Tail Moment Values Research Articles

CD276 enhances sunitinib resistance in clear cell renal cell carcinoma by promoting DNA damage repair and activation of FAK-MAPK signaling pathway.

This study aimed to explore the effect of CD276 expression on the sunitinib sensitivity of clear cell renal cell carcinoma (ccRCC) cell and animal models and the potential mechanisms involved. CD276 expression levels of ccRCC and normal samples were analyzed via online databases and real-time quantitative PCR (RT-qPCR). CD276 was knocked down in ccRCC cell models (sunitinib-resistant 786-O/R cells and sunitinib-sensitive 786-O cells) using shRNA transfection, and the cells were exposed to a sunitinib (2 µM) environment. Cells proliferation was then analyzed using MTT assay and colony formation experiment. Alkaline comet assay, immunofluorescent staining, and western blot experiments were conducted to assess the DNA damage repair ability of the cells. Western blot was also used to observe the activation of FAK-MAPK pathway within the cells. Finally, a nude mouse xenograft model was established and the nude mice were orally administered sunitinib (40mg/kg/d) to evaluate the in vivo effects of CD276 knockdown on the therapeutic efficacy of sunitinib against ccRCC. CD276 was significantly upregulated in both ccRCC clinical tissue samples and cell models. In vitro experiments showed that knocking down CD276 reduced the survival rate, IC50 value, and colony-forming ability of ccRCC cells. Knocking down CD276 increased the comet tail moment (TM) values and γH2AX foci number, and reduced BRCA1 and RAD51 protein levels. Knocking down CD276 also decreased the levels of p-FAK, p-MEK, and p-ERK proteins. Knocking down CD276 effectively improved the sensitivity of ccRCC cell and animal models to sunitinib treatment.

Assessment of genotoxicity biomarkers in gasoline station attendants due to occupational exposure.

Gasoline station attendants are exposed to numerous chemicals that might have genotoxic and carcinogenic potential, such as benzene in fuel vapor and particulate matter and polycyclic aromatic hydrocarbons in vehicle exhaust emission. According to IARC, benzene and diesel particulates are Group 1 human carcinogens, and gasoline has been classified as Group 2A "possibly carcinogenic to humans." At gas stations, self-service is not implemented in Turkey; fuel-filling service is provided entirely by employees, and therefore they are exposed to those chemicals in the workplace during all working hours. Genetic monitoring of workers with occupational exposure to possible genotoxic agents allows early detection of cancer. We aimed to investigate the genotoxic damage due to exposures in gasoline station attendants in Turkey. Genotoxicity was evaluated by the Comet, chromosomal aberration, and cytokinesis-block micronucleus assays in peripheral blood lymphocytes. Gasoline station attendants (n = 53) had higher tail length, tail intensity, and tail moment values than controls (n = 61). In gasoline station attendants (n = 46), the frequencies of chromatid gaps, chromosome gaps, and total aberrations were higher compared with controls (n = 59). Increased frequencies of micronuclei and nucleoplasmic bridges were determined in gasoline station attendants (n = 47) compared with controls (n = 40). Factors such as age, duration of working, and smoking did not have any significant impact on genotoxic endpoints. Only exposure increased genotoxic damage in gasoline station attendants independently from demographic and clinical characteristics. Occupational exposure-related genotoxicity risk may increase in gasoline station attendants who are chronically exposed to gasoline and various chemicals in vehicle exhaust emissions.

Oxidative stress, DNA damage, and gene expression in earthworms (Eisenia fetida) exposure to ethiprole.

To evaluate the potential ecotoxicity of ethiprole and early warning to earthworms (Eisenia fetida), different concentrations (0 mg·kg-1, 416mg·kg-1, 625 mg·kg-1, and 1000 mg·kg-1) of ethiprole were added to artificial soil. The key bioindicators were measured and screened at 3 days, 7 days, 14 days, 21 days, and 28 days. The results show that the activity of catalase (CAT) was inhibited for all treatments during the whole exposure period. Besides, the olive tail moment (OTM) value increased gradually as the concentration got higher, which exhibited a dose-time-dependent relationship. Superoxide dismutase (SOD) gene reached the maximum on the 7th day. Mitochondrial large ribosomal RNA (l-rRNA) subunit gene was always in a downregulated state as the concentration increased. Our results show that different concentrations of ethiprole induced certain oxidative stress, DNA damage, and genotoxicity in earthworms. The CAT activity, OTM, and SOD gene could be the most sensitive biomarkers to monitor the toxicity of ethiprole in the soil.

