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Mushroom extracts have been shown to contain bioactive compounds that confer health benefits to humans and animals, including honey bees. Honey bees are often challenged simultaneously with several abiotic (e.g., pesticides and poor nutrition) and biotic (e.g., parasites and pathogens) stressors. Mushroom extracts, for example, those made from Agaricus bisporus, may hold the potential to mitigate the negative effects of these stressors through actions on the honey bee's immune system, metabolism, and other physiological processes. Exploring the health benefits of mushroom extracts for honey bees requires a basic understanding of their bioactive properties at the cellular level. The aim of the present study was to evaluate, for the first time, the genotoxic and antigenotoxic potential of A. bisporus on the honey bee cell line AmE-711 using the comet assay. Three increasing concentrations of A. bisporus water extract (100, 200, and 400 µg/mL) were tested. The cells in the negative control group were not treated either with A. bisporus extract or with H2O2. In the positive control group, DNA damage was induced with 100 µM H2O2. For antigenotoxic effect, tested extracts were mixed and incubated with H2O2. None of the tested concentrations exerted genotoxic potential, but all showed antigenotoxic effects against H2O2-induced DNA damage. The concentration of 200 μg/mL A. bisporus extract was the most effective in its action against DNA damage. To conclude, A. bisporus extract did not demonstrate genotoxic effects but showed promising antigenotoxic properties. AmE-711 cell line may serve as a cell culture system for genotoxicity investigations. [ABSTRACT FROM AUTHOR]

The present study was undertaken to evaluate cypermethrin (CYP)-induced oxidative stress [reactive oxygen species (ROS) and lipid peroxidation (LPO) in gills, muscles, brain, and liver tissues] and DNA damage/genotoxicity (peripheral blood erythrocytes) in a freshwater teleost rohu (Labeo rohita) and the protective role of vitamin C. The LC50 of CYP against rohu was found to be 4.5 µg/L in a semi-static culture system through probit analysis. Fingerlings of rohu were distributed into four groups (Group 1st served as a control, fed 35% protein basal diet and was not exposed to CYP; Group 2nd was fed a basal diet and exposed to CYP; Group 3rd and Group 4th were fed diets supplemented with vitamin C at the rate of 100 and 200 mg/kg diet, respectively, and exposed to CYP). Fingerlings were reared on a basal and vitamin C-supplemented diet for 28 days prior to exposure to CYP. The results indicate a time-dependent significant increase in ROS and LPO (indicated by time course increase in TBARS level) as well as DNA damage in terms of number of comets, % DNA in tail, tail moment, tail length, and olive tail moment after exposure to LC50 of CYP. However, statistically comparable results in both Groups 1st and 4th indicate the protective role of vitamin C. The results reveal the effectiveness of vitamin C as a feed additive for countering pesticides toxicity in Labeo rohita. The current study indicates CYP as a potential genotoxicant for fish and classifies SCGE as a reliable and sensitive tool for assessing DNA damage. [ABSTRACT FROM AUTHOR]

The present study was undertaken to evaluate cypermethrin (CYP)-induced oxidative stress [reactive oxygen species (ROS) and lipid peroxidation (LPO) in gills, muscles, brain, and liver tissues] and DNA damage/genotoxicity (peripheral blood erythrocytes) in a freshwater teleost rohu (Labeo rohita) and the protective role of vitamin C. The LC50 of CYP against rohu was found to be 4.5 µg/L in a semi-static culture system through probit analysis. Fingerlings of rohu were distributed into four groups (Group 1st served as a control, fed 35% protein basal diet and was not exposed to CYP; Group 2nd was fed a basal diet and exposed to CYP; Group 3rd and Group 4th were fed diets supplemented with vitamin C at the rate of 100 and 200 mg/kg diet, respectively, and exposed to CYP). Fingerlings were reared on a basal and vitamin C-supplemented diet for 28 days prior to exposure to CYP. The results indicate a time-dependent significant increase in ROS and LPO (indicated by time course increase in TBARS level) as well as DNA damage in terms of number of comets, % DNA in tail, tail moment, tail length, and olive tail moment after exposure to LC50 of CYP. However, statistically comparable results in both Groups 1st and 4th indicate the protective role of vitamin C. The results reveal the effectiveness of vitamin C as a feed additive for countering pesticides toxicity in Labeo rohita. The current study indicates CYP as a potential genotoxicant for fish and classifies SCGE as a reliable and sensitive tool for assessing DNA damage.

Copper (Cu) toxicity is a pressing concern for several soils, especially in organic viticulture. The objective of this work was to assess Cu toxicity on the non-target organism Eisenia fetida, employing both traditional and novel tools for early identification of Cu-induced damages. In addition to traditional tests like avoidance and reproductive toxicity experiments, other tests such as the single cell gel electrophoresis (SCGE) and gut microbiome analysis were evaluated to identify early and more sensitive pollution biomarkers. Four sub-lethal Cu concentrations were studied, and the results showed strong dose-dependent responses by the earthworm avoidance test and the exceeding of habitat threshold limit at the higher Cu doses. An inverse proportionality was observed between reproductive output and soil Cu concentration. Bioaccumulation was not detected in earthworms; soil concentrations of potentially bioavailable Cu were not affected by E. fetida presence or by time. On the contrary, the SCGE test revealed dose-dependent genotoxicity for the 'tail length' parameter already at the second day of Cu exposition. Gut microbiome analysis a modulation of microbial composition, with the most aboundant families being Pectobateriaceae, Comamonadaceae and Microscillaceae. Bacillaceae increased over time and showed adaptability to copper up to 165 mg/kg, while at the highest dose even the sensitive Acetobacteriaceae family was affected. The research provided new insights into the ecotoxicity of Cu sub-lethal doses highlighting both alterations at earthworms' cellular level and changes in their gut microbiota. [ABSTRACT FROM AUTHOR]

In criminal investigations, postmortem interval （PMI） is important information to be inferred in homicide investigations, as well as the focus and the difficulty in forensic pathology research. Because the DNA content in different tissues is relatively constant and shows changes regularly with the extension of PMI, it has become a research hotspot of PMI estimation. This paper reviews the recent progress of PMI estimation technologies including DNA-based single cell gel electrophoresis, image analysis, flow cytometry, real-time fluorescence quantitative PCR and high-throughput sequencing, hoping to provide references for forensic medicine practice and scientific research.

