The effects of mercury exposure on Amazonian fishes: An investigation of potential biomarkers.

Exposure to mercury can interfere with the expression of proteins and enzymes, compromise important pathways, such as apoptosis and glucose metabolism, and even induce the expression of metallothioneins. In this study, analytical techniques were used to determine the concentration of total mercury (THg) in muscle and liver tissue, protein pellets, and spots [using graphite furnace atomic absorption spectrometry (GFAAS)], and molecular techniques were used to identify metalloproteins present in mercury-associated protein spots. Thirty individuals from three different fish species, Cichla sp. (n = 10), Brachyplatystoma filamentosum (n = 10), and Semaprochilodus sp. (n = 10) from the Brazilian Amazon were used. Oxidative stress indicators [such as glutathione peroxidase (GSH-Px), catalase (CAT), superoxide dismutase (SOD), a marker of lipid peroxidation (LPO)] and the possible expression of metallothioneins in muscle and liver tissues were investigated. The two piscivorous species, Cichla sp. and B. filamentosum, presented the highest concentrations of mercury in their hepatic tissue, 1219 ± 15.00 and 1044 ± 13.6 µg kg-1, respectively, and in their muscle tissue, 101 ± 1.30 µg kg-1 and 87.4 ± 0.900 µg kg-1, respectively. The non-carnivorous species Semaprochilodus sp. had comparatively low concentrations of mercury in both its hepatic (852 ± 11.1 µg kg-1) and muscle (71.4 ± 0.930 µg kg-1) tissues. The presence of mercury was identified in 24 protein spots using GFAAS; concentrations ranged from 11.5 to 787 µg kg-1, and mass spectrometry identified 21 metal-binding proteins. The activities of GSH-Px, CAT, and SOD, related to oxidative stress, decreased proportionally as tissue Hg concentrations increased, while the levels of LPO markers increased, indicating the presence of stress. Our study results demonstrate possible mercury interference in oxidative stress markers (GSH-Px, CAT, SOD, and LPO), in addition to the identification of 21 metal-binding proteins as possible biomarkers of mercury exposure in fish.

Investigation of Protein Biomarkers and Oxidative Stress in Pinirampus pirinampu Exposed to Mercury Species from the Madeira River, Amazon-Brazil.

In recent decades, the scientific community has widely debated the contamination of fish in the Amazon region by mercury species. As the diet of riverside populations in the Amazon region is based mainly on fish, these populations are exposed to mercurial species that can cause serious and irreversible damage to their health. The risks of consuming fish exposed to mercurial species in the Amazon region have motivated toxicological investigations. However, the effect of mercurial species on protein and enzyme levels is still controversial. In this work, analytical and bioanalytical techniques Two-dimensional polyacrylamide gel electrophoresis [2D-PAGE] Graphite Furnace Atomic Absorption Spectrometry [GFAAS], and Mass Spectrometry in Sequence with Electrospray Ionization [ESI-MS/MS] were used to identify proteins associated with mercury (metal-binding protein) in muscle and liver tissues of the fish species Pinirampus pirinampu from the Madeira River, in the Brazilian Amazon. Enzymatic and lipid peroxidation analyses were also used to assess changes related to oxidative stress. Determinations of total mercury by GFAAS indicated higher concentrations in liver tissue (555 ± 19.0 µg kg-1) when compared to muscle tissue (60 ± 2.0 µg kg-1). The fractionation process of tissue proteomes by 2D-PAGE and subsequent mapping of mercury by GFAAS in the protein spots of the gels identified the presence of mercury in three spots of the liver tissue (concentrations in the range of 0.800 to 1.90 mg kg-1). The characterization of protein spots associated with mercury by ESI-MS/MS identified the enzymes triosephosphate isomerase A, adenylate kinase 2 mitochondrial, and glyceraldehyde-3-phosphate dehydrogenase as possible candidates for mercury exposure biomarkers. The muscle tissue did not show protein spots associated with mercury. Enzymatic activity decreased proportionally to the increase in mercury concentrations in the tissues.

Influence of environmental conditions on the mercury levels of the sediment along the Balbina Reservoir, Brazilian Amazon.

Mercury dynamics in hydroelectric reservoirs have been studied worldwide. In tropical reservoirs, especially in those of the Amazon, the influence of geochemistry on Hg levels along this aquatic system is not well understood. The main objective of the present study was to assess the influence of environmental conditions (physical and chemical water parameters, trace element concentrations of sediment and sediment geochemistry) on mercury levels of sediment along the Balbina Reservoir (Amazon basin, Brazil). Sediment was collected along the reservoir and in the main tributaries in May 2015 (n = 10). These samples were assessed for labile iron (LFe), manganese (LMn), aluminum (LAl) and mercury (LHg) concentrations, total mercury (THg) concentrations, organic matter (OM) content, and granulometry. Concentrations in the sediment were 4-212 (LFe), 2-460 (LAl), 180-613 (LMn), < detection limit-256 (LHg), and 12-307 µg kg-1 dry weight (THg). In general, these concentrations decreased along the reservoir from upstream to downstream, reaching the lowest concentrations in the middle of the lake, and they increased in the sampling points near the dam. The lability and concentrations of mercury were influenced by the concentrations of LFe, LMn, LAl, and the granulometry and OM content of the sediment. Altogether, THg concentrations of sediments of the Balbina Reservoir encompass the range of concentrations of other reservoirs or natural lakes in the Amazon basin (from ten to a few hundred µg kg-1 dry weight).

Metalloproteomic approach of mercury-binding proteins in liver and kidney tissues of Plagioscion squamosissimus (corvina) and Colossoma macropomum (tambaqui) from Amazon region: Possible identification of mercury contamination biomarkers.

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.

Study of proteins with mercury in fish from the Amazon region.

The high concentrations of mercury found in Amazon have been intensively studied by the scientific community in the last decades. These mercurial species bind preferentially to proteins. Therefore, this work proposal sought to obtain the fractionation, identification and study of mercury - bound proteins present in samples of muscular and hepatic tissue from fish collected in the reservoir of the Jirau Hydroelectric Power Plant - on the Madeira River. Two-dimensional electrophoresis (2D-PAGE) for protein fractionation, graphite furnace atomic absorption spectrometry (GFAAS) for the quantification of mercury and Mass Spectrometry (ESI-MS/MS) were used for the identification of proteins. Concluding the work with analysis of graphics from the Blast2go program. Two mercury - bound proteins were identified as triosephosphate isomerase A and Protein FAM45A. The data generated by the bioinformatics programs confirm the tendency of these proteins to be linked to mercury and elucity the possibles existing physiological and cellular interactions.

Physiological and functional aspects of metal-binding protein associated with mercury in the liver tissue of pirarucu (Arapaima gigas) from the Brazilian Amazon.

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.

Correction to: Mercury Exposure: Protein Biomarkers of Mercury Exposure in Jaraqui Fish from the Amazon Region.

In the affiliation section, Luiz Fabricio Zara's affiliation "Pontifical Catholic University of Goiás (PUC), Goiânia, GO, Brazil" was incorrect. The correct affiliation is College of Planaltina, UnB - University of Brasília, Distrito Federal, Brazil.

Mercury Exposure: Protein Biomarkers of Mercury Exposure in Jaraqui Fish from the Amazon Region.

This study presents data on the extraction and characterization of proteins associated with mercury in the muscle and liver tissues of jaraqui (Semaprochilodus spp.) from the Madeira River in the Brazilian Amazon. Protein fractionation was carried out by two-dimensional electrophoresis (2D-PAGE). Mercury determination in tissues, pellets, and protein spots was performed by graphite furnace atomic absorption spectrometry (GFAAS). Proteins in the spots that showed mercury were characterized by electrospray ionization tandem mass spectrometry (ESI-MS/MS). The highest mercury concentrations were found in liver tissues and pellets (426 ± 6 and 277 ± 4 µg kg-1), followed by muscle tissues and pellets (132 ± 4 and 86 ± 1 µg kg-1, respectively). Mercury quantification in the protein spots allowed us to propose stoichiometric ratios in the range of 1-4 mercury atoms per molecule of protein in the protein spots. The proteins characterized in the analysis by ESI-MS/MS were keratin, type II cytoskeletal 8, parvalbumin beta, parvalbumin-2, ubiquitin-40S ribosomal S27a, 39S ribosomal protein L36 mitochondrial, hemoglobin subunit beta, and hemoglobin subunit beta-A/B. The results suggest that proteins such as ubiquitin-40S ribosomal protein S27a, which have specific domains, possibly zinc finger, can be used as biomarkers of mercury, whereas mercury and zinc present characteristics of soft acids.

A Metalloproteomics Study on the Association of Mercury With Breast Milk in Samples From Lactating Women in the Amazon Region of Brazil.

This study aimed to identify metalloproteins that lose their metal ions in the presence of mercury (Hg) and bind to Hg in breast milk samples collected from the riverine population of the Madeira River, a tributary of the Amazon River. Initially, total Hg was determined from the hair of lactating women to identify individuals who were contaminated followed by a proteomic analysis of breast milk samples through two-dimensional polyacrylamide gel electrophoresis after acetone precipitation. The presence of Hg in the obtained protein spots was determined by cold vapor atomic absorption spectrometry and cold vapor atomic fluorescence spectrometry. These determinations indicated the presence of Hg in one protein spot, which was then characterized through electrospray ionization tandem mass spectrometry. Based on searches in the UniProt database, this protein spot was identified as lysozyme C.

