Pub Date : 2023-08-01DOI: 10.1016/j.mrgentox.2023.503668
Durre Shahwar , Zeba Khan , Mohammad Yunus Khalil Ansari , Younghoon Park
Mutagenic effectiveness and efficiency are the utmost vital indices to determine the effective and efficient mutagenic concentrations for the generation of high frequency of desirable mutation in mutation breeding. Nevertheless, there are meager study are available for employing effective and efficient concentration of caffeine, EMS, Pb(NO3)2 and Cd(NO3)2 for the crop improvement. Thus, the current study was performed to assess the mutagenic effectiveness and efficiency of caffeine, EMS and heavy metals [Pb(NO3)2 Cd(NO3)2] and to determine the genetic variability of M2 and M3 lentil mutant lines. The frequency of different chlorophyll and chromophyll mutation was found highest at moderate and higher concentrations of chemical mutagens and heavy metals in M1 and M2 generation. The highest effectiveness was in 20 ppm Cd(NO3)2, followed by 20 ppm Pb(NO3)2, 0.10% EMS, and 0.10% caffeine. The present investigation also showed lower doses of caffeine, EMS, Pb(NO3)2, and Cd(NO3)2 were more efficient than higher concentrations, and caffeine was found more efficient followed by EMS, Pb(NO3)2, and Cd(NO3)2. Furthermore, a broad spectrum of viable mutations affecting different morphological characters of the plants viz., leaves, plant height, growth habits, flowers, pods, and seeds in M2 and M3 generation were recorded. Ten morphological mutants showing acceptable agronomic and horticultural features were identified, as genetic resources for further breeding.
{"title":"Relative mutagenic effectiveness and efficiency of chemical mutagens (Caffeine and EMS) and heavy metals [(Pb(NO3)2 and Cd(NO3)2)] in developing chlorophyll and morphological mutants in lentil","authors":"Durre Shahwar , Zeba Khan , Mohammad Yunus Khalil Ansari , Younghoon Park","doi":"10.1016/j.mrgentox.2023.503668","DOIUrl":"https://doi.org/10.1016/j.mrgentox.2023.503668","url":null,"abstract":"<div><p>Mutagenic effectiveness and efficiency are the utmost vital indices to determine the effective and efficient mutagenic concentrations for the generation of high frequency of desirable mutation in mutation breeding. Nevertheless, there are meager study are available for employing effective and efficient concentration of caffeine, EMS, Pb(NO<sub>3</sub>)<sub>2</sub> and Cd(NO<sub>3</sub>)<sub>2</sub> for the crop improvement. Thus, the current study was performed to assess the mutagenic effectiveness and efficiency of caffeine, EMS and heavy metals [Pb(NO<sub>3</sub>)<sub>2</sub> Cd(NO<sub>3</sub>)<sub>2</sub>] and to determine the genetic variability of M<sub>2</sub> and M<sub>3</sub> lentil mutant lines. The frequency of different chlorophyll and chromophyll mutation was found highest at moderate and higher concentrations of chemical mutagens and heavy metals in M<sub>1</sub> and M<sub>2</sub> generation. The highest effectiveness was in 20 ppm Cd(NO<sub>3</sub>)<sub>2</sub>, followed by 20 ppm Pb(NO<sub>3</sub>)<sub>2</sub>, 0.10% EMS, and 0.10% caffeine. The present investigation also showed lower doses of caffeine, EMS, Pb(NO<sub>3</sub>)<sub>2</sub>, and Cd(NO<sub>3</sub>)<sub>2</sub> were more efficient than higher concentrations, and caffeine was found more efficient followed by EMS, Pb(NO<sub>3</sub>)<sub>2</sub>, and Cd(NO<sub>3</sub>)<sub>2</sub>. Furthermore, a broad spectrum of viable mutations affecting different morphological characters of the plants viz., leaves, plant height, growth habits, flowers, pods, and seeds in M<sub>2</sub> and M<sub>3</sub> generation were recorded. Ten morphological mutants showing acceptable agronomic and horticultural features were identified, as genetic resources for further breeding.</p></div>","PeriodicalId":18799,"journal":{"name":"Mutation research. Genetic toxicology and environmental mutagenesis","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49839480","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-08-01DOI: 10.1016/j.mrgentox.2023.503660
Diene da Silva Schlickmann , Patrícia Molz , Gabriela Cristina Uebel , Caroline Santos , Caroline Brand , Renato Alberto Weber Colombelli , Thalia Gama da Silva , Juliana Priebe Steffens , Eduarda da Silva Limberger Castilhos , Pedro J. Benito , Alexandre Rieger , Silvia Isabel Rech Franke
In a cross-sectional study of gymnasium users (both sexes, ages = 41.9 ± 14.8 years), we examined the moderating role of macronutrient intake in relation to body composition and genotoxicity. A questionnaire was administered to evaluate characteristics of the participants. To assess macronutrient consumption, we used 24-h food recalls on three non-consecutive days. Body composition (body fat percentage and muscle mass) was evaluated with a bioimpedance scale. Genotoxicity was assessed with the buccal micronucleus cytome assay. Multiple linear regression models were applied, adjusting for age; sex; tobacco and alcohol consumption; and (with regard to exercise habits) frequency, training time, intensity, and types. Micronucleus frequency was directly associated with body fat and inversely associated with muscle mass. Our study shows that carbohydrate and fat intakes affect body fat percentage and micronucleus frequency in gymnasium users.