Simple Summary: The aim of the present study was to evaluate, for the first time, the genotoxic and antigenotoxic potential of thymol on the honey bee continuous cell line AmE-711 using the Comet assay. Thymol did not show antigenotoxic effect on honey bee cells in any of the tested concentrations (10, 100, and 1000 μg/mL). Two concentrations (100 and 1000 μg/mL) expressed genotoxic effects on cultured honey bee cells, thus suggesting the careful application of thymol in beekeeping practice to avoid possible negative effects on honey bees. Thymol is a natural essential oil derived from the plant Thymus vulgaris L. It is known to be beneficial for human and animal health and has been used in beekeeping practice against Varroa mite for years. In this study, the genotoxic and antigenotoxic potential of thymol were evaluated on the honey bee (Apis mellifera L.) continuous cell line AmE-711 for the first time. Using the Comet assay, three increasing concentrations (10, 100, and 1000 µg/mL) of thymol were tested. Negative control (non-treated cells) and positive control (cells treated with 100 µM H2O2) were also included. The absence of thymol cytotoxicity was confirmed with the Trypan blue exclusion test. Thymol in the concentration of 10 µg/mL did not increase DNA damage in AmE-711 honey bee cells, while 100 and 1000 µg/mL concentrations showed genotoxic effects. For testing the antigenotoxic effect, all concentrations of thymol were mixed and incubated with H2O2. The antigenotoxic effect against was absent at all concentrations (10, 100, 1000 μg/mL) tested. Moreover, thymol enhanced the H2O2-induced DNA migration in the Comet assay. The obtained results indicate genotoxic effects of thymol on cultured honey bee cells suggesting its careful application in beekeeping practice to avoid possible negative effects on honey bees. [ABSTRACT FROM AUTHOR]

Accurate verification of the time passed since death is a challenging task in daily forensic investigations, chiefly in homicidal or unwitnessed death. The study evaluated the applicability of the time-dependent changes in histological structure, DNA content,and oxidant/antioxidant parameters in rat's skeletal muscles in an attempt to reach a reliable method for post-mortem interval (PMI) estimation. We used 56 adult male albino rats. These animals were sacrificed then and allocated randomly to seven groups (eight rats each). First group: rats were dissected to obtain samples from quadriceps femoris muscles promptly after death. While the rats in the second, third, fourth, fifth, sixth and seventh groups were dissected to get muscle samples from quadriceps femoris at 6, 24, 48, 72, 96 and 144 hours post-mortem (hpm). The results showed that a significant gradual histopathological alteration of muscle structure started at 48 hpm and progressed afterwards. Also, the DNA degradation percentage increased significantly parallel to the increased oxidant parameter levels from 6 to 144 hpm. Our results concluded that changes in oxidant/antioxidant parameters and DNA degradation, as well as the histological examination of muscle tissues, showed a strong correlation with post-mortem intervals. Consequently, the integration of all these parameters can provide a valuable method of post-mortem intervals estimation. [ABSTRACT FROM AUTHOR]

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Thymol is a natural essential oil derived from the plant Thymus vulgaris L. It is known to be beneficial for human and animal health and has been used in beekeeping practice against Varroa mite for years. In this study, the genotoxic and antigenotoxic potential of thymol were evaluated on the honey bee (Apis mellifera L.) continuous cell line AmE-711 for the first time. Using the Comet assay, three increasing concentrations (10, 100, and 1000 µg/mL) of thymol were tested. Negative control (non-treated cells) and positive control (cells treated with 100 µM H2O2) were also included. The absence of thymol cytotoxicity was confirmed with the Trypan blue exclusion test. Thymol in the concentration of 10 µg/mL did not increase DNA damage in AmE-711 honey bee cells, while 100 and 1000 µg/mL concentrations showed genotoxic effects. For testing the antigenotoxic effect, all concentrations of thymol were mixed and incubated with H2O2. The antigenotoxic effect against was absent at all concentrations (10, 100, 1000 μg/mL) tested. Moreover, thymol enhanced the H2O2-induced DNA migration in the Comet assay. The obtained results indicate genotoxic effects of thymol on cultured honey bee cells suggesting its careful application in beekeeping practice to avoid possible negative effects on honey bees.

Background: Most plants encounter water stress at one or more different stages of their life cycle. The maintenance of genetic stability is the integral component of desiccation tolerance that defines the storage ability and long-term survival of seeds. Embryonic axes of desiccation-sensitive recalcitrant seeds of Acer pseudoplatnus L. were used to investigate the genotoxic effect of desiccation. Alkaline single-cell gel electrophoresis (comet assay) methodology was optimized and used to provide unique insights into the onset and repair of DNA strand breaks and 8-oxo-7,8-dihydroguanine (8-oxoG) formation during progressive steps of desiccation and rehydration. Results: The loss of DNA integrity and impairment of damage repair were significant predictors of the viability of embryonic axes. In contrast to the comet assay, automated electrophoresis failed to detect changes in DNA integrity resulting from desiccation. Notably, no significant correlation was observed between hydroxyl radical (٠OH) production and 8-oxoG formation, although the former is regarded to play a major role in guanine oxidation. Conclusions: The high-throughput comet assay represents a sensitive tool for monitoring discrete changes in DNA integrity and assessing the viability status in plant germplasm processed for long-term storage. [ABSTRACT FROM AUTHOR]

Single cell gel electrophoresis was used to observe the effect of bifenthrin and chlorpyrifos on Labeo rohita. The present work was planned to study the genotoxic effect of pesticide mixture on the major indian carp, L. rohita at sub-lethal concentration 33% LC50. First of all, 96-hr LC50 was determined for L. rohita, then fish was given sub-lethal environment for 70 days. Erythrocytes from blood were taken after each fortnight for the assessment of time dependent DNA damage (% damage), aggregate tail length of comets and genomic destruction. There was increase in damage up to 56 days and after that a slight decrease was observed in next fourteen days. After statistical analysis, it was concluded that the Single Cell Gel Electrophoresis may be implemented in ecofriendly observer courses in evaluation of toxicity.

Abstract Time since death (TSD) estimation is a crucial issue in death investigation. In majority of homicide/suicide cases deceased body recovered within the first 48 h, therefore, it is critically important to determine the time of death quickly and precisely. In the present scenario, TSD estimation still remains difficult even for experienced pathologists undertaken with extreme caution. After death numerous factors may be of cadaveric or environmental origin can influence the ‘normal’ rate of postmortem changes. Therefore, it could be more difficult to estimate longer TSD. However, these environmental influencing factors can also effect on DNA in the form of degradation, which start after 1–2 h of death. This DNA degradation could be work as molecular clock which help to estimate early TSD. Single-cell gel electrophoresis (SCGE), also known as the Comet assay, is the technique by which DNA degradation can be visualized and measured qualitatively as well as quantitatively. Using this assay not only early TSD estimated but many other challenges such as time since deposition of biological fluids, repair of genetic material from degraded biological sample could also be resolved. With the help of this paper an attempt was made to introduce a well-known cytogenetic technique that is SCGE assay, which could be a versatile technique for forensic science.

