Micronucleated Polychromatic Erythrocytes Research Articles

Luteolin mitigates potassium dichromate-induced nephrotoxicity, cardiotoxicity and genotoxicity through modulation of Kim-1/Nrf2 signaling pathways.

Environmental and occupational exposure to chromium compounds has become potential aetiologic agent for kidney disease with excessive generation of free radicals, apoptosis, and inflammatory. These pathophysiologic mechanisms of potassium dichromate (K2 Cr2 O7 ) have been well correlated with nephrotoxicity and cardiotoxicity. The cardioprotective and nephroprotective effects of Luteolin, a known potent antioxidant were evaluated in this study with 40 healthy rats in four experimental groups: Group A (normal saline), Groups B (30 mg/kg K2 Cr2 O7 ), Group C (Luteolin 100 mg/kg and K2 Cr2 O7 30 mg/kg), and Group D (Luteolin 200 mg/kg and K2 Cr2 O7 30 mg/kg), respectively. Markers of antioxidant defense system, oxidative stress, blood pressure and micronucleated polychromatic erythrocytes (MnPEs), immunohistochemistry of Kidney, injury molecule (Kim-1), nuclear factor erythroid 2-related factor 2 (Nrf2), and cardiac troponin I were determined. Administration of K2 Cr2 O7 increased blood pressure parameters in systolic, diastolic and mean arterial blood pressures, markers of oxidative stress, and frequency of micronucleated polychromatic erythrocytes, together with reduction in serum nitric oxide level. Renal Kim-1 and cardiac troponin I expressions were higher, but lower expressions of renal and cardiac Nrf2 were recorded with immunohistochemical analysis. Pre-treatment with Luteolin restored blood pressure parameters, with concomitant reduction in oxidative stress indicators, augmented antioxidant mechanisms and serum Nitric oxide level, lowered the expressions of Kim-1, cardiac troponin I and up-regulated of both cardiac and renal Nrf2, reduced the frequency of micronucleated polychromatic erythrocytes. Taken together, this study therefore demonstrates the cardioprotective, nephro protective and antigenotoxic effects of Luteolin through antioxidantive and radical scavenging mechanisms.

Investigation of genotoxic effects of rhododendron honey using three mammalian bioassays in vivo

Rhododendron honey (RH) is obtained from the rhododendron plants are grown in many regions around the world, causes poisoning in humans due to the grayanotoxin (GTX) compound in its structure. It is used by the public as a therapeutic for some diseases. It was aimed to study the genotoxic and cytotoxic effects of RH in mouse bone-marrow and sperm cells by using three mammalian bioassays. 25, 50 and 75 mg kg−1 concentrations of RH given to male mice via gavage for 24 and 48 h treatment periods and its active ingredient Grayanatoxin (GTX-III) 0.01 mg kg−1 by i.p. injection. Chromosome aberrations (CA), polychromatic erythrocytes (PCE)/normochromatic erythrocytes (NCE), micronucleated polychromatic erythrocytes (MNPCE) and sperm abnormalities were investigated. The results demonstrated that all the tested concentrations of RH significantly induced total abnormal cell frequency including chromosomal breaks for two time periods. In the MN assay, 75 mg kg−1 RH and 0.01 mg kg−1 GTX-III significantly increased % MNPCE and significantly reduced PCE/NCE ratios after 24 and 48 h treatments on mice demonstrating potential genotoxic and cytotoxic effect. Although there was a concentration-related increase in the percentage of total sperm abnormalities, this increase was not statistically significant compared to control. As a result, microscopic genotoxicity and cytotoxicity marker tests showed that RH and its active ingredient GTX-III have potential genotoxic and cytotoxic effect on mice bone marrow cells. It is understood that RH that is used to treat some diseases by public, should be handled carefully and used in a controlled manner. Highlights Chromosome aberration, micronucleus and sperm morphology assays are recommended as reliable biological indicators. RH and its active ingredient GTX-III have potential genotoxic and cytotoxic effect on mice bone marrow cells. Significant changes were observed upon the treatment of 75 mg kg−1 MH for MN assay.