{"title":"The moderating role of macronutrient intake in relation to body composition and genotoxicity: A study with gym users","authors":"Diene da Silva Schlickmann , Patrícia Molz , Gabriela Cristina Uebel , Caroline Santos , Caroline Brand , Renato Alberto Weber Colombelli , Thalia Gama da Silva , Juliana Priebe Steffens , Eduarda da Silva Limberger Castilhos , Pedro J. Benito , Alexandre Rieger , Silvia Isabel Rech Franke","doi":"10.1016/j.mrgentox.2023.503660","DOIUrl":"10.1016/j.mrgentox.2023.503660","url":null,"abstract":"<div><p>In a cross-sectional study of gymnasium users (both sexes, ages = 41.9 ± 14.8 years), we examined the moderating role of macronutrient intake in relation to body composition and genotoxicity. A questionnaire was administered to evaluate characteristics of the participants. To assess macronutrient consumption, we used 24-h food recalls on three non-consecutive days. Body composition (body fat percentage and muscle mass) was evaluated with a bioimpedance scale. Genotoxicity was assessed with the buccal micronucleus cytome assay. Multiple linear regression models were applied, adjusting for age; sex; tobacco and alcohol consumption; and (with regard to exercise habits) frequency, training time, intensity, and types. Micronucleus frequency was directly associated with body fat and inversely associated with muscle mass. Our study shows that carbohydrate and fat intakes affect body fat percentage and micronucleus frequency in gymnasium users.</p></div>","PeriodicalId":18799,"journal":{"name":"Mutation research. Genetic toxicology and environmental mutagenesis","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9980005","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Fetal development can be altered by DNA damage caused by maternal exposure to chemical, physical, or biological agents during gestation. One method of assessing genotoxicity is to detect micronuclei (MNs) and/or nuclear abnormalities. This can be performed in vivo and requires only frequently dividing tissues, such as amniotic tissue (AT), which is in contact with the fetal environment and is composed of very thin layers of cells. This study evaluated the presence of MNs, nucleoplasmic bridges, and nuclear buds (NBs) in the fetal AT following maternal exposure to cyclophosphamide (CP) during pregnancy. Pregnant Wistar rats were divided into a negative control group and an experimental group that was orally administered CP (10 mg/kg). Daily blood smears were obtained from pregnant rats on days 14–19 of gestation. The rats were dissected, and fetal ATs were obtained on the 19th day of gestation. The MN and NB frequencies in AT cells were analyzed using a fluorescence microscope (100 ×). Micronucleated erythrocytes in the peripheral blood of the control rats were also assessed. Micronucleated polychromatic erythrocyte frequencies were significantly higher than those in the controls. Polychromatic erythrocyte frequencies were lower in CP-treated rats than in controls at 48–120 h. Fetuses in the CP-treated group also showed a significant increase in MNs and NBs in AT cells. In conclusion, AT could be used for analyzing MNs and NBs in rats following maternal exposure to a genotoxic agent and as a viable alternative for analyzing the integrity of fetal DNA during gestation.
{"title":"Micronuclei and nuclear buds in amniotic tissue of rats treated with cyclophosphamide","authors":"Ramón Guillermo Ortiz-García , Belinda Claudia Gómez-Meda , Juan Ernesto Gutiérrez-Sevilla , Martha Patricia Gallegos-Arreola , Ana Lourdes Zamora-Perez , Yveth Marlene Ortiz-García , Víctor Eduardo García-Arias , Blanca Miriam Torres-Mendoza , Guillermo Moisés Zúñiga-González","doi":"10.1016/j.mrgentox.2023.503659","DOIUrl":"10.1016/j.mrgentox.2023.503659","url":null,"abstract":"<div><p>Fetal development can be altered by DNA damage caused by maternal exposure to chemical, physical, or biological agents during gestation. One method of assessing genotoxicity is to detect micronuclei (MNs) and/or nuclear abnormalities. This can be performed in vivo and requires only frequently dividing tissues, such as amniotic tissue (AT), which is in contact with the fetal environment and is composed of very thin layers of cells. This study evaluated the presence of MNs, nucleoplasmic bridges, and nuclear buds (NBs) in the fetal AT following maternal exposure to cyclophosphamide (CP) during pregnancy. Pregnant Wistar rats were divided into a negative control group and an experimental group that was orally administered CP (10 mg/kg). Daily blood smears were obtained from pregnant rats on days 14–19 of gestation. The rats were dissected, and fetal ATs were obtained on the 19th day of gestation. The MN and NB frequencies in AT cells were analyzed using a fluorescence microscope (100 ×). Micronucleated erythrocytes in the peripheral blood of the control rats were also assessed. Micronucleated polychromatic erythrocyte frequencies were significantly higher than those in the controls. Polychromatic erythrocyte frequencies were lower in CP-treated rats than in controls at 48–120 h. Fetuses in the CP-treated group also showed a significant increase in MNs and NBs in AT cells. In conclusion, AT could be used for analyzing MNs and NBs in rats following maternal exposure to a genotoxic agent and as a viable alternative for analyzing the integrity of fetal DNA during gestation.</p></div>","PeriodicalId":18799,"journal":{"name":"Mutation research. Genetic toxicology and environmental mutagenesis","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9980008","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-07-01DOI: 10.1016/j.mrgentox.2023.503652