DNA damage is one of the most important consequences of oxidative stress in the cells. If DNA repair is unable to modify these inducible DNA damages, genomic instability may lead to mutation, cancer, aging and many other diseases. Single cell gel electrophoresis or comet assay is a common and versatile method to quantify these types of DNA damages. DNA damages induced by hydrogen peroxide (H2O2) are one of the proper models for measurement of protective ability of different compounds. So the main aim of this review is to provide an overview about protection ability of medicinal plants and their potential mechanism against H2O2 induced DNA damages. In this review, relevant researches on the effect of medicinal plants on DNA damages induced by H2O2 and possible molecular mechanisms are discussed. It seems that, medicinal plants are considered as therapeutic key factors to protect DNA from consequences caused by oxidative stress. Sufficient in vitro evidences introduce them as DNA protective agents through different mechanisms including antioxidant activity and some other cellular mechanisms. Moreover, in order to correlate the antigenotoxicity effects with their potential antioxidant property, most of medicinal plants were evaluated in term of antioxidant activity using standard methods. This review highlights the preventive effects of herbal medicine against oxidative DNA damages as well as provides rational possibility to engage them in animal studies and future clinical investigations.

• The comet assay in assessing genomic stability in relation to folate status and human cancer. • Impact of folate deficiency/supplementation on genomic stability in vitro , in rodents and humans. • The impact of MTHFR genotype on folate status and genomic stability in human carcinogenesis. • Adaptation of the comet assay in measuring DNA methylation and impact on diet and epigenetics. Intake of folate (vitamin B 9) is strongly inversely linked with human cancer risk, particularly colon cancer. In general, people with the highest dietary intake of folate or with high blood folate levels are at a reduced risk (approx. 25%) of developing colon cancer. Folate acts in normal cellular metabolism to maintain genomic stability through the provision of nucleotides for DNA replication and DNA repair and by regulating DNA methylation and gene expression. Folate deficiency can accelerate carcinogenesis by inducing misincorporation of uracil into DNA, by increasing DNA strand breakage, by inhibiting DNA base excision repair capacity and by inducing DNA hypomethylation and consequently aberrant gene and protein expression. Conversely, increasing folate intake may improve genomic stability. This review describes key applications of single cell gel electrophoresis (the comet assay) in assessing genomic instability (misincorporated uracil, DNA single strand breakage and DNA repair capacity) in response to folate status (deficient or supplemented) in human cells in vitro , in rodent models and in human case-control and intervention studies. It highlights an adaptation of the SCGE comet assay for measuring genome-wide and gene-specific DNA methylation in human cells and colon tissue. [ABSTRACT FROM AUTHOR]

We employed the comet assay (single cell gel electrophoresis) to evaluate induced DNA damage in nuclei isolated from tobacco leaves (Nicotiana tabacum var. xanthi) inoculated with Potato virus X (PVX). The highest DNA damage, expressed by the tail moment value, was observed in the inoculated leaves and decreased in the 1st to 4th systemic leaves. DNA damage increased with the time after the inoculation (from day 3 to day 21) and was higher in nuclei isolated from a part of the leaf at the petiole compared to nuclei isolated from the leaf tip. A Pearson moment correlation (r = 0.94) between the induced DNA damage and the PVX titres expressed by ELISA absorbance values was observed. The PVX infection did not induce a significant increase in the rate of somatic mutations evaluated by appearance of dark green, yellow, and double green/yellow sectors on the heterozygous pale green leaves of N. tabacum var. xanthi.

According to global estimates, at least 107,000 people die each year from asbestos-related lung cancer, mesothelioma, and asbestosis resulting from occupational exposure. Chrysotile accounts for approximately 90% of asbestos used worldwide. Artificial substitutes can also be cytotoxic to the same degree as chrysotile. But only a few researchers focused on their genetic effects and mutagenicity information which is useful in evaluating the carcinogenicity of chemicals. In this study, chrysotile from Mangnai, Qinghai, China, and an artificial substitute, rock wool fiber were prepared as suspensions and were tested at concentrations of 50, 100, and 200 μg/ml in V79 lung fibroblasts. Chromosome aberrations were detected by micronucleus assay after exposure for 24 h, and DNA damage were estimated by single cell gel electrophoresis after exposure for 12, 24, or 48 h. According to the results, chrysotile and rock wool fibers caused micronuclei to form in a dose-dependent manner in V79 cells; olive tail moment values increased in a dose- and time-dependent manner. When V79 cells were exposed to a concentration of 200 μg/ml, the degree of DNA damage induced by chrysotile fibers was greater than rock wool fibers. Our study suggests that both chrysotile and rock wool fibers could induce chromosome aberrations and DNA damage. These materials are worthy of further study. [ABSTRACT FROM AUTHOR]

DNA damage is one of the most important consequences of oxidative stress in the cells. If DNA repair is unable to modify these inducible DNA damages, genomic instability may lead to mutation, cancer, aging and many other diseases. Single cell gel electrophoresis or comet assay is a common and versatile method to quantify these types of DNA damages. DNA damages induced by hydrogen peroxide (H2O2) are one of the proper models for measurement of protective ability of different compounds. So the main aim of this review is to provide an overview about protection ability of medicinal plants and their potential mechanism against H2O2 induced DNA damages. In this review, relevant researches on the effect of medicinal plants on DNA damages induced by H2O2 and possible molecular mechanisms are discussed. It seems that, medicinal plants are considered as therapeutic key factors to protect DNA from consequences caused by oxidative stress. Sufficient in vitro evidences introduce them as DNA protective agents through different mechanisms including antioxidant activity and some other cellular mechanisms. Moreover, in order to correlate the antigenotoxicity effects with their potential antioxidant property, most of medicinal plants were evaluated in term of antioxidant activity using standard methods. This review highlights the preventive effects of herbal medicine against oxidative DNA damages as well as provides rational possibility to engage them in animal studies and future clinical investigations. [ABSTRACT FROM AUTHOR]

Time since death (TSD) estimation is a crucial issue in death investigation. In majority of homicide/suicide cases deceased body recovered within the first 48 h, therefore, it is critically important to determine the time of death quickly and precisely. In the present scenario, TSD estimation still remains difficult even for experienced pathologists undertaken with extreme caution. After death numerous factors may be of cadaveric or environmental origin can influence the 'normal' rate of postmortem changes. Therefore, it could be more difficult to estimate longer TSD. However, these environmental influencing factors can also effect on DNA in the form of degradation, which start after 1-2 h of death. This DNA degradation could be work as molecular clock which help to estimate early TSD. Single-cell gel electrophoresis (SCGE), also known as the Comet assay, is the technique by which DNA degradation can be visualized and measured qualitatively as well as quantitatively. Using this assay not only early TSD estimated but many other challenges such as time since deposition of biological fluids, repair of genetic material from degraded biological sample could also be resolved. With the help of this paper an attempt was made to introduce a well-known cytogenetic technique that is SCGE assay, which could be a versatile technique for forensic science. [ABSTRACT FROM AUTHOR]

