose group (217.6 mg/kg BW). Additionally, two pregnant rats died in this group. The study has established the no observed adverse effect level (NOAEL) as 21.76 mg/kg BW, while the lowest observed adverse effect level (LOAEL) was 217.6 mg/kg BW.

Comparative evaluation of the genotoxicity of carbendazim technical grade active ingredients in the Ames test and micronucleus in vivo test

Introduction. Carbendazim is a systemic benzimidazole fungicide used against a wide range of crop diseases. The ability of carbendazim to induce the incidence of chromosomal aberrations and micronuclei in mammalian cells by influencing the processes of mitotic spindle formation in the cell cycle have been shown in various in vitro and in vivo tests. Contradictory data were obtained in the bacterial test system Salmonella/microsomes, indicating both the absence and the presence of mutagenic activity of carbendazim. The discrepancy in the results may stem from the presence of impurities. The aim of the study was a comparative evaluation of the genotoxicity of various technical products of carbendazim. Materials and methods. The genotoxicity of carbendazim was studied using the plate incorporation version of the Ames test on 5 strains of Salmonella typhimurium in the presence and the absence of metabolic activation system (+S9/-S9) and in a micronucleus test in CD-1 mice. Two technical grade active ingredients (TGAI) and an analytical standard for carbendazim were tested. Results. In the Ames test, the analytical standard of carbendazim possessed no mutagenic activity (±S9) on any of the strains. The most pronounced mutagenic effect was observed for the TGAI I in TA98 strain, the number of revertants at the maximum concentration was 5-7 times higher than that in the negative control. The positive effects of carbendazim TGAIs in the Ames test are likely mediated by the presence of impurities. Under in vivo conditions, all tested TGAIs of carbendazim induced a statistically significant and dose-dependent formation of micronuclei in polychromatic erythrocytes (PCE) of mouse bone marrow. The mean frequency of PCE with micronuclei at the maximum dose exceeded this rate in the negative control by 21-24 times. Research limitations. The study is limited to testing the mutagenic activity of two samples of carbendazim technical products and one sample of its analytical standard in both in vivo and in vitro tests. Conclusion. Taking into account the high content of the active substance in the tested TGAIs, the bacterial reverse mutation test is a highly sensitive method for assessment of the equivalence of carbendazim generic products. The use of a micronucleus test for evaluating of the equivalence of carbendazim TGAIs to the original substance is inappropriate due to the pronounced aneugenic effect.

Characterization and mechanism of aflatoxin degradation by a novel strain of Trichoderma reesei CGMCC3.5218.

Aflatoxins, which are produced mainly by Aspergillus flavus and A. parasiticus, are recognized as the most toxic mycotoxins, which are strongly carcinogenic and pose a serious threat to human and animal health. Therefore, strategies to degrade or eliminate aflatoxins in agro-products are urgently needed. We investigated 65 Trichoderma isolates belonging to 23 species for their aflatoxin B1 (AFB1)-degrading capabilities. Trichoderma reesei CGMCC3.5218 had the best performance, and degraded 100% of 50 ng/kg AFB1 within 3 days and 87.6% of 10 μg/kg AFB1 within 5 days in a liquid-medium system. CGMCC3.5218 degraded more than 85.0% of total aflatoxins (aflatoxin B1, B2, G1, and G2) at 108.2–2323.5 ng/kg in artificially and naturally contaminated peanut, maize, and feed within 7 days. Box–Behnken design and response surface methodology showed that the optimal degradation conditions for CGMCC3.5218 were pH 6.7 and 31.3°C for 5.1 days in liquid medium. Possible functional detoxification components were analyzed, indicating that the culture supernatant of CGMCC3.5218 could efficiently degrade AFB1 (500 ng/kg) with a ratio of 91.8%, compared with 19.5 and 8.9% by intracellular components and mycelial adsorption, respectively. The aflatoxin-degrading activity of the fermentation supernatant was sensitive to proteinase K and proteinase K plus sodium dodecyl sulfonate, but was stable at high temperatures, suggesting that thermostable enzymes or proteins in the fermentation supernatant played a major role in AFB1 degradation. Furthermore, toxicological experiments by a micronucleus assay in mouse bone marrow erythrocytes and by intraperitoneal injection and skin irritation tests in mice proved that the degradation products by CGMCC3.5218 were nontoxic. To the best of our knowledge, this is the first comprehensive study on Trichoderma aflatoxin detoxification, and the candidate strain T. reesei CGMCC3.5218 has high efficient and environment-friendly characteristics, and qualifies as a potential biological detoxifier for application in aflatoxin removal from contaminated feeds.