Luíza Araújo da Costa Xavier , Julio Alejandro Navoni , Viviane Souza do Amaral
Radon gas inhalation is the main source of exposure to ionizing radiation by humans. There is still lack in knowledge concerning the chronic and indirect effects of exposure to this carcinogenic factor. Therefore, the aim of this work is to analyze the levels of oxidative genomic damage in inhabitants of a medium-high background radiation area (HBRA) (N = 82) in Northeastern Brazil and compare them with people living in a low background radiation area (LBRA) (N = 46). 8-hydroxy-2-deoxyguanosine (8-OHdG) was quantified in urine, Ser326Cys polymorphism was determined in the hOGG1 gene and indoor radon was measured. HBRA houses had 6.5 times higher indoor radon levels than those from LBRA (p-value < 0.001). The 8-OHdG mean (95% confidence interval) were significantly different, 8.42 (5.98–11.9) ng/mg creatinine and 29.91 (23.37–38.30) ng/mg creatinine for LBRA and HBRA, respectively. The variables representing lifestyle and environmental and occupational exposures did not have a significant association with oxidized guanosine concentrations. On the other hand, lower 8-OHdG values were observed in subjects that had one mutant allele (326Cys) in the hOGG1 gene than those who had both wild alleles (Ser/Ser (p-value < 0.05). It can be concluded that high radon levels have significantly influenced the genome oxidative metabolism and hOGG1 gene polymorphism would mediate the observed biological response.
{"title":"Oxidative genomic damage in humans exposed to high indoor radon levels in Northeast Brazil","authors":"Luíza Araújo da Costa Xavier , Julio Alejandro Navoni , Viviane Souza do Amaral","doi":"10.1016/j.mrgentox.2023.503652","DOIUrl":"10.1016/j.mrgentox.2023.503652","url":null,"abstract":"<div><p>Radon gas inhalation is the main source of exposure to ionizing radiation by humans. There is still lack in knowledge concerning the chronic and indirect effects of exposure to this carcinogenic factor. Therefore, the aim of this work is to analyze the levels of oxidative genomic damage in inhabitants of a medium-high background radiation area (HBRA) (N = 82) in Northeastern Brazil and compare them with people living in a low background radiation area (LBRA) (N = 46). 8-hydroxy-2-deoxyguanosine (8-OHdG) was quantified in urine, Ser326Cys polymorphism was determined in the <em>hOGG1</em> gene and indoor radon was measured. HBRA houses had 6.5 times higher indoor radon levels than those from LBRA (p-value < 0.001). The 8-OHdG mean (95% confidence interval) were significantly different, 8.42 (5.98–11.9) ng/mg creatinine and 29.91 (23.37–38.30) ng/mg creatinine for LBRA and HBRA, respectively. The variables representing lifestyle and environmental and occupational exposures did not have a significant association with oxidized guanosine concentrations. On the other hand, lower 8-OHdG values were observed in subjects that had one mutant allele (326Cys) in the <em>hOGG1</em> gene than those who had both wild alleles (Ser/Ser (p-value < 0.05). It can be concluded that high radon levels have significantly influenced the genome oxidative metabolism and <em>hOGG1</em> gene polymorphism would mediate the observed biological response.</p></div>","PeriodicalId":18799,"journal":{"name":"Mutation research. Genetic toxicology and environmental mutagenesis","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9929461","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The aim of our study was to assess the oxidative stress and inflammatory status in critically ill patients with sepsis as well as their relationship with the level of DNA damage. The study also evaluated the influence of all analyzed parameters on the outcome of the patients. The study included 27 critically ill patients with sepsis and 20 healthy subjects. Comet Assay was used for the measurement of the level of DNA damage, expressed as genetic damage index (GDI). Both oxidative stress parameters and the antioxidant parameters were obtained spectrophotometrically. The standard laboratory methods and the appropriate autoanalyzers were performed for determination the parameters of inflammation. A higher level of oxidative stress and more pronounced inflammation were found in the patients with sepsis compared to healthy subjects. The activity of the antioxidant enzymes was statistically declined in patients with sepsis, so that the most notable differences between two groups of participants were found for the activity of superoxide dismutase (SOD) (p = 0.004). Comet assay indicated that patients with sepsis had significantly higher GDI compared to healthy subjects (p < 0.001), which positively correlated with the concentration of superoxide anion radical (О2-) (r = 0.497, p = 0.010), and nitrites (NО2-) (r = 0.473, p = 0.015), as well with the concentration of C reactive protein (CRP) (r = 0.460, p = 0.041). Regression analysis confirmed that patients' age (p = 0.033), the level of О2- (p = 0.007), CRP concentration (p = 0.029) and GDI (p = 0.001) increased the risk of lethal outcome in critically ill patients with sepsis. In conclusion, critically ill patients with sepsis have a higher degree of oxidative stress and inflammation which contribute to a higher level of DNA damage. Consequently, above mentioned parameters, including patients' age, adversely affect the outcome of critically ill patients with sepsis.