Decrease of the DXR-induced genotoxicity and nongenotoxic effects of Theobroma cacao revealed by micronucleus assay.

This study evaluated the genotoxicity of lyophilized glycolic extract of Theobroma cacao Linné seeds (TCL), using the micronucleus assay in bone marrow of mice. The interaction between TCL and doxorubicin (DXR) was also analyzed. Experimental groups were evaluated 24-48 h after treatment with N-Nitroso-N-ethylurea (NEU: 50 mg/kg), DXR (5 mg/kg), NaCl (145 mM), TCL (0.5-2 g/kg), and TCL (2 g/kg) in combination with DXR (antigenotoxic assays). Analysis of micronucleated polychromatic erythrocytes (MNPCEs) showed no significant differences between all the treatment doses of TCL and NaCl control. Mice experimentally treated with DXR and NEU significantly induced MNPCEs. However, a significant reduction of MNPCEs was also observed when TCL was administered in combination with the chemotherapeutic agent DXR. The analysis of the PCE/NCE ratio revealed no significant differences between the NaCl control, all doses of TCL, and DXR. However, there were significant differences in the PCE/NCE ratio between positive NEU control and all other treatments. The PCE/NCE ratio observed after treatment with TCL and DXR showed significant differences and intermediate values to controls (NaCl and NEU). This study suggests absence of genotoxicity and cytotoxicity of TCL, regardless of dose, sex, and time. TCL reduced genotoxic effects induced by DXR, suggesting potential antigenotoxic effects.

Clastogenic, anti-clastogenic profile and safety assessment of Camel urine towards the development of new drug target

Camel Urine (CU) is composed of components that have antitumor properties and other therapeutic benefits. Regardless of short-term preliminary CU genotoxicity is reported, comprehensive genotoxic studies are limited. In this study, sensitive in vitro and in vivo genotoxic bioassays such as mitotic index (MI), chromosomal aberrations (CA), micronucleated polychromatic erythrocytes (MPE), and analysis of primary spermatocytes were employed. The adventitious roots of Allium cepa L. and mice (Mus musculus), as an experimental mammalian system, were employed to assess the MI and CA of CU induced by sodium nitrate and cyclophosphamide respectively. In contrast, other clastogenic assays were studied in mice (Mus musculus). Twenty-eight days of four repeated doses (2.5, 5, 25, and 50 mL/kg BW) of CU were tested and compared with three doses (10, 25, and 50 mg/kg BW) cyclophosphamide as a positive control and deionized water as the negative control. The results proved that cytological examination of CU was cytotoxic since a decrease in mitotic activity (16.8–1.1) was observed, since the significant reduction in cell proliferation in A. cepa L. and also in mice bone marrow cells. On the other hand, CU did not induce a clastogenic effect since no significant stickiness, fragment, multinucleoli were observed compared to the control group. Additionally, the data showed that CU decreased the CA when mice had received cyclophosphamide (25 mg BW) followed by CU doses. CU was found to be cytotoxic but no clastogenic effect. Furthermore, it possesses anticlastogenic properties. The observed results suggest that CU in whole or the metabolites present in CU could be a potent drug target. Further research is warranted to study the complete metabolites profiling and to study the molecular mechanisms.

Micronucleated erythrocytes in peripheral blood from neonate rats fed by nursing mothers exposed to X-rays.

Most women with breast cancer can become pregnant and give birth while undergoing radiation therapy and breastfeeding is generally not contraindicated. The induction of long-lived reactive species in proteins, such as casein by X-ray radiation and DNA damage to unexposed organisms, has been shown when ingesting irradiated cheese. To determine whether exposing lactating rats to X-rays increases the number of micronucleated erythrocytes (MNEs) in peripheral blood of their unexposed or breastfeeding rat pups, 15 female Wistar rats were divided into three groups: Negative control; Experimental group exposed to X-rays, and group exposed to X-rays plus vitamin C. The mothers of groups 2 and 3 were irradiated for three consecutive days after giving birth, returning them to their respective cages each time to continue lactation. A blood sample was taken from the mothers and pups at 0, 24, and 48 hr. Blood smears were stained with acridine orange to analyze MNEs. In mother rats, the frequency of micronucleated polychromatic erythrocytes (MNPCEs) increased significantly at 24 and 48 hr in both study groups exposed to radiation. Likewise, in rat pups the MNPCE and MNE frequencies increased in both groups with radiation and radiation plus vitamin C at 24 and 48 hr, and a protection from vitamin C was observed. In conclusion, the genotoxic damage produced in rat pups that were lactated by mothers irradiated with X-rays is possibly due to the effect of long-lived reactive species that were formed in the breast milk of female Wistar rats during the irradiation process.