Cinnamaldehyde protects cardiomyocytes from oxygen-glucose deprivation/reoxygenation-induced lipid peroxidation and DNA damage via activating the Nrf2 pathway.

Rapid restoration of perfusion in ischemic myocardium is the most direct and effective treatment for coronary heart disease but may cause myocardial ischemia/reperfusion injury (MIRI). Cinnamaldehyde (CA, C9H8O), a key component in the well-known Chinese medicine cinnamomum cassia, has cardioprotective effects against MIRI. This study aimed to observe the therapeutic effect of CA on MIRI and to elucidate its potential mechanism. H9C2 rat cardiomyocytes were pretreated with CA solution at 0, 10, and 100 μM, respectively and subjected to oxygen-glucose deprivation/reoxygenation (OGD/R). Then the cell viability, the NF-κB and caspase3 gene levels, the reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio, superoxide dismutase (SOD) level, reactive oxygen species (ROS) generation, 4-hydroxynonenal (4-HNE), and malondialdehyde (MDA) were detected. The severity of DNA damage was assessed by tail moment (TM) values using alkaline comet assay. Besides, the DNA damage-related proteins and the key proteins of the Nrf2 pathway were detected by western blot. CA treatment increased the cell viability, GHS/GSSG ratio, SOD level, PARP1, Nrf2, PPAR-γ, and HO-1 protein levels of H9C2 cardiomyocytes, while reducing NF-κB, caspase3, ROS level, 4-HNE and MDA content, γ-H2AX protein level, and TM values. Inhibition of the Nrf2 pathway reversed the effect of CA on cell viability and apoptosis of OGD/R induced H9C2 cardiomyocytes. Besides, 100 μM CA was more effective than 10 μM CA. In the OGD/R-induced H9C2 cardiomyocyte model, CA can protect cardiomyocytes from MIRI by attenuating lipid peroxidation and repairing DNA damage. The mechanism may be related to the activation of the Nrf2 pathway.

Cytotoxicity induced by abamectin in hepatopancreas cells of Chinese mitten crab, Eriocheir sinensis: An in vitro assay

Toxic effects of abamectin on non-target aquatic organisms have been well documented due to its extensive use in both agricultural and aquacultural areas. However, knowledge of the abamectin induced cytotoxicity in crustacean hepatopancreas is still incomplete. In this study, we investigated the cytotoxic effects of abamectin on hepatopancreas cells of Chinese mitten crab, Eriocheir sinensis by an in vitro assay. The results showed that abamectin inhibited cell viability with elevated reactive oxygen species (ROS) and malondialdehyde (MDA) levels in a dose-dependent manner. Increased olive tail moment (OTM) values and 8-hydroxy-2′-deoxyguanosine (8-OHdG) contents indicate the DNA damage under abamectin exposure. The up-regulation of the typical apoptosis-related protein BCL2-associated X protein (Bax) and the down-regulation of B cell leukemia/lymphoma 2 (Bcl-2) demonstrate apoptosis in hepatopancreas cells. Meanwhile, the activities of both caspase-3 and caspase-9 were increased, indicating caspase-mediated apoptosis. In addition, qRT-PCR results showed the up-regulation of antioxidant genes superoxide dismutase (SOD) and catalase (CAT). The mRNA expression of Cap ‘n′ Collar isoform-C (CncC) and c-Jun NH2-terminal kinases (JNK) was also significantly increased, implying the involvement of the Nrf2/MAPK pathway in the antioxidative response. The alteration of innate immune-associated genes Toll-like receptor (TLR) and myeloid differentiation primary response gene 88 (Myd88) also indicates the influence of abamectin on immune status. In summary, the present study reveals the cytotoxicity of abamectin on hepatopancreas cells of E. sinensis and this in vitro cell culture model could be used for further assessment of pesticide toxicity.

Genotoxicity of waterpipe smoking in young adults from Sarajevo, Bosnia & Herzegovina

BackgroundWaterpipe, also known as a hookah or narghile, is a type of tobacco products consumption device. Recently it has been increasingly popular in Bosnia and Herzegovina and the region. Waterpipe consumers are predominantly adolescents and young adults. Many of them believe in slighter harmful effects of waterpipes, compared to cigarettes. We aimed to determine the DNA damage in oral leukocytes and buccal cells of young individuals who have smoked a waterpipe for more than one year. MethodsThe study group consisted of 40 cigarette non-smokers who regularly smoked a waterpipe on average of once per week. As a control, 40 non-smoking individuals were selected to match smokers for age. All participants in the study were healthy male and female adults from Bosnia and Herzegovina, 18–30 years of age. Before sampling, detailed survey and informed consent have been provided by each participant. Comet assay in oral leukocytes and buccal micronucleus cytome (BMCyt) assay in exfoliated buccal cells were applied. ResultsAlmost half of waterpipe smokers (WPS) tasted waterpipe at 15–16 years of age. Comet assay analysis showed increased tail intensity, tail length, and tail moment values among WPS compared to non-smokers (NS) (p = 0.0001, p = 0.0067, and p = 0.0001, respectively). Frequencies of the micronucleated (p = 0.0004), binucleated (p = 0.01), karyorrhectic, (p = 0.0036), and pycnotic cells (p = 0.03) were significantly higher in WPS compared to NS group. ConclusionsGenotoxicity and DNA damage biomarkers were increased in oral leukocytes and exfoliated buccal cells of young waterpipe smokers from Bosnia and Herzegovina, compared to NS group.

Genotoxic effects of oxyclozanide on hemocytes of Galleria mellonella (Lepidoptera: Pyralidae) larvae

Oxyclozanide is a salicylanilide derivative anthelmintic drug with a well-known effect on parasites that cause infections in humans and animals. In this study, the effect of oxyclozanide on DNA damage in hemocytes of Galleria mellonella (L.) (Lepidoptera: Pyralidae) larvae, which has been used as a model organism in many fields, was investigated. Hemolymph was collected from the last instar larvae (7th instar) reared on artificial diets containing oxyclozanide at different concentrations (0.003%, 0.03%, 0.3%, and 1.5%) under laboratory conditions and then hemocytes suspension was prepared. Genotoxic damage in hemocytes was determined by the comet assay which enables microscopically detecting DNA damage and is a very sensitive assay in chemical genotoxicity. When compared to the control group, tail length, tail DNA percent, and tail moment values were significantly increased parallel with increasing oxyclozanide concentrations. While the tail length was determined as 5.11 ± 0.46 μm in the control group, it was significantly increased in all tested groups to 13.17 ± 0.53, 27.98 ± 1.08, 98.44 ± 0.77, and 137.67 ± 0.74 μm, respectively. Similarly to tail length, tail DNA percentage and tail moment levels were also significantly increased from 12.86 ± 0.74 to 91.96 ± 0.31 at the highest concentration of oxyclozanide. These results showed that oxyclozanide caused DNA damage in the hemocytes of G. mellonella. It is also known that hemocytes are an important bioindicator in determining the genotoxicity of anthelmintics to be used as insecticides within environmentally friendly limits. It is thought that our results will contribute to the studies in this field.