{"title":"Oxidative stress and DNA damage in critically ill patients with sepsis","authors":"Olgica Mihaljevic , Snezana Zivancevic-Simonovic , Danijela Jovanovic , Svetlana Miletic Drakulic , Jovana Tubic Vukajlovic , Aleksandra Markovic , Marijana Stanojevic Pirkovic , Ivan Srejovic , Vladimir Jakovljevic , Olivera Milosevic-Djordjevic","doi":"10.1016/j.mrgentox.2023.503655","DOIUrl":"10.1016/j.mrgentox.2023.503655","url":null,"abstract":"<div><p>The aim of our study was to assess the oxidative stress and inflammatory status in critically ill patients with sepsis as well as their relationship with the level of DNA damage. The study also evaluated the influence of all analyzed parameters on the outcome of the patients. The study included 27 critically ill patients with sepsis and 20 healthy subjects. Comet Assay was used for the measurement of the level of DNA damage, expressed as genetic damage index (GDI). Both oxidative stress parameters and the antioxidant parameters were obtained spectrophotometrically. The standard laboratory methods and the appropriate autoanalyzers were performed for determination the parameters of inflammation. A higher level of oxidative stress and more pronounced inflammation were found in the patients with sepsis compared to healthy subjects. The activity of the antioxidant enzymes was statistically declined in patients with sepsis, so that the most notable differences between two groups of participants were found for the activity of superoxide dismutase (SOD) (p = 0.004). Comet assay indicated that patients with sepsis had significantly higher GDI compared to healthy subjects (p < 0.001), which positively correlated with the concentration of superoxide anion radical (О<sub>2</sub><sup>-</sup>) (r = 0.497, p = 0.010), and nitrites (NО<sub>2</sub><sup>-</sup>) (r = 0.473, p = 0.015), as well with the concentration of C reactive protein (CRP) (r = 0.460, p = 0.041). Regression analysis confirmed that patients' age (p = 0.033), the level of О<sub>2</sub><sup>-</sup> (p = 0.007), CRP concentration (p = 0.029) and GDI (p = 0.001) increased the risk of lethal outcome in critically ill patients with sepsis. In conclusion, critically ill patients with sepsis have a higher degree of oxidative stress and inflammation which contribute to a higher level of DNA damage. Consequently, above mentioned parameters, including patients' age, adversely affect the outcome of critically ill patients with sepsis.</p></div>","PeriodicalId":18799,"journal":{"name":"Mutation research. Genetic toxicology and environmental mutagenesis","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9929463","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-07-01DOI: 10.1016/j.mrgentox.2023.503649
Eunnara Cho , Carol D. Swartz , Andrew Williams , Miriam V. Rivas , Leslie Recio , Kristine L. Witt , Elizabeth K. Schmidt , Jeffry Yaplee , Thomas H. Smith , Phu Van , Fang Yin Lo , Charles C. Valentine III , Jesse J. Salk , Francesco Marchetti , Stephanie L. Smith-Roe , Carole L. Yauk
Error-corrected duplex sequencing (DS) enables direct quantification of low-frequency mutations and offers tremendous potential for chemical mutagenicity assessment. We investigated the utility of DS to quantify induced mutation frequency (MF) and spectrum in human lymphoblastoid TK6 cells exposed to a prototypical DNA alkylating agent, N-ethyl-N-nitrosourea (ENU). Furthermore, we explored appropriate experimental parameters for this application, and assessed inter-laboratory reproducibility. In two independent experiments in two laboratories, TK6 cells were exposed to ENU (25–200 µM) and DNA was sequenced 48, 72, and 96 h post-exposure. A DS mutagenicity panel targeting twenty 2.4-kb regions distributed across the genome was used to sample diverse, genome-representative sequence contexts. A significant increase in MF that was unaffected by time was observed in both laboratories. Concentration-response in the MF from the two laboratories was strongly positively correlated (r = 0.97). C:G>T:A, T:A>C:G, T:A>A:T, and T:A>G:C mutations increased in consistent, concentration-dependent manners in both laboratories, with high proportions of C:G>T:A at all time points. The consistent results across the three time points suggest that 48 h may be sufficient for mutation analysis post-exposure. The target sites responded similarly between the two laboratories and revealed a higher average MF in intergenic regions. These results, demonstrating remarkable reproducibility across time and laboratory for both MF and spectrum, support the high value of DS for characterizing chemical mutagenicity in both research and regulatory evaluation.
