Search results for: micronucleus
Commenced in January 2007
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Edition: International
Paper Count: 16

Search results for: micronucleus

16 Mutagenic in vitro Activity and Genotoxic Effect of Zygophyllum Cornutun Methanolic Extract

Authors: Awatif Boumaza, Abderraouf Hilali, Hayat Talbi, Houda Sbayou

Abstract:

The methanolic extract of Zygophyllum cornutun coss, an Algerian medicinal plant, was screened to the presence of mutagenic activity and genotoxic effect using the Ames test (Salmonella/microsome) and the micronucleus assay respectively. Positive results were obtained with both tests. The Ames test showed mutagenic activity in the presence of microsomal activation, while negative result was observed without microsomal activation. In the micronucleus test, two parameters were evaluated: the frequency of the micronucleus that increased in a dose dependent way and the proliferation index that decreased according to the micronucleus frequency. Even that further studies must be carried out, the mutagenic activity and the genotoxic effect of Zygophyllum cornutum should be taken in consideration when used as therapeutic plant.

Keywords: ames test, micronucleus test, mutagenic activity, genotoxicity, Zygophyllum cornutum

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15 Genotoxicity Induced by Nanoparticles on Human Lymphoblast Cells (TK6)

Authors: Piyaporn Buaklang, Narisa Kengtrong Bordeerat

Abstract:

The use of nanoparticles is increasing worldwide and there are many nanotech-based daily products available in the market. The toxicity of nanoparticles results from their extremely small size which can be transported easily into the blood stream and other organs. We aimed to study the genotoxicity of two nanoparticles, Titanium dioxide (TiO2-NPs) and Zinc oxide (ZnO-NPs), in TK6 cells by micronucleus assay. The cells were tested at 8, 24, and 48 hours after exposed to 0.10, 0.25, 0.50 and 1.00 µg/mL of TiO2-NPs particles size < 25 nm and < 100 nm and to ZnO-NPs at 1, 10, 50, and 100 µg/mL, particles size < 50 nm and < 100 nm. At 24 hours of incubation transmission electron microscope (TEM) revealed that the nanoparticles TiO2-NPs at 1.00 µg/mL and ZnO-NPs at 10 µg/mL were able to be taken into the cells and induced the production of increasing amount of micronucleus in dose-dependent manner. The effect of the two nanoparticles on chromosome aberration indicated that TiO2-NPs and ZnO-NPs are genotoxic. In addition, the toxicity of TiO2-NPs was found to be 10 times more toxic than ZnO-NPs after 24 hours exposure. Analysis showed that the TiO2-NPs induced formation of micronucleus was both time and dose dependent, whereas the genotoxicity of ZnO-NPs was only dose dependent. In conclusion, TiO2-NPs and ZnO-NPs were able to transport through the cells membrane and directly genotoxic to TK6 cells in dose-dependent manner.

Keywords: nanoparticles, genotoxicity, human lymphoblast cells (TK6), micronucleus

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14 In vivo Genotoxicity Testing of Sesbania Grandiflora (Katuray) Flower Methanolic Extract

Authors: Levylee Bautista, Dawn Grace Santos, Aishwarya Veluchamy, Jesusa Santos, Ghafoor, Jr. I Haque, Rodolfo Rafael