Toxicological Assessments of a Proprietary Blend of Punica granatum Fruit Rind and Theobroma cacao Seed Extracts: Acute, Subchronic, and Genetic Toxicity Studies.

LN18178 (Tesnor®) is a standardized, proprietary composition of aqueous ethanol extracts of Punica granatum fruit rind and Theobroma cacao seeds. The present study demonstrates a broad-spectrum toxicological evaluation of LN18178 utilizing in vitro and in vivo preclinical models following the Organization for Economic Cooperation and Development (OECD) guidelines for testing chemicals. Wistar rats did not show any clinical signs of toxicity and morbidity in acute oral and dermal toxicity tests with the median lethal dose (LD50) values of at least 5000 mg/kg and 2000 mg/kg body weight, respectively. LN18178 was nonirritating to the skin and eyes of the treated rabbits. In a ninety-day subchronic repeated oral dose toxicity study, the LN18178-treated Wistar rats did not show dose-related signs of toxicity on their body weight, food consumption, organ weights, hematology, and clinical chemistry parameters. The estimated no-observed-adverse-effect level (NOAEL) of LN18178 in male and female rats was 2500 mg/kg body weight. The observations from the bacterial reverse mutation test, in vitro chromosomal aberration assay, micronucleus assay in mouse bone marrow erythrocytes, and in vitro mouse lymphoma TK+/− gene mutation assay suggest that LN18178 is neither mutagenic nor clastogenic. In summary, the present study demonstrates that oral consumption of the herbal blend LN18178 does not show signs of toxicity; also it does not elicit genetic toxicity in the standard preclinical models.

Safety assessment of a novel water-soluble extract of Boswellia serrata gum resin: acute toxicity, 90-day sub-chronic toxicity, Ames’ bacterial reverse mutation, and in vivo micronucleus assays

Boswellia serrata gum resin extracts have demonstrated potential benefits in alleviating joint pain and discomfort of osteoarthritis. The major objective of the present study was to assess the safety of a water-soluble B. serrata gum resin extract (LI51202F1) in diverse models of acute oral, acute dermal, primary dermal irritation, eye irritation, and 90-day sub-chronic repeated dose toxicity studies, as well as Ames’ bacterial reverse mutation assay and in vivo micronucleus assay. The acute oral and dermal toxicity studies in Sprague Dawley (SD) rats demonstrated that the median lethal dose (LD50) of LI51202F1 is >2000 mg/kg body weight (BW). The acute dermal and eye irritation tests in New Zealand white rabbits exhibited that LI51202F1 is non-irritating to the skin and mildly irritating to the eyes, respectively. The 90-days sub-chronic repeated oral dose study demonstrated that the LI51202F1-treated male and female SD rats did not show signs of toxicity on their BW, food intake, organ weights, thyroid hormones, and on the clinical pathology, gross pathology, and histopathological assessments. In male and female rats, the no-observed-adverse-effect level (NOAEL) of LI51202F1 was 500 mg/kg/day, the highest tested dose in the study. The results of the bacterial reverse mutation assay in Salmonella typhimurium TA98, TA100, TA1535, TA1537, and Escherichia coli WP2uvrA (pKM101) strains in the presence or absence of S9 metabolic activation system and a micro-nucleus assay in mouse bone marrow erythrocytes demonstrated that LI51202F1 is neither mutagenic nor clastogenic. In conclusion, under the conditions of these studies, LI51202F1 demonstrated broad-spectrum safety.