Combined Effect of Pesticides Containing Active Ingredients of Chlorpyrifos and Mancozeb on the DNA Damage of Chlorella Sorokiniana Shihira and Krauss

The intensive use of pesticides in agricultural areas can leave residues of pesticide mixtures on the soil surface. Surface run-off can carry pesticide residues, enter water bodies, and then may affect non-target organisms. Chlorpyrifos and mancozeb are active ingredients commonly contained in pesticides applied in shallot farming. This study aims to evaluate the combined effect of pesticides containing active ingredients of chlorpyrifos and mancozeb on the growth and DNA damage of the microalgae Chlorella sorokiniana. The test organism was exposed to the combined concentration of chlorpyrifos:mancozeb, i.e., 0:0, 20%:20%, 20%:80%, 80%:20%, and 80%:80% of the individual EC50 of each pesticide with sampling time at hours 0, 6, 24, and 48. Microalgae growth was estimated by cell counting method, and DNA damage was analyzed by alkaline comet assay method with parameters, i.e., Tail Intensity (TI%), Head Intensity (HI%), Tail Moment (TM), Olive Tail Moment (OTM), and Tail Factor (TF). The results showed that the combined pesticides inhibited the growth of C. sorokiniana, with the highest growth inhibition being at a combined concentration of 80%:80%. The TM and OTM values of C. sorokiniana increased with the increase of combined concentrations at an exposure period of up to 24 hours. In conclusion, the combined exposure could induce growth inhibition and DNA damage of Chlorella sorokiniana, mainly in the first 24 hours. The TM and OTM can be used as sensitive biomarkers for biomonitoring pesticide pollution.

Resolving male infertility that is induced by β-and ca channel antagonist drugs: Propranolol/Verapamil using an optimized nanohybrid formula: Experimental and computational studies

Two major objectives of the current study, the first was to study theoretically the electronic structure of Chitosan-Sodium alginate, Epigallocatechin-gallate (EGCG) and Sodium alginate-EGCG complex using density-functional theory (DFT) calculations. The binding energy (ΔEbind), the atoms in molecules (AIM) theory and the Natural bond orbital (NBO) analysis of Chitosan-Sodium alginate-EGCG complex have been studied in gas and water phases. The second was to investigate the treatment effect of EGCG in normal form and EGCG-loaded chitosan-sodium alginate nanoparticles (EGCG-CS-SA NPs) on the male infertility that was induced in male albino rats by the oral administration of Propranolol Hydrochloride and Verapamil Hydrochloride. Full characterization of the synthesized EGCG-CS-SA NPs were carried out using different techniques. Then eight groups of rats were classified as follows: control negative (A); Propranolol Hydrochloride (B); Verapamil Hydrochloride (C); EGCG normal (D); EGCG nano (EGCG-CS-SA NPs) (E); Propranolol Hydrochloride + EGCG (F); Propranolol Hydrochloride + EGCG-CS-SA NPs (G); and Verapamil Hydrochloride + EGCG-CS-SA NPs (H). The study registered the body weight gain percentage as well the weights of the liver, testes, and seminal vesicles. Malondialdehyde (MDA), glutathione reduced (GSH), and testosterone levels were assessed. The semen analysis was performed, the deoxyribonucleic acid (DNA) damage was detected, and the histopathological sections were examined. The computational study identified that the interaction between chitosan-sodium alginate and EGCG is covalent and electrostatic interactions in nature in the gas and water phases. The results recorded an increase in body weight gain percentage in Propranolol Hydrochloride + EGCG-CS-SA NPs and Verapamil Hydrochloride + EGCG-CS-SA NPs groups but no significant difference (P› 0.05) in organs weight of experimental groups (B, C, D, E, F, G, and H) as compared to control group. In the treated rats (F, G, and H), the MDA decreased significantly, while the GSH increased.The results showed that the formula improved the negative effects of Propranolol Hydrochloride and Verapamil Hydrochloride administration on serum testosterone level, sperm count, motility, percentage of live sperms, and histological structures in the testes and epididymides. Post-administration of EGCG normal and EGCG-CS-SA NPs to Propranolol Hydrochloride and Verapamil Hydrochloride caused no significant difference (P› 0.05) in tail moment value in testes cells as compared to Propranolol Hydrochloride and verapamil Hydrochloride groups. Finally, the herbal nanohybrid formula EGCG-CS-SA NPs was found to be safe and could be used to treat male infertility caused by antihypertensives.

Evaluation of the cytotoxic and genotoxic/antigenotoxic effects of resveratrol in human limbal explant cultures.

