RESUMO
Proteomics studies can be used to identify proteins that affect feed efficiency traits, related to cost and profitability of meat production. We used a proteomic approach based on two-dimensional electrophoresis (2D-PAGE) in combination with mass spectrometry (ESI-MS) to study liver samples of Nellore bulls divergently ranked according to residual feed intake (RFI). The study showed that 71 protein spots were expressed differentially (P < 0.05) among RFI groups and 47 were identified by ESI-MS. In RFI, efficient animals (low RFI) eat less than predictions, based on their weights and growth rate, while inefficient animals (high RFI) that eat more than predicted. Data from 18 animals (9 high vs. 9 low RFI) aged 24-26 months in feedlot finishing were used. Immediately after slaughter, liver samples were collected and protein extracts were separated. The gels of RFI groups were scanned and the images analyzed, whereby we found 279 and 215 liver protein spots in high and low RFI bulls, respectively. The proteins identified were related to the following biological functions: (I) oxygen transport and blood flow; (II) mitochondrial function and energy metabolism; (III) amino acid metabolism, ion transport, and cell survival. The study suggests hemoglobin subunit beta and heat shock protein 71 kDa and as molecular markers to study FE in Nellore cattle. Moreover, proteins such as 3-ketoacyl-CoA thiolase and glutamate dehydrogenase 1 were found in liver from high and low RFI animals, respectively. Such protein expression could be associated with changes in the oxidative capacity of RFI phenotypes.
Assuntos
Ração Animal/análise , Fígado/química , Proteômica/métodos , Criação de Animais Domésticos , Animais , Brasil , Bovinos , Ingestão de Alimentos , