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1.
J Hazard Mater ; 416: 125889, 2021 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-34492827

RESUMO

Green toxicology is a novel approach increasingly applied for the development of materials and chemicals that are more benign to the environment and human health than their conventional counterparts. It includes predictive eco-toxicological assessments of chemicals during the early developmental process to exclude adverse effects. In the present study, two guanidine zinc catalysts for the ring-opening polymerization of lactide were investigated using eco-toxicological tools. Namely, the fish embryo toxicity assay for teratogenic effects, the ER (α) CALUX assay for endocrine activity and the Ames fluctuation assay for mutagenic potential were applied. Both complexes showed no endocrine activity, mutagenicity or acute aquatic toxicity, however a delayed hatch could be observed, therefore suggesting potential effects on a molecular level. This proof-of-concept study aims to assess the toxicity of guanidine zinc catalysts and is a first step towards the incorporation of toxicological assessments into chemical developmental processes to achieve a sustainable and safe production of catalysts.


Assuntos
Bioensaio , Zinco , Animais , Catálise , Humanos , Testes de Mutagenicidade , Polimerização , Zinco/toxicidade
2.
J Hazard Mater ; 416: 126053, 2021 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-34492893

RESUMO

Various genotoxic substances in household effluents have not been sufficiently studied. The purpose of this study is to evaluate them using the umu test after dividing them based on the acid-base properties of their functional groups by solid-phase extraction cartridges. The results of the samples concentrated with reverse-phase cartridges showed that the substances with acid functional groups had stronger genotoxicity as 4.1-12.1 ng-4-NQO/mL without S9 enzyme and 17.4-51.8 ng-2-AA/mL with S9 enzyme, while the basic substances also showed a certain degree of toxicity. The results of dividing the effluents by acid-base properties using ion-exchange cartridges showed that chemical substances with strong acid functional groups did not demonstrate genotoxicity. It was found that the genotoxicity of chemicals with functional groups of weak acids was half of that of the total amount. The genotoxicity of the neutral substance was not strong, and the genotoxicity of the weak basic substances was negligible. The zwitterions and substances with strong basic functional groups showed about half the total genotoxicity. This is the first report that has investigated the genotoxicity of zwitterions in effluents.


Assuntos
Poluentes Químicos da Água , Dano ao DNA , Testes de Mutagenicidade , Mutagênicos/toxicidade , Extração em Fase Sólida , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidade
3.
Artigo em Inglês | MEDLINE | ID: mdl-34454692

RESUMO

Kirkland et al. [Mutation Research/Genetic Toxicology and Environmental Mutagenesis 847 (2019) 403035, https://doi.org/10.1016/j.mrgentox.2019.03.008; Mutation Research/Genetic Toxicology and Environmental Mutagenesis 839 (2019): 21-35, https://doi.org/10.1016/j.mrgentox.2019.01.007] made recommendations on the use of the in vivo comet and transgenic rodent (TGR) gene mutation assays to screen for in vivo mutagenicity. Although it is not directly stated in either of these publications, we are concerned that the reports could potentially be used to support assertions that it is equally acceptable to follow up a positive bacterial reverse mutation (Ames) finding for an investigational drug with either the in vivo TGR mutation assay or an in vivo comet assay. For regulatory genotoxicity assessment, the in vivo follow-up for an in vitro bacterial mutation-positive drug, drug-related metabolite, or impurity should be based upon evaluating a similar endpoint (i.e., mutagenicity) as the intent is to determine if the findings of in vitro gene mutation correlate with findings of in vivo gene mutation (i.e., biologically relevant to the in vitro results). Thus, the most scientifically appropriate in vivo assays would be the TGR mutation assay or, in some circumstances, the in vivo Pig-a assay. An in vivo rodent comet assay or combination of the in vivo micronucleus and in vivo rodent comet assays would generally not be an appropriate follow-up test.