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The booming interest in using natural compounds as an alternative to conventional medications has paved way to focus the attention on plants that provide rich sources of bioactive phytochemicals. For regulatory purposes, evaluation of the genotoxic effects of such alternatives is therefore empirical as part of the plant’s hazard assessment. Sesbania grandiflora is among the plants used as a traditional remedy in folk medicine and a subject of research for its medicinal benefits. This study aimed to evaluate the genotoxic potential induced by S. grandiflora flower methanol extract (SGFME) in terms of the frequency of micronucleus (MN) in polychromatic erythrocyte (PCE) (MNPCE) and PCE ratio employing the micronucleus assay. The frequency of MN was examined in bone marrow cells (BMCs) obtained from male Swiss albino mice exposed in vivo to four different concentrations (11.25, 22.5, 40, and 90 mg/kg) of SGFME and MMC (70 mg/kg; positive control) and sacrificed 24 hours post-intraperitoneal injection. Results showed a significant (p < 0.01) rate of MNPCEs for 11.25 and 22.5 tested concentrations of SGFME and is comparable with the MMC-treated mice. Although PCE ratio values in all doses of SGFME-treated mice were over 0.20, it is worth noting that 40 and 90 tested concentrations of SGFME-treated mice exhibited the lowest value, i.e., 0.22 and 0.28, respectively. The present study has demonstrated that S. grandiflora possesses genotoxic potential for murine BMCs. Such activity could be ascribed from the bioactive compounds present in S. grandiflora that require further isolation and characterization of the active molecules. Likewise, findings of this study warrant a caution of the use of S. grandiflora insomuch as further investigations do not demonstrate their safety.

Keywords: genotoxicity, micronucleus, phytochemicals, Sesbania grandiflora

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13 Zooplankton Health Status Monitoring in Bir Mcherga Dam (Tunisia)

Authors: Sabria Barka, Imen Gdara, Zouhour Ouanès, Samia Mouelhi, Monia El Bour, Amel Hamza-Chaffai

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Because dams are large semi-closed reservoirs of pollutants originating from numerous anthropogenic activities, they represent a threat to aquatic life and they should be monitored. The present work aims to use freshwater zooplankton (Copepods and Cladocerans) in order to evaluate the environmental health status of Bir M'cherga dam in Tunisia. Animals were collected in situ monthly between October and August. Genotoxicity (micronucleus test), neurotoxicity (acetylcholinesterase, AChE) and oxidative stress (catalase, CAT and malondialdehyde, MDA) biomarkers were analyzed in zooplankton. High frequencies of micronucleus were observed in zooplankton cells during summer. AChE activities were inhibited during early winter and summer. CAT and MDA biomarker levels showed high seasonal variability, suggesting that animals are permanently exposed to multiple oxidative stress. The results of this study suggest that the Bir Mcherga dam is subject to continuous multi-origin stress, probably amplified by abiotic parameters. It is then recommended to urgently monitor freshwater environments in Tunisia, especially those used for irrigation and consumption.

Keywords: Biomonitoring, Bir Mcherga Dam, cladocerans, copepods, freshwater zooplankton, genotoxicity, neurotoxicity, oxidative stress, Tunisia

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12 Evaluation of Airborne Particulate Matter Early Biological Effects in Children with Micronucleus Cytome Assay: The MAPEC_LIFE Project

Authors: E. Carraro, Sa. Bonetta, Si. Bonetta, E. Ceretti, G. C. V. Viola, C. Pignata, S. Levorato, T. Salvatori, S. Vannini, V. Romanazzi, A. Carducci, G. Donzelli, T. Schilirò, A. De Donno, T. Grassi, S. Bonizzoni, A. Bonetti, G. Gilli, U. Gelatti

Abstract:

