Ratio Of Polychromatic Erythrocytes Research Articles

Evaluation of Genotoxic and Cytotoxic Effects of Gluten in Male Albino Mice

Gluten is a protein commonly found in daily diets in the form of wheat, barley, rye, and other grains. It serves as the structural component in flour, providing the binding qualities that maintain the shape and texture of food items. This study aimed to investigate the genotoxic and cytotoxic effects of gluten on bone marrow chromosomes and DNA of male albino mice. The animals were divided into four groups: a control group, a negative control group that received an oral dose of 0.02M glacial acetic acid, and two groups that were treated with gluten dissolved in 0.02M glacial acetic acid at doses of 1.5 g/kg and 3.0 g/kg body weight. The treated animals received oral doses with non-consecutively three times a week for a period of four weeks. The study evaluated chromosomal aberrations in the bone marrow, micronucleus test, and DNA damage using the comet assay. The results of the study showed that treatment with 1.5 and 3.0g/kg body weight of gluten induced chromosomal aberrations and damage in DNA content, with an increase in the severity of effects at a higher dose of gluten. The chromosomal aberrations seen included deletion, fragment, centromeric attenuation, centric fusion, ring formation, end to end association, chromosomal gap, beaded chromosomes, and polyploidy. The micronucleus test revealed toxicity in the bone marrow, as shown by appearance of micronuclei in polychromatic erythrocytes and a reduction in the ratio of polychromatic erythrocytes. The comet assay showed a significant increase of DNA damage in the tail length of the comet cells. This study concluded that the treatment with gluten has detrimental effects on the bone marrow chromosomes and DNA of mice, as demonstrated by the increased chromosomal aberrations, micronuclei, and DNA damage observed in the treated mice. So, the use of gluten should be within an acceptable and safe range.

A pilot in vivo evaluation of Sb(III) and Sb(V) genotoxicity using comet assay and micronucleus test on the freshwater fish, silver perch Bidyanus bidyanus (Mitchell, 1838)

Antimony (Sb) is a priority water pollutant known to be toxic to aquatic organisms at high concentrations. Environmental exposure, however, occurs most often at sub-lethal concentrations but very limited information is available on effects of sub-lethal, chronic exposure to Sb, which hinders reliable risk assessment and the setting of protective guidelines. In this pilot study, in vivo screening for Sb genotoxicity in the erythrocytes of the freshwater fish, silver perch (Bidyanus bidyanus) was conducted where fish were exposed to environmentally relevant and sub-lethal Sb concentrations of 0.4, 0.9 and 1.8 mg L−1 Sb(III), and 0.9, 2 and 5 mg L−1 Sb(V), for 14 d. Genotoxicity was assessed by both a single cell gel electrophoresis (SCGE) assay and a micronucleus (MN) test. The SCGE assay showed that all Sb(III) exposure concentrations induced a statistically significant non-dose-related increase in DNA damage after 2 d of exposure after which there was no further increase in DNA damage evident in relation to the control. Mortality of fish was 100 % in all Sb(III) exposures by 14 d. Clastogenic and/or aneugenic effects were not observed. The 1.8 mg L−1 Sb(III) exposure was the only Sb concentration at which a significant increase in the cytotoxicity index as measured by the ratio of polychromatic erythrocytes (PCEs) to normochromatic erythrocytes (NCE) was induced at 2 d exposure. For Sb(V) exposures, no significant genotoxic effects were observed using either assay, nor was the PCE/NCE altered. This pilot investigation has indicated that sub-lethal waterborne Sb(III) exposure manifests in genotoxic effects in freshwater fish species, which may have consequences for resilience and survival. Further study is needed for deeper insight into the relationship between Sb(III) and genotoxicity and the multiple biomarker responses that need assessment to evidence effects.