СРАВНИТЕЛЬНАЯ ОЦЕНКА ПРОТИВОЛУЧЕВОЙ ЭФФЕКТИВНОСТИ ФЛАГЕЛЛИНА ПО ВЫЖИВАЕМОСТИ И МИКРОЯДЕРНОМУ ТЕСТУ

Purpose: Comparative evaluation of the effectiveness of the domestic drug flagellin (development of the State Research Institute of High-Purity Biological Products, St. Petersburg) in an extended range of drug administration periods before and after irradiation, as well as evaluation of the possibility of using the micronucleus test as a biomarker of its effectiveness.
 Material and methods: The work was performed on male ICR CD1 mice weighing 20–22g. The radioprotective effectiveness of flagellin was evaluated by the 30-day survival rate of experimental animals in comparison with control groups. The cytogenetic effect was evaluated by a micronucleus test in polychromatophilic erythrocytes (MJ-PCE) of mouse bone marrow. Irradiation was performed on the RUST M1 X-unit at a dose rate of 1.1 Gy/min in the dose range from 7 to 10 Gy for survival assessment and at a dose of 1 Gy for the micronucleus test. Flagellin was administered intraperitoneally at 0.2 mg/kg for 18 h and 30 min before irradiation, 10 and 30 min after irradiation. The animals of the control groups were injected with a solvent – phosphate-albumin buffer at the same time and in the same volume.
 Results: The most effective was the use of flagellin for 30 min and 10 min in relation to radiation (survival at 8.5 Gy 92 % and 78 %, at 9 Gy 81 % and 55 %, respectively) with survival in the control 29 % and 0. In the most studied version of the introduction of flagellin 30 minutes before exposure, the FID values at the level of generally accepted estimates LD16, LD50, LD84, were 1.3; 1.2 and 1.2, respectively.
 Conclusions: It is of interest to further expand the time range of the use of flagellin, especially after irradiation. The use of the method of estimating the proportion of polychromatophilic erythrocytes with micronuclei in the bone marrow allowed us to identify the optimal timing of drug administration much faster than in terms of survival (MJ-PCR test was performed 24 hours after irradiation). The MJ-PHE indicator can be considered as a potential biomarker of drug-induced increased radioresistance (this was previously shown in our studies for betaleukin, indralin, and riboxin).

Assessment of the mutagenicity of the technical product
 of N-(1-ethylpropyl)-2,6-dinitro -3,4-xylidinen

Introduction. Evaluation of genotoxicity of the pesticide technical products is one of the mandatory requirements for their toxicological and hygienic assessment. The data about mutagenic property is ambiguous for some pesticides. This may be due to the use of various active ingredients of technical products of the pesticide for testing, as they may have different profiles of relevant impurities, some of which may be potentially genotoxic. Material and methods. A technical product of N-(1-ethylpropyl)-2,6-dinitro-3,4-xylidine was tested using the bacterial reverse mutation method with Salmonella typhimurium (Ames test) and the in vivo mammalian micronucleus analysis in mouse bone marrow erythrocytes. Results. Statistically significant dose-dependent mutagenic effects of the technical product of N-(1-ethylpropyl)-2,6-dinitro-3,4-xylidine were revealed for TA97 (+S9 / -S9); TA100 (+S9 / -S9); TA102 (+S9 / -S9) and TA98 (+S9 / -S9) strains. In all cases, the fold increase of the revertant numbers mediated by the tested substance compared with the concurrent negative control was > 2 except TA98 in the presence of S9. In the micronucleus test, the technical product did not induce a statistically significant increase in the frequency of the micronucleated polychromatophilic erythrocytes in CD-1 mouse bone marrow up to 2000 mg/kg bw. Conclusion. The data suggest all technical products of pesticides entering the market should be tested for the potential genotoxicity. In such a case it is necessary to use at least two methods on different test systems for obtaining reliable results.

Assessment of the mutagenicity of the technical product
 of N-(1-ethylpropyl)-2,6-dinitro -3,4-xylidinen

Introduction. Evaluation of genotoxicity of the pesticide technical products is one of the mandatory requirements for their toxicological and hygienic assessment. The data about mutagenic property is ambiguous for some pesticides. This may be due to the use of various active ingredients of technical products of the pesticide for testing, as they may have different profiles of relevant impurities, some of which may be potentially genotoxic. Material and methods. A technical product of N-(1-ethylpropyl)-2,6-dinitro-3,4-xylidine was tested using the bacterial reverse mutation method with Salmonella typhimurium (Ames test) and the in vivo mammalian micronucleus analysis in mouse bone marrow erythrocytes. Results. Statistically significant dose-dependent mutagenic effects of the technical product of N-(1-ethylpropyl)-2,6-dinitro-3,4-xylidine were revealed for TA97 (+S9 / -S9); TA100 (+S9 / -S9); TA102 (+S9 / -S9) and TA98 (+S9 / -S9) strains. In all cases, the fold increase of the revertant numbers mediated by the tested substance compared with the concurrent negative control was > 2 except TA98 in the presence of S9. In the micronucleus test, the technical product did not induce a statistically significant increase in the frequency of the micronucleated polychromatophilic erythrocytes in CD-1 mouse bone marrow up to 2000 mg/kg bw. Conclusion. The data suggest all technical products of pesticides entering the market should be tested for the potential genotoxicity. In such a case it is necessary to use at least two methods on different test systems for obtaining reliable results.