In 2013, air pollution and particulate matter were classified as carcinogenic to human by the IARC. At present, PM is Europe's most problematic pollutant in terms of harm to health, as reported by European Environmental Agency (EEA) in the EEA Technical Report on Air quality in Europe, 2015. A percentage between 17-30 of the EU urban population lives in areas where the EU air quality 24-hour limit value for PM10 is exceeded. Many studies have found a consistent association between exposure to PM and the incidence and mortality for some chronic diseases (i.e. lung cancer, cardiovascular diseases). Among the mechanisms responsible for these adverse effects, genotoxic damage is of particular concern. Children are a high-risk group in terms of the health effects of air pollution and early exposure during childhood can increase the risk of developing chronic diseases in adulthood. The MAPEC_LIFE (Monitoring Air Pollution Effects on Children for supporting public health policy) is a project founded by EU Life+ Programme (LIFE12 ENV/IT/000614) which intends to evaluate the associations between air pollution and early biological effects in children and to propose a model for estimating the global risk of early biological effects due to air pollutants and other factors in children. This work is focused on the micronuclei frequency in child buccal cells in association with airborne PM levels taking into account the influence of other factors associated with the lifestyle of children. The micronucleus test was performed in exfoliated buccal cells of 6–8 years old children from 5 Italian towns with different air pollution levels. Data on air quality during the study period were obtained from the Regional Agency for Environmental Protection. A questionnaire administered to children’s parents was used to obtain details on family socio-economic status, children health condition, exposures to other indoor and outdoor pollutants (i.e. passive smoke) and life-style, with particular reference to eating habits. During the first sampling campaign (winter 2014-15) 1315 children were recruited and sampled for Micronuclei test in buccal cells. In the sampling period the levels of the main pollutants and PM10 were, as expected, higher in the North of Italy (PM10 mean values 62 μg/m3 in Torino and 40 μg/m3 in Brescia) than in the other towns (Pisa, Perugia, Lecce). A higher Micronucleus frequency in buccal cells of children was found in Brescia (0.6/1000 cells) than in the other towns (range 0.3-0.5/1000 cells). The statistical analysis underlines a relation of the micronuclei frequency with PM concentrations, traffic level near child residence, and level of education of parents. The results suggest that, in addition to air pollution exposure, some other factors, related to lifestyle or further exposures, may influence micronucleus frequency and cellular response to air pollutants.

Keywords: air pollution, buccal cells, children, micronucleus cytome assay

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11 Assessment of Airborne PM0.5 Mutagenic and Genotoxic Effects in Five Different Italian Cities: The MAPEC_LIFE Project

Authors: T. Schilirò, S. Bonetta, S. Bonetta, E. Ceretti, D. Feretti, I. Zerbini, V. Romanazzi, S. Levorato, T. Salvatori, S. Vannini, M. Verani, C. Pignata, F. Bagordo, G. Gilli, S. Bonizzoni, A. Bonetti, E. Carraro, U. Gelatti

Abstract:

Air pollution is one of the most important worldwide health concern. In the last years, in both the US and Europe, new directives and regulations supporting more restrictive pollution limits were published. However, the early effects of air pollution occur, especially for the urban population. Several epidemiological and toxicological studies have documented the remarkable effect of particulate matter (PM) in increasing morbidity and mortality for cardiovascular disease, lung cancer and natural cause mortality. The finest fractions of PM (PM with aerodynamic diameter <2.5 µm and less) play a major role in causing chronic diseases. The International Agency for Research on Cancer (IARC) has recently classified air pollution and fine PM as carcinogenic to human (1 Group). The structure and composition of PM influence the biological properties of particles. The chemical composition varies with season and region of sampling, photochemical-meteorological conditions and sources of emissions. The aim of the MAPEC (Monitoring Air Pollution Effects on Children for supporting public health policy) study is to evaluate the associations between air pollution and biomarkers of early biological effects in oral mucosa cells of 6-8 year old children recruited from first grade schools. The study was performed in five Italian towns (Brescia, Torino, Lecce, Perugia and Pisa) characterized by different levels of airborne PM (PM10 annual average from 44 µg/m3 measured in Torino to 20 µg/m3 measured in Lecce). Two to five schools for each town were chosen to evaluate the variability of pollution within the same town. Child exposure to urban air pollution was evaluated by collecting ultrafine PM (PM0.5) in the school area, on the same day of biological sampling. PM samples were collected for 72h using a high-volume gravimetric air sampler and glass fiber filters in two different seasons (winter and spring). Gravimetric analysis of the collected filters was performed; PM0.5 organic extracts were chemically analyzed (PAH, Nitro-PAH) and tested on A549 by the Comet assay and Micronucleus test and on Salmonella strains (TA100, TA98, TA98NR and YG1021) by Ames test. Results showed that PM0.5 represents a high variable PM10 percentage (range 19.6-63%). PM10 concentration were generally lower than 50µg/m3 (EU daily limit). All PM0.5 extracts showed a mutagenic effect with TA98 strain (net revertant/m3 range 0.3-1.5) and suggested the presence of indirect mutagens, while lower effect was observed with TA100 strain. The results with the TA98NR and YG1021 strains showed the presence of nitroaromatic compounds as confirmed by the chemical analysis. No genotoxic or oxidative effect of PM0.5 extracts was observed using the comet assay (with/without Fpg enzyme) and micronucleus test except for some sporadic samples. The low biological effect observed could be related to the low level of air pollution observed in this winter sampling associated to a high atmospheric instability. For a greater understanding of the relationship between PM size, composition and biological effects the results obtained in this study suggest to investigate the biological effect of the other PM fractions and in particular of the PM0.5-1 fraction.

