| Author | Lopes, Renato Matos | |
| Author | Bezerra, Adriana L. A. | |
| Author | Hauser-Davis, Rachel A. | |
| Author | Oliveira, Manildo Marcião de | |
| Author | Salles, João B. | |
| Author | Bastos, Vera L. F. Cunha | |
| Author | Bastos, Jayme Cunha | |
| Access date | 2018-09-20T12:30:59Z | |
| Available date | 2018-09-20T12:30:59Z | |
| Document date | 2018 | |
| Citation | LOPES, Renato Lopes; et al. The importance of an efficient extraction protocol for the use of fish muscle cholinesterases as biomarkers. Talanta, v.179,p.769-774, 2018. | pt_BR |
| ISSN | 0039-9140 | pt_BR |
| URI | https://www.arca.fiocruz.br/handle/icict/28927 | |
| Language | eng | pt_BR |
| Publisher | Elsevier | pt_BR |
| Rights | restricted access | pt_BR |
| Subject in Portuguese | Acetylcholinesterase | pt_BR |
| Subject in Portuguese | método de validação | pt_BR |
| Subject in Portuguese | Biomarcador | pt_BR |
| Subject in Portuguese | Pesticida | pt_BR |
| Subject in Portuguese | músculo | pt_BR |
| Subject in Portuguese | Peixe | pt_BR |
| Title | The importance of an efficient extraction protocol for the use of fish muscle cholinesterases as biomarkers | pt_BR |
| Type | Article | pt_BR |
| DOI | 10.1016/j.talanta.2017.12.013 | |
| Abstract | Esterase activity found in muscle extracts is useful to evaluate harmful effects of anticholinesterase pollutants. Yet, most procedures applied in the extraction of fish muscle esterases in order to investigate their activity as a biomarker of environmental exposure comprise the homogenization of muscle tissue in low-salt solutions, followed by centrifugation to separate the supernatant as the enzyme source. However, acetylcholinesterase (AChE), the main target in these monitoring efforts, is a membrane-bound protein and is only present in muscle extracts if homogenization is carried out using chaotropic high-salt solutions. In this context, four extraction procedures using muscle tissue from six fish species were evaluated in order to establish a reproducible and reliable AChE assay for the determination of this biomarker. Results indicate that over 80% of AChE activity might be lacking in low-salt supernatants, and that the highest activities are obtained after extraction with solutions containing either 1molL-1 NaCl or 1molL-1 NaCl plus 3% Triton X-100, preserving almost 100% esterase activity over acetylthiocholine as substrate after centrifugation. Thus, many studies in the literature suffer from theoretical flaws and report erroneous AChE activity, since typical muscle AChE activity, the end-point biomarker for anticholinesterase pollutants, may have not been consistently assayed. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Comunicação Celular. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Escola Nacional de Saúde Pública Sergio Arouca. Centro de Estudos da Saúde do Trabalhador e Ecologia Humana. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Escola Nacional de Saúde Pública Sergio Arouca. Centro de Estudos da Saúde do Trabalhador e Ecologia Humana. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Instituto Federal de Educação, Ciência e Tecnologia Fluminense. Laboratório de Ecotoxicologia e Microbiologia Ambiental. Cabo Frio, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Centro Universitário Estadual da Zona Oeste. Departamento de Bioquímica. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Universidade do Estado do Rio de Janeiro. Instituto de Biologia Roberto Alcântara Gomes. Departamento de Bioquímica. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Universidade do Estado do Rio de Janeiro. Instituto de Biologia Roberto Alcântara Gomes. Departamento de Bioquímica. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Subject | Inibidores da Colinesterase | pt_BR |
| Subject | Method validation | pt_BR |
| Subject | Biomarker | pt_BR |
| Subject | Pesticide | pt_BR |
| Subject | Muscle | pt_BR |
| Subject | Fish | pt_BR |
| DeCS | Inibidores da Colinesterase | pt_BR |
| e-ISSN | 1873-3573 | |
| Embargo date | 2030-01-01 | |
| xmlui.metadata.dc.subject.ods | 14 Vida na água | |