Assessment of water quality of selected rivers in the Niger Delta region of Nigeria using biomarkers in Clarias gariepinus.

Petroleum exploration has continued to impact negatively the Niger Delta region of Nigeria. Oil spills from petroleum exploration contaminate both the aquatic and non- aquatic environments and destroy the ecosystem which makes people in these regions to be prone to diseases such as cancer, cardiovascular diseases, neurodegenerative disease, infertility, and sterility and also poverty. Some oxidative stress markers of the liver and muscle of fish (Clarias gariepinus) from Apata, Araromi, and Mahin rivers (located in the Niger Delta region) were compared with those of fish samples from a clean fish farm (control) in order to assess the level of pollution of the rivers. A significant increase in malondialdehyde level and myeloperoxidase (MPO) activity as well as alterations in antioxidant status (glutathione peroxidase, superoxide dismutase, reduced glutathione, and glutathione S-transferase) was observed in the organs of fish samples from Apata, Araromi, and Mahin rivers compared with the control. A significant increase in the frequency of micronucleated polychromatic erythrocytes (MnPCEs) was also observed in the fish samples from these rivers in comparison with the control. Based on results of our research, we can conclude that these rivers are polluted mainly by oil spillage and other industrial activities; therefore, the consumption of aquatic organisms from these rivers and the use of water from the rivers for domestic activities might be unsafe.

Genotoxicity Evaluation of Capsaicin-Containing (CP) Pharmacopuncture, in an In Vivo Micronucleus Test.

ObjectivesCapsaicin-containing (CP) pharmacopuncture was developed to treat neuropathic pain. This study was conducted to assess the toxicity of CP extract for pharmacopuncture, using a micronucleus test.MethodsFirst, a dose range finding study was conducted. Then an in vivo micronucleus test was performed to determine the induction of micronuclei in mouse bone marrow cells after intramuscular administration of CP twice with a 24-hour interval to 8-week-old ICR mice. A high dose of 0.2 mL/animal was selected, and this was sequentially diluted by applying a geometric ratio of 2 to produce two lower dose levels (0.1 and 0.05 mL/animal). In addition, negative and positive control groups were set up, and an HPLC analysis was conducted to confirm the capsaicin content of CP.ResultsThe incidence of micro-nucleated polychromatic erythrocytes in polychromatic erythrocytes in the CP-treated group was similar to that in the negative-control group, while that in the positive-control group was significantly greater. In addition, the ratio of polychromatic erythrocytes to total erythrocytes in the CP treatment group and the positive control group was not significantly different from the negative control group. In the HPLC analysis, capsaicin in the CP was identified through a comparison with the retention time of the capsaicin standard of 27 min.ConclusionCP did not show any indication of any potential to induce micronuclei formation in bone marrow cells of ICR mice under the conditions of this study. Further toxicity studies are necessary to ensure the safety of the use of CP in clinical practice.

A study of the genetic and prenatal developmental toxicity potential of lithothamnion sp