AIM: To observe the situation of rat retinal tissue DNA damage at early diabetic period, discuss the role of the blood glucose fluctuations, and provide a new method for studying the pathogenesis of diabetic retinopathy(DR).METHODS: SD rats were randomly divided into four groups: normal control group(NC), normal fluctuation group(NF), diabetes group(DM)and diabetes fluctuation group(DF). Diabetic models were established through intraperitoneal injection of STZ. A certain amount of glucose was injected in the rats of group NF and DF in an intraperitoneal mode three times a day after the model was established, thereby causing blood glucose fluctuations. Rats were killed and the retinal tissues were taken in the 8th week. Single cell gel electrophoresis(SCGE)technique was adopted for detecting DNA injury extent in the retina tissue. RESULTS:Groups NF and DF showed significant and regular fluctuations. The curve of blood glucose fluctuations was relatively stable. All values of MBG, SDBG, LAGE and M were significantly increased compared with group NC. Group DF was increased more significantly. It was statistically significant(PPPCONCLUSION: Rat retinal tissues have DNA injury during early diabetic period. DNA injury is gradually aggravated with blood glucose fluctuation. It indicates that high blood glucose and blood glucose fluctuation are involved in the mechanism of cell DNA injury, and they may be one of DR early event, have played a certain role in the incidence of DR.

Cyclosativene (CSV) is a tetracyclic sesquiterpene found in the essential oils of Centaurea cineraria (Asteraceae) and Abies magnifica A. Murray (Pinaceae) plants. To the best of our knowledge, its cytotoxic, genotoxic and oxidant effects have never been studied on any cell lines. Therefore, we aimed to investigate the in vitro antiproliferative and/or cytotoxic properties, antioxidant/oxidant activity and genotoxic damage potential of CSV in healthy neurons and N2a neuroblastoma (N2a-NB) cell cultures. After treatment with 10-400 μg/ml of CSV for 24 h, cell proliferation was measured by the MTT (3,(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The antioxidant activity was assessed by the total antioxidant capacity (TAC) and total oxidative stress (TOS) assays. To evaluate the level of DNA damage, single cell gel alkaline electrophoresis (SCGE) was used. The MTT assay showed that the application of CSV significantly reduced cell viability in both cell types. CSV treatments at higher doses led to decreases of TAC levels and increases of TOS levels in neuron and N2a-NB cells. The mean values of the total scores of cells showing DNA damage were not found to be significantly different from the control values in both cells. In conclusion, this study suggests that CSV has weak anticancer potential.

Reactive species such as free radicals are constantly generated in vivo and DNA is the most important target of oxidative stress. Oxidative DNA damage is used as a predictive biomarker to monitor the risk of development of many diseases. The comet assay is widely used for measuring oxidative DNA damage at a single cell level. The analysis of comet assay output images, however, poses considerable challenges. Commercial software is costly and restrictive, while free software generally requires laborious manual tagging of cells. This paper presents OpenComet, an open-source software tool providing automated analysis of comet assay images. It uses a novel and robust method for finding comets based on geometric shape attributes and segmenting the comet heads through image intensity profile analysis. Due to automation, OpenComet is more accurate, less prone to human bias, and faster than manual analysis. A live analysis functionality also allows users to analyze images captured directly from a microscope. We have validated OpenComet on both alkaline and neutral comet assay images as well as sample images from existing software packages. Our results show that OpenComet achieves high accuracy with significantly reduced analysis time.

Perfluorinated compounds (PFC s ) such as PFOS and PFOA, are xenobiotics that can be detected worldwide in the environment and humans. PFOS (C 8 F 17 SO 3 − ) is a fluorinated organic compound has been used for decades in industrial and commercial products. We investigated the genotoxic and apoptotic impact of PFOS in rat liver using comet assay, micronucleus test and apoptotic gene expression methods for caspase 3, caspase 8 and the protective role of curcumin on the PFOS- induced damage under chronic exposure. In this study, rats were treated either with three different PFOS doses only (0.6, 1.25 and 2.5 mg/kg) or one dose of curcumin (80 mg/kg) or three different doses of PFOS combined with 80 mg/kg dose of curcumin by gavage for 30 days at 48 h intervals. We evaluated the DNA damage via comet assay and micronucleus test. Doses of PFOS increased micronucleus frequency (p < 0.05) and strongly induced DNA damage in liver in two different parameters; i: the damaged cell percentage and ii: genetic damage index. Curcumin prevented the formation of DNA damage induced by PFOS and curcumin substance applied with PFOS caused a decrease in the micronucleus frequency. PFOS increased apoptotic gene expression but curcumin decreased the expression levels of caspase 3 and 8. [ABSTRACT FROM AUTHOR]

The present study explores the in vivo and in vitro genotoxic effects of lead nitrate, [Pb(NO)] a recognized environmental pollutant and cobalt chloride (CoCl), an emerging environmental pollutant in polychaete Perinereis cultrifera using comet assay . Despite widespread occurrence and extensive industrial applications, no previous published reports on genotoxicity of these compounds are available in polychaete as detected by comet assay. Polychaetes were exposed in vivo to Pb(NO) (0, 100, 500, and 1000 μg/l) and CoCl (0, 100, 300, and 500 μg/l) for 5 days. At 100 μg/l Pb(NO) concentration, tail DNA (TDNA) values in coelomocytes were increase by 1.16, 1.43, and 1.55-fold after day 1, day 3, and day 5, whereas, OTM showed 1.12, 2.33, and 2.10-fold increase in in vivo. Pb(NO) showed a concentration and time-dependent genotoxicity whereas CoCl showed a concentration-dependent genotoxicity in in vivo. A concentration-dependent increase in DNA damage was observed in in vitro studies for Pb(NO) and CoCl. DNA damage at 500 μg/L showed almost threefold increase in TDNA and approximately fourfold increase in OTM as compared to control in in vitro. Our studies suggest that Pb(NO) and CoCl have potential to cause genotoxic damage, with Pb(NO) being more genotoxic in polychaete and should be used more carefully in industrial and other activities. [ABSTRACT FROM AUTHOR]

