Toxicogenetic study of omeprazole and the modulatory effects of retinol palmitate and ascorbic acid on Allium cepa.

Omeprazole (OME) is a proton pump inhibitor used for the treatment of various gastric and intestinal disease; however, studies on its effects on the genetic materials are still restricted. The present study aimed to evaluate possible toxicogenic effects of OME in Allium cepa meristems with the application of cytogenetic biomarkers for DNA damage, mutagenic, toxic and cytotoxic effects. Additionally, retinol palmitate (RP) and ascorbic acid (AA) were also co-treated with OME to evaluate possible modulatory effects of OME-induced cytogenetic damages. OME was tested at 10, 20 and 40 µg/mL, while RP and AA at 55 µg/mL and 352.2 µg/mL, respectively. Copper sulphate (0.6 µg/mL) and dechlorinated water were used as positive control and negative control, respectively. The results suggest that OME induced genotoxicity and mutagenicity in A. cepa at all tested concentrations. It was noted that cotreatment of OME with the antioxidant vitamins RP and/or AA significantly (p < 0.05) inhibited and/or modulated all toxicogenic damages induced by OME. These observations demonstrate their antigenotoxic, antimutagenic, antitoxic and anticitotoxic effects in A. cepa. This study indicates that application of antioxidants may be useful tools to overcome OME-induced toxic effects.

Guia para a análise transcriptômica: potencial toxicológico e mecanismos de ação de produtos de origem natural / Guide to transcriptomic analysis: toxicological potential and mechanisms of natural products action

Nas últimas décadas houve um grande avanço nos ensaios in vitro, na tentativa de prever resultados de tratamentos in vivo e, com isso, reduzir a utilização de animais de laboratório. A toxicogenômica, que é a integração entre a toxicologia, a bioinformática e as tecnologias ômicas, tem se mostrado preditiva para os mecanismos de ação e toxicidade. Esta ferramenta possibilita a investigação da expressão de milhares de genes, permitindo a visualização de diferentes perfis de alteração e de assinatura gênica. A utilização da toxicogenômica se torna adequada para a compreensão de mecanismos genéticos de resposta de organismos expostos a formulações de origem natural que são capazes de modular múltiplos genes. Este trabalho objetivou destacar os pontos críticos das etapas que devem ser seguidas para se realizar, apropriadamente, uma análise toxicogenômica. Dentre essas etapas estão a análise das características desejáveis para os compostos, a escolha apropriada das linhagens celulares para realizar a exposição, a seleção das concentrações das amostras a serem testadas e dos controles que devem ser utilizados nos ensaios biológicos, a escolha do princípio a ser utilizado para o ensaio de viabilidade celular, a seleção dos procedimentos para tratar os dados gerados e avaliação dos resultados encontrados. As sugestões apresentadas têm a intenção de divulgar a técnica de toxicogenômica, bem como, estimular e facilitar o desenvolvimento de um estudo que utilize a análise toxicogenômica como ferramenta alternativa na investigação da toxicidade e de mecanismos/modo de ação de produtos de origem natural.

In the last decades there has been a great advance in the in vitro tests, in the attempt to predict results of treatments in vivo and, with that, to reduce the use of laboratory animals. Toxicogenomics, which is the integration between toxicology, bioinformatics and atomic technologies, has been shown to be predictive of mechanisms of action and toxicity. This tool allows the investigation of the expression of thousands of genes, allowing the visualization of different profiles of gene signature and change. The use of toxicogenomics becomes suitable for the understanding of genetic mechanisms of response of organisms exposed to formulations of natural origin that are able to modulate multiple genes. This work aimed to highlight the critical points of the steps that must be followed to carry out, properly, a toxicogenomic analysis. Among these steps are the analysis of the desirable characteristics for the compounds, the appropriate choice of the cell lines to carry out the exposure, the selection of the concentrations of the samples to be tested and the controls to be used in the biological assays, the choice of principle to be used for the cell viability assay, the selection of procedures to treat the generated data and evaluation of the results found. The suggestions presented are intended to stimulate and facilitate the developmentof a study that uses toxicogenomic analysis as an alternative tool in the investigation of the toxicity and mechanisms / mode of action of products of natural origin.