Resveratrol (RSV) is a natural polyphenol phytoalexin compound and has long been considered to possess antioxidant and anti-inflammatory effects. In order to exploit the protective potential of RSV in anterior segment diseases, we investigated the possible cytotoxic, genotoxic/antigenotoxic effects of human limbal explant cultures to RSV and MMC or H2O2 alone and in combination. A total of 18 limbal explant tissues obtained from three corneal donors were placed on the 12 well tissue culture polystyrene plates and cultured for 14days. Cell growth from limbal explants was observed by inverted phase contrast microscopy. The cytotoxic effects of RSV was studied by neutral red uptake assay. For the assessment of the genotoxic and antigenotoxic effects, basic alkaline technique of comet assay was performed. It was found that the concentrations of RSV up to 100μM did not significantly affect the viability of outgrowth cells of limbal explant during 24h exposure. When compared to negative control, all concentrations of RSV alone caused an increase in DNA strand breakage. Interestingly, 10μg/mL MMC alone caused similar tail intensity and tail moment values with RSV alone. On the other hand, RSV treatment in all doses seemed to decrease the DNA damage induced by either H2O2 or MMC. RSV is an attractive natural compound for the purpose of oxidative stress reduction in ocular surface and can be utilized as a supplement to promote ocular surface regeneration in vivo.

Evaluation of the biological response of propofol in zebrafish (Danio rerio): Focusing on biochemical, transcriptional, and molecular level

Propofol, one of the most widely used intravenous anesthetic in clinical practice, has been reported to impair cognitive and memory function. However, the toxicological effects of propofol on aquatic organisms are still poorly understood. This study explored the toxic effects of chronic propofol exposure (0.008, 0.04, and 0.2 mg L−1) on adult zebrafish from biochemical, transcriptional, and molecular level after 7, 14, 21 and 28 days of exposure. Results indicated that the reactive oxygen species (ROS) levels were significantly upregulated during the 28 days exposure period, and excessive ROS caused lipid peroxidation, resulting in increased malondialdehyde (MDA) contents in the zebrafish brain. In order to relieve the oxidative damage induced by the excessive ROS, the activities of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT)) were significantly activated, and detoxification enzyme (glutathione S-transferase, GST) activities showed an “activation-inhibition” trend. However, the antioxidant enzymes and detoxification enzyme system could not eliminate the excessive ROS in time and thus caused DNA damage in zebrafish brain. The olive tail moment (OTM) values displayed a “dose-response” relationship with propofol concentrations. Meanwhile, the transcription of related genes of Nrf2-Keap1 pathway was activated. Further molecular simulation experiments suggested that propofol could directly combine with SOD/CAT to change the activity of its biological enzyme. These findings indicated that zebrafish could regulate antioxidant capacity to combat oxidative stress at the early exposure stage, but the activity of antioxidant enzymes were significantly inhibited with the increase of propofol exposure time. Our results are of great importance for understanding toxicological effects of propofol on aquatic organisms.

Investigation of Genotoxic Damage in Overweight Individuals Using the Comet Assay

AbstractObjective: Research has revealed that obesity changed the repair mechanism of DNA chain breaks. Also, the increase in body mass index is found out to be associated with genomic instability. In this study, it was aimed to investigate the possible genotoxic damage of overweight individuals.Material and Methods: In the present study the level of genotoxic damage was calculated in Turkish adult peripheral blood samples using Single Cell Gel Electrophoresis assay. The results of possible DNA damage levels belonging to overweight people were compared statistically by SPSS analysis program with the results of normal-weight people.Results: Fifty five volunteers (21 normal weight and 34 overweight); 23 women, mean age=30.13±7.97 and 32 men, mean age=38.13±10.63 participated in the study. Tail moment is an average of 1.21±0.45 in all individuals. Tail moment value was found of overweight people as 1.29±0.46. When this value was compared with the results of individuals with normal weight (1.09±0.40), statistically no significant difference was determined (p>0.05). According to results, no significant difference was found between the increase in body mass index and DNA damage (p>0.05).Conclusion: The present study is the first study which gives information about the level of DNA damage relation with being overweight in adults of Turkey. In the presented study, it was determined that there was no relationship between body mass index and genotoxic damage according to the findings of the comet assay, and we recommended that new studies should be conducted in the future to investigate the level of DNA damage with different genotoxicity tests.