Genotoxicity and glucose tolerance induction by acetyltriethylcitrate, substitute plasticizer compared to di(2-ethylhexyl)phthalate

As di(2-ethylhexyl) phthalate (DEHP), one of phthalates, is classified as probable human carcinogens in EPA, acetyltriethyl citrate(ATEC), one of aliphatic esters, could be applied to DEHP substitute. ATEC is used as plasticizers in cosmetics and nail products. Here, we studied whether ATEC might have genotoxic potential and induce glucose tolerance as compared to DEHP. Genotoxicity was determined by Ames test with histidine-requiring Salmonella typhimurium (TA98, TA100, TA1535 and TA1537) and tryptophan-requiring Escherichia coli (WP2uvrA(pKM101)) strains, chromosomal aberration assay with Chinese hamster lung(CHL/IU) cells, and micronucleus test with bone marrow cells of CD-1 mice. The number of revertants was not significantly changed in Ames test. The frequency of cells with chromosome aberrations was less than 5% in ATEC- or DEHP-treated cells for 6 or 24 h. In addition, no statistically significant increase was observed for the incidence of micronucleated polychromatic erythrocytes (MNPCE) in polychromatic erythrocytes (PCE) and for the ratio of PCE among total erythrocytes at 24 or 48 h after the treatment of mice with ATEC or DEHP. In the meanwhile, blood glucose level (BGL) was increased by the treatment of mice with DEHP or ATEC for 5 consecutive days. Additional 7 days later, BGL by DEHP was recovered to normal level, but not that by ATEC. Then, taken together, our results suggest that ATEC could disrupt glucose metabolism under our experimental conditions. Therefore, although DEHP and ATEC may not be genotoxic, our data should be helpful for persons with the problem in glucose metabolism to choose products containing DEHP or ATEC.

An Analysis of Micronuclei and DNA Damage Induced by Methotrexate Treatment of Male Albino Mice

Background: Methotrexate is an antineoplastic, antipsoriatic and antirheumatic agent belongs to the group of antimetabolites and inhibits folic acid metabolism. Materials and methods: To investigate its possible effect, sixty male mice were randomly assigned to one of four groups (one control and three treated groups with different doses of methotrexate). Mice of groups 1, 2 and 3 were intraperitoneally injected with 2.5, 5 and 10 mg /kg b.wt. methotrexate respectively. All the control and treated animals were sacrificed at 24, 48 and 72 hour by cervical dislocation post treatment. Results:Micronucleus assay results showed that methotrexate treatment induced genotoxicity in bone marrow cells, the number of micronucleated polychromatic erythrocytes (MNPCEs) and the ratio of polychromatic erythrocytes / normochromatic erythrocytes was gradually increased significantly (P < 0.001) by increasing dose and time of treatment in methotrexate treated groups in comparison with the control group. An analysis of randomly amplified polymorphism DNA-polymerase chain reaction (RAPD-PCR) showed different ranges of DNA modifications in the treated groups after 24, 48 and 72 hour of treatment in comparison with the control group. Results of this study indicate that methotrexate treatment induced cytotoxic and genotoxic effect on bone marrow cells and DNA content of male albino mice even after a low dose and single treatment. Conclusion: Therefore, the therapeutic uses of methotrexate should be restricted to a very narrow range border.

Evaluation of the cytogenotoxic damage in immature and mature rats exposed to 900 MHz radiofrequency electromagnetic fields

Purpose: One of the most important issues regarding radiofrequency electromagnetic fields (RF-EMF) is their effect on genetic material. Therefore, we investigated the cytogenotoxic effects of 900 MHz radiofrequency electromagnetic fields (RF-EMF) and the effect of a recovery period after exposure to RF-EMF on bone marrow cells of immature and mature rats.Materials and methods: The immature and mature rats in treatment groups were exposed to RF-EMF for 2 h/day for 45 days. Average electrical field values for immature and mature rats were 28.1 ± 4.8 V/m and 20.0 ± 3.2 V/m, respectively. Whole-body specific absorption rate (SAR) values for immature and mature rats were in the range of 0.38–0.78 W/kg, and 0.31–0.52 W/kg during the 45 days, respectively. Two recovery groups were kept for 15 days after RF-EMF exposure.Results: Significant differences were observed in chromosome aberrations (CA), micronucleus (MN) frequency, mitotic index (MI) and ratio of polychromatic erythrocytes (PCE) in all treatment and recovery groups. The cytogenotoxic damage in immature rats was statistically higher than the mature rats. The recovery period did not reduce the damage to the same extent as the corresponding control groups.Conclusions: The exposure of RF-EMF leads to cytotoxic and genotoxic damage in immature and mature rats. More sensitive studies are required to elucidate the possible carcinogenic risk of EMF exposure in humans, especially children.