Assuntos
Bioensaio/métodos , Drogas em Investigação/química , Drogas em Investigação/metabolismo , Mutação/efeitos dos fármacos , Animais , Animais Geneticamente Modificados/genética , Carcinógenos/toxicidade , Ensaio Cometa/métodos , Seguimentos , Testes para Micronúcleos/métodos , Testes de Mutagenicidade/métodos , Mutagênicos/toxicidade , Roedores
4.
Artigo em Inglês | MEDLINE | ID: mdl-34454694

RESUMO

Formal requirements for genotoxicity testing of drug candidates to support clinical entry have been in place since the issue of initial regulatory guidance over 25 years ago and subsequent update a decade ago. An evaluation of such testing, supporting first clinical entry of 108 small molecule drug candidates over the last decade, showed that the most common approach (75 % of tested compounds) was for a Good Laboratory Practice test battery in the form of 2 in vitro (a bacterial reverse mutation and a mammalian cell) assays and one in vivo assay. The majority of other tested compounds involved in vitro testing only in bacterial reverse mutation and mammalian cell assays. Testing using a bacterial reverse mutation assay and an in vivo assessment of genotoxicity with 2 different tissues was limited to 2 occasions. For in vitro mammalian cell testing, the chromosome aberration test was most commonly used (70 % occasions), followed by a micronucleus test (16 % occasions) or a mouse lymphoma assay (14 % occasions). For in vivo evaluation, the most common test was a rodent bone marrow micronucleus test (87 % occasions). A positive in vitro mammalian cell assay result was seen on 13 % occasions but was not confirmed with further in vivo testing and the drug candidates were taken into the clinic. In conclusion, the present evaluation showed that the current test battery paradigm for genotoxicity testing has an integral part in supporting clinical entry to confirm candidate drugs taken into the clinic are unlikely to have genotoxic activity.


Assuntos
Desenvolvimento de Medicamentos/métodos , Testes para Micronúcleos/métodos , Testes de Mutagenicidade/métodos , Bibliotecas de Moléculas Pequenas/toxicidade , Animais , Medula Óssea/efeitos dos fármacos , Carcinógenos/toxicidade , Aberrações Cromossômicas/induzido quimicamente , Dano ao DNA/efeitos dos fármacos , Drogas em Investigação/toxicidade , Feminino , Humanos , Técnicas In Vitro/métodos , Linfoma/induzido quimicamente , Masculino , Camundongos , Mutação/efeitos dos fármacos , Ratos , Roedores
5.
Artigo em Inglês | MEDLINE | ID: mdl-34360388

RESUMO

Pesticide adjuvants (PAs) denote the general term for auxiliaries in pesticide preparations except for the active components. Toluene, chloroform, and trichloroethylene are the three most commonly used PAs as organic solvents. The residues of the three chemicals in the process of production and application of pesticides may endanger the ecosystem. In the present study, the mutagenicity of toluene, chloroform, and trichloroethylene as well the mixture of the three chemicals was tested by the Salmonella typhimurium reverse mutation test (Ames test) with TA97, TA98, TA100, and TA102 strains in the system with and without rat liver microsomal preparations (S9). The four tester strains have been used for more than 40 years to detect mutagenic compounds in chemicals, cosmetics, and environmental samples. The mutagenicity was detected on tester strains in the separated experiment from the three chemicals. The addition of S9 decreased the mutation ratios of toluene to four strains, except for the TA100 strain, but increased the mutation ratios of chloroform to four strains except for the TA98 strain. Trichloroethylene caused positive mutagenicity to become negative on the TA102 strain. In the mixed experiment, positive effects were detected only on the TA102 strain in the absence of S9. The addition of S9 increased the mutagenicity except for the TA102 strain. The mixture of toluene, chloroform, and trichloroethylene showed antagonism in mutagenicity to tester strains, except for the TA102 strain without S9. However, the mixture showed a synergistic effect to tester strains after adding S9 except for the TA98 strain.