The Mitigating Effect of Jatropha curcas L. Latex against Genotoxicity Induced by Doxorubicin

Jatropha curcas (Euphorbiaceae) is a multiple purpose lacticiferous plant with potential for biodiesel production and medicinal uses. There is in the literature different analyses about the toxic and cytogenotoxic effects of J. curcas extracts, but few information about latex toxicity. In addition, few models were employed to evaluate the toxicity response to J. curcas latex, and the toxicity in in vivo mammal’s model has not been tested yet. The cytotoxic, mutagenic and antimutagenic potential of J. curcas latex were investigated using mouse bone marrow erythrocytes. The results indicated a cytotoxic and mutagenic potential of this latex to mammalian cells. But, when J. curcas latex was co-administrated with doxorubicin (DXR – chemotherapy medication), a reduction in the number of micronuclei was observed, indicating an interaction between J. curcas latex and DXR. The interaction of latex with DXR can cause a reduction in the activity of this drug and impair the treatment of its users. Moreover, there is a lack of data on herb–drug interactions, what should be more investigated to safeguard the wellbeing of patients.

Toxicological Assessment of a Standardized Boswellia serrata Gum Resin Extract.

The acidic and non-acidic fractions of Boswellia serrata gum resin extracts were combined to prepare a unique product, LI13019F1 (Serratrin). The present series of studies evaluated LI13019F1 for acute and subchronic (28-day) toxicity in Wistar rats and acute dermal and eye irritation in New Zealand white rabbits. The mutagenicity and clastogenicity of LI13019F1 were evaluated in bacteria and mouse bone marrow erythrocytes, respectively. All studies were performed following the Organization for Economic Co-operation and Development guidelines. Acute oral and acute dermal toxicity studies did not show mortality or signs of toxicity in Wistar rats at a limit dose of 2,000 mg/kg LI13019F1. LI13019F1 did not cause irritation to the skin or the eyes of New Zealand white rabbits. In a repeated dose 28-day oral toxicity study, LI13019F1-treated Wistar rats did not show dose-related signs of toxicity on their body weights, organ weights, and on the hematology and clinical chemistry parameters. The estimated no observed adverse effect level for LI13019F1 was 1,000 mg/kg/day in both male and female rats. The bacterial reverse mutation test and a micronucleus assay in mouse bone marrow erythrocytes revealed that LI13019F1 was neither mutagenic nor clastogenic. Together, the present observations demonstrate a broad-spectrum safety of LI13019F1.

APPLICABILITY OF THE AMES TEST AND MICRONUCLEUS TEST IN VIVO FOR THE EVALUATION OF THE EQUIVALENCE OF PESTICIDE TECHNICAL GRADE ACTIVE INGREDIENTS COMPARED TO ORIGINAL ACTIVE SUBSTANCES

Introduction. Analogs of pesticides may differ from the original products in their properties because of the elevated level or the modified composition of the impurities. Therefore, it is necessary to determine the equivalence of such analogs using a number of criteria, including mutagenicity, to ensure their safety. The article compares the results of the research of genotoxic effects of technical grade active ingredients of pesticides in vitro and in vivo conditions to assess the applicability of such methods for equivalence determination of analogs of pesticides to patented products. Material and methods. The genotoxicity of 99 technical grade active ingredients of pesticides (59 names) was studied in vitro (Ames test) and in vivo. Results. In the Ames test mutagenic dose-dependent effects were revealed in the study of technical products of mesotrione, dimethoate, and pendimethalin both in the presence and in the absence of a metabolic activation system.In the in vivo test, a statistically significant dose-dependent increase in the frequency of micronucleated polychromatophilic erythrocytes in mouse bone marrow was detected after administration of six technical products mesotrione, pyrimiphos-methyl, dimethoate, glyphosate (2 products), isoproturon. Furthermore, different levels of genotoxic effects were found with technical materials of the same active ingredient from various productions, probably due to differences in the qualitative and quantitative composition of impurities. Conclusion. The present study indicated that in vitro and in vivo tests do not always demonstrate the same results of the genotoxicity assessment. Therefore, the use of only one bacterial reverse mutation test may not be sufficient to determine the equivalence of technical grade active ingredients of pesticides to the original active substances. To obtain а reliable evidence for the safe use of analogs of pesticides, it is necessary to usе at least two methods on different test objects.