Due to its calcium-rich and diverse multimineral profile, Aquamin (derived from the red seaweed Lithothamnion sp.) is used globally as a dietary food supplement. Published reports on the genetic and prenatal developmental toxicity of Lithothamnion sp. do not exist. In accordance with the standardized protocols set by the Ministry of Health of the People’s Republic of China (GB-15193), the following studies were performed: the Ames test, the mammalian erythrocyte micronucleus test, the mammalian spermatocyte chromosome test, and prenatal developmental toxicity testing. The results showed that Lithothamnion sp. did not induce a significant increase in the following: revertant colony numbers for Salmonella typhimurium strains TA 97, 98, 100, 102 and 1535; frequency of micronucleated polychromatic erythrocytes (MNPCE); spermatocyte chromosomal aberration rate. In the prenatal developmental toxicity study, no mortality, no abnormal changes in behavior and activities, and the absence of toxic symptoms and abnormalities in macroscopic autopsy were observed in each dam/all pups. Compared to the negative control group, Lithothamnion sp. at all tested doses had no effects on body weight gain, number of corpora lutea and implantations, fetal body weight and length, external, visceral and skeletal malformations. In conclusion, Lithothamnion sp. did not cause genetic toxicity. Furthermore, the prenatal developmental toxicity no observed adverse effect level (NOAEL) was determined to be greater than 2000 mg/kg.bw.

Protective effect of Moringa oleifera and green tea aqueous extract in combination against beta-cyfluthrin induced mutagenicity in albino rats

The present work was designed to investigate the mutagenicity effect of beta-cyfluthrin in bone marrow of male albino rats and the protective role of the combined aqueous extract of dried leaves of Moringa oleifera (MO) and green tea (GT). Twenty adult male rats were divided randomly into 4 groups, each containing five rats per group. The route of administration was orally gavage for 28 consecutive days. Rats in Group (a) were given distilled water and served as control, rats in Group (b) were given beta-cyfluthrin at a dose of 6.75 mg/kg b.wt (1/20 of estimated LD50). Rats in Group (c) were given aqueous extract of (MO+GT) as the only source of drinking fluid, for 28 days. Rats in Group (d) were given the same doses of group (b) and enabled continuously to an aqueous of (MO+GT) extract as the only source of drinking fluid. Animals were sacrificed humanely at the end of study. The micronucleus and chromosomal aberrations assays were used as mutagenicity tests. The results showed that beta-cyfluthrin administration alone increased the frequency of micronucleated polychromatic erythrocytes (MNPCEs) and induced several of chromosomal aberrations (CAs) in bone marrow cells of treated rats compared to control group. While the aqueous extract of (Mo+GT) in combination against the mutagenic potential of beta-cyfluthrin significantly decrease the (MNPCEs) and (CAs). In conclusion, this study confirmed the genotoxic potential of beta-cyfluthrin but also revealed the strong protective effect of aqueous leaves extract of Moringa oleifera and Green tea in combination as anti-mutagenic agents.

The protective effect of herniarin on genotoxicity and apoptosis induced by cisplatin in bone marrow cells of rats

Herniarin is a member of simple coumarins, which are a group of common secondary metabolites in plants. The aim of the present study was to investigate the effects of herniarin on genotoxicity and apoptosis induced by cisplatin in rat bone marrow cells. The experimental rats were treated with four different doses of herniarin (50, 100, 200, and 400 mg/kg.) for seven consecutive days. The cisplatin (5 mg/kg, i.p.) was injected into mice 1 h after the last oral herniarin administration on the seventh day. The protective effects of herniarin were investigated by hematological test, flow cytometry, micronucleus assay, and reactive oxygen species (ROS) level analysis. Herniarin caused a marked reduction in the frequencies of micronucleated polychromatic erythrocytes (MnPCEs) and micronucleated normochromatic erythrocytes (MnNCEs) 24 h after exposure to cisplatin at doses of 200 and 400 mg/kg. Furthermore, herniarin significantly increased the levels of both red and white blood cells in peripheral blood. Treatment of rats with herniarin before cisplatin, significantly decreased the percentage of apoptotic and necrotic cells and the ROS level in bone marrow cells. This study indicated that herniarin can be introduced as a new chemoprotective agent against cisplatin-induced genotoxicity in the future.