Espectroscopia Atômica Identifica Diferenças nos Parâmetros de Elementos Traço na Saliva de Pacientes Portadores de Fibrose Cística / Atomic Spectroscopy Identifies Differences in Trace Element Parameters in the Saliva of Patients with Cystic Fibrosis

Objetivo: Determinar elementos traço na saliva de indivíduos com fibrose cística.Métodos: Trinta e cinco indivíduos com fibrose cística e trinta e nove indivíduos saudáveis participaram deste estudo. A distribuição de 24 elementos traço na saliva total estimulada foi avaliada por espectrometria de massa com plasma de argônio acoplado indutivamente (ICP-MS), espectrometria de emissão óptica com plasma de argônio acoplado indutivamente (ICP-OES) e espectrometria de absorção atômica (AAS).Resultados: Verificou-se uma diferença significativa na distribuição de elementos traço na saliva de indivíduos com fibrose cística quando comparado a indivíduos sem a doença. Sódio; Potássio; Vanádio; Cromo; Arsênio e Selênio apresentaram valores estatisticamente diferentes entre os grupos, enquanto Alumínio, Boro, Bário, Cadmo, Cobre, Ferro, Mercúrio, Magnésio, Manganês, Níquel, Titânio, Estrôncio e Zinco não apresentaram diferenças significativas entre os grupos. As análises estatísticas indicaram que os elementos Sódio e Potássio apresentaram maiores teores, enquanto que os elementos traço Vanádio, Cromo, Selênio e Arsênio apresentaram menores teores nos indivíduos com fibrose cística.Conclusões: A espectroscopia atômica mostrou um potencial considerável no desenvolvimento de metodologia destinada a detectar a fibrose cística neonatal. A saliva deve ser considerada como material biológico para análise de diagnóstico

Objective: To determine trace elements in the saliva of individuals with cystic fibrosis.Method: Thirty-five individuals with cystic fibrosis and thirty-nine healthy individuals participated in this study. The distribution of 24 trace elements in stimulated whole saliva was evaluated by inductivelycoupled plasma mass spectrometry (ICP-MS), inductively coupled plasma optical emission spectrometry (ICP-OES) and atomic absorption spectrometry (AAS).Results: There was a statistically significant difference in the distribution of trace elements in the saliva of individuals with cystic fibrosis compared with individuals without the disease. Sodium, potassium, vanadium, chromium, arsenic and selenium presented significantly different values between the groups, while no significant differences was found between the groups for aluminum, boron, barium, cadmium, copper, iron, mercury, magnesium, manganese, nickel, titanium, strontium and zinc. Statistical analyses indicated higher contents of the trace elements sodium and potassium and lower contents of the trace elements vanadium, chromium, selenium and arsenic in the individuals with cystic fibrosis.Conclusion: Atomic spectroscopy showed a considerable potential in the development of a methodology to detect a neonatal cystic fibrosis. The saliva should be considered as biological material for diagnostic analysis

Risco de câncer cutâneo associado à radiação ultravioleta incidente no Distrito Federal / Skin cancer risk correlation with incident ultraviolet radiation in the Distrito Federal

Recentes estudos indicam que aproximadamente 90 por cento das incidências dos casos de câncer estão associadas a fatores ambientais. Em especial, a radiação ultravioleta apresenta forte correlação com câncer cutâneo. Este trabalho mostra o monitoramento dos índices de raios ultravioleta incidentes no Distrito Federal durante o período de março de 2003 a abril de 2004. Foram utilizadas como referência a estação meteorológica instalada no Campus I da Universidade Católica de Brasília. As análises das correlações entre o índice de radiação ultravioleta e a temperatura possibilitaram verificar que não há relação proporcional entre esses critérios ambientais. A localização geográfica do Distrito Federal possibilita incidência de elevados valores de índices de radiação ultravioleta. Tais informações justificam a importância da conscientização, sobre o risco de câncer, de pessoas diretamente expostas a essa radiação, sendo necessárias ações e medidas preventivas de proteção contra esta.

Reduction of mercury(II) by tropical river humic substances (Rio Negro)-Part II. Influence of structural features (molecular size, aromaticity, phenolic groups, organically bound sulfur).

The influence of structural features of tropical river humic substances (HS) on their capability to reduce mercury(II) in aqueous solutions was studied. The HS investigated were conventionally isolated from Rio Negro water-Amazonas State/Brazil by means of the collector XAD 8. In addition, the isolated HS were on-line fractionated by tangential-flow multistage ultrafiltration (nominal molecular-weight cut-offs: 100, 50, 30, 10, 5 kDa) and characterized by potentiometry and UV/VIS spectroscopy. The reduction of Hg(II) ions to elemental Hg by size-fractions of Rio Negro HS was assessed by cold-vapor AAS (CVAAS). UV/VIS spectrometry revealed that the fractions of high molecular-size (F(1)>100 kDa and F(2): 50-100 kDa) have a higher aromaticity compared to the fractions of small molecular-size (F(5): 5-10 kDa, F(6): <5 kDa). In contrast, the potentiometric study showed different concentration of functional groups in the studied HS fractions. The reduction of Hg(II) by aquatic HS fractions at pH 5 proceeded in two steps (I, II) of slow first order kinetics (t(1/2) of I: 160 min, t(1/2) of II: 300 min) weakly influenced by the molecular-size, in contrast to the differing degree of Hg(II) reduction (F(5)>F(2)>>F(1)>F(3)>F(4)>>F(6)). Accordingly, Hg(II) ions were preferably reduced by HS molecules having a relatively high ratio of phenolic/carboxylic groups and a small concentration of sulfur. From these results a complex 'competition' between reduction and complexation of mercury(II) by aquatic HS occurring in tropical rivers such as the Rio Negro can be suggested.