Lack of micronuclei induction by fumonisin B1 mycotoxin in BALB/c mice

The genotoxic potential of fumonisin B(1) (FB(1)) in vivo in BALB/c mice (male and female) was assessed by induction of micronuclei (MN) formation in bone marrow polychromatic erythrocytes. The ratio of polychromatic erythrocytes to normochromatic erythrocytes (PCE/NCE) was also determined. Mice were injected intraperitoneally (i.p.) with FB(1) at a low dose (0.1mg kg(-1) body mass), middle dose (1.0mg kg(-1) body mass) and high dose (10mg kg(-1) body mass) as single and multiple doses in normal saline to test the genotoxicity. Mitomycin C, a known clastogen, was used as positive control. The frequency of MN and the PCE/NCE value in animals treated with FB(1) at low, middle, and high doses in single dose studies, and the frequency of MN in multiple dose studies, were statistically non-significant from that of the controls injected with saline only. The multiple dose studies at all doses revealed that the PCE/NCE value was found to be reduced upon exposure to FB(1) as compared to the controls. In animals injected with multiple low doses of FB(1), the PCE/NCE value was found to be 0.66 in males and 0.82 in the females; at multiple middle doses the value was 0.30 in males and 0.41 in the females and was statistically significant (p < 0.001); however, at multiple high doses, the ratio was found to be 0.36 in both males and females. The present study confirms that FB(1) is non-genotoxic in nature while the reduced ratio of PCE/NCE suggests the cytotoxic nature of FB(1).

Relieved residual damage in the hematopoietic system of mice rescued by radiation-induced adaptive response (Yonezawa Effect)

Existence of adaptive response (AR) was previously demonstrated in C57BL/6J mice. Irradiations were performed by delivering a priming low dose of X-rays (0.50 Gy) in combination with a challenge high dose of accelerated carbon or neon ion particles. AR was characterized by significantly decreased mortality in the 30-day survival test. This mouse AR model (‘Yonezawa Effect’) was originally established by using X-rays as both the priming and challenge irradiations. The underlying mechanism was due to radio-resistance occurring in blood-forming tissues. In this study, we verified the existence of AR and further investigated residual damage in the hematopoietic system in surviving animals. Results showed that the priming low dose of X-rays could relieve the detrimental effects on the hematopoietic system. We observed both an improvement in the blood platelet count and the ratio of polychromatic erythrocytes (PCEs) to the sum of PCEs and normochromatic erythrocytes (NCEs) and a marked reduction of the incidences of micronucleated PCEs and micronucleated NCEs. These findings suggest that the priming low dose of low linear energy transfer (LET) X-rays induced a protective effect on the hematopoietic system, which may play an important role in both rescue from acute lethal damage (mouse killing) and prevention of late detrimental consequences (residual anhematopoiesis and delayed genotoxic effects) caused by exposure to a high challenge dose from low-LET (X-ray) or high-LET (carbon and neon ion) irradiations. These findings provide new knowledge of the characterization of the Yonezawa Effect by providing new insight into the mechanistic study of AR in vivo.

Cytotoxic and genotoxic effects of high-frequency electromagnetic fields (GSM 1800 MHz) on immature and mature rats

We investigated the cytogenotoxic effects of high frequency electromagnetic fields (HF-EMF) for 45 day and the effect of a recovery period of 15 day after exposure to EMF on bone marrow cells of immature and mature rats. The animals in treatment groups were exposed to 1800MHz EMF at SAR of 0.37W/kg and 0.49W/kg for 2h/day for 45 day. Two recovery groups were kept for a recovery period of 15 day without EMF after exposure to HF-EMF. Two control groups for both immature and mature rats were also included. Significant differences were also observed in chromosome aberrations (CA), micronucleus (MN) frequency, mitotic index (MI) and ratio of polychromatic erythrocytes (PCEs) in all treatment groups. The cytogenotoxic damage was more remarkable in immature rats and, the recovery period did not improve this damage in immature rats. Because much higher and irreversible cytogenotoxic damage was observed in immature rats than in mature rats, further studies are needed to understand effects of EMF on DNA damage and DNA repair, and to determine safe limits for environment and human, especially for children.