PROTECTIVE EFFECT OF VITAMIN C ON LEACHATE INDUCED GENOTOXICITY AND OXIDATIVE STRESS IN THE LIVER AND OVARY OF RAT

IntroductionThis study investigated the protective effect of vitamin C on leachate induced genotoxicity and oxidative stress in the liver and ovary of rats exposed to Ajakanga landfill leachate. Methods Sixty female rats(150-200g) were equally divided into ten groups(A-I). Group A animals(control) were given distilled water, groups B – E animals were given 12.5%, 25%, 50%, and 100% leachate respectively via drinking water for twenty-eight days. Animals in group F were given distilled water plus 100mg/kg vitamin C while groups G – I animals were given 100mg/kg vitamin C plus 12.5%, 25%, 50%, and 100% leachate respectively. Activities of alanine amino transferase(ALT), aspartate aminotransferase(AST) and Gamma-glutamyltransferase(GGT) were determined in the serum of rats using enzyme kits. Oxidative stress markers were determined in the liver and ovary of the rats by spectrophotometry. Genotoxicity study was carried out using micronucleus assay.Results There was increase in the activities of ALT, AST and GGT in serum of rats exposed to leachate compared with control but ALT and AST activity decreased in rats treated with vitamin C. There was increase in malondialdehyde concentration (index of lipid peroxidation) in the liver (102.3-302.7mol/mg protein) and ovary (1.1-2.1mol/mg protein) of rats exposed to leachate compared with control (72.6mol/mg protein;0.7mol/mg protein) but there was a significant decrease in animals treated with vitamin C. There was decrease in the levels of reduced glutathione and super oxide dismutase in the ovary of rats in groups B-E, but vitamin C treatment increased the levels of these antioxidants. A significant increase in micronucleated polychromatic erythrocytes(MPCE) was observed in bone marrow of rats exposed to leachate (74.7-113 MPCE/1000PCE) but there was a decrease in MPCEs in rats treated with vitamin C (66.8-78.8 MPCE/1000PCE).Conclusion Vitamin C has a protective effect on leachate induced genotoxicity and oxidative stress in the liver and ovary of the rats.

Genotoxicity evaluation using micronucleus test in Rattus norvegicus captured in urban ecosystems of Buenos Aires, Argentina.

The overall goal of the present study was to evaluate and compare the existence of genotoxic damage in Rattus norvegicus captured in sites with different levels of chemical mixtures along the Matanza-Riachuelo river basin(MRRB). For this, thirty-six wild rats (R. norvegicus) were captured from six different sites in the MRRB. Capture sites were characterized with concentrations of 20 parameters in surface water, including concentrations of metals and its soluble state and physicochemical parameters. To evaluate genotoxic damage in the rats, the bone marrow micronucleus test was performed. For this, the frequency of micronucleated polychromatic erythrocytes (MNPCE) was calculated based on the observation of 2000 polychromatic erythrocytes (PCE) per animal. Also, to detect possible cytotoxic effects, the PCE/TE (total erythrocytes) ratio in 1000 erythrocytes/animal was calculated. The frequency of MNPCE was positively associated with the highest concentration of chrome, lead, copper, biological oxygen demand, and chemical oxygen demand. Also, the decrease on PCE/TE ratio in bone marrow was related with increase of practically all metals and physicochemical parameters in surface water. The results of this study reveal that rats that live in the MRRB were exposed to concentrations of pollutants that can cause genotoxic damage, even though the concentrations of these pollutants are mostly below the safety standards. Therefore,this work highlights the importance of using methods that allowto integrate all environmental parametersin risk assessment, such as sentinel species.

Evaluation of Anti-Cytotoxic and Anti-Genotoxic Effects of Nigella sativa through a Micronucleus Test in BALB/c Mice.

Nigella sativa (N. sativa) is a medicinal plant used for its therapeutic pharmacological effects such as anti-inflammatory, antioxidant, anticancer, antidiabetic, and immunomodulation. This study explored the anti-cytotoxic and anti-genotoxic effect of N. sativa through a micronucleus test (MNT) of BALB/c mice peripheral blood. Using 6-to-8-week-old healthy male BALB/c mice, four groups were formed: (1) Control (sterile water), single-dose 2 mg/kg/intraperitoneal (i.p); (2) N. sativa oil, 500 mg/kg/24 h/7 days/i.p; (3) Cisplatin (CP), single-dose 2 mg/kg/subcutaneous (s.c); (4) N. sativa + CP with their respective dosage. When evaluating polychromatic erythrocytes (PCE), a biomarker of cytotoxicity, the group treated with N. sativa + CP experienced an increase in the frequency of PCE, which demonstrated the recovery of bone marrow and modulation of cell proliferation. The analysis of micronucleated polychromatic erythrocytes (MNPCE), an acute genotoxicity biomarker, showed similar frequency of MNPCE within the groups except in CP, but, in the N. sativa + CP group, the frequency of MNPCE decreased and then regulated. Finally, the frequency of micronucleated erythrocytes (MNE), a biomarker of genotoxicity, the supplementation of N. sativa oil did not induce genotoxic damage in this model. Thus, we conclude that N. sativa has both cytoprotective, genoprotective effects and modulates cell proliferation in BALB/c mice.