Keywords: airborne PM, ames test, comet assay, micronucleus test

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10 Monitoring the Pollution Status of the Goan Coast Using Genotoxicity Biomarkers in the Bivalve, Meretrix ovum

Authors: Avelyno D'Costa, S. K. Shyama, M. K. Praveen Kumar

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The coast of Goa, India receives constant anthropogenic stress through its major rivers which carry mining rejects of iron and manganese ores from upstream mining sites and petroleum hydrocarbons from shipping and harbor-related activities which put the aquatic fauna such as bivalves at risk. The present study reports the pollution status of the Goan coast by the above xenobiotics employing genotoxicity studies. This is further supplemented by the quantification of total petroleum hydrocarbons (TPHs) and various trace metals (iron, manganese, copper, cadmium, and lead) in gills of the estuarine clam, Meretrix ovum as well as from the surrounding water and sediment, over a two-year sampling period, from January 2013 to December 2014. Bivalves were collected from a probable unpolluted site at Palolem and a probable polluted site at Vasco, based upon the anthropogenic activities at these sites. Genotoxicity was assessed in the gill cells using the comet assay and micronucleus test. The quantity of TPHs and trace metals present in gill tissue, water and sediments were analyzed using spectrofluorometry and atomic absorption spectrophotometry (AAS), respectively. The statistical significance of data was analyzed employing Student’s t-test. The relationship between DNA damage and pollutant concentrations was evaluated using multiple regression analysis. Significant DNA damage was observed in the bivalves collected from Vasco which is a region of high industrial activity. Concentrations of TPHs and trace metals (iron, manganese, and cadmium) were also found to be significantly high in gills of the bivalves collected from Vasco compared to those collected from Palolem. Further, the concentrations of these pollutants were also found to be significantly high in the water and sediments at Vasco compared to that of Palolem. This may be due to the lack of industrial activity at Palolem. A high positive correlation was observed between the pollutant levels and DNA damage in the bivalves collected from Vasco suggesting the genotoxic nature of these pollutants. Further, M. ovum can be used as a bioindicator species for monitoring the level of pollution of the estuarine/coastal regions by TPHs and trace metals.

Keywords: comet assay, metals, micronucleus test, total petroleum Hydrocarbons

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9 Aloe vera Prevents Injuries Induced by Whole Body X-ray Irradiation in Rodents

Authors: Shashi Bala, Neha A. Chugh, Subhash C. Bansal, Mohal L. Garg, Ashwani Koul