Micronucleus Test of Picrorrhiza Rhizoma Aqueous Extract in Bone Marrow Cells of Male ICR Mice.

In this research, the genotoxic effect of Picrorrhiza Rhizoma (PR) aqueous extract was evaluated using the mouse micronucleus test. PR extract was administered once a day for 2 continuous days by oral gavage to male ICR mice at doses of 2000, 1000 and 500 mg/kg. Cyclophosphamide was used as a known genotoxic agent in a positive control. The appearance of a micronucleus (MN) in polychromatic erythrocyte (PCE) is used as an index for genotoxic potential, and PCE ratio is used as an index of cytotoxicity. Although significant (p < 0.01) increase of the number of PCE with one or more nuclei (MNPCE) was detected in cyclophosphamide treated groups, no significant increases of MNPCE numbers were observed in all three different dosages of PR extracts treated mice with over 0.39 of the individual polychromatic erythrocyte ratio in all mice used in this study. The results obtained indicated that PR extract shows no genotoxicity effects up to 2000 mg/kg dosing levels.

Induction of micronuclei and nuclear abnormalities in erythrocytes of mosquito fish (Gambusia affinis) following exposure to the pyrethroid insecticide lambda-cyhalothrin

In the present study the induction of micronuclei (MN) and nuclear abnormalities (NA) in erythrocytes of mosquitofish (Gambusia affinis) (Baird & Girard 1853) was studied. Fish were exposed to three different concentrations of lambda-cyhalothrin (LCT) (1×10−4μg/l, 2×10−4μg/l, 4×10−4μg/l) for periods of 6, 12, 24, and 48h. NA (notched, lobed, blebbed nuclei), MN, bi-nucleated cells, and the ratio of polychromatic erythrocytes (PCEs) to normochromatic erythrocytes (NCEs) were evaluated to assess genotoxicity and cytotoxicity. LCT significantly induced MN and NA in erythrocytes of G. affinis. The PCE/NCE ratio was also decreased after 24- and 48-h treatments of 4×10−4μg/l LCT. The results show that LCT has genotoxic and cytotoxic potential on G. affinis.

Micronuclei, hemoglobin adducts and respiratory tract irritation in mice after inhalation of toluene diisocyanate (TDI) and 4,4′-methylenediphenyl diisocyanate (MDI)

Toluene diisocyanate (TDI) and 4,4′-methylenediphenyl diisocyanate (MDI), used in the production of polyurethane foam, are well known for their irritating and sensitizing properties. Contradictory results have been obtained on their genotoxicity. We investigated the genotoxicity and protein binding of inhaled TDI and MDI in mice by examining micronucleated polychromatic erythrocytes (PCEs) in bone marrow and peripheral blood and TDI- and MDI-derived adducts in hemoglobin. Male C57Bl/6J mice (8 per group) were exposed head-only to TDI vapour (mean concentrations 1.1, 1.5, and 2.4 mg/m 3; the mixture of isomers contained, on the average, 63% 2,4-TDI and 37% 2,6-TDI) or MDI aerosol (mean concentrations 10.7, 20.9 and 23.3 mg/m 3), during 1 h/day for 5 consecutive days. Bone marrow and peripheral blood were collected 24 h after the last exposure. Inhalation of TDI caused sensory irritation (SI) in the upper respiratory tract, and cumulative effects were observed at the highest exposure level. Inhalation of MDI produced SI and airflow limitation, and influx of inflammatory cells into the lungs. Hemoglobin adducts detected in the exposed mice resulted from direct binding to globin of 2,4- and 2,6-TDI and MDI, and dose-dependent increases were observed especially for 2,4-TDI-derived adducts. Adducts originating from the diamines of TDI (toluene diamine) or MDI (methylene dianiline) were not observed. No significant increase in the frequency of micronucleated PCEs was detected in the bone marrow or peripheral blood of the mice exposed to TDI or MDI. The ratio of PCEs and normochromatic erythrocytes (NCEs) was reduced at the highest concentration of MDI, and a slight reduction of the PCE/NCE ratio, dependent on cumulative inhaled dose, was also seen with TDI. Our results indicate that inhalation of TDI or MDI (1 h/day for 5 days), at levels that induce toxic effects and formation of TDI- or MDI-specific adducts in hemoglobin, does not have detectable genotoxic effects in mice, as studied with the micronucleus assay.

