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2.
Sci Rep ; 10(1): 2190, 2020 02 10.
Artigo em Inglês | MEDLINE | ID: mdl-32042077

RESUMO

Understanding the effect of pesticides on the survival of honeybee colonies is important because these pollinators are reportedly declining globally. In the present study, we examined the changes in the head proteome of nurse honeybees exposed to individual and combined pesticides (the fungicide pyraclostrobin and the insecticide fipronil) at field-relevant doses (850 and 2.5 ppb, respectively). The head proteomes of bees exposed to pesticides were compared with those of bees that were not exposed, and proteins with differences in expression were identified by mass spectrometry. The exposure of nurse bees to pesticides reduced the expression of four of the major royal jelly proteins (MRJP1, MRJP2, MRJP4, and MRJP5) and also several proteins associated with carbohydrate metabolism and energy synthesis, the antioxidant system, detoxification, biosynthesis, amino acid metabolism, transcription and translation, protein folding and binding, olfaction, and learning and memory. Overall, when pyraclostrobin and fipronil were combined, the changes in protein expression were exacerbated. Our results demonstrate that vital proteins and metabolic processes are impaired in nurse honeybees exposed to pesticides in doses close to those experienced by these insects in the field, increasing their susceptibility to stressors and affecting the nutrition and maintenance of both managed and natural colonies.


Assuntos
Abelhas/metabolismo , Praguicidas/efeitos adversos , Proteoma/efeitos dos fármacos , Animais , Abelhas/efeitos dos fármacos , Conservação dos Recursos Naturais/métodos , Ácidos Graxos/metabolismo , Fungicidas Industriais/efeitos adversos , Proteínas de Insetos/metabolismo , Inseticidas/efeitos adversos , Proteoma/metabolismo , Proteômica/métodos , Pirazóis/efeitos adversos , Estrobilurinas/efeitos adversos
3.
Sci Total Environ ; 711: 134547, 2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-31812405

RESUMO

Fish is an important source of protein, vitamins, and minerals. However, this food is also a major source of human exposure to toxic contaminants such as mercury. Thus, this paper aimed to evaluate mercury-binding proteins for possible application as biomarkers of mercury contamination in hepatic and renal tissues of Plagioscion squamosissimus (carnivorous fish) and Colossoma macropomum (omnivorous fish) from the Amazon region using metalloproteomic approach. The proteome of hepatic and renal tissues of fish species was separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and the mercury concentrations in protein spots were determined by graphite furnace atomic absorption spectrometry (GFAAS). Finally, the protein spots associated to mercury were characterized by electrospray ionization mass spectrometry (ESI-MS/MS). The activity of antioxidant enzymes (SOD, CAT, GPx, and GST) and lipid peroxidation (LPO) were also determined. The results showed that the highest concentrations of mercury were found in the carnivorous species (P. squamosissimus) and that the accumulation pattern of this metal was higher in hepatic tissues than in renal tissues for both species. A tendency was observed for greater enzymatic activity in the hepatic and renal tissues of P. squamosissimus, the species with the highest concentration of mercury. Only GPx activity in the kidney and GST in the liver were lower for the P. squamosissimus species, and this finding can be explained by the interaction of mercury with these enzymes. The data obtained by ESI-MS/MS allowed for the characterization of the protein spots associated to mercury, revealing proteins involved in energy metabolism, biomolecules transport, protein synthesis and degradation, cell differentiation, gene regulation, and the antioxidant system. The results obtained in the present study can contribute to understanding the physiological processes underlying mercury toxicity and have provided new perspectives on possible candidates for mercury contamination biomarkers in fish.


Assuntos
Fígado , Animais , Biomarcadores , Proteínas de Transporte , Humanos , Mercúrio , Proteômica , Espectrometria de Massas em Tandem , Poluentes Químicos da Água
4.
Biol Trace Elem Res ; 195(2): 648-657, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31473899

RESUMO

This study aimed to evaluate the quality of the royal jelly produced by Apis mellifera bees in the presence of different iron concentrations (ferrous sulfate heptahydrate-0, 25, 50, and 100 mg L-1). Two-dimensional electrophoresis was used for the fractionation of royal jelly proteins, and iron level was quantified using flame atomic absorption spectrometry technique. The proteins were identified using electrospray ionisation mass spectrometry. Analysis of variance followed by the Tukey test (P < 0.05) was utilised. Dietary supplementation with mineral Fe affected the protein content and number of proteins in the experimental period. Further, the diet containing the highest iron concentration showed a greater number of spots containing iron, as well as in the abdomen of the bees. The most protein containing Fe were classified as major royal jelly proteins. These results showed that Fe influenced the quality of royal jelly and can improve its nutritional value.