Assuntos
Praguicidas , Tricloroetileno , Animais , Clorofórmio/toxicidade , Ecossistema , Testes de Mutagenicidade , Mutagênicos , Ratos , Tolueno/toxicidade , Tricloroetileno/toxicidade
6.
Sci Total Environ ; 795: 148806, 2021 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-34243001

RESUMO

Previous studies have demonstrated the presence of precursors and coupling agents in wastewater from hair dyeing processes. The complex reaction involved in the oxidation of these compounds can generate extremely hazardous sub-products, leading to an increase in the mutagenicity and toxicity of wastewater. Without proper treatment, this highly toxic wastewater may find its way into the drinking water treatment plant. The present work aimed to investigate the main products generated after the oxidation reaction involving p-toluenediamine (PTD) and p-aminophenol (PAP) - precursors that widely used in the composition of commercial permanent hair dyes, under experimental conditions close to the routine hair dyeing process (in the presence and absence of hydrogen peroxide in ammoniacal medium), using spectroscopic techniques. The study also investigated the mutagenicity and toxicity of the products formed in the hairdressing wash water and conducted detection analysis to determine the presence of the precursors and Bandrowski's Base Derivative (BBD) in samples of wastewater, surface and drinking water using HPLC-DAD and linear voltammetry techniques. Based on this investigation, we identified several PTD and PAP self-oxidation products and eleven sub-products derived from the reaction between PTD and PAP. Assays conducted using Salmonella typhimurium YG1041, with and without activation-induced rat liver metabolism (S9), indicated mutagenicity of the reaction products in concentrations above 10.0 µg µL-1. The concentrations of PTD, PAP, and several reactions and oxidation products of these precursors were detected in wastewater and water samples.


Assuntos
Tinturas para Cabelo , Aminofenóis , Animais , Tinturas para Cabelo/toxicidade , Testes de Mutagenicidade , Mutagênicos/toxicidade , Estresse Oxidativo , Fenilenodiaminas , Ratos
7.
J Toxicol Environ Health A ; 84(18): 743-760, 2021 09 17.
Artigo em Inglês | MEDLINE | ID: mdl-34120581

RESUMO

Jatropha mollissima is used in folk medicine as antimicrobial, antiparasitic, and larvicidal. However, few toxicogenetic studies have been carried out. Therefore, the aim of this study was to determine the phytochemical profile of ethanolic leaf extract of J. mollissima (EEJM) as well as potential cytotoxic, mutagenic, and antimutagenic properties. The EEJM was subjected to successive fractionation for the isolation of secondary metabolites, and five concentrations (0.01; 0.1; 1; 10 and 100 mg/ml) of extract were investigated using Allium cepa assay and the Somatic Mutation and Recombination (SMART) test. The mitotic index and % damage reduction were analyzed for A. cepa and the frequency of mutant hair for SMART. The presence of coumarins, alkaloids, flavonoids, saponins, and tannins was detected, while spinasterol and n-triacontane were the isolates identified for the first time for this species. EEJM did not exhibit cytotoxicity and was not mutagenic at 1 or 10 mg/ml using A. cepa and all concentrations of EEJM were not mutagenic in the SMART test. A cytoprotective effect was found at all concentrations. At 1 or 10 mg/ml EEJM exhibited antimutagenicity in A. cepa. In SMART, the protective effect was observed at 0.1 to 100 mg/ml EEJM. Our results demonstrate the important chemopreventive activity of EEJM, a desired quality in the search for natural anticarcinogenic compounds.


Assuntos
Jatropha/química , Testes de Mutagenicidade , Cebolas/efeitos dos fármacos , Compostos Fitoquímicos/farmacologia , Extratos Vegetais/farmacologia , Substâncias Protetoras/farmacologia , Relação Dose-Resposta a Droga , Compostos Fitoquímicos/química , Extratos Vegetais/química , Folhas de Planta/química
8.
Expert Opin Drug Metab Toxicol ; 17(8): 987-1005, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34078212

RESUMO

Introduction: Genotoxicity is an imperative component of the human health safety assessment of chemicals. Its secure forecast is of the utmost importance for all health prevention strategies and regulations.Areas covered: We surveyed several types of alternative, animal-free approaches ((quantitative) structure-activity relationship (Q)SAR, read-across, Adverse Outcome Pathway, Integrated Approaches to Testing and Assessment) for genotoxicity prediction within the needs of regulatory frameworks, putting special emphasis on data quality and uncertainties issues.Expert opinion: (Q)SAR models and read-across approaches for in vitro bacterial mutagenicity have sufficient reliability for use in prioritization processes, and as support in regulatory decisions in combination with other types of evidence. (Q)SARs and read-across methodologies for other genotoxicity endpoints need further improvements and should be applied with caution. It appears that there is still large room for improvement of genotoxicity prediction methods. Availability of well-curated high-quality databases, covering a broader chemical space, is one of the most important needs. Integration of in silico predictions with expert knowledge, weight-of-evidence-based assessment, and mechanistic understanding of genotoxicity pathways are other key points to be addressed for the generation of more accurate and trustable results.