In vivo Genotoxicity of Silver Nanoparticles after 90-day Silver Nanoparticle Inhalation Exposure

ObjectivesThe antimicrobial activity of silver nanoparticles has resulted in their widespread use in many consumer products. Yet, despite their many advantages, it is also important to determine whether silver nanoparticles may represent a hazard to the environment and human health.MethodsThus, to evaluate the genotoxic potential of silver nanoparticles, in vivo genotoxicity testing (OECD 474, in vivo micronuclei test) was conducted after exposing male and female Sprague-Dawley rats to silver nanoparticles by inhalation for 90 days according to OECD test guideline 413 (Subchronic Inhalation Toxicity: 90 Day Study) with a good laboratory practice system. The rats were exposed to silver nanoparticles (18 nm diameter) at concentrations of 0.7 × 106 particles/cm3 (low dose), 1.4 × 106 particles/cm3 (middle dose), and 2.9 × 106 particles/cm3 (high dose) for 6 hr/day in an inhalation chamber for 90 days. The rats were killed 24 hr after the last administration, then the femurs were removed and the bone marrow collected and evaluated for micronucleus induction.ResultsThere were no statistically significant differences in the micronucleated polychromatic erythrocytes or in the ratio of polychromatic erythrocytes among the total erythrocytes after silver nanoparticle exposure when compared with the control.ConclusionThe present results suggest that exposure to silver nanoparticles by inhalation for 90 days does not induce genetic toxicity in male and female rat bone marrow in vivo.

Antinociceptive, genotoxic and histopathological study of Heliopsis longipes S.F. Blake in mice

Ethnopharmacological relevance H. longipes S.F. Blake (Asteraceae) is a Mexican plant, whose roots are traditionally used as a condiment, as a mouth anesthetic, and as an antiparasitic. Affinin is the alkamide present in higher amounts in the roots of H. longipes. Aim of the study To date, there are no published studies regarding the relation between the analgesic properties, in vivo cytotoxicity, and DNA-damaging potential of H. longipes ethanol extract (HLEE). Materials and methods The HLEE was chromatographically fingerprinted to validate its affinin contents. Biological evaluation was conducted in sets of 6–8 CD1 + mice. Antinociceptive effect was evaluated using the writhing and hot-plate tests, and mutagenic and cytotoxic effects were evaluated with micronucleous test in CD1 + mice. For histopathological studies, biological samples from liver, heart, kidneys, spleen, lung, and brain were collected and stained. Results Oral administration of HLEE (3–100 mg/kg) produced a dose-dependent antinociceptive effect in both assays. In micronucleus assay, the variability in the number of micronucleated polychromatic erythrocytes (MNPE) induced, and PE/NE index, the ratio of polychromatic erythrocytes with respect to the number of normochromatic erythrocytes induced by HLEE in the evaluated schedule, were small and nonsignificant. After histopathological results, HLEE showed polioencephalomalacia with 1000 mg/kg dose. Conclusions This work provides evidence that HLEE exerts analgesic effects, with no genotoxic effects in vivo. These findings would be an important contribution to explain the use of H. longipes root as an effective analgesic in traditional medicine, and to establish for the first time the absence of genotoxic and cytotoxic effects of the root in bioactive doses in vivo.

Micronucleus Test of DHU001, a Polyherbal Formula, in Bone Marrow Cells of Male ICR Mice.