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Purpose: The present study was designed to evaluate the radioprotective efficacy of Aloe vera from whole body X-ray exposure in rodents. Materials and Methods: For this purpose, after on week’s acclimatization, male balb/c mice procured from Central Animal House, Panjab University, Chandigarh (India), were divided into four groups: Group I mice served as control. Group II mice were orally administrated Aloe vera pulp extract (50 mg/ kg body weight) on alternate days for 30 days. Group III mice were subjected to whole body X-ray irradiation to cumulative dose of 2Gy (0.258Gy twice a day for four days in the last week). Group IV animals were pretreated with Aloe vera pulp extract on alternate days as in Group II and in the last week of the study, they were exposed to X-ray as in Group III. Results: Spleen of X-ray irradiated mice showed histopathological alterations accompanied with enhanced activity of lactate dehydrogenase (LDH) in serum. Elevated levels of reactive oxygen species (ROS), lipid peroxidation (LPO), enhanced activities in Glutathione based enzymes such as Glutathione peroxidase (GSH-Px), Glutathione reductase (GR), Catalase (CAT), Superoxide dismutase (SOD) associated with depletion in reduced Glutathione (GSH) concentration were observed after X-ray exposure in blood plasma and spleen.. Pro-inflammatory cytokines like tumor necrosis factors (TNF-α) and Inteleukin-6 (IL-6) levels were also found to be enhanced in serum of irradiated mice. Irradiation-induced significant elevation in Total leucocyte counts (TLC), neutrophil counts and decline in platelet counts, associated with unaltered levels of red blood cell counts (RBC’s) and haemoglobin (Hb) in various treatment groups. Clastogenic damage and apoptosis was also found to be increase in splenic tissue of X-ray exposed mice as assessed by micronucleus and TUNEL assay. However, X-ray irradiated animals administered with Aloe vera revealed significant improvement in levels of ROS/ LPO, LDH activity, and antioxidant mechanism. Aloe vera pretreated animals exhibited less severe damage, and early recovery in micronucleated cells, hematological parameters, apoptotic cells and inflammatory markers as compared to X-ray exposed mice. Conclusion: These results indicate that the radioprotective potential of Aloe vera against X-ray induced damage. This may be due to its free radical scavenging, antioxidant, anti-apoptotic and anti-inflammatory properties.

Keywords: aloe vera, antioxidant defense system, lactate dehydrogenase (LDH), micronucleus assay, x-ray

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8 Forced Swim Stress Does Not Induce Structural Chromosomal Aberrations in Rat Bone Marrow

Authors: Mohammad Y. Alfaifi

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Anything that poses a challenge or a threat to our well-being is a stress. Understanding the genetic material and cellular response of rats threatened with Repeated swimming stress provides insights that can influence human health. The aim of the present study was to assess the genetical damage and cytological changes caused by exposure of the test organism (Rattus rattus) to forced swimming stress. For this purpose, animals have been submerged in water path 15 minutes daily for 2 weeks. Following that, we performed a micronuclei (MN) test using MNNCE (Micronucleated normocromatic erythrocytes) and MNPCE (Micronucleated polychromatic erythrocytes), NDI (Nuclear division index) and cytological parameters using NDCI (nuclear division cytotoxicity index), necrotic and apoptotic cells in rat's bone marrow samples. Results showed that there was a slightly but not significant increase in the frequency of micronucleated as well as in cytological parameters in bone marrow cells.

Keywords: submergence stress, micronucleus, NDI, NDCI, toxicity, chromosomal aberrations

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7 Assessment of Cytogenetic Damage as a Function of Radiofrequency Electromagnetic Radiations Exposure Measured by Electric Field Strength: A Gender Based Study