{"title":"Error-corrected duplex sequencing enables direct detection and quantification of mutations in human TK6 cells with strong inter-laboratory consistency","authors":"Eunnara Cho , Carol D. Swartz , Andrew Williams , Miriam V. Rivas , Leslie Recio , Kristine L. Witt , Elizabeth K. Schmidt , Jeffry Yaplee , Thomas H. Smith , Phu Van , Fang Yin Lo , Charles C. Valentine III , Jesse J. Salk , Francesco Marchetti , Stephanie L. Smith-Roe , Carole L. Yauk","doi":"10.1016/j.mrgentox.2023.503649","DOIUrl":"10.1016/j.mrgentox.2023.503649","url":null,"abstract":"<div><p>Error-corrected duplex sequencing (DS) enables direct quantification of low-frequency mutations and offers tremendous potential for chemical mutagenicity assessment. We investigated the utility of DS to quantify induced mutation frequency (MF) and spectrum in human lymphoblastoid TK6 cells exposed to a prototypical DNA alkylating agent, <em>N</em>-ethyl-<em>N</em>-nitrosourea (ENU). Furthermore, we explored appropriate experimental parameters for this application, and assessed inter-laboratory reproducibility. In two independent experiments in two laboratories, TK6 cells were exposed to ENU (25–200 µM) and DNA was sequenced 48, 72, and 96 h post-exposure. A DS mutagenicity panel targeting twenty 2.4-kb regions distributed across the genome was used to sample diverse, genome-representative sequence contexts. A significant increase in MF that was unaffected by time was observed in both laboratories. Concentration-response in the MF from the two laboratories was strongly positively correlated (<em>r</em> = 0.97). C:G>T:A, T:A>C:G, T:A>A:T, and T:A>G:C mutations increased in consistent, concentration-dependent manners in both laboratories, with high proportions of C:G>T:A at all time points. The consistent results across the three time points suggest that 48 h may be sufficient for mutation analysis post-exposure. The target sites responded similarly between the two laboratories and revealed a higher average MF in intergenic regions. These results, demonstrating remarkable reproducibility across time and laboratory for both MF and spectrum, support the high value of DS for characterizing chemical mutagenicity in both research and regulatory evaluation.</p></div>","PeriodicalId":18799,"journal":{"name":"Mutation research. Genetic toxicology and environmental mutagenesis","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10395007/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9971696","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Tributyltin (TBT) is used in many commercial applications, including pesticides and antifouling paints, due to its biocidal properties. We examined the cytotoxicity and genotoxicity of TBT in the early chick embryo (Gallus gallus domesticus). Chick embryos (11 days) were treated with various doses of TBT to measure LD50 values for 24, 48, and 72 h exposures, which were determined to be 110, 54, and 18 μg/egg, respectively. The embryos were exposed to sub-lethal doses of TBT for evaluation of cytotoxicity and genotoxicity. An increase in the incidence of micronuclei (MN) was observed but it was not statistically significant. Induction of other nuclear abnormalities (ONA) after 72 h TBT exposure was significant. A significant increase in comet assay tail DNA content was also detected in TBT-exposed embryos. Cytotoxicity was also evidenced by alteration in the polychromatic erythrocytes (PCE) to normochromatic erythrocytes (NCE) ratio and by an increase in the erythroblast population in treated organisms. The cytotoxicity and genotoxicity of TBT may have long-term complications in later stages of the life cycle.