The genotoxic effects of DHU001, a polyherbal formula were evaluated using the mouse micronucleus test. DHU001 was administered once a day for 2 continuous days by oral gavage to male ICR mice at doses of 2000, 1000 and 500 mg/kg. Cyclophosphamide was used as a known geno-toxic agent in a positive control. The appearance of a micronucleus is used as an index for genotoxic potential. In addition, the changes on the total white blood cells and differential counts on the lymphocytes, neutrophils, eosinophils, basophils and monocytes in the prepared blood smears were also conducted to observe the possible immunosuppression. The results indicats that DHU001 showed no genotoxicity effects up to 2000 mg/kg dosing levels and did not influenced on the total white blood cells and differential counts. In addition, it is also considered that there were no problems from cytotoxicity of DHU001 tested in this study because the polychromatic erythrocyte ratio was detected as > 0.41 in all tested groups.

Evaluation of genotoxicity of pan masala employing chromosomal aberration and micronucleus assay in bone marrow cells of the mice

Pan masala is commonly consumed in south-east Asian and other oriental countries as an alternate of tobacco chewing and smoking. Genotoxic potential of pan masala (pan masala plain and pan masala with tobacco known as gutkha) was evaluated employing chromosome aberration (CA) and micronucleus (MN) assay in vivo. Animals were exposed to three different doses (0.5%, 1.5% and 3%) of pan masala plain (PMP) and gutkha (PMT) through feed for a period of 6 months and micronucleus and chromosomal aberrations were studied in the bone marrow cells. Induction of mean micronuclei in polychromatic erythrocytes (MNPCE) and normochromatic erythrocyte (MNNCE) was higher in both types of pan masala treated groups with respect to control group. Both pan masala plain and gutkha treatment significantly induced the frequency of MNPCE and MNNCE in the bone marrow cells, indicating the genotoxic potential. Furthermore, slight decline in the ratio of polychromatic erythrocytes to normochromatic erythrocytes was also noticed, suggesting the cytotoxic potential even though the ratio was statistically non significant. A dose-dependent, significant increase in chromosome aberration was observed in both types of pan masala treated mice with respect to control. However, no significant difference in micronucleus and chromosomal aberration induction was noticed between two types of pan masala exposed (PMP and PMT) groups. Results suggest that both types of pan masala, i.e. plain and gutkha, have genotoxic potential.

Assessment of Cadmium Genotoxicity in Peripheral Blood and Bone Marrow Tissues of Male Wistar Rats

In this study, Cadmium (Cd) genotoxicity was investigated in both bone marrow and peripheral blood treatment using rat micronucleus technique as genotoxicity test at acute and chronic treatment in the same animals. This study evaluated the frequency of micronuclei in the peripheral blood and bone marrow of male rats treated with unique cadmium dose (15 mg/kg. body w/day) by gavage for 60 days and acute treatment for 24 h, respectively. Mitomycin C (MMC) 2 mg/kg body wt was used as a positive control. This study shows that cadmium chloride treatment significantly induced the frequency of micronucleus in polychromatic erythrocytes in both tibia bone marrow and peripheral blood (p < 0.001, p < 0.01, respectively). This increase in micronucleus frequency shows that cadmium has a genotoxic effect on bone marrow and peripheral blood at this level. Also, in order to determine cytotoxicity in bone marrow and peripheral blood, the ratio of polychromatic erythrocytes to normochromatic erythrocytes was calculated in bone marrow and peripheral blood. Cd treatment decreased this ratio in only bone marrow. The results of this study demonstrate that Cd has both toxic and genotoxic potential in bone marrow and only genotoxic potential in peripheral blood. There is a significant difference between the control group and exposed group, including acute and chronic treatment for blood Cd level (p < 0.001). No significant difference was found between acute and chronic exposure group (p > 0.05).

Effect of N-acetylcysteine on Radiation-induced Genotoxicity and Cytotoxicity in Rat Bone Marrow