Assuntos
Simulação por Computador , Testes de Mutagenicidade/métodos , Mutagênicos/toxicidade , Alternativas aos Testes com Animais/métodos , Animais , Bases de Dados Factuais , Humanos , Relação Quantitativa Estrutura-Atividade , Reprodutibilidade dos Testes
9.
Ecotoxicol Environ Saf ; 221: 112421, 2021 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-34147865

RESUMO

A wide variety of organic micropollutants in drinking water pose a serious threat to human health. This study was aimed to reveal the characteristics of organic micropollution profiles in water from a drinking water treatment plant (DWTP) in the Yangtze River Delta, China and investigate the mutagenicity, health risk and disease burden through mixed exposure to micropollutants in water. The presence of organic micropollutants in seven categories in organic extracts (OEs) of water from the DWTP was determined, and Ames test was conducted to test the mutagenic effect of OEs. Meanwhile, health risk of exposure to organic micropollutants in finished water through three exposure routes (ingestion, dermal absorption and inhalation) was assessed with the method proposed by U.S. EPA, and disability-adjusted life years (DALYs) were combined to estimate the disease burden of cancer based on the carcinogenic risk (CR) assessment. The results showed that 28 organic micropollutants were detected in the raw and finished water at total concentrations of 967.28 ng/L and 1073.45 ng/L, respectively, of which phthalate esters (PAEs) were the dominant category (95.79% in the raw water and 96.61% in the finished water). Although the results of the Ames test for OEs were negative and the non-carcinogenic hazard index of the organic micropollutants in the finished water was less than 1 in all age groups, the total CR was 2.17 × 10-5, higher than the negligible risk level (1.00 × 10-6). The total DALYs caused by the organic micropollutants in the finished water was 2945.59 person-years, and the average individual DALYs was 2.21 × 10-6 per person-year (ppy), which was 2.21 times the reference risk level (1.00 × 10-6 ppy) defined by the WHO. Exposure to nitrosamines (NAms) was the major contributor to the total CR (92.06%) and average individual DALYs (94.58%). This study demonstrated that despite the negative result of the mutagenicity test with TA98 and TA100 strains, the health risk of exposure to organic micropollutants in drinking water should not be neglected.


Assuntos
Água Potável/análise , Mutagênicos/análise , Compostos Orgânicos/análise , Poluentes Químicos da Água/análise , China , Efeitos Psicossociais da Doença , Monitoramento Ambiental , Humanos , Testes de Mutagenicidade , Mutagênicos/toxicidade , Compostos Orgânicos/toxicidade , Medição de Risco , Rios , Poluentes Químicos da Água/toxicidade , Purificação da Água
10.
Molecules ; 26(11)2021 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-34072474

RESUMO

Ficus deltoidea var. deltoidea is used as traditional medicine for diabetes, inflammation, and nociception. However, the antimutagenic potential and cytoprotective effects of this plant remain unknown. In this study, the mutagenic and antimutagenic activities of F. deltoidea aqueous extract (FDD) on both Salmonella typhimurium TA 98 and TA 100 strains were assessed using Salmonella mutagenicity assay (Ames test). Then, the cytoprotective potential of FDD on menadione-induced oxidative stress was determined in a V79 mouse lung fibroblast cell line. The ferric-reducing antioxidant power (FRAP) assay was conducted to evaluate FDD antioxidant capacity. Results showed that FDD (up to 50 mg/mL) did not exhibit a mutagenic effect on either TA 98 or TA 100 strains. Notably, FDD decreased the revertant colony count induced by 2-aminoanthracene in both strains in the presence of metabolic activation (p < 0.05). Additionally, pretreatment of FDD (50 and 100 µg/mL) demonstrated remarkable protection against menadione-induced oxidative stress in V79 cells significantly by decreasing superoxide anion level (p < 0.05). FDD at all concentrations tested (12.5-100 µg/mL) exhibited antioxidant power, suggesting the cytoprotective effect of FDD could be partly attributed to its antioxidant properties. This report highlights that F. deltoidea may provide a chemopreventive effect on mutagenic and oxidative stress inducers.