The aim of the present study was to investigate the effects of eicosapentaenoic acid (EPA) supplementation during in vitro maturation (IVM) of bovine oocytes. The concentrations tested in all experiments were 1 nM, 1 μM, and 1 mM EPA. The effect of EPA was evaluated on cumulus-oocyte complexes (COC) by oocyte maturation (cumulus expansion area and oocyte nuclear maturation), genotoxicity [single cell gel electrophoresis (SCGE)], and cytotoxicity (apoptosis, viability, and MTT assays) end points. The maturation parameters were affected by exposure of COC to different EPA concentrations in the IVM medium. Cumulus expansion area increased in the presence of 1 nM EPA ( P < 0.05) whereas addition of 1 nM EPA ( P < 0.05) decreased cumulus expansion after 24 h of IVM. Moreover, the maturation rate significantly decreased when 1 mM of EPA was assayed ( P < 0.001). EPA at 1 nM induced genotoxic and cytotoxic effects on bovine cumulus cells (CC) and primary DNA lesions ( P < 0.001). A significant increase in the frequency of apoptotic ( P < 0.01) and necrotic ( P < 0.001) cells was observed after 24 h of treatment with 1 nM, 1 μM, and 1 mM EPA. Mitochondrial activity was altered with 1 mM EPA ( P < 0.001). We inferred that optimal oocyte quality was partially dependent on the presence of adequate EPA concentrations; EPA could be beneficial to improve oocyte quality in the maturation process, because low concentration tested (1 nM EPA) improved cumulus expansion. [ABSTRACT FROM AUTHOR]

Due to the widespread use of silver nanoparticles in consumer products, the toxicity of silver nanoparticles has also been studied in relation to their application. However, most genotoxicity studies of silver nanoparticles have been performed in vitro. Therefore, this study evaluated the DNA damage to lung cells caused by repeated inhalation of silver nanoparticles. Male Sprague Dawley rats were exposed to silver nanoparticles for 12 weeks in a whole-body inhalation chamber. The animals were divided into one control group and three dose groups that were exposed to silver nanoparticles (14–15 nm diameter) at concentrations of 0.66 × 106 particles/cm3 (49 μg/m3, low dose), 1.41 × 106 particles/cm3 (117 μg/m3, middle dose), and 3.24 × 106 particles /cm3 (381 μg/m3, high dose), respectively, for six hours/day over 12 weeks. The rats were sacrificed after the 12-week exposure period and the DNA damage assessed using a Comet assay of cells obtained from the right lungs. The olive tail moment values were 2.93 ± 0.19, 3.81 ± 0.23, 3.40 ± 0.22, and 5.16 ± 0.32 for the control, low-, middle-, and high-dose groups, respectively. Although no dose-dependent results were observed, a significant increase in the level of DNA damage was noted for the high-dose group.

Introduction: Drug delivery systems could induce cellular toxicity as side effect of nanomaterials. The mechanism of toxicity usually involves DNA damage. The comet assay or single cell gel electrophoresis (SCGE) is a sensitive method for detecting strand damages in the DNA of a cell with applications in genotoxicity testing and molecular epidemiology as well as fundamental research in DNA damage and repair. Methods: In the current study, we reviewed recent drug delivery researches related to SCGE. Results: We found that one preference for choosing the assay is that comet images may result from apoptosis-mediated nuclear fragmentation. This method has been widely used over the last decade in several different areas. Overall cells, such as cultured cells are embedded in agarose on a microscope slide, lysed with detergent, and treated with high salt. Nucleoids are supercoiled DNA form. When the slide is faced to alkaline electrophoresis any breakages present in the DNA cause the supercoiling to relax locally and loops of DNA extend toward the anode as a ‘‘comet tail’’. Conclusion: This article provides a relatively comprehensive review upon potentiality of the comet assay for assessment of DNA damage and accordingly it can be used as an informative platform in genotoxicity studies of drug delivery systems.

DNA damage is recognised as the first step in the process of carcinogenesis, induced by endogenous or environmental agents. At low levels, it is best detected with the comet assay (single cell gel electrophoresis), a method based on electrophoresis of lysed cells embedded in agarose; the amount of DNA migrating out to form a comet-like tail indicates the frequency of DNA breaks. Including a digestion with lesion-specific enzymes allows measurement of oxidised or alkylated bases, and UV-induced damage. The comet assay has applications in human biomonitoring, genotoxicity testing, basic research in DNA damage and repair, and environmental monitoring for effects of pollutants.

The comet assays or single cell gel electrophoresis is a simple, rapid, relatively inexpensive and very sensitive method that has been recently widely used to evaluate genomic integrity (or DNA damage) in aquatic animals exposed to diverse physical and chemical agents. However, the accuracy to evaluate comets assay has been the major obstacle in order to determine specific damage undergone cellular stress. The aim of this study was to evaluate DNA fragmentation in sperm cells based in fluorescence image analysis. The species used was the giant mussel Choromytilus chorus. The in vitro experiments were carried out through oxidative stress with five hydrogen peroxide concentrations (0, 50, 100, 500 and 1000 mM). To evaluate DNA damage two groups of parameters were used: (1) Morphological comet parameters as tail length, nuclear diameter (head) and tail moment, and (2) Quantification of DNA strand breaks grade by fluorescence image analysis. The sperm DNA damage was possitively correlated with the morphological parameters, as they were exposed to an increasing dose range of H2O2 and negatively correlated with nuclear diameter. Additionally, fluorescence analysis showed that fragment sizes and the fluorescence intensity could be identified and related the level of DNA damage independently the intercellular variation of DNA strand breaks. The large fragment sizes decrease with H2O2 doses, while medium and small fragments increase in treatments with highest peroxide concentration. This study supported future research to examine comet assay conjugated with molecular approaches, especially in reference to integrate FISH to evaluate if the DNA fragmentation includes specific genes.El ensayo cometa o gel de electroforesis de única célula es un procedimiento sencillo, rápido, relativamente barato y un método muy sensible para evaluar la integridad del genoma (ADN o daños) en los animales acuáticos expuestos a diversos agentes físicos y químicos. Sin embargo, la precisión para evaluar los ensayos cometas han sido el principal obstáculo para determinar el daño específico sufrido por estrés celular. El objetivo de este estudio fue evaluar la fragmentación del ADN en espermatozoides basado en análisis de imágenes fluorescente. La especie utilizada fue el choro zapato, Choromytilus chorus. Los experimentos in vitro se llevaron a cabo por estrés oxidativo con cinco concentraciones de peróxido de hidrógeno (0, 50, 100, 500 y 1000 mM). Para evaluar el daño del ADN se utilizaron dos grupos de parámetros: (1) parámetros morfológicos de los cometas como la longitud de la cola, el diámetro nuclear (cabeza) y el tail moment, y (2) Cuantificación de la fragmentación de ADN mediante análisis de imagen fluorescentes. El daño del ADN en el esperma se correlacionó positivamente con los parámetros morfológicos, a medida que fue expuesto a una mayor dosis de H2O2 y negativamente correlacionado con el diámetro nuclear. Además, el análisis de fluorescencia mostró que los tamaños de los fragmentos y la intensidad de la fluorescencia pueden ser relacionados con el nivel de daño en el ADN independiente de la variación intercelular. Los fragmentos grandes disminuyen con la dosis de H2O2, mientras que los medianos y pequeños aumentan a mayor concentración de peróxido. Este estudio apoya futuras investigaciones para examinar el ensayo cometa en conjunto con enfoques moleculares, especialmente en referencia a la integración de FISH para evaluar si la fragmentación del ADN incluye genes específicos.