Potent radioprotective effect of herbal immunomodulator drug (IMOD) on mouse bone marrow erythrocytes as assayed with the micronucleus test

Background : Although numerous natural or synthec drugs have been tested for their radioprotecve capacity, yet no suitable drug has been introduced for roune clinical use. In this study the radioprotecve effect of new herbal commercially known as IMOD, specifically made to decrease the side effects of HIV virus, was invesgated on mouse bone marrow cells. Materials and Methods : Female NMRI mice (in a group of five) were exposed to 2 Gy gamma radiaon following three days of intravenously injecon (IV) of IMOD at various doses. Mice were sacrificed 48 and 72 h a1er irradiaon. Bone marrow was flushed and slides for bone marrow smears were prepared according to standard method. A1er st aining slides in May Grunwald and Giemsa, polychromac erythrocytes (PCE) and normochromac erythrocytes (NCE) were scored for presence of micronucleus (MN). Results: The results showed that gamma irradiaon increased the frequency of micronuclei dramacally and excreted cytotoxic effect of cell proliferaon. Injecon of various dos es of IMOD before irradiaon however, led to a considerable reducon in the frequency of micronuclei in bone marrow erythrocytes as well as cellular toxicity. Conclusion: Results indicated radioprotecve capability of IMOD with a dose reducon factor (DRF) of about 2.3 at a dose of 20 mg/kg body weight. The considerably high DRF might be indicave that IMOD besides being an immunomodulator might also posses' anoxidant property.

English

This study evaluated the chomosomal damage potential of the extract from leaves of Calophyllum brasiliense Cambess. (Clusiaceae) in mouse bone-marrow cells. The extract was analyzed by high-performance liquid chromatography (HPLC) to assess the concentration of coumarin (-) mammea A/BB, a compound with antileishmanial activity. The chromosomal damage potential of the extract was evaluated by the micronucleus test in erythrocytes of mouse bone marrow. The animals were treated with cyclophosphamide (50 mg) as positive control, DMSO (1%) as negative control, or the crude extract from C. brasiliense (100 or 200 mg/kg). The HPLC analyses showed that the extract contained 25.97 ± 0.91 µg of (-) mammea A/BB per mg of extract. The extract of C. brasiliense did not show chromosomal damage potential at the concentrations used for the preparation of creams with antileishmanial activity. This study may contribute to the registration of a topical phytomedicine containing the extract of C. brasiliense to treat cutaneous leishmaniasis. Key words: Calophyllum brasiliense, genotoxicity assay, (-) mammea A/BB, high-performance liquid chromatography (HPLC), antileishmanial activity, medicinal plants.

Potent radioprotective effects of combined regimens of famotidine and vitamin C against radiation-induced micronuclei in mouse bone marrow erythrocytes.

To investigate the radioprotective effect of the combination of famotidine and vitamin C against radiation-induced micronucleus formation in mouse bone marrow erythrocytes, various doses of famotidine or vitamin C or combinations thereof were administered intraperitoneally to adult male NMRI mice 2 h before 2 and 4 Gy γ-irradiation. The frequency of micronucleated polychromatic erythrocytes (MnPCEs) was scored in 5,000 polychromatic erythrocytes (PCEs), and the cell proliferation ratio [PCE/(PCE + NCE); NCE = normochromatic erythrocytes] was also calculated for each treatment group. Data were statistically evaluated using one-way ANOVA test. The results show that pretreatment with various doses of famotidine and vitamin C before γ-irradiation significantly reduced the frequency of MnPCEs with a protection factor (PF) of 2 and 1.7, respectively. Pretreatment with vitamin C also significantly increased the cell proliferation ratio, while famotidine had no effect. Combination of famotidine and vitamin C was more effective in reducing MnPCEs than each compound alone, leading to a PF of 4.3 after irradiation. Cell proliferation ratio was also significantly improved by the combination compared with the irradiated control groups. Both famotidine and vitamin C are potent scavengers of free radicals and reactive oxygen species, especially OH(·). The combination of the two compounds probably further enhances this activity, thus leading to high bone marrow protection.