Evaluation of the antioxidant activity and mutagenicity of Brazil nut (Bertholletia excelsa Bonpl.).

This study evaluated the antioxidant capacity and the genotoxic and antigenotoxic effects of the fixed oil of B. excelsa Bonpl. in peripheral blood of Swiss mice. For the antioxidant capacity, were used the β-carotene/linoleic acid and DPPH methods. In the genotoxicity assay, were used healthy, 6-7 week old male Swiss mice, and there were six animals per group. For the genotoxic test, animals were treated with different concentrations of B. excelsa (500, 1.000 and 2.000 mg/kg body weight bw) in 0.5 mL orally. For the antigenotoxic test, animals were treated with predetermined concentrations, followed by intraperitoneal injection of doxorubicin (DXR 15 mg/kg bw) in 0.3 ml, in addition to the negative group (water) and dimethylsulfoxide (200 μL). Peripheral blood samples were collected 24 and 48 hours after the treatments. The frequency of micronucleated polychromatic erythrocytes (MNPCEs) was obtained from the analysis of 2.000 MNPCEs/animal. After data analysis, the conclusion was that the fixed oil of B. excelsa showed excellent protective activity by the β-carotene/linoleic acid method, demonstrated absence of genotoxic effect, and significant antigenotoxic effect according to the protocols and treatments performed in this study.

Antimicrobial potential, phytochemical profile, cytotoxic and genotoxic screening of Sedum praealtum A. DC. (balsam)

BackgroundSedum praealtum has been used for a long time in traditional medicine as an analgesic and anti-inflammatory agent. Its beneficial effects have been known since ancient times, when Latinos used it to treat sore and swollen eyes. This research evaluated the antimicrobial potential, the cytotoxic and genotoxic effects, and some chromatographic profiles of the hydroethanolic extract of leaves, stems and roots of S. praealtum.MethodsThe antimicrobial activities were carried out by broth microdilution and agar diffusion. In vitro cytotoxicity was evaluated by cell cultures of Aedes albopictus and the selectivity index (SI) was estimated: SI=CI50/MIC. Genotoxic and systemic toxic effects of S. praealtum leaves were analyzed by micronucleus assay in mice bone marrow. Chromatographic profiles and mass spectra were investigated by GC-MS.ResultsGram-positive (B. subtilis, B. cereus, M. luteus, E. faecalis and S. aureus) and gram-negative (E. coli, E. aerogenes, S. marcescens, P. aeruginosa, P. mirabilis and S. typhimurium) bacteria exhibited MICs ranging from 12.5–50 and 0–50 mg/ml, respectively. Sedum praealtum showed no efficacy against M. tuberculosis and M. bovis. Cytotoxicity (CI50) of S. praealtum was 4.22 and 5.96 mg/ml for leaves and stems, respectively, while its roots showed no cytotoxicity. Micronucleated polychromatic erythrocytes (MNPCEs) analyzes showed no differences between treatment doses (0.5–2 g/kg) and negative control (NaCl), but the PCE/NCE ratio (polychromatic erythrocyte/normochromatic erythrocyte) showed significant differences. Phytochemical screening identified thirteen compounds in the leaves, stems and roots of S. praealtum potentially associated with their biological activities.ConclusionsThis research comprises a first scientific study on genotoxicity, cytotoxicity and antimicrobial effects of S. praealtum (Balsam), and it provides an initial theoretical foundation for its comprehensive use. Results showed antibacterial action of S. praealtum against gram-positive bacteria and some gram-negative species (depending on the plant anatomical part), but ineffective antimycobacterial action. However, S. praealtum leaves and stems display potential cytotoxicity, contributing to the SI < 1 values. In addition, S. praealtum leaves exhibit no clastogenic and/or aneugenic effects, but it has systemic toxicity dose-independent.