Assuntos
Ácidos Graxos/química , Compostos Ferrosos/análise , Proteínas de Insetos/análise , Animais , Abelhas , Suplementos Nutricionais , Ácidos Graxos/biossíntese , Compostos Ferrosos/administração & dosagem , Compostos Ferrosos/metabolismo , Proteínas de Insetos/metabolismo
5.
Chemosphere ; 236: 124320, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31323548

RESUMO

High concentrations of mercury found in soils, sediments, fish, and humans of the Amazon region have gained prominence in scientific studies during the last decade. However, studies related to the elucidation of mercury toxicity mechanisms in ichthyofauna at the molecular and metallomic levels that seek to elucidate physiological and functional aspects, as well as the search for biomarkers of mercury exposure, are still sparse. In the search for these answers, the present study analyzed the hepatic tissue proteome of the Arapaima gigas (pirarucu) fish species collected in the Jirau hydroelectric power plant reservoir in the state of Rondônia state, Brazil, in order to identify mercury-related metal-binding proteins and to elucidate their physiological and functional aspects. The proteomic profile of the hepatic tissue of Arapaima gigas was obtained by two-dimensional electrophoresis (2D-PAGE) and the presence of mercury was mapped in the protein SPOTS by graphite furnace atomic absorption spectrometry(GFAAS). Mercury was detected in 18 protein SPOTS with concentrations ranging from 0.13 ±â€¯0.003 to 131.00 ±â€¯3 mg kg-1. The characterization of the protein SPOTS associated with mercury was performed by electrospray ionisation tandem mass spectrometry (ESI-MS/MS), and 10 proteins were identified. Bioinformatics analyses showed that most of the proteins found linked to mercury were involved in cellular component processes and biological processes. For the most part, protein sequences have cellular functions comprising catalytic, binding, sense of localization, and metabolic processes.


Assuntos
Proteínas de Transporte/química , Mercúrio/química , Proteômica/métodos , Animais , Brasil , Peixes , Humanos
6.
Food Chem ; 276: 247-254, 2019 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-30409591

RESUMO

Mercury has the ability to bind to a variety of biomolecules, which can compromise its structure and functionality and thus promote its toxic effects. The aim of this study is to identify possible mercury biomarkers in muscle samples of Plagioscion squamosissimus (carnivorous fish) and Colossoma macropomum (omnivorous fish), from the Amazon region. The muscle proteome of fish species was separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and the total mercury concentrations in protein spots were determined by graphite furnace atomic absorption spectrometry (GFAAS). The protein spots containing mercury were characterized by electrospray ionization tandem mass spectrometry (ESI-MS/MS). The mercury concentrations in the protein spots were in the range of 1.10 ±â€¯0.02-23.90 ±â€¯0.33 µg g-1. The proteins phosphoglycerate mutase 2 (P. squamosissimus), hemoglobin ß and cytochrome P450scc (C. macropomum), identified by ESI-MS/MS and showing the highest values of mercury concentration, may be considered possible mercury biomarkers.


Assuntos
Caraciformes/metabolismo , Poluentes Ambientais/toxicidade , Mercúrio/toxicidade , Músculos/efeitos dos fármacos , Músculos/metabolismo , Perciformes/metabolismo , Animais , Biomarcadores/metabolismo , Proteômica
7.
Biol Trace Elem Res ; 184(2): 517-522, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29196873

RESUMO

This paper presents a slurry sampling method for total mercury determination by graphite furnace atomic absorption spectrometry (GFAAS) in tissue of fish from the Amazon. The tissue samples were lyophilized and macerated, and then the slurry samples were prepared by putting 20 mg of tissue, added to a solution containing Triton X-100, Suprapur HNO3, and zirconium nitrate directly in sampling vials of a spectrometer. Mercury standard solutions were prepared under the same conditions as the slurry samples. The slurry samples and the mercury standard solutions were sonicated for 20 s. Twenty microliters of slurry samples were injected into the graphite tube, which contained an internal wall lined with tungsten carbide. Under these conditions, it was possible to thermally stabilize the mercury up to an atomization temperature of 1700 °C. The method was validated by mercury determination in reference materials DORM-4 and DOLT-4. The LOD and LOQ were 0.014 and 0.045 mg kg-1, respectively, and recovery percentages in relation to the concentration values were certified in the order of 98%.


Assuntos
Peixes , Fígado/química , Mercúrio/análise , Músculos/química , Animais , Brasil , Grafite/química , Temperatura Alta , Mercúrio/normas , Padrões de Referência , Reprodutibilidade dos Testes , Espectrofotometria Atômica/métodos
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