Guia para a análise transcriptômica: potencial toxicológico e mecanismos de ação de produtos de origem natural / Guide to transcriptomic analysis: toxicological potential and mechanisms of natural products action

Nas últimas décadas houve um grande avanço nos ensaios in vitro, na tentativa de prever resultados de tratamentos in vivo e, com isso, reduzir a utilização de animais de laboratório. A toxicogenômica, que é a integração entre a toxicologia, a bioinformática e as tecnologias ômicas, tem se mostrado preditiva para os mecanismos de ação e toxicidade. Esta ferramenta possibilita a investigação da expressão de milhares de genes, permitindo a visualização de diferentes perfis de alteração e de assinatura gênica. A utilização da toxicogenômica se torna adequada para a compreensão de mecanismos genéticos de resposta de organismos expostos a formulações de origem natural que são capazes de modular múltiplos genes. Este trabalho objetivou destacar os pontos críticos das etapas que devem ser seguidas para se realizar, apropriadamente, uma análise toxicogenômica. Dentre essas etapas estão a análise das características desejáveis para os compostos, a escolha apropriada das linhagens celulares para realizar a exposição, a seleção das concentrações das amostras a serem testadas e dos controles que devem ser utilizados nos ensaios biológicos, a escolha do princípio a ser utilizado para o ensaio de viabilidade celular, a seleção dos procedimentos para tratar os dados gerados e avaliação dos resultados encontrados. As sugestões apresentadas têm a intenção de divulgar a técnica de toxicogenômica, bem como, estimular e facilitar o desenvolvimento de um estudo que utilize a análise toxicogenômica como ferramenta alternativa na investigação da toxicidade e de mecanismos/modo de ação de produtos de origem natural.(AU)

In the last decades there has been a great advance in the in vitro tests, in the attempt to predict results of treatments in vivo and, with that, to reduce the use of laboratory animals. Toxicogenomics, which is the integration between toxicology, bioinformatics and atomic technologies, has been shown to be predictive of mechanisms of action and toxicity. This tool allows the investigation of the expression of thousands of genes, allowing the visualization of different profiles of gene signature and change. The use of toxicogenomics becomes suitable for the understanding of genetic mechanisms of response of organisms exposed to formulations of natural origin that are able to modulate multiple genes. This work aimed to highlight the critical points of the steps that must be followed to carry out, properly, a toxicogenomic analysis. Among these steps are the analysis of the desirable characteristics for the compounds, the appropriate choice of the cell lines to carry out the exposure, the selection of the concentrations of the samples to be tested and the controls to be used in the biological assays, the choice of principle to be used for the cell viability assay, the selection of procedures to treat the generated data and evaluation of the results found. The suggestions presented are intended to stimulate and facilitate the developmentof a study that uses toxicogenomic analysis as an alternative tool in the investigation of the toxicity and mechanisms / mode of action of products of natural origin.(AU)

Use of Copaifera multijuga for acute corneal repair after chemical injury: A clinical, histopathological and toxicogenetic study.

Copaiba oil is widely used in medicine, but there are no reports regarding its application in ophthalmology. Therefore, the objective of this study was to evaluate the clinical, histopathological and toxicogenetic effects of eye drops containing 0.1 and 0.5% of Copaifera multijuga Hayne oil on superficial corneal ulcers induced with alkali in the left eye of rats. For histological analysis, the percent reduction in ulcers and thickness of the corneal epithelium and stroma were evaluated 48 and 72 h after ulcer induction. Additionally, neovascularization and polymorphonuclear infiltration were classified in the stroma. The bone marrow micronucleus test was used for toxicogenetic assessment. None of the animals exhibited clinical signs of immediate ocular discomfort after instillation and the eye drops were harmless to the ocular surface. There was a significant difference in percent ulcer reduction and corneal stroma thickness between animals treated with the C. multijuga eye drops and untreated animals with corneal injury and the negative control, respectively, suggesting a healing effect of the oleoresin. Analysis of the thickness of the corneal epithelium at the two time points showed that the eye drops formulated did not significantly reduce the damage caused by alkali. The same was observed for the treatments with the reference drugs. No difference in stromal neovascularization or inflammatory infiltration was observed between the treated groups. The toxicogenetic results revealed the absence of cytotoxicity and genotoxicity of the treatments. In conclusion, the C. multijuga eye drops did not cause damage to the ocular surface under the present experimental conditions and corneal epithelization was similar to the conventional treatments. These results indicate that eye drops containing C. multijuga oleoresin are a promising option for the treatment of superficial keratitis.

De novo transcriptome assembly, functional annotation and differential gene expression analysis of juvenile and adult E. fetida, a model oligochaete used in ecotoxicological studies.