Evaluation Of The Protective Activity Of Phythanthis Fruit Extract On Adriamycin Induced Micronuclei In Bone Marrow Erythrocytes Of Mice

Ariamycin (Doxorubin) is a potent antitumor agent used worldwide for the treatment of many cancers. It has been demonstrated to have a potential for initiating genetic events in non tumor cells in human and in animal models. Due to the importance of adriamycin in chemotherapy for the treatment of cancer, it is important to reduce its toxicity in normal cells. This objective can be achieved by concurrent administration of free radical scavenging agent such as antioxidants Phythanthus emblica fruit extract was used to evaluate its ability to reduce the induction of genetic damage (MNPCEs) by anticancer drug in bone marrow cells of male mice. Two experiments wee conducted. In the first experiment the animals were fed with 100, 250 and 500 mg/kg of PFE for two weeks to analyse the antimutagenicity of fruit extract on micronuclei incidence. In the second experiment Group I were given only saline, G – II animals were administred with single dose of 16 mg/kg adriamycin and G – III the animals were fed with 500 mg/kg PFE for two weeks G – IV were administered with 500 mg/kg for two weeks and adriamycin 16 mg/kg as single dose, one day before the last treatment. Results indicated that the treatment with PFE, co-concurrent administration with adriamycin reduced in the percentage of MNPCEs. No significant difference was observed with treated group of animals. Treatment of mice with PFE did not induce any significant variation in the incidence of MNPCEs as compared to control values. Therefore it is concluded that PFE may act as positive modulators of cytotoxic anticancer agents.

Benzophenone guttiferone A from Garcinia achachairu Rusby (Clusiaceae) Presents Genotoxic Effects in Different Cells of Mice

Benzophenones from natural sources and those of synthetic analogues present several reports of potent biological properties, and Guttiferone A represents a promising medicinal natural compound with analgesic and gastroprotective profiles. Considering that there are no reports that assess the genetic toxicity of Guttiferone A, the present study was undertaken to investigate the genotoxic potential of this benzophenone isolated from seeds of Garcinia achachairu in terms of DNA damage in different cells of Swiss albino mice using the comet assay, and its clastogenic/aneugenic effects in bone marrow cells in vivo by the micronucleus test. Cytotoxicity was assessed by scoring polychromatic (PCE) and normochromatic (NCE) erythrocytes ratio. Guttiferone A was administered by oral gavage at doses of 15, 30 and 60 mg/kg. The results showed that Guttiferone A produced genotoxic effects in leukocytes, liver, bone marrow, brain and testicle cells and clastogenic/aneugenic effects in bone marrow erythrocytes of mice. The PCE/NCE ratio indicated no cytotoxicity. Since guttiferone A is harmful to the genetic material we suggest caution in its use by humans.

Coking wastewater increases micronucleus frequency in mouse in vivo via oxidative stress

Coking wastewater has caused serious health risk in coal-producing areas of China, however its toxic effects have not been well understood. The genotoxicity induced by coking wastewater on mice in vivo and its possible oxidative mechanisms were investigated via observing the induction of micronuclei in polychromatic erythrocytes of mouse bone marrow, and subsequently determining the antioxidative enzyme activities (superoxide dismutase Cu, Zn-SOD, Se-dependent glutathione peroxidase, and catalase), thiobarbituric acid reactive substance contents and protein carbonyl levels in brains and livers of mice. Results showed that the tested coking wastewater caused a significant increase of micronucleus frequencies in a concentration-dependent manner. Also, the sample increased lipid peroxidation and protein oxidation levels, which was accompanied by changes in antioxidative status. Interestingly, pre-treatment with an antioxidant (vitamin C) led to a statistical reduction in the micronucleus frequency caused by coking wastewater. This implies that coking wastewater induces evident genetic damage in mammalian cells, and exposure to polluted areas might pose a potential genotoxic risk to human beings; in the process, oxidative stress played a crucial role.