Assuntos
Antimutagênicos/química , Antioxidantes/química , Ficus/metabolismo , Extratos Vegetais/química , Animais , Ânions , Linhagem Celular , Cricetulus , Diabetes Mellitus , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Glutationa , Camundongos , Mutagênese/efeitos dos fármacos , Testes de Mutagenicidade , Mutagênicos , Estresse Oxidativo , Salmonella typhimurium/efeitos dos fármacos , Sais de Tetrazólio/química , Tiazóis/química , Vitamina K 3/química , Água
11.
Mutat Res Rev Mutat Res ; 787: 108363, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34083041

RESUMO

Dr. Bruce Ames turned 92 on December 16, 2020. He considers his most recent work linking adequate consumption of 30 known vitamins and minerals with successful aging to be his most important contribution. With the passage of time, it is not uncommon for the accomplishments of a well-known scientist to undergo a parsimonious reductionism in the public mind - Pasteur's vaccine, Mendel's peas, Pavlov's dogs, Ames' test. Those of us in the research generation subsequent to Dr. Ames' are undoubtedly affected by our own unconscious tendencies toward accepting the outstanding achievements of the past as commonplace. In doing so, seminal advances made by earlier investigators are often inadvertently subsumed into common knowledge. But having followed Ames' work since the mid-1970s, we are cognizant that the eponymous Ames Test is but a single chapter in a long and rich narrative. That narrative begins with Ames' classic studies on the histidine operon of Salmonella, for which he was elected to the National Academy of Sciences. A summary of the historical progression of the understanding of chemical carcinogenesis to which Ames and his colleagues contributed is provided. Any summary of a topic as expansive and complex as the ongoing unraveling of the mechanisms underlying chemical carcinogenesis will only touch upon some of the major conceptual advances to which Ames and his colleagues contributed. We hope that scientists of all ages familiar with Ames only through the eponymous Ames Test will further investigate the historical progression of the conceptualization of cancer caused by chemical exposure. As the field of chemical carcinogenesis gradually moves away from primary reliance on animal testing to alternative protocols under the rubric of New Approach Methodologies (NAM) an understanding of where we have been might help to guide where we should go.


Assuntos
Bioensaio/métodos , Animais , Bases de Dados de Ácidos Nucleicos , Humanos , Testes de Mutagenicidade , Mutação/genética
12.
Mutat Res Rev Mutat Res ; 787: 108364, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34083043

RESUMO

The purpose of this review is to evaluate the literature on the genotoxicity of cumene (CAS # 98-82-8) and to assess the role of mutagenicity, if any, in the mode of action for cumene-induced rodent tumors. The studies reviewed included microbial mutagenicity, DNA damage/ repair, cytogenetic effects, and gene mutations. In reviewing these studies, attention was paid to their conformance to applicable OECD test guidelines which are considered as internationally recognized standards for performing these assays. Cumene was not a bacterial mutagen and did not induce Hprt mutations in CHO cell cultures. In the primary rat hepatocyte cultures, cumene induced unscheduled DNA synthesis in one study but this response could not be reproduced in an independent study using a similar protocol. In a study that is not fully compliant to the current OECD guideline, no increase in chromosomal aberrations was observed in CHO cells treated with cumene. The weight of the evidence (WoE) from multiple in vivo studies indicates that cumene is not a clastogen or aneugen. The weak positive response in an in vivo comet assay in the rat liver and mouse lung tissues is of questionable significance due to several study deficiencies. The genotoxicity profile of cumene does not match that of a classic DNA-reactive molecule and the available data does not support a conclusion that cumene is an in vivo mutagen. As such, mutagenicity does not appear to be an early key event in cumene-induced rodent tumors and alternate hypothesized non-mutagenic modes-of-action are presented. Further data are necessary to rule in or rule out a particular MoA.