Authors: Ramanpreet, Gursatej Gandhi

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Background: Dependence on electromagnetic radiations involved in communication and information technologies has incredibly increased in the personal and professional world. Among the numerous radiations, sources are fixed site transmitters, mobile phone base stations, and power lines beside indoor devices like cordless phones, WiFi, Bluetooth, TV, radio, microwave ovens, etc. Rather there is the continuous emittance of radiofrequency radiations (RFR) even to those not using the devices from mobile phone base stations. The consistent and widespread usage of wireless devices has build-up electromagnetic fields everywhere. In fact, the radiofrequency electromagnetic field (RF-EMF) has insidiously become a part of the environment and like any contaminant may pose to be health-hazardous requiring assessment. Materials and Methods: In the present study, cytogenetic damage was assessed using the Buccal Micronucleus Cytome (BMCyt) assay as a function of radiation exposure after Institutional Ethics Committee clearance of the study and written voluntary informed consent from the participants. On a pre-designed questionnaire, general information lifestyle patterns (diet, physical activity, smoking, drinking, use of mobile phones, internet, Wi-Fi usage, etc.) genetic, reproductive (pedigrees) and medical histories were recorded. For this, 24 hour-personal exposimeter measurements (PEM) were recorded for unrelated 60 healthy adults (40 cases residing in the vicinity of mobile phone base stations since their installation and 20 controls residing in areas with no base stations). The personal exposimeter collects information from all the sources generating EMF (TETRA, GSM, UMTS, DECT, and WLAN) as total RF-EMF uplink and downlink. Findings: The cases (n=40; 23-90 years) and the controls (n=20; 19-65 years) matched for alcohol drinking, smoking habits, and mobile and cordless phone usage. The PEM in cases (149.28 ± 8.98 mV/m) revealed significantly higher (p=0.000) electric field strength compared to the recorded value (80.40 ± 0.30 mV/m) in controls. The GSM 900 uplink (p=0.000), GSM 1800 downlink (p=0.000),UMTS (both uplink; p=0.013 and downlink; p=0.001) and DECT (p=0.000) electric field strength were significantly elevated in the cases as compared to controls. The electric field strength in the cases was significantly from GSM1800 (52.26 ± 4.49mV/m) followed by GSM900 (45.69 ± 4.98mV/m), UMTS (25.03 ± 3.33mV/m), DECT (18.02 ± 2.14mV/m) and was least from WLAN (8.26 ± 2.35mV/m). The higher significantly (p=0.000) increased exposure to the cases was from GSM (97.96 ± 6.97mV/m) in comparison to UMTS, DECT, and WLAN. The frequencies of micronuclei (1.86X, p=0.007), nuclear buds (2.95X, p=0.002) and cell death parameter (condensed chromatin cells) were significantly (1.75X, p=0.007) elevated in cases compared to that in controls probably as a function of radiofrequency radiation exposure. Conclusion: In the absence of other exposure(s), any cytogenetic damage if unrepaired is a cause of concern as it can cause malignancy. Larger sample size with the clinical assessment will prove more insightful of such an effect.

Keywords: Buccal micronucleus cytome assay, cytogenetic damage, electric field strength, personal exposimeter

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6 Genotoxic and Cytotoxic Effects of Methidathion Pesticide

Authors: Mohammad Y. Alfaifi

Abstract:

Methidathion (MTD) (Trade name Supracide®) is a non-systemic organophosphorus insecticide used intensively worldwide including Saudi Arabia. However, there is a lack in published studies about it's genotoxicity. In this study we evaluated MTD toxicity in rat bone marrow cells (in vivo) and in lymphocytes (in vitro) using different doses based on LD50. MNNCE (Micronucleated normocromatic erythrocytes) and MNPCE (Micronucleated polychromatic erythrocytes), NDI (Nuclear division index) and NDCI (nuclear division cytotoxicity index), necrotic and apoptotic cells were recorded in rat's bone marrow samples. CA, MI (number of cells undergoing mitosis) necrotic, and apoptotic cells recorded in lymphocytes. Results showed that there was a slight increase in the frequency of micronucleated bone marrow cells. However, no structural chromosomal aberrations were detected in vivo or in vitro. On the other hand, the results showed significant increase in necrotic and apoptotic cells following MTD administration in a dose-dependent manner comparing to positive and negative control groups. In light of these results, MTD can be considered highly cytotoxic and moderate genotoxic, and precaution should be taken when using MTD.

Keywords: methidathion, micronucleus, NDI, NDCI, toxicity, chromosomal aberrations

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5 Genotoxicity of 4-Nonylphenol (4NP) on Oreochromus spilurs Fish

Authors: M. M. Alsharif

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4-Nonylphenol Compound is widely used as an element of detergents, paints, insecticides and many others products. It is known that the existence of this compound may lead to the emission of estrogenic responses in mammals, birds and fish. It is described as pollutant since it causes disorder of endocrine glands. In previous studies, it was proven that this compound exists in water and in the materials precipitated in Red Sea coast in Jeddah near the drains of processed drainage water and near the drainage site of the residuals of paper factories. Therefore, this study aimed to evaluate the cytogenetic aberrations caused by 4-nonylphenol through exposing Talapia Fishes to aquatic solution of the compound with 0, 15, 30 microgram/liter for one month. Samples of gills and liver were collected for micronuclei, nuclear abnormalities and measuring DNA and RNA amount in the treated fish. The results pointed out that there is a significant increase in the numbers of micronuclei in the fish exposed to the former concentrations as compared to the control group. Exposing fishes to 4-nonylphenol resulted in an increased amount of both DNA and RNA, compared to the control group. There is a positive correlation between the amount of the compound (i.e. dosage dependent effect) and the inspiring for cytogenetic effect on Talapia fishes in Jeddah. Therefore, micronucleus test, DNA and RNA contents can be considered as an index of cumulative exposure, which appear to be a sensitive model to evaluate genotoxic effects of 4-Nonylphenol compound on fish.