{"title":"Cytotoxicity and genotoxicity of tributyltin in the early embryonic chick, Gallus gallus domesticus","authors":"Abhijit Mandal, Malaya Ghosh, Doli Talukdar, Pubali Dey, Aparajita Das, Sarbani Giri","doi":"10.1016/j.mrgentox.2023.503656","DOIUrl":"10.1016/j.mrgentox.2023.503656","url":null,"abstract":"<div><p>Tributyltin (TBT) is used in many commercial applications, including pesticides and antifouling paints, due to its biocidal properties. We examined the cytotoxicity and genotoxicity of TBT in the early chick embryo <em>(Gallus gallus domesticus)</em>. Chick embryos (11 days) were treated with various doses of TBT to measure LD<sub>50</sub> values for 24, 48, and 72 h exposures, which were determined to be 110, 54, and 18 μg/egg, respectively. The embryos were exposed to sub-lethal doses of TBT for evaluation of cytotoxicity and genotoxicity. An increase in the incidence of micronuclei (MN) was observed but it was not statistically significant. Induction of other nuclear abnormalities (ONA) after 72 h TBT exposure was significant. A significant increase in comet assay tail DNA content was also detected in TBT-exposed embryos. Cytotoxicity was also evidenced by alteration in the polychromatic erythrocytes (PCE) to normochromatic erythrocytes (NCE) ratio and by an increase in the erythroblast population in treated organisms. The cytotoxicity and genotoxicity of TBT may have long-term complications in later stages of the life cycle.</p></div>","PeriodicalId":18799,"journal":{"name":"Mutation research. Genetic toxicology and environmental mutagenesis","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9929466","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Many fungal genera such as Aspergillus, Penicillium, Fusarium and Alternaria are able to produce, among many other metabolites, the aflatoxins, a group of toxic and carcinogenic compounds. To reduce their formation, synthetic fungicides are used as an effective way of intervention. However, the extensive use of such molecules generates long-term residues into the food and the environment. The need of new antifungal molecules, with high specificity and low off-target toxicity is worth. The aim of this study was to evaluate: i) the toxicity and genotoxicity of newly synthesized molecules with a good anti-mycotoxic activity, and ii) the suitability of the Allium cepa multi-endpoint assay as an early screening method for chemicals. Eight compounds were tested for toxicity by using the A. cepa bulb root elongation test and for genotoxicity using the A. cepa bulb mitotic index, micronuclei and chromosome aberrations tests. Three molecules showed no toxicity, while two induced mild toxic effects in roots exposed to the highest dose (100 µM). A more pronounced toxic effect was caused by the other three compounds for which the EC50 was approximately 50 μM. Furthermore, all molecules showed a clear genotoxic activity, both in terms of chromosomal aberrations and micronuclei. Albeit the known good antifungal activity, the different molecules caused strong toxic and genotoxic effects. The results indicate the suitability of experiments with A. cepa as a research model for the evaluation of the toxic and genotoxic activities of new molecules in plants before they are released into the environment.
{"title":"Allium cepa tests: A plant-based tool for the early evaluation of toxicity and genotoxicity of newly synthetized antifungal molecules","authors":"Carlotta Alias , Donatella Feretti , Gaia V.C. Viola , Ilaria Zerbini , Franco Bisceglie , Giorgio Pelosi , Claudia Zani","doi":"10.1016/j.mrgentox.2023.503654","DOIUrl":"10.1016/j.mrgentox.2023.503654","url":null,"abstract":"<div><p>Many fungal genera such as <em>Aspergillus</em>, <em>Penicillium</em>, <em>Fusarium</em> and <em>Alternaria</em> are able to produce, among many other metabolites, the aflatoxins, a group of toxic and carcinogenic compounds. To reduce their formation, synthetic fungicides are used as an effective way of intervention. However, the extensive use of such molecules generates long-term residues into the food and the environment. The need of new antifungal molecules, with high specificity and low off-target toxicity is worth. The aim of this study was to evaluate: i) the toxicity and genotoxicity of newly synthesized molecules with a good anti-mycotoxic activity, and ii) the suitability of the <em>Allium cepa</em> multi-endpoint assay as an early screening method for chemicals. Eight compounds were tested for toxicity by using the <em>A. cepa</em> bulb root elongation test and for genotoxicity using the <em>A. cepa</em> bulb mitotic index, micronuclei and chromosome aberrations tests. Three molecules showed no toxicity, while two induced mild toxic effects in roots exposed to the highest dose (100 µM). A more pronounced toxic effect was caused by the other three compounds for which the EC50 was approximately 50 μM. Furthermore, all molecules showed a clear genotoxic activity, both in terms of chromosomal aberrations and micronuclei. Albeit the known good antifungal activity, the different molecules caused strong toxic and genotoxic effects. The results indicate the suitability of experiments with <em>A. cepa</em> as a research model for the evaluation of the toxic and genotoxic activities of new molecules in plants before they are released into the environment.</p></div>","PeriodicalId":18799,"journal":{"name":"Mutation research. Genetic toxicology and environmental mutagenesis","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9929467","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-07-01DOI: 10.1016/j.mrgentox.2023.503650
S.M. Attia , S.F. Ahmad , A. Nadeem , M.S.M. Attia , M.A. Ansari , N.B. Alsaleh , A.F. Alasmari , M.A. Al-Hamamah , A. Alanazi , A.A. Alshamrani , S.A. Bakheet , G.I. Harisa
Multiple sclerosis (MS) is a demyelinating disorder in which the myelin sheath covering the central nervous system axons is damaged or lost, disrupting action potential conduction and leading to various neurological complications. The pathogenesis of MS remains unclear, and no effective therapies are currently available. MS is triggered by environmental factors in genetically susceptible individuals. DNA damage and DNA repair failure have been proposed as MS genetic risk factors; however, inconsistent evidence has been found in multiple studies. Therefore, more investigations are needed to ascertain whether DNA damage/repair is altered in this disorder. In this context, therapies that prevent DNA damage or enhance DNA repair could be effective strategies for MS treatment. The overactivation of the extracellular-signal-related kinase 1 and 2 (Erk1/2) pathway can lead to DNA damage and has been linked to MS pathogenesis. In our study, we observed substantially elevated oxidative DNA damage and slower DNA repair rates in an experimentally autoimmune encephalomyelitis animal model of MS (EAE). Moreover, statistical decreases in oxidative DNA strand breaks and faster repair rates were observed in EAE animals injected with the Erk1/2 inhibitor PD98059 (PD). Moreover, the expression of several genes associated with DNA strand breaks and repair changed in EAE mice at both the mRNA and protein levels, as revealed by the RT2 Profiler PCR array and verified by RT-PCR and protein analyses. The treatment with PD mitigated these changes and improved DNA repair gene expression. Our results demonstrate clear associations between Erk1/2 activation, DNA damage/repair, and MS pathology, and further suggest that PD therapy may be a promising adjuvant therapeutic strategy.