Cell culture and variations in the comet assay do not affect the genomic integrity of adipose-derived stem cells

The application of quality control tests, such as the comet assay, are essential when adipose-derived stem cells are cultured for therapeutic purposes. However, the steps involved in the development of this assay should be investigated, in order to reduce their influence on genomic damage in cells. The study aimed to evaluate if the cell culture process causes DNA damage, and if variations in the lysis time and pH of the electrophoresis buffer interfere in the genotoxicity results. Four different comet assay protocols were evaluated, and the effects of lysis time and pH conditions of the electrophoresis buffer solution were stated as follows: 2 hours and pH 12; 24 hours and pH 12; 2 hours and pH ≥ 13 and 24 hours and pH ≥ 13. The tail moment was analyzed, and results indicated that at the time cells were detached from the flasks, there was little damage to the DNA in the adipose-derived stem cells, which was confirmed by evaluation of the expression of mRNA genes involved in damage and repair processes of genetic material. Also, the tail moment values ​​did not show significant differences among the four evaluated protocols (p < 0.05), with no indication of damage when compared to the positive control (p < 0.05). Thus, any of the tested protocols can be applied in genotoxicity tests with adipose-derived stem cells, without causing damage to them.

Adaptive Response of Peripheral Blood Lymphocytes in Medical Radiation Workers using the Comet Assay: A Preliminary Studyomet Assay: A Preliminary Study

Background: Gamma irradiation can cause DNA damage in single and double-strand breaks (SSBs & DSBs), especially in peripheral blood lymphocytes. Radiotherapy medical radiation workers can be exposed to gamma radiation related to their daily work. The comet assay is a sensitive method for analyzing DNA damage, especially SSBs. This study explores DNA damage in medical radiation workers’ peripheral blood lymphocytes as an adaptive response using the comet assay.Methods: Blood samples were obtained from four radiotherapy medical radiation workers as a case study (MRW) and two non-medical radiation workers as controls, and then irradiated with various doses of 0, 1, 1.5, and 2 Gy. Lymphocytes were isolated by histopaque and processed by comet assay on the slide under alkaline conditions. The imaging results were analyzed using the Casplab_1.2.3b2 software. The comet assay parameters observed were Tail Length (TL), % Tail DNA (T.DNA), Tail Moment (TM), and Olive Tail Moment (OTM). The one-way ANOVA method was used to analyze statistically between treatment groups. Results: Based on the study results, an increase in TL, T.DNA, TM, and OTM values in all samples was directly proportional to the increase in radiation dose. However, there was no significant difference (P > 0.05) between the MRW group and the control group on each parameter of the comet assay.Conclusions: From this study, it can be concluded that the level of DNA damage of lymphocyte cells as part of the adaptive response in the MRW and control groups was relatively similar after exposure at doses of 0, 1, 1.5, and 2 Gy.

Oxidative stress, DNA damage, and gene expression induced by flufiprole enantiomers in the earthworms (Eisenia fetida)

To evaluate flufiprole enantiomers ecotoxicology and early warning to earthworms in the soil, the effect of flufiprole enantiomers on the activity of antioxidative enzymes (superoxide dismutase, SOD; catalase, CAT; peroxidase, POD), malondialdehyde (MDA) content, DNA damage and expression level of target genes (SOD, TCTP, l-rRNA, and HSP90) have been investigated. Compared to the controls, the activities of SOD, CAT, POD, and MDA content were first inhibited and then stimulated by all concentrations of flufiprole enantiomers in 28 days. The olive tail moment (OTM) value was maximum on the 14th day, then decreased gradually, and the higher the concentration, the slower the decrease. The expression level of the SOD and TCTP genes increased first and then decreased. In conclusion, the toxicity of flufiprole enantiomers to earthworms is: R-(+)- flufiprole >Rac-(±)- flufiprole >S-(−)- flufiprole. The SOD could be the key biomarker for monitoring the risk of flufiprole enantiomers.