Effect of High-Dose Topical Minoxidil on Erythrocyte Quality in SKH1 Hairless Mice.

Simple SummaryIn any animal species, involuntary exposure to unknown agents may increase genetic material damage. This genetic damage can also induce the appearance of diseases such as cancer or other pathologies, including problems that can be passed on to the offspring of the damaged individual. For instance, living organisms may be affected due to the use of medications or exposure to certain chemical, physical, or biological agents which cause cell failure. This impaired function acts as an indicator that helps identify and evaluate damage in order to avoid or minimize it. In this work, excessive doses of a cosmetic drug for topical use in dermatological treatments, known as minoxidil, produced defects in the blood of hairless mice, particularly in red cells, indicating loss of DNA, a situation that may compromise life or the offspring by causing damage to their genetic material. It is important to consider that compounds may be tissue- or species-specific, although we cannot rule out the possibility that similar damage could occur in other animal species. Thus, excessive exposure to this compound should be prevented.SKH1 hairless mice are widely used in carcinogenesis and dermatology research due to their bare skin, as exposure to different agents is facilitated. Minoxidil is a cosmetic drug that is recognized as a mitogenic agent, and mitogens are suggested to have carcinogenic and mutagenic potential by inducing cell division and increasing the possibility of perpetuating DNA damage. Therefore, we hypothesized that the application of high doses of minoxidil to the skin of hairless mice would increase the number of micronucleated erythrocytes (MNEs) in peripheral blood. The objective of this study was to evaluate the topical administration of high doses of minoxidil on peripheral blood erythrocytes of SKH1 mice by means of micronucleus assay. Minoxidil was administered on the entire body surface of mice every 12 or 24 h. Minoxidil dosing every 24 h increased the number of micronucleated polychromatic erythrocytes (MNPCEs), and dosing every 12 h increased the number of MNEs and MNPCEs, as compared to baseline and the negative control group. No decrease in polychromatic erythrocyte frequencies was observed in the minoxidil groups. Therefore, topical application of high minoxidil doses to mice can produce DNA damage, as observed through an increase in the number of MNEs, without producing cytotoxicity, possibly due to its mitogenic effect.

Genotoxic potential of a novel PDE-4B inhibitor Apremilast by chromosomal aberration and micronucleus assay in mice

ObjectiveResearchers have confirmed that chronic administration of drugs at high doses causes genotoxicity which serve as first step in development of cancers. Apremilast, a phosphodiesterase-4 inhibitor is Food and Drug Administration (FDA) approved drug for Psoriatic Arthritis. The present study designed to conduct genotoxicity testing using the genotoxic study which give simple, sensitive, economical and fast tools for the assessment of damage of genetic material. MethodsTo conduct genotoxicity study of Apremilast, 60 Swiss albino male mice divided into 6 groups (n = 10). Group1 served as a normal control group without any treatment, Group 2 treated as a disease control and administered with cyclophosphamide 40 mg/kg, IP. Group 3, 4, 5 and 6 treated as test groups and received 10, 20, 40 and 80 mg/kg/day Apremilast respectively. The total duration of study was 13 weeks. At termination day animals were sacrificed and chromosomal aberration assay (BMCAA) and micronucleus assay (BMMNA) were performed to know the genotoxicity potential of Apremilast. ResultsThe results indicates significant rise in chromosomal aberrations (CA) frequency in bone marrow cells and decrease in the MI of the disease control animals as well as Apremilast treated groups. Further significant (p < 0.001; p < 0.0001) increase in score of micronucleated polychromatic erythrocytes (MNPCEs) and percentage of micronucleated PCEs per 1000 PCEs and decrease in the ratio of polychromatic/normochromatic erythrocytes (PCE/NCE) was observed in micronucleus assay. Genotoxic effect increases with the increase of Apremilast dose. Conclusion: Finding of present indicates that Apremilast shows genotoxic potential on high administration although further detailed toxicity studies required for confirmations.