{"title":"The small molecule Erk1/2 signaling pathway inhibitor PD98059 improves DNA repair in an experimental autoimmune encephalomyelitis SJL/J mouse model of multiple sclerosis","authors":"S.M. Attia , S.F. Ahmad , A. Nadeem , M.S.M. Attia , M.A. Ansari , N.B. Alsaleh , A.F. Alasmari , M.A. Al-Hamamah , A. Alanazi , A.A. Alshamrani , S.A. Bakheet , G.I. Harisa","doi":"10.1016/j.mrgentox.2023.503650","DOIUrl":"10.1016/j.mrgentox.2023.503650","url":null,"abstract":"<div><p>Multiple sclerosis (MS) is a demyelinating disorder in which the myelin sheath covering the central nervous system axons is damaged or lost, disrupting action potential conduction and leading to various neurological complications. The pathogenesis of MS remains unclear, and no effective therapies are currently available. MS is triggered by environmental factors in genetically susceptible individuals. DNA damage and DNA repair failure have been proposed as MS genetic risk factors; however, inconsistent evidence has been found in multiple studies. Therefore, more investigations are needed to ascertain whether DNA damage/repair is altered in this disorder. In this context, therapies that prevent DNA damage or enhance DNA repair could be effective strategies for MS treatment. The overactivation of the extracellular-signal-related kinase 1 and 2 (Erk1/2) pathway can lead to DNA damage and has been linked to MS pathogenesis. In our study, we observed substantially elevated oxidative DNA damage and slower DNA repair rates in an experimentally autoimmune encephalomyelitis animal model of MS (EAE). Moreover, statistical decreases in oxidative DNA strand breaks and faster repair rates were observed in EAE animals injected with the Erk1/2 inhibitor PD98059 (PD). Moreover, the expression of several genes associated with DNA strand breaks and repair changed in EAE mice at both the mRNA and protein levels, as revealed by the RT<sup>2</sup> Profiler PCR array and verified by RT-PCR and protein analyses. The treatment with PD mitigated these changes and improved DNA repair gene expression. Our results demonstrate clear associations between Erk1/2 activation, DNA damage/repair, and MS pathology, and further suggest that PD therapy may be a promising adjuvant therapeutic strategy.</p></div>","PeriodicalId":18799,"journal":{"name":"Mutation research. Genetic toxicology and environmental mutagenesis","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9929462","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-07-01DOI: 10.1016/j.mrgentox.2023.503651
Xiaoqiang Hu , Chuncao He , Lijun Zhang , Yunheng Zhang , Liangjing Chen , Chuan Sun , Jun Wei , Lei Yang , Xiaohua Tan , Jun Yang , Yan Zhang
Stem cell-derived exosomes (SC-Exos) have been shown to protect cells from chemical-induced deoxyribonucleic acid (DNA) damage. However, there has been no systematic comparison of the efficacy of exosomes against different types of DNA damage. Therefore, in this study, we assessed the protective effect of exosomes derived from human embryonic stem cell-induced mesenchymal stem cells (hESC-MSC-Exos) on two types of DNA damage, namely, intra-/inter-strand crosslinks and DNA double-strand breaks induced by cisplatin (Pt) and bleomycin (BLM), respectively, in HeLa cells. The alkaline comet assay demonstrated that hESC-MSC-Exos effectively inhibited Pt- and BLM-induced DNA damage in a dose-dependent manner. When the concentration of hESC-MSC-Exos reaches 2.0 × 106 and 4.0 × 106 particles/mL in Pt- and BLM-treated groups, respectively, there was a significant decrease in tail DNA percentage (Pt: 20.80 ± 1.61 vs 9.40 ± 1.14, p < 0.01; BLM: 21.80 ± 1.31 vs 6.70 ± 0.60, p < 0.01), tail moment (Pt: 10.00 ± 1.21 vs 2.08 ± 0.51, p < 0.01; BLM: 12.00 ± 0.81 vs 2.00 ± 0.21, p < 0.01), and olive tail moment (Pt: 6.01 ± 0.55 vs 2.09 ± 0.25, p < 0.01; BLM: 6.03 ± 0.37 vs 1.53 ± 0.13, p < 0.01). Phospho-histone H2AX (γH2AX) immunofluorescence and western blotting showed an over 50 % decrease in γH2AX expression when the cells were pretreated with hESC-MSC-Exos. As reactive oxygen species (ROS) are important mediators of Pt- and BLM-induced DNA damage, dichloro-dihydro-fluorescein diacetate staining indicated that hESC-MSC-Exos inhibited the increase in intracellular ROS in drug-treated cells. In conclusion, our findings suggest that hESC-MSC-Exos can protect cells from the two types of DNA-damaging drugs and that reduced intracellular ROS is involved in this effect.