The comet assay is a versatile and sensitive method for measuring strand breaks in DNA. The first paper on this single-cell gel electrophoresis assay was published in 1984 by Ostling and Johanson (1984). This assay (with and without inclusion of lesion-specific enzymes) is widely used as a biomarker assay in human population studies - primarily to measure DNA damage, but increasingly also to assess the capacity of cells for DNA repair. Ostling and Johanson were also the first to report experiments to measure DNA repair, by simply following the decrease of DNA damage over time after challenging cells with ionising radiation. However, this approach is time-consuming and laborious, and therefore not ideal for biomonitoring studies, which typically require high-throughput processing of many samples. As an alternative approach, the in vitro comet-based repair assay was developed: a cell extract is incubated with a DNA substrate containing specific lesions, and DNA incisions accumulate. The in vitro comet-based repair assay was first devised to measure base excision repair in lymphocytes (Collins et al., 2001). However, over the past decade it has been modified and improved to study incision of other lesions and thus other repair pathways, as well as being applied to tissue samples in addition to cell suspensions. The application of the in vitro repair assay in dietary intervention, environmental biomonitoring and animal studies will be discussed.

Levels of Reactive Oxygen Species (ROS) molecules during aerobic metabolism are often regulated by unique endogenous antioxidant system. During hyperglycaemic condition, accumulation of excess fatty acids & glucose in adipose tissue (Wright Jr E., 2006) results in increased levels of ROS. When ROS molecules overwhelms the cells antioxidant defence system, it ends up in cellular oxidative stress; which in turn is reported to cause oxidative DNA damage & intervene damage to macromolecules & cellular membranes (Ahmad et al., 2013). Our novel anti-hyperglycaemic polyherbal formulation (ADPHF6) had already illustrated significant inhibitory activity against α-amylase & α-glucosidase enzymes and also scavenging free radicals (in vitro models). The present study demonstrates the protective effect of formulation against H2O2 induced DNA damage in human lymphocytes by Single Cell Gel Electrophoresis (SCGE) assay. Experimental procedures were approved by Institutional Human Ethics Committee of Frontier Lifeline Hospital, Chennai, India (FLL/IEC/02/2014). Peripheral human lymphocytes were isolated (Duthie et.al, 2002) and subjected for Cell viability by Trypan blue exclusion method. The alkaline SCGE assay was carried out to determine the level of DNA damage in ADPHF6 treated cells with minor modifications from Singh et al., 1988. Frosted microscopic slides were pre-coated with 1% NMA followed by 1% LMA and incubated for 15 min at 15-20o C. 100 μL of freshly prepared cell suspension (2 x 104 cells) was mixed with 0.5% LMA & casted on microscopic slide. The cells were immersed in lysing solution for 2 hours at 4O C and washed in TBE buffer for 5 min at RT. All the slides were treated with fresh alkaline solution for 20 minutes for expression of alkali-labile damage. Electrophoresis was performed at 24 V for 20 min at RT. Slides were washed in neutralizing buffer for 5 min at RT. All the groups were stained with Acridine Orange (20µg/ml) & Propidium Iodide (20µg/ml) and visualized under microscope (Nikon Eclipse 80i). Reaction mixture of Lymphocytes (suspended in RPMI-1640) with H2O2 (50 µM) served as Positive Control (PC); Lymphocytes in RPMI-1640 served as Vehicle control; Lymphocytes with H2O2 (50 µM) & Quercetin (50 mM) served as Standard Control. Lymphocytes treated with ADPHF6 aqueous extract (50 µg/ml) & H2O2 (50 µM) in various concentrations served as Test group. In untreated group (Vehicle control), % of tail DNA found to be null. Cells treated with 50 µM H2O2 (Positive control) showed maximum % of tail DNA (>90%) compared to Standard control group (< 30%). ADPHF6 treated cells (50 µg/ml) with 50 µM H2O2 showed significant decrease in % of tail DNA (< 20%). Though mammalian cells are equipped with impressive range of antioxidant enzymes & molecules, but in oxidative stress conditions these agents fails to normalize the redox status of cell. Various scientific investigations in recent times have stated that natural based antioxidants proved to be an alternative therapy for ROS mediated effects. To support this hypothesis, findings from our present study based on SCGE assay clearly suggests that, our anti-hyperglycaemic polyherbal formulation possesses strong antioxidant property in scavenging Free Radical molecules which restore the redox homeostasis in cells. Further studies are required to confirm the potential role of ADPHF6 against Type II Diabetes Mellitus involving in vivo models.

Potential genotoxic/cytotoxic effects of the epoxiconazole/fenpropimorph-based fungicide were investigated using single cell gel electrophoresis and cytogenetic assays: chromosomal aberrations, sister chromatid exchanges, micronuclei and fluorescence in situ hybridization in cultured bovine lymphocytes. No statistically significant elevations of DNA damage and increases in cytogenetic endpoints were seen. However, evident cytotoxic effect presented as a decrease in mitotic and proliferation indices were recorded after exposure of bovine lymphocytes to the fungicide for 24 and 48 h at concentrations ranging from 3 to 15 µg mL−1(P< 0.05,P< 0.01,P< 0.001). Similarly, for 24 h an inhibition in the cytokinesis block proliferation index (CBPI) was obtained after exposure to the fungicide at concentrations ranging from 1.5 to 15 µg mL−1(P< 0.01,P< 0.001) in each donor. [ABSTRACT FROM PUBLISHER]

High intensity exercise can enhance the production of reactive oxygen and nitrogen free radical species, which may cause a number of perturbations to cellular integrity, including deoxyribonucleic acid (DNA) modification. In the absence of adequate DNA repair, it is theoretically possible that several biological disorders may ensue, in addition to premature aging. This striking hypothesis and supposition can only be realized in the presence of sound methodology for the quantification of DNA damage and repair. The alkaline single-cell gel electrophoresis or "comet assay" is a simple and reliable method for measuring the components of DNA stability in eukaryotic cells. The assay is commonly used in research associated with genotoxicology and in human bio-monitoring studies concerned with gene-environment interactions; but is currently less appreciated and under-utilized in the domain of exercise science. No exercise related study for example, has incorporated the comet assay combined with fluorescent in situ hybridization methodology to detect and investigate whole genome, telomeric DNA, or gene region-specific DNA damage and repair in cells. Our laboratory and others have used the comet assay in conjunction with lesion-specific endonucleases to measure DNA strand breaks and oxidized bases to confirm that high intensity exercise can damage and destabilize DNA. Thus, the primary function of this review is to highlight recent advances and innovation with the comet assay, in order to enhance our future understanding of the complex interrelationship between exercise and DNA modification in eukaryotic cells. A brief synopsis of the current literature addressing DNA stability as a function of continuous aerobic exercise is also included. [ABSTRACT FROM AUTHOR]