Assuntos
Dano ao DNA/fisiologia , Animais , Células CHO , Ensaio Cometa , Cricetulus , Dano ao DNA/genética , Humanos , Mutagênese/genética , Mutagênese/fisiologia , Testes de Mutagenicidade , Mutação/genética , Ratos
14.
Methods Mol Biol ; 2326: 275-285, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34097276

RESUMO

The Ames assay is a classic and robust method for identifying and evaluating chemical mutagens that reverse the mutations of Salmonella typhimurium and/or Escherichia coli bacteria strains with amino acid synthesis defects. It is also called the bacterial reverse mutation assay. Ames assay has been widely used for detecting genetic toxicity of many chemicals and gained increased applications in risk assessment of emerging environmental pollutants such as nanomaterials. In this chapter, we presented a detailed step-by-step method using the Ames assay to detect potential mutagenicity of metal oxide nanoparticles. The strategy to use the liver S9 fraction for bioactivation and a preincubation procedure is recommended. This method is easy to use to test genetic toxicity of other environmental contaminants and new chemicals.


Assuntos
Testes de Mutagenicidade/métodos , Mutagênicos/toxicidade , Nanopartículas/toxicidade , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Metais/toxicidade , Mutação/efeitos dos fármacos , Óxidos/toxicidade , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/genética
15.
J Toxicol Environ Health A ; 84(19): 769-782, 2021 10 02.
Artigo em Inglês | MEDLINE | ID: mdl-34176449

RESUMO

Kavain, kavalactone, present in Piper methysticum exhibits anticonvulsive, analgesic, anxiolytic, antiepileptic, antithrombotic, anti-inflammatory and antioxidant properties. Given its importance, the aim of the present study was to assess (1) the mutagenic and carcinogenicity of kavain administered alone and (2) the antimutagenic and anticarcinogenic potential when administered simultaneously with the chemotherapeutic drug doxorubicin (DXR) using the Somatic Mutation and Recombination Test (SMART) and Epithelial Tumor Test (ETT) using Drosophila melanogaster as a model system. Third-stage larvae from a standard (ST) and high metabolic bioactivation (HB) crosses were treated with different kavain concentrations (32, 64 or 128 µg/ml), alone or in conjunction with DXR (0.125 mg/ml). In ST descendants, kavain produced no significant mutagenic or recombinogenic effects. In the HB cross, mutagenic activity was observed at kavain concentrations of 64 and 128 µg/ml. In the DXR and kavain co-treatment, a modulating effect of the DXR-mediated mutagenic response dependent upon the concentration was detected in both crosses. In ETT, no marked carcinogenic or anticarcinogenic activity was noted for kavain. However, when kavain was combined with DXR synergistic induction of tumors by the chemotherapeutic drug occurred indicating that kavain enhanced the carcinogenic action of DXR.


Assuntos
Doxorrubicina/toxicidade , Drosophila melanogaster/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Pironas/farmacologia , Animais , Carcinogênese , Carcinógenos/toxicidade , Drosophila melanogaster/crescimento & desenvolvimento , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Testes de Mutagenicidade , Mutagênicos/toxicidade
16.
DNA Repair (Amst) ; 105: 103156, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34139663