BACKGROUND: Earthworms are sensitive to toxic chemicals present in the soil and so are useful indicator organisms for soil health. Eisenia fetida are commonly used in ecotoxicological studies; therefore the assembly of a baseline transcriptome is important for subsequent analyses exploring the impact of toxin exposure on genome wide gene expression. RESULTS: This paper reports on the de novo transcriptome assembly of E. fetida using Trinity, a freely available software tool. Trinotate was used to carry out functional annotation of the Trinity generated transcriptome file and the transdecoder generated peptide sequence file along with BLASTX, BLASTP and HMMER searches and were loaded into a Sqlite3 database. To identify differentially expressed transcripts; each of the original sequence files were aligned to the de novo assembled transcriptome using Bowtie and then RSEM was used to estimate expression values based on the alignment. EdgeR was used to calculate differential expression between the two conditions, with an FDR corrected P value cut off of 0.001, this returned six significantly differentially expressed genes. Initial BLASTX hits of these putative genes included hits with annelid ferritin and lysozyme proteins, as well as fungal NADH cytochrome b5 reductase and senescence associated proteins. At a cut off of P = 0.01 there were a further 26 differentially expressed genes. CONCLUSION: These data have been made publicly available, and to our knowledge represent the most comprehensive available transcriptome for E. fetida assembled from RNA sequencing data. This provides important groundwork for subsequent ecotoxicogenomic studies exploring the impact of the environment on global gene expression in E. fetida and other earthworm species.

Toxicogenetic study of Persea americana fruit pulp oil and its effect on genomic instability.

Persea americana Mill., commonly known as avocado, is a tree native to Central America that is widely used as a food source and for the treatment of diseases. This plant has various biological properties such as analgesic, anti-inflammatory and total cholesterol-lowering activity. In view of its pharmacological potential, we conducted a toxicogenetic study of the fruit pulp oil of P. americana (PAO) and investigated its influence on genotoxicity induced by methyl methanesulfonate (MMS) and doxorubicin. V79 cells and Swiss mice were used for the assays. The results showed no genotoxic effects of PAO in the in vitro or in vivo test systems. However, the highest PAO dose tested led to an increase in the levels of aspartate aminotransferase, indicating hepatic/tissue damage. This effect may be related to high concentrations of palmitic acid, the main component of PAO. Furthermore, PAO was effective in reducing the chromosome damage induced by MMS and doxorubicin. These results contribute to the safety assessment of PAO as a medicinal plant for human use.

De novo transcriptome assembly, functional annotation and differential gene expression analysis of juvenile and adult E. fetida, a model oligochaete used in ecotoxicological studies

BACKGROUND: Earthworms are sensitive to toxic chemicals present in the soil and so are useful indicator organisms for soil health. Eisenia fetida are commonly used in ecotoxicological studies; therefore the assembly of a baseline transcriptome is important for subsequent analyses exploring the impact of toxin exposure on genome wide gene expression. RESULTS: This paper reports on the de novo transcriptome assembly of E. fetida using Trinity, a freely available software tool. Trinotate was used to carry out functional annotation of the Trinity generated transcriptome file and the transdecoder generated peptide sequence file along with BLASTX, BLASTP and HMMER searches and were loaded into a Sqlite3 database. To identify differentially expressed transcripts; each of the original sequence files were aligned to the de novo assembled transcriptome using Bowtie and then RSEM was used to estimate expression values based on the alignment. EdgeR was used to calculate differential expression between the two conditions, with an FDR corrected P value cut off of 0.001, this returned six significantly differentially expressed genes. Initial BLASTX hits of these putative genes included hits with annelid ferritin and lysozyme proteins, as well as fungal NADH cytochrome b5 reductase and senescence associated proteins. At a cut off of P = 0.01 there were a further 26 differentially expressed genes. CONCLUSION: These data have been made publicly available, and to our knowledge represent the most comprehensive available transcriptome for E. fetida assembled from RNA sequencing data. This provides important groundwork for subsequent ecotoxicogenomic studies exploring the impact of the environment on global gene expression in E. fetida and other earthworm species.

Etanol e isopropanol: genotoxicidad y teratogénesis evaluada aplicando el modelo de Drosophila melanogaster / Ethanol and isopropanol: genotoxicity and teratogenicity evaluated using Drosophila melanogaster as a model system

El etanol y el isopropanol son, de los alcoholes alifáticos de cadena corta, los más frecuentemente asociados a la actividad humana tanto a nivel industrial como en el entorno doméstico. En este trabajo se presentan los principales hallazgos reportados en la literatura para ensayos de genotoxicidad y teratogénesis en modelos experimentales de distinto nivel de complejidad, con especial énfasis en Drosophila melanogaster. El metabolismo de estos alcoholes es semejante en Drosophila y en humanos por lo cual la mosca es un buen modelo in vivo para la evaluación de sus potenciales efectos tóxicos, genotóxicos y teratogénicos.

Ethanol and isopropanol are two of the short chain aliphatic alcohols more frequently associated to the human environment, both in the industrial and domestic conditions. The aim of this work was to present the main findings reported in the literature about their genotoxicity and teratogenicity in experimental models of different level of complexity, with special emphasis in Drosophila melanogaster. Taking into account that the metabolism of both alcohols in Drosophila and humans is similar, the fly is a good model for the evaluation of their potentially toxic, genotoxic and teratogenic effects.