Objective To investigate the effect of sub-acute exposure to benzene on hematopoietic system in rats and to explore its related mechanism. Methods Thirh male SD rats were raixlomly divided into the blank control group, solvent control group, low-dose benzene group, medium-dose benzene group and high-dose benzene group, with 6 rats in each group. Benzene exposure groups were given 200, 400 and 800 mg/kg benzene solution respectively according to the body mass, the blank control group was not treated, and the solvent control group was given equal volume of corn oil, once a day and for 28 days. Routine blood test was analyzed by automatic blood-counter system. We killed the rats, then calculated the coefficient of liver and spleen, and observed the pathological changes of the femur and the morphological changes of bone marrow and peripheral blood cells. DNA damage of bone marrow and peripheral blood cells were detected by single cell gel electrophoresis (SCGE), and we recorded the tail moment (TM) and Olive tail moment (OTM). Results The WBC of low-dose, medium-dose and high-dose benzene groups were lower than that of the blank control group and the solvent control group (P <0.05). The RBC-in the high-dose benzene group was lower than the other groups, and ihe hemoglobin (Hb) in the medium-dose and high-dose benzene groups was lower than that of the other groups (P < 0.05). There was no significant difference in PLT among these groups (all P > 0.05). There were no significant differences in liver and spleen coefficients among these groups (all P >0.05). In the blank control group and solvent control group, the morphological changes of the bone marrow and peripheral blood cells and the palhologie changes of the bone marrow were basically consistent. The pathological changes showed that in the three benzene groups, the area of hematopoietic tissue was decreased, there were more toxic particles and vacuoles in the bone marrow and peripheral blood cells, meanwhile, with the increased dose of benzene, the above changes were more significant. Compared with the blank control group and solvent control group, TM and OTM were increased in turn in the three benzene groups (P<0. 01). Conclusion Aub-acute ex­posure to benzene has a toxic effect on bone marrow of rats with a dose-dependent manner, which may be rated to DNA damage in peripheral blood and bone marrow cells. [ABSTRACT FROM AUTHOR]

The single cell gel electrophoresis is a common technology for the detection of DNA damage. It has the advantages of high sensitivity, economic, easy and fast. In the past 30 years three decades, this technology has been widely used in genetic toxicology, environmental monito-ring, medical diagnostics and nutrition research. Recently, single-cell gel was widely used in many fields by combining with fluorescence in situ hybridization. This article reviewed the applications of single cell gel electrophoresis in various fields, and discussed its development prospect. [ABSTRACT FROM AUTHOR]

Perfluorooctane sulfonate (PFOS) is a man-made fluorosurfactant and global pollutant. PFOS a persistent and bioaccumulative compound, and it is widely distributed in humans and wildlife. Therefore, it was added to Annex B of the Stockholm Convention on Persistent Organic Pollutants in May 2009. Curcumin is a natural polyphenolic compound abundant in the rhizome of the perennial herb turmeric. It is commonly used as a dietary spice and coloring agent in cooking and anecdotally as an herb in traditional Asian medicine. In this study, male rats were treated with three different PFOS doses (0.6, 1.25, and 2.5 mg/kg) and one dose of curcumin, fromCurcuma longa(80 mg/kg), and combined three doses of PFOS with 80 mg/kg dose of curcumin by gavage for 30 d at 48 h intervals. Here, we investigated the DNA damage via single-cell gel electrophoresis/comet assay and micronucleus test in rat peripheral bloodin vivo. It is found that all doses of PFOS increased micronucleus frequency (p < 0.05) and strongly induced DNA damage in peripheral blood in two different parameters; the damaged cell percent and genetically damage index, and curcumin prevented the formation of DNA damage induced by PFOS. Results showed that curcumin inhibited DNA damage including GDI at certain levels at statistical manner, 30.07%, 54.41%, and 36.99% for 0.6 mg/kg, 1.25 mg/kg, and 2.5 mg/kg. [ABSTRACT FROM PUBLISHER]

The study of DNA damage and its repair is critical to our understanding of human aging and cancer. This review reflects on the development of a simple technique, now known as the comet assay, to study the accumulation of DNA damage and its repair. It describes my journey into aging research and the need for a method that sensitively quantifies DNA damage on a cell-by-cell basis and on a day-by-day basis. My inspirations, obstacles and successes on the path to developing this assay and improving its reliability and sensitivity are discussed. Recent modifications, applications, and the process of standardizing the technique are also described. What was once untried and unknown has become a technique used around the world for understanding and monitoring DNA damage. The comet assay’s use has grown exponentially in the new millennium, as emphasis on studying biological phenomena at the single-cell level has increased. I and others have applied the technique across cell types (including germ cells) and species (including bacteria). As it enters new realms and gains clinical relevance, the comet assay may very well illuminate human aging and its prevention. [ABSTRACT FROM AUTHOR]

The main objective of this study is to investigate the cytotoxic, genotoxic and antioxidant properties of zingiberene (ZBN) in an in vitro rat brain cell culture study. The cytotoxic effect was determined against the rat neuron and N2a neuroblastoma (N2a-NB) cell lines using the 3,(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, while the antioxidant activity was assessed using the total antioxidant capacity (TAC) and total oxidative stress (TOS) assays. The effects on DNA damage were also evaluated in this study by the single cell gel electrophoresis assay. The results indicated that ZBN has an anti-proliferative activity suppressing the proliferation of N2a-NB cells at concentrations over 50 mg L and neuron cells at concentrations over 150 mg L. In addition, ZBN treatments at higher doses (≤50 mg L) led to increases of TOS levels in N2a-NB cell cultures. However 25 mg L of ZBN treatment caused increases of TAC levels in cultured neuron and N2a-NB cell cultures while ZBN at doses of 10-400 mg L did not increase the number of total damage score in both cell lines. This study clearly indicates that ZBN has a significant potential to be used as a natural anticancer agent in cultured N2a-NBs. [ABSTRACT FROM AUTHOR]