RESUMO

Nuclear reorganization, including the localization of proteins into discrete subnuclear foci, is a hallmark of the cellular response to DNA damage and replication stress. These foci are thought to represent transient environments or repair factories, in which the lesion is sequestered with molecules and co-factors that catalyze repair. For example, nuclear foci contain signaling proteins that recruit transducer proteins. One important class of transducers is the structure-selective endonucleases, such as SLX1-SLX4, MUS81-EME1, and XPF-ERCC1, which remove branched DNA structures that form during repair. The relocalization of structure-selective endonucleases into subnuclear foci provides a visual read-out for the presence of direct DNA damage, replication barriers, or DNA entanglements and can be monitored using fluorescence microscopy. By simultaneously probing for two or more fluorescent signals, fluorescence microscopy can also provide insights into the proximal association of proteins within a local environment. Here, we report an open-source and semi-automated method to detect and quantify subnuclear foci, as well as foci colocalization and the accompanying pixel-based colocalization metrics. We use this pipeline to show that pre-mitotic nuclei contain a basal threshold of foci marked by SLX1-SLX4, MUS81, or XPF. Some of these foci colocalize with FANCD2 and have a high degree of correlation and co-occurrence. We also show that pre-mitotic cells experiencing replication stress contain elevated levels of foci containing SLX1-SLX4 or XPF, but not MUS81. These results point towards a role for SLX1-SLX4 and XPF-ERCC1 in the early cellular response to replication stress. Nevertheless, most of the foci that form in response to replication stress contain either FANCD2 or one of the three endonucleases. Altogether, our work highlights the compositional heterogeneity of subnuclear foci that form in response to replication stress. We also describe a user-friendly pipeline that can be used to characterize these dynamic structures.


Assuntos
Núcleo Celular/metabolismo , Dano ao DNA , Reparo do DNA , Replicação do DNA , Testes de Mutagenicidade/métodos , Software , Linhagem Celular Tumoral , DNA/metabolismo , Proteínas de Ligação a DNA/metabolismo , Endodesoxirribonucleases/metabolismo , Endonucleases/metabolismo , Proteína do Grupo de Complementação D2 da Anemia de Fanconi/metabolismo , Humanos , Recombinases/metabolismo
17.
J Hazard Mater ; 419: 126422, 2021 10 05.
Artigo em Inglês | MEDLINE | ID: mdl-34182426

RESUMO

The chemical constituents in food contact materials (FCMs) may transfer into food during the contact, which may pose potential risk to humans. So, it is important to evaluate the safety of FCMs. Due to the advantages of cost-effectiveness and high throughput, (Q)SAR tools have been gradually used for risk assessment. In this work, a risk classification strategy for migrants of food contact materials combined with three (Q)SAR tools was developed based on a single endpoint (Mutagenicity) assessment and risk matrix approach, respectively. 419 migrants existing in a self-built toxicology database beneficial from Python crawler technology were evaluated. 5 toxic hazard ranks and 4 risk ranks were obtained for single endpoint assessment and risk matrix respectively, with 21 substances assigned as Toxic hazard Class I and 43 substances assigned as RISK Ⅰ which need the highest safety concern. Besides, for the Toxic hazard Class I substances assessed by the single endpoint, 19 of them were confirmed experimentally, and all of them were overlapped in the RISK Ⅰ substances, which suggests the effectiveness and reliability of this strategy.


Assuntos
Embalagem de Alimentos , Mutagênicos , Simulação por Computador , Humanos , Testes de Mutagenicidade , Reprodutibilidade dos Testes , Medição de Risco
18.
Artigo em Inglês | MEDLINE | ID: mdl-34067860

RESUMO

The aim of this paper was to investigate the relationship between micronuclei and DNA damage in children's buccal mucosa cells and the genotoxicity and mutagenicity of the different sized fractions of particulate matter as well as the concentration of PAHs (polycyclic aromatic hydrocarbons) and metals in particulate matter. Air particulate matter was collected by high volume samplers located near the schools attended by the children on the same days of biological samplings. The mutagenic activity was assessed in different cells in in vitro tests (Ames test on bacteria and comet test on leukocytes). Our study showed weak positive correlations between (a) the mutagenicity of the PM0.5 fraction and PAHs and (b) the micronuclei test of children's buccal cells and PAHs detected in PM0.5 and PM0.5-3 fractions. A positive correlation was also found between in vitro comet test on leukocytes and PAHs in the PM3-10 fraction. No correlation was observed for metal concentrations in each PM fraction.