Evaluation of Genotoxic and Cytotoxic Activity of Ganoderma Lucidum In Rat Bone Marrow Induced By Lipopolysaccharide

Objective: It was aimed to investigate the antigenotoxic effect of Ganoderma lucidum against the damage caused by lipopolysaccharide (LPS) in rat bone marrow cells by micronucleus test and comet analysis.
 Method: In the experiment, 28 Wistar albino female rats were used and 4 groups were formed. Only physiological saline was applied to the control group. A single dose of 7.5 mg/kg LPS was administered intraperitoneally to the LPS group. LPS+ Ganoderma lucidum (GL): Ganoderma lucidum was given by gavage at a dose of 50 mg/kg per day for one week. Then, on the 8th day, LPS was applied intraperitoneally. Ganoderma lucidum group: GL was given by gavage at 50 mg/kg per day for 1 week. After 24 hours, the rats were euthanized by cervical dislocation under anesthesia. Analyzes were performed for percentage of micronucleated polychromatic erythrocytes (MNPCE) and Polychromatic erythrocytes/Normochromatic erythrocytes (PCE/NCE) ratio and DNA damage in rat femur bone marrow cells.
 Results: When the percentage of micronucleus and PCE/NCE ratio were examined, no statistical significance was observed when the groups were compared both with the control and among themselves. Compared to the control group, it was observed that the comet parameters of the experimental groups (excluding the head DNA) increased significantly. Length tail (L tail), tail DNA, olive tail moment (OTM) values were significantly different both compared to the control and among themselves. While there was a significant difference between LPS+GL group and LPS and GL groups in Tail Moment (TM) value and Length comet (L comet) value, no difference was observed between LPS and GL groups.
 Conclusion: When all of the findings is considered, Ganoderma lucidum cannot be classified as a genotoxic agent. We can say, however, that it reverses the genotoxic damage in issue.

HPRT gene mutation, sister chromatid exchange and comet assay in peripheral lymphocytes of radiation laborers at Al-Amal Cancer Hospital in Baghdad

A Study performed on 30 Iraqi radiation Labors had been exposed tо a low dose tо ionizing radiation at Al-Amal Cancer Hospital in Baghdad the use of three genetic endpoints. Covered 12 females and 18 males between the aged tо (22-57) years. In addition to 20 Healthy individuals from the population, dwelling Baghdad, covered, 7 females and 13 males, aged (19 - 55) years which are non-smokers, non- alcoholic. The cytogenetic analysis, including, HPRT gene mutation¸ comet assay and SCE have been carried out on peripheral blood lymphocytes of labors and control groups. The HPRT gene mutation assay showed a significant increase (p<0.05) in the radiation labors. There were found a significant increase (p<0.05) in comet tail length and tail moment values in thе human lymphocyte in these radiation Labors compared with thе control group. Also the SCE in human lymphocyte for radiation Laborers was significantly (p<0.05) higher than in a control group. Results obtained confirmed the cytogenetic analysis; including, HPRT gene mutation¸ comet assay and SCE are useful as biodosimetric markers for the detection of human exposure to low doses of ionizing radiation. It reflects the simultaneous exposure and actual levels of DNA damage present in radiological laborers' peripheral blood leukocytes by detecting temporary DNA damage and / or repair activity.

The effects of glipizide on DNA damage and nuclear transport in differentiated 3T3-L1 adipocytes.