In the context of reducing CO 2 emissions to the atmosphere, chemical absorption with amines is emerging as the most advanced technology for post-combustion CO 2 capture from exhaust gases of fossil fuel power plants. Despite amine solvent recycling during the capture process, degradation products are formed and released into the environment, among them aliphatic nitramines, for which the environmental impact is unknown. In this study, we determined the acute and chronic toxicity of two nitramines identified as important transformation products of amine-based carbon capture, dimethylnitramine and ethanolnitramine, using a multi-trophic suite of bioassays. The results were then used to produce the first environmental risk assessment for the marine ecosystem. In addition, the in vivo genotoxicity of nitramines was studied by adapting the comet assay to cells from experimentally exposed fish. Overall, based on the whole organism bioassays, the toxicity of both nitramines was considered to be low. The most sensitive response to both compounds was found in oysters, and dimethylnitramine was consistently more toxic than ethanolnitramine in all bioassays. The Predicted No Effect Concentrations for dimethylnitramine and ethanolnitramine were 0.08 and 0.18 mg/L, respectively. The genotoxicity assessment revealed contrasting results to the whole organism bioassays, with ethanolnitramine found to be more genotoxic than dimethylnitramine by three orders of magnitude. At the lowest ethanolnitramine concentration (1 mg/L), 84% DNA damage was observed, whereas 100 mg/L dimethylnitramine was required to cause 37% DNA damage. The mechanisms of genotoxicity were also shown to differ between the two compounds, with oxidation of the DNA bases responsible for over 90% of the genotoxicity of dimethylnitramine, whereas DNA strand breaks and alkali-labile sites were responsible for over 90% of the genotoxicity of ethanolnitramine. Fish exposed to > 3 mg/L ethanolnitramine had virtually no DNA left in their red blood cells. [ABSTRACT FROM AUTHOR]

To detect the genetic toxicity of heavy metals( Pb2+、Cd2+) in bone marrow cell DNA of mice, and to provide the reference for the study of genetic toxicity of lead and cadmium. We infected the mice by gavage with the exposure dose of 1 /32LD50,1 /16LD50,1 /8LD50,1 /4LD50 and 1 /2LD50 respectively, namely Pb( NO3)2( 3. 75 mg /L,7. 5mg / L,15 mg / L,30 mg / L,60 mg / L) and Cd Cl2( 5. 84 mg /L,11. 69 mg /L,23. 38 mg /L,46. 75 mg /L,93. 50 mg / L), and used distilled water to compare with them. Single cell gel electrophoresis( SCGE) was used to detect DNA damage, the results were analyzed by CASP comet image analysis software. The results showed that Pb2+and Cd2+can both increase proportions of the comet cell in mice bone marrow cells, and each damage index of the comet cell of exposed groups was significantly different from that of a control group. The experiment showed some dose effect relationship of genetic-toxicology, and indicated that the heavy metal Pb2+and Cd2+can cause a certain degree of DNA injury of mouse bone marrow cells and genetic toxicity of Cd is higher than that of Pb. Microelectrophoresis bath was used to improve the SCGE, and got significant effects. [ABSTRACT FROM AUTHOR]

Introduction: The present study was designed to investigate the in vitro antioxidant and macromolecule damage protective effects of Asparagus racemosus water (AWE) and methanolic (AME) fractions of roots. Methods: The in vitro antioxidant activity of AWE/AME was estimated by free radical scavenging assays. The DNA damage of HT29 and C2C12 cells was analyzed by comet assay. The plasmid DNA damage and protein oxidation were carried out by agarose gel electrophoresis and SDS-PAGE analysis respectively, where as lipid peroxidation was performed by TBARS assay. Results: Both the extracts showed scavenging activity with IC50 values of 417.4 ± 19.5 / 298 ± 13.5, 381 ± 18.2 / 235 ± 11.9, 54.8 ± 2.95 / 31.6 ± 1.52, 28.9 ± 1.73 / 19.7 ± 1.55 μg/mL for DPPH, metal chelating, ABTS and Nitric oxide scavenging activities respectively. Similarly the methanolic extract showed more potent reducing power and total antioxidant activities over water fraction. The AME showed 56.8% and 41.2% protection against H2O2 (Hydrogen peroxide) induced DNA damage of HT29 human colon cells and C2C12 murine myoblasts. The extract also showed protection against H2O2 induced plasmid DNA damge, AAPH induced protein oxidation of bovine serum albumin (BSA) and lipid peroxidation of rat hepatic tissue. Conclusion: Over all this study showed remarkable antioxidant and macromolecule damage protective effects of A.racemosus. The observed biological properties may be attributed to the high content phenols and flavonoids in the methanolic extract A. racemosus over water extract. [ABSTRACT FROM AUTHOR]

Fine particles with a characteristic size smaller than 100 nm (i.e. nanoparticlesspread out in nowadays life. Silicon or Si, is one of the most abundant chemical elements found on the Earth. Its oxide forms, such as silicate (SiO4) and silicon dioxide, also known as silica (SiO2), are the main constituents of sand and quartz contributing to 90% of the Earth's crust. In this work, three genotoxicity systems 'sister chromatid exchange, cytokinesis block micronucleus test and single cell gel electrophoresis (comet) assay' were employed to provide further insight into the cytotoxic and mutagenic/genotoxic potential of SiO2 nanoparticules (particle size 6 nm, 20 nm, 50 nm) in cultured peripheral blood lymphocytes as in vitro. It was observed that there is a significant decrease in Mitotic index (MI), Cytokinesis block proliferation index (CBPI), proliferation index (PRI) values expressed as Cell Kinetic parameters compared with negative control ( p < 0.05). There is a statistically significant difference between negative control culture and culture exposed to SiO2 (6 nm, 20 nm, 50 nm) ( p < 0.01, p < 0.01, p < 0.05, respectively). It is found that SiO2 nanoparticles at different size (6, 20, 50 nm) progressively increased the SCE frequency and DNA damage on the basis the AU values compared with negative control ( p < 0.05). Results showed that the genotoxic/mutagenic and cytotoxic effects of SiO2 nanoparticules is dependent to particule size. [ABSTRACT FROM AUTHOR]

In this study, the genotoxic effects of commonly used eco-friendly pesticides selected from herbicide (tribenuron methyl), fungicide (triadimenol) and insecticide (lambdacyhalothrin) were evaluated on mononuclear leukocytes isolated from higher cells using the single cell gel electrophoresis (SCGE) assay. Mononuclear cells were exposed for a period of 30 min to 50-, 25-, 15-, 10- and 5 µg ml-1 doses of lambda-cyhalothrin and triadimenol while tribenuron methyl was applied at 10-, 7.5-, 5-, 2.5- and 1 µg ml-1 doses on those cells. DNA damages characterized with Olive tail moment (OTM), comet length and % Tail DNA (% TDNA) increased while % Head DNA (% H-DNA) values significantly decreased with the increase of concentrations of all pesticides used. The results showed that the pesticide lambda-cyhalothrin was found more genotoxic than triadimenol and tribenuron methyl at all doses. However, the lowest doses of triadimenol and tribenuron methyl (last two doses) had only marginal DNA damaging effects on the leukocytes while lambda-cyhalothrin had a remarkable effect at 10 µg ml-1. We conclude that the pesticide lambda-cyhalothrin had a toxic effect on human and could have significant impact on environmental health even if they are applied at low doses. It would be crucially important to determine the effects of genotoxicity and oxidative stress of those pesticides with descending doses on higher cells as well as on plants and animals with extended incubation period if time creates extensive DNA damage. [ABSTRACT FROM AUTHOR]