Assuntos
Poluentes Atmosféricos , Poluição do Ar , Hidrocarbonetos Policíclicos Aromáticos , Poluentes Atmosféricos/análise , Poluentes Atmosféricos/toxicidade , Poluição do Ar/efeitos adversos , Criança , Dano ao DNA , Humanos , Mucosa Bucal , Testes de Mutagenicidade , Mutagênicos/toxicidade , Material Particulado/análise , Material Particulado/toxicidade , Hidrocarbonetos Policíclicos Aromáticos/análise , Hidrocarbonetos Policíclicos Aromáticos/toxicidade
19.
J Toxicol Environ Health A ; 84(17): 689-701, 2021 09 02.
Artigo em Inglês | MEDLINE | ID: mdl-34034641

RESUMO

Nicotiana tabacum is the most cultivated tobacco species in the state of Rio Grande do Sul, Brazil. Workers who handle the plant are exposed to the leaf components during the harvesting process and when separating and classifying the dried leaves. In addition to nicotine, after the drying process, other components may be found including tobacco-specific nitrosamines, polycyclic aromatic hydrocarbons, as well as pesticides residues. The objective of this study was to examine the genotoxicity attributed to the aqueous extract of dried tobacco leaves obtained from tobacco barns using Chinese hamster lung fibroblast cells (V79) as a model system by employing alkaline comet assay, micronucleus (MN) and Ames test. MTT assay was used to assess cytotoxicity and establish concentrations for this study. Data demonstrated cell viability > 85% for concentrations of 0.625-5 mg/ml while the comet assay indicated a significant increase in DNA damage at all concentrations tested. A significant elevation of MN and nuclear buds (NBUD) was found for 5 mg/ml compared to control and other dry tobacco leaves concentrations (0.625-2.5 mg/ml). Mutagenicity was not found using the Salmonella/Microsome test (TA98, TA100, and TA102 strains) with and without metabolic activation. The concentration of inorganic elements was determined employing the PIXE technique, and 13 inorganic elements were detected. Using CG/MS nicotine amounts present were 1.56 mg/g dry tobacco leaf powder. Due to the observed genotoxicity in V79 cells, more investigations are needed to protect the health of tobacco workers exposed daily to this complex mixture of toxic substances present in dry tobacco leaves.


Assuntos
Mutagênicos/toxicidade , Folhas de Planta/química , Tabaco/química , Animais , Linhagem Celular , Ensaio Cometa , Cricetulus , Testes para Micronúcleos , Testes de Mutagenicidade
20.
Int J Biol Macromol ; 181: 1196-1206, 2021 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-33991555

RESUMO

Loading propolis by a simple process using genipin as a crosslinking agent and fabrication of a novel PVA/Chitosan-Propolis membrane scaffolds were reported for wound dressing applications. The research is focused on the effects of propolis on characterization properties of membrane such as chemical structure, surface morphology, degradation ratio, crystallinity, hydrophilicity, water uptake capacity, water vapour transmission rate and mechanical aspect. It was noticed that water uptake capacity and hydrophilicity properties of membrane considerably affected by the propolis. By addition of (0.50, % v/v) propolis, the contact angle of the PVA/Chitosan membrane was remarkably decreased from 86.29° ± 3 to 45 ± 2°. 3-(4,5-dimethylthiazoyl-2-yl)-2,5-diphenylte-trazolium (MTT) bromide test and SEM were used to analyse the cytocompatibility of the membranes and morphology of cells on membrane. The propolis incorporated membrane showed cell proliferation rate 176 ± 13%, 775 ± 1%, and 853 ± 23%, at 24 h, 27 h and 120 h, respectively. SEM images also supported the cell behaviour on membrane. DNA fragmentation was also investigated with genotoxicity test. The studies on the interactions between membranes and MEF cells revealed that the incorporation of propolis into membrane promoted cell proliferation. These overall results presented that propolis incorporated membranes could have potentially appealing application as scaffolds for wound healing applications.


Assuntos
Materiais Biocompatíveis/química , Quitosana/química , Própole/química , Cicatrização/efeitos dos fármacos , Materiais Biocompatíveis/farmacologia , Proliferação de Células/efeitos dos fármacos , Quitosana/farmacologia , Fragmentação do DNA/efeitos dos fármacos , Humanos , Iridoides/química , Membranas Artificiais , Testes de Mutagenicidade , Própole/farmacologia
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