Despite commonly use for treatment of type II diabetes, possible effects of glipizide on nuclear transport and DNA damage in cells are unknown. Since clinical response of glipizide may change with aging, the aim of the study was to investigate the effect of glipizide by comparing mature and senescent adipocytes. The effects of glipizide were investigated in 3T3-L1 adipocytes. Effective and lethal doses were determined by real-time monitoring iCELLigence system. Comet assay was performed to determine DNA damage and quantitative PCR was conducted to detect gene expression levels. RAN expressions were found to be up regulated in mature 180µM glipizide treated adipocytes compared to control group (p < 0.05); whereas down regulated in senescent 180µM glipizide treated adipocytes compared to their control adipocytes (p < 0.05). Olive Tail Moment values were significantly higher in mature 180µM glipizide treated adipocytes (MTG) and senescent 180µM glipizide treated adipocytes (STG) comparing their untreated controls (p < 0.001 and p < 0.001 respectively). Also class 5 comets that shows severe DNA damage were found to be higher in both MTG and STG groups than their controls (p < 0.001 and p < 0.001, respectively). OTM values were higher in STG than MTG (p < 0.001). This is the first study that reports glipizide caused DNA damage increasing with senescence in adipocytes. As a response to glipizide treatment Ran gene expression increased in mature; and decreased in senescent adipocytes. Further studies are needed to reveal the effect of glipizide on DNA and nuclear interactions in molecular level.

Effect of Imidocarb Application on Oxidative DNA Damage Caused by Anaplasmosis

This study was aimed to evaluate DNA fragmentation by using Comet assay in naturally infected sheep with Anaplasmosis before and after treatment with the Comet method, which shows DNA damage specifically. In the study, blood samples were collected from 10 Anaplosmosis infected and 10 healthy sheep. The anaplosmosis was diagnosed by clinical signs and symptoms. The infection was confirmed by Giemsa staining. The blood was collected from control group and infected group before and after the treatment, from the vena jugularis with the appropriate method. The DNA fragmentation was checked by using the Comet assay of blood cells. The data were analysed throught ANNOVA one-way. The result showed higher DNA fragmentation in sick animals diagnosed with anaplasmosis; tail length and tail moment values were found to be statistically significantly higher than the control group. When the data obtained after imidocarb (IMD) application were compared with obtained during the disease, a decreased DNA damage and tail moment was determined, however, these values higher than control. In this study, DNA damage and the extent of this damage were investigated by the Comet assay method using a healthy control group before and after treatment in animals with Anaplasmosis. When the findings obtained from the study were evaluated, it was seen that Anaplasma agents caused DNA damage and with the imidocarb application given for treatment, DNA damage was reduced and results close to healthy individuals were obtained.

Toxic effects of benzovindiflupyr, a new SDHI-type fungicide on earthworms (Eisenia fetida).

Benzovindiflupyr has received increasing attention as a new novel succinate dehydrogenase inhibitor (SDHI)-type fungicide. Nonetheless, its traces remaining in soil potentially trigger an ecotoxicological threat to soil organisms including earthworms. This paper evaluates the eco-toxicity of different benzovindiflupyr doses (0.1, 1, 5, and 10 mg kg-1) on earthworms (Eisenia fetida) after long-term exposure. Consequently, benzovindiflupyr at higher doses significantly inhibited the activities of respiratory chain complex II and succinate dehydrogenase (SDH) in E. fetida. Besides, the reactive oxygen species (ROS) and lipid peroxidation (LPO) were significantly induced in earthworms when treated with this fungicide. After benzovindiflupyr exposure, activities of antioxidant enzymes including catalase, peroxidase, and superoxide dismutase were activated. However, glutathione S-transferase activity in E. fetida was initially induced then inhibited in earthworms after treatment. Furthermore, benzovindiflupyr exposure induced the protein carbonylation (PCO) level in cells indicating oxidative damage to the cellular protein. Due to the destruction of the normal function in the coelomocytes, the phagocytic activity was initially activated, then inhibited when earthworms were treated at 5 and 10 mg kg-1 concentrations. Additionally, DNA damage was induced (larger olive tail moment (OTM) values) with the increase of benzovindiflupyr doses and exposure time. The weight was significantly decreased after benzovindiflupyr exposure on days 21 and 28. Benzovindiflupyr at higher doses significantly decreased the reproduction (number of cocoons and juveniles) of E. fetida. These findings reveal that benzovindiflupyr potentially induces a potential toxicological risk to earthworms when applied in the mentioned above dosages.