Acute oral toxicity and genotoxicity of Dryopteris crassirhizoma

Ethnopharmacological relevanceDryopteris crassirhizoma has been traditionally used for the treatment of tapeworm infestation, the common cold and cancer in Korea, China and Japan. Despite various pharmacological properties of Dryopteris crassirhizoma, there is no available information about the safety of Dryopteris crassirhizoma. Aim of this studyTo ensure more information about the safety of Dryopteris crassirhizoma, we performed the acute oral toxicity and genotoxicity tests of Dryopteris crassirhizoma. Materials and methodsThe acute oral toxicity test of Dryopteris crassirhizoma was performed in rats. Genotoxicity of Dryopteris crassirhizoma was evaluated by bacterial reverse mutation, chromosomal aberration and bone marrow micronucleus tests. ResultsIn acute toxicity test, Dryopteris crassirhizoma exhibited no mortality, body weight and behavioral changes and adverse effects in male and female rats. Dryopteris crassirhizoma did not significantly increase the number of the bacterial revertant and chromosomal aberration in both in vitro assays. Moreover, the Dryopteris crassirhizoma-related increases of micronucleated polychromatic erythrocytes (MNPCE) in mouse bone marrow were not observed. ConclusionTherefore, Dryopteris crassirhizoma is non-genotoxic in a three standard battery of tests and the oral LD50 of Dryopteris crassirhizoma is >2000mg/kg.

Genotoxicity of the cancer chemopreventive drug candidates CP-31398, SHetA2, and phospho-ibuprofen

The genotoxic activities of three cancer chemopreventive drug candidates, CP-31398 (a cell permeable styrylquinazoline p53 modulator), SHetA2 (a flexible heteroarotinoid), and phospho-ibuprofen (PI, a derivative of ibuprofen) were tested. None of the compounds were mutagenic in the Salmonella/Escherichia coli/microsome plate incorporation test. CP-31398 and SHetA2 did not induce chromosomal aberrations (CA) in Chinese hamster ovary (CHO) cells, either in the presence or absence of rat hepatic S9 (S9). PI induced CA in CHO cells, but only in the presence of S9. PI, its parent compound ibuprofen, and its moiety diethoxyphosphoryloxybutyl alcohol (DEPBA) were tested for CA and micronuclei (MN) in CHO cells in the presence of S9. PI induced CA as well as MN, both kinetochore-positive (Kin+) and -negative (Kin−), in the presence of S9 at ≤100μg/ml. Ibuprofen was negative for CA, positive for MN with Kin+ at 250μg/ml, and positive for MN with Kin− at 125 and 250μg/ml. DEPBA induced neither CA nor MN at ≤5000μg/ml. The induction of chromosomal damage in PI-treated CHO cells in the presence of S9 may be due to its metabolites. None of the compounds were genotoxic, in the presence or absence of S9, in the GADD45α-GFP Human GreenScreen assay and none induced MN in mouse bone marrow erythrocytes.

In vitro and in vivo evaluation of the genotoxicity of Gumiganghwal-tang, a traditional herbal prescription

Ethnopharmacological relevanceGumiganghwal-tang (GGT, known as Kumi-Kyokatsu-to in Japanese) is a traditional herbal prescription made from nine different herbs that is used for the treatment of the common cold, pain, and inflammatory diseases. Aim of the studyThis study evaluated the potential genotoxicity of an aqueous GGT extract using three standard battery of tests as part of a safety evaluation. Materials and methodsWe prepared GGT using a water extraction method and subsequently extracted six compounds from GGT by high performance liquid chromatography. GGT extract genotoxicity was assayed using three standard tests including the in vitro bacterial reverse mutation test, the in vitro chromosomal aberration test with Chinese hamster lung cells, and the in vivo micronucleus test using ICR mouse bone marrow recommended by the Korean Food and Drug Administration. ResultsThe bacterial reverse mutation assay showed that GGT extract doses ranging from 333.3 up to 5000mg/plate induced a greater than 2-fold increase in the number of revertant TA1537 strain colonies exhibiting metabolic activation (with S-9 mix), when compared with the vehicle control. The chromosomal aberration test showed that GGT extract induced an increase in the number of chromosomal aberrations after treatment for 6h with the S-9 mix and 22h without the S-9 mix, when compared with vehicle control. In contrast, the micronucleus test showed that GGT extract did not significantly increase the number of micronucleated polychromatic erythrocytes (MNPCEs) in ICR mouse bone marrow. ConclusionsBased on these results, it was concluded that GGT extract acted as a genotoxic material in our experimental conditions. We did not identify the compounds responsible for the induction of genotoxic effects, but it was significant that we provided a basic genotoxicity profile for GGT.