Keywords: genotoxic, 4-nonylphenol, micronuclei, fish, DNA, RNA

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4 In vivo Evidence of Protective Effect of Hyparrhenia Hirta against Nitrate-Induced Genotoxicity

Authors: H. Bouaziz-Ketata, G. Ben Salah, Z. Aidi, C. Kallel, H. Kammoun, F. Fakhfakh, N. Zeghal

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The present study was performed to evaluate the potential protective effect of Hyparrhenia hirta methanolic extract in NaNO3-induced genotoxic and hematotoxic effects. Male Wistar rats were randomly divided into three groups: a control group and two treated groups during 50 days with NaNO3 administered at a dose of 400 mg kg-1 bw either alone in drinking water or co-administered with Hyparrhenia hirta at a dose of 200 mg kg-1 bw. NaNO3 treatment showed a significant increase in the frequencies of total chromosomal aberrations, aberrant metaphases and micronucleus in bone-marrow cells. In parallel, the NaNO3-treated group showed a significant decrease in red blood cell count, hemoglobin and hematocrit and a significant increase in total white blood cell, in neutrophil and eosinophil counts. Platelet count, mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration remained unchanged in treated groups compared to those of controls. Hyparrhenia hirta methanolic extract appeared to be effective against genotoxic and hematotoxic changes induced by nitrate, as evidenced by the improvement of the markers cited above.

Keywords: Hyparrhenia hirta, sodium nitrate, erythrocytes, genotoxicity

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3 Radioprotective Effects of Selenium and Vitamin-E against 6Mv X-Rays in Human Volunteers Blood Lymphocytes by Micronuclei Assay

Authors: Vahid Changizi, Aram Rostami, Akbar Mosavi

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Purpose of study: Critical macromolecules of cells such as DNA are in exposure to damage of free radicals that induced from interaction of ionizing radiation with biological systems. Selenium and vitamin-E are natural compound that has been shown to be a direct free radical scavenger. The aim of this study was to investigate the in vivo/in vitro radioprotective effect of selenium and vitamin-E separately and synergistically against genotoxicity induced by 6MV x-rays irradiation in cultured blood lymphocytes from 15 human volunteers. Methods: Fifteen volunteers were divided in three groups include A, B and C. These groups were given slenium(800 IU), vitamin-E(100 mg) and selenium(400 IU) + vitamin-E(50 mg), respectively. Peripheral blood samples were collected from each group before(0 hr) and 1, 2 and 3 hr after selenium and vitamin-E administration (separately and synergistically). Then the blood samples were irradiated to 200 cGy of 6 Mv x-rays. After that, lymphocyte samples were cultured with mitogenic stimulation to determine the chromosomal aberrations wih micronucleus assay in cytokinesis-blocked binucleated cells. Results: The lymphocytes in the blood samples collected at 1 hr after ingestion selenium and vitamin-E, exposed in vitro to x-rays exhibited a significant decrease in the incidence of micronuclei, compared with control group at 0 hr. The maximum protection and decrease in frequency of micronuclei(50%) was observed at 1 hr after administration of selenium and vitamin-E synergistically. Conclusion: The data suggest that ingestion of selenium and vitamin-E as a radioprotector substances before exposures may reduce genetic damage caused by x-rays irradiation.