干细胞衍生的外泌体(SC-Exos)已被证明可以保护细胞免受化学诱导的脱氧核糖核酸(DNA)损伤。然而,目前还没有系统比较外泌体对不同类型DNA损伤的疗效。因此,在本研究中,我们评估了来源于人类胚胎干细胞诱导的间充质干细胞(hESC-MSC-Exos)的外泌体对HeLa细胞中两种类型的DNA损伤的保护作用,即分别由顺铂(Pt)和博来霉素(BLM)诱导的链内/链间交联和DNA双链断裂。碱性彗星试验表明,hESC-MSC-Exos以剂量依赖的方式有效抑制Pt和BLM诱导的DNA损伤。当Pt-和BLM处理组的hESC-MSC-Exos浓度分别达到2.0×106和4.0×106粒子/mL时,尾部DNA百分比显著降低(Pt:28.0±1.61 vs 9.40±1.14,p<;0.01;BLM:21.80±1.31 vs 6.70±0.60,p<;0.01),尾力矩(Pt:10.00±1.21 vs 2.08±0.51,p<;0.01;BLM:12.00±0.81 vs 2.00±0.21,p&lgt;0.01),和橄榄尾力矩(Pt:6.01±0.55vs2.09±0.25,p<;0.01;BLM:6.03±0.37vs1.53±0.13,p&lgt;0.01)。由于活性氧(ROS)是Pt-和BLM诱导的DNA损伤的重要介质,二氯二氢荧光素双乙酸酯染色表明hESC-MSC-Exos抑制了药物处理细胞中细胞内ROS的增加。总之,我们的研究结果表明,hESC-MSC-Exos可以保护细胞免受两种类型的DNA损伤药物的影响,并且减少的细胞内ROS参与了这种作用。
{"title":"Mesenchymal stem cell-derived exosomes attenuate DNA damage response induced by cisplatin and bleomycin","authors":"Xiaoqiang Hu , Chuncao He , Lijun Zhang , Yunheng Zhang , Liangjing Chen , Chuan Sun , Jun Wei , Lei Yang , Xiaohua Tan , Jun Yang , Yan Zhang","doi":"10.1016/j.mrgentox.2023.503651","DOIUrl":"10.1016/j.mrgentox.2023.503651","url":null,"abstract":"<div><p>Stem cell-derived exosomes (SC-Exos) have been shown to protect cells from chemical-induced deoxyribonucleic acid (DNA) damage. However, there has been no systematic comparison of the efficacy of exosomes against different types of DNA damage. Therefore, in this study, we assessed the protective effect of exosomes derived from human embryonic stem cell-induced mesenchymal stem cells (hESC-MSC-Exos) on two types of DNA damage, namely, intra-/inter-strand crosslinks and DNA double-strand breaks induced by cisplatin (Pt) and bleomycin (BLM), respectively, in HeLa cells. The alkaline comet assay demonstrated that hESC-MSC-Exos effectively inhibited Pt- and BLM-induced DNA damage in a dose-dependent manner. When the concentration of hESC-MSC-Exos reaches 2.0 × 10<sup>6</sup> and 4.0 × 10<sup>6</sup> particles/mL in Pt- and BLM-treated groups, respectively, there was a significant decrease in tail DNA percentage (Pt: 20.80 ± 1.61 vs 9.40 ± 1.14, <em>p</em> < 0.01; BLM: 21.80 ± 1.31 vs 6.70 ± 0.60, <em>p</em> < 0.01), tail moment (Pt: 10.00 ± 1.21 vs 2.08 ± 0.51, <em>p</em> < 0.01; BLM: 12.00 ± 0.81 vs 2.00 ± 0.21, <em>p</em> < 0.01), and olive tail moment (Pt: 6.01 ± 0.55 vs 2.09 ± 0.25, <em>p</em> < 0.01; BLM: 6.03 ± 0.37 vs 1.53 ± 0.13, <em>p</em> < 0.01). Phospho-histone H2AX (γH2AX) immunofluorescence and western blotting showed an over 50 % decrease in γH2AX expression when the cells were pretreated with hESC-MSC-Exos. As reactive oxygen species (ROS) are important mediators of Pt- and BLM-induced DNA damage, dichloro-dihydro-fluorescein diacetate staining indicated that hESC-MSC-Exos inhibited the increase in intracellular ROS in drug-treated cells. In conclusion, our findings suggest that hESC-MSC-Exos can protect cells from the two types of DNA-damaging drugs and that reduced intracellular ROS is involved in this effect.</p></div>","PeriodicalId":18799,"journal":{"name":"Mutation research. Genetic toxicology and environmental mutagenesis","volume":null,"pages":null},"PeriodicalIF":1.9,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9929464","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}