The aim of this study is to evaluate the potential radioprotective effects of N-acetylcysteine (NAC) against genotoxicity and cytotoxicity. The effect of WR-2721, as a representative of clinically used radioprotector, was compared with that of NAC, using the chromosomal aberration (CA) and micronucleus (MN) test systems in the irradiated rat's femoral bone marrow cells. We also investigated the mitotic index (MI), and the ratio of polychromatic erythrocytes (PCEs) to normochromatic erythrocytes (NCEs). The rats (n = 16) were divided randomly and equally into four groups: Control (C), Radiation (R), R+NAC (received irradiation and 1000 mg/kg NAC) and R+WR-2721 (received irradiation and 200 mg/kg WR-2721) rats. All the irradiated groups received whole-body gamma irradiation as a single dose of 6 Gy. Group R showed higher CA and MN formation when compared to C. Group R showed higher frequency of MN formation when compared to both R+NAC and R+WR-2721. The mean MI and PCE/NCE ratios were lower in Group R when compared to those of Group C. The mean MI and PCE/NCE ratios of both R+NAC and R+WR-2721 groups were lower when compared to those of Group C. The MI in Group R was lower when compared to that of both R+NAC and R+WR-2721 groups. In this study, the results give clues about the beneficial effects of NAC against radiation-induced genotoxicity and cytotoxicity in rat bone marrow and its effect may be comparable to that observed for WR-2721.

Micronucleus Test of Kong-Jin-Dan, a Polyherbal Formula, in Bone Marrow Cells of Male ICR Mice

In this research, the genotoxic effects of Kong-Jin-Dan (KJD), a polyherbal formula were evaluated using the mouse micronucleus test. KJD was administered once a day for 2 continuous days by oral gavage to male ICR mice at doses of 2000, 1000 and 500 mg/kg. Cyclophosphamide was used as a known genotoxic agent in a positive control. The appearance of a micronucleus is used as an index for genotoxic potential. In addition, the changes on the total white blood cells and differential counts on the lymphocytes, neutrophils, eosinophils, basophils and monocytes in the prepared blood smears were also conducted to observe the possible immunosuppress. The results obtained indicated that KJD shows no genotoxicity effects up to 2000 mg/kg dosing levels, but KJD shows slight increased trends in the blood total leukocyte numbers as pharmacological effects of immune stimulation. In addition, it is also considered that there were no problems from cytotoxicity of KJD tested in this study because the polychromatic erythrocyte ratio was detected as > 0.42 in all tested groups.

Genotoxicity studies on licorice flavonoid oil (LFO)

Licorice flavonoid oil (LFO) is a new functional food ingredient. In this study, the genotoxicity of LFO was investigated using a test battery of three different methods. In a reverse mutation assay using four Salmonella typhimurium strains and Escherichia coli, LFO did not increase the number of revertant colonies in any tester strain with or without metabolic activation by rat liver S9 mix. In a chromosomal aberration test using Chinese hamster lung (CHL/IU) cells, LFO did not induce any chromosomal aberrations either in the short period test without rat liver S9 mix or in the continuous treatment (24 h or 48 h) test. However, in the short-period test with rat liver S9 mix, LFO induced structural chromosomal aberrations at concentrations higher than 0.6 mg/mL. A bone marrow micronucleus test using male F344 rats was initially conducted. The animals were dosed by oral gavage at doses up to 5000 mg/kg/day. No significant or dose-dependent increases in the frequency of micronucleated polychromatic erythrocytes (MNPCE) were observed and the high dose suppressed the ratio of polychromatic erythrocytes (PCE) to total erythrocytes. Subsequently, a liver and peripheral blood micronucleus test using male F344 rats was conducted. No micronuclei induction either in hepatocytes or PCE was observed even at the highest dose of 5000 mg/kg/day. From the findings obtained from the genotoxicity assays performed in this study and the published pharmacokinetic studies of LFO, it appears unlikely that dietary consumption of LFO will present any genotoxic hazard to humans.

Micronucleus Test of Polycan™, β-Glucan Originated from Aureobasidium, in Bone Marrow Cells of Male ICR Mice.

In this research the genotoxic effect of Polycan™ β-glucans originated from Aureobasidium pullulans SM-2001, was evaluated using the mouse micronucleus test. Polycan™ was administered once a day for 2 days by oral gavage to male ICR mice at doses of 1000, 500 and 250 mg/kg. Cyclophosphamide was used as a known genotoxic agent in a positive control group. The appearance of a micronucleus is used as an index for genotoxic potential. The results obtained indicated that Polycan™ shows no genotoxicity effect up to 1000 mg/kg dosing levels. In addition, it is also considered that there were no problems from cytotoxicity of Polycan™ tested in this study because the polychromatic erythrocyte ratio was detected as > 0.47 in all tested groups.