Keywords: x-rays, selenium, vitamin-e, lymphocyte, micronuclei

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2 Genotoxic and Cytotoxic Effects of Salvia officinals Extracts on Rat Bone Marrow

Authors: Mohammed A. Alshehri

Abstract:

Salvia officinalis is an aromatic plant member of the mint (Labiatae) family. It is popular kitchen herb. Not surprise to find that the name of this herb related to cure, in Latin language Salvia means to cure where officinalis means medicinal which answer why the sage has a top place in the list of medicinal plants. The aim of the present study was to assess the genetic damage and cytological changes caused by exposure of the test organism (Rattusrattus) to Salvia officinals. For this purpose, adult female rats, weighing 200–250 g, were used as donors. A total of 36 adult Wister male rats were randomly assigned to five groups: the experimental groups (rats were intraperitonealy injected with Salvia officinalis pure extract at (0.1, 0.2, 0.5, 0.1mg/kg body weight, the same dose was administered once a day. Control group (rats were injected intraperitonealy physiological saline. And positive control were injected with Cyclophosphamide. On the 21st days following Salvia officinalis pure extract exposure, rats were sacrificed, and samples of bone marrow were collected. Following that, we performed a micronuclei (MN) test using MNNCE (Micro-nucleated normocromatic erythrocytes) and MNPCE (Micronucleated polychromatic erythrocytes), NDI (Nuclear division index), and cytological parameters using NDCI (nuclear division cytotoxicity index), necrotic, and apoptotic cells in rat's bone marrow samples. Results showed that there was a no significant increase in the frequency of micro-nucleatedas well as in cytological parameters in bone marrow cells. In light of these results, if Salvia officinalis pure extract may considered to be safe from the stand point of genotoxicity and cytotoxicity effects.

Keywords: Salvia officinalis, micronucleus, NDI, NDCI, toxicity, chromosomal aberrations

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1 The Usefulness of Premature Chromosome Condensation Scoring Module in Cell Response to Ionizing Radiation

Authors: K. Rawojć, J. Miszczyk, A. Możdżeń, A. Panek, J. Swakoń, M. Rydygier

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Due to the mitotic delay, poor mitotic index and disappearance of lymphocytes from peripheral blood circulation, assessing the DNA damage after high dose exposure is less effective. Conventional chromosome aberration analysis or cytokinesis-blocked micronucleus assay do not provide an accurate dose estimation or radiosensitivity prediction in doses higher than 6.0 Gy. For this reason, there is a need to establish reliable methods allowing analysis of biological effects after exposure in high dose range i.e., during particle radiotherapy. Lately, Premature Chromosome Condensation (PCC) has become an important method in high dose biodosimetry and a promising treatment modality to cancer patients. The aim of the study was to evaluate the usefulness of drug-induced PCC scoring procedure in an experimental mode, where 100 G2/M cells were analyzed in different dose ranges. To test the consistency of obtained results, scoring was performed by 3 independent persons in the same mode and following identical scoring criteria. Whole-body exposure was simulated in an in vitro experiment by irradiating whole blood collected from healthy donors with 60 MeV protons and 250 keV X-rays, in the range of 4.0 – 20.0 Gy. Drug-induced PCC assay was performed on human peripheral blood lymphocytes (HPBL) isolated after in vitro exposure. Cells were cultured for 48 hours with PHA. Then to achieve premature condensation, calyculin A was added. After Giemsa staining, chromosome spreads were photographed and manually analyzed by scorers. The dose-effect curves were derived by counting the excess chromosome fragments. The results indicated adequate dose estimates for the whole-body exposure scenario in the high dose range for both studied types of radiation. Moreover, compared results revealed no significant differences between scores, which has an important meaning in reducing the analysis time. These investigations were conducted as a part of an extended examination of 60 MeV protons from AIC-144 isochronous cyclotron, at the Institute of Nuclear Physics in Kraków, Poland (IFJ PAN) by cytogenetic and molecular methods and were partially supported by grant DEC-2013/09/D/NZ7/00324 from the National Science Centre, Poland.

Keywords: cell response to radiation exposure, drug induced premature chromosome condensation, premature chromosome condensation procedure, proton therapy

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