Genotoxicity assessment of saline extract from Pilosocereus gounellei (Cactaceae) and its chemopreventive effect against cyclophosphamide-induced DNA damage

Pilosocereus gounellei (Cactaceae) is used to treat wounds and inflammation. In this study, we evaluated whether the saline extract from its stem would have genotoxic or anti-genotoxic effects. In the genotoxicity evaluation, mice received the extract (500, 1,000, or 2,000 mg/kg) orally while negative and positive controls were treated with saline solution (0.9% NaCl) per os and cyclophosphamide (CPA, 80 mg/kg i.p.), respectively. In the anti-genotoxicity assay, using other animals, treatments were carried out by administering the extract (500, 1,000 or 2,000 mg/kg) or saline solution (negative control) per os and then CPA (80 mg/kg i.p.) 1 h later. Genotoxic effects were evaluated by micronucleus test and comet assay using peripheral blood and bone marrow cells. Oral administration of only the extract at 500 and 1,000 mg/kg did not result in genotoxicity. A slight increase in the incidence of micronucleus was observed at the highest dose (2,000 mg/kg). Administration of the extract before CPA reduced the micronucleated polychromatic erythrocytes (MNPCE) number by 49.07–71.43%, and DNA fragmentation in peripheral blood (85.04–94.44%) and bone marrow (87.43–92.70%) cells also decreased. In conclusion, when administered orally at the tested doses, the extract is genotoxically safe, being cautious in doses above 1,000 mg/kg, and has a protective effect against CPA-induced DNA damage.

In vivo assessment of genotoxic potential of brown shammah (smokeless tobacco) in bone marrow cells of mice

This study was aimed to assess the genotoxicity of brown shammah (BS), a local form of smokeless tobacco, popular in Middle East countries including Yemen, Saudi Arabia and Sudan. The genotoxicity was explored using in vivo chromosomal aberration (CA), micronucleus (MN) and sperm abnormality (SA) assays. In addition, oxidative stress was also determined using various hepatic markers. Swiss albino mice were selected for the study, which were divided in to 5 groups of six animals each. They include, negative control (NC, received only vehicle) as well as positive control group (PC, received vehicle for 2 weeks followed by administration of cyclophosphamide, CP). Depending upon their dose, three BS treated animal groups were BS-100, 300 and 900 mg/kg. Doses of BS were obtained by suspending BS in 0.5% CMC (carboxy methyl cellulose) and orally administered once a day for 2 weeks. Significant augmentation of the average percentage of aberrant metaphase (AM), CA per cells and suppressed mitotic activity was observed on post administration of BS. In addition, BS increased the occurrence of MNPCEs (micronucleated polychromatic erythrocytes) formation, induced cytotoxicity and increased percentage of abnormal sperms as compared to NC. Moreover, BS also induced oxidative stress as the activities of hepatic superoxide dismutase (SOD) and glutathione (GSH) were reduced and malondialdehyde (MDA) content were increased by BS. Cyclophosphamide was utilized as clastogen, showed anticipated positive results and confirmed the sensitivity of test system. Therefore, it may be deduced from the study that the BS possesses genotoxic effects on mice bone marrow and germ cells in vivo.

Solid-state fermentation of ammoniated corn straw to produce feed protein and toxicological assessment of the product.

Solid-state fermentation (SSF) of ammoniated corn straw was used to produce feed protein, followed by a toxicological assessment of the fermentation product. Results showed that through ammonification at 35°C for 9days and the subsequent SSF by the two fungi Penicillium sp. and Torula allii at 30°C for 5days, the contents of real protein and crude protein of the corn straw reached 29.66% and 35.41%, respectively. Toxicological assessment in mice showed that there were no significant differences (P > 0.05) for micronucleated polychromatic erythrocytes (Mn-PCEs) and sperm abnormality between dose groups and the control group. Malondialdehyde (MDA) levels and activities of superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) also showed no significant difference (P > 0.05) between tissues (heart, liver, spleen, stomach, kidney, and brain), which indicates that the fermentation product did not induce toxic effects and is safe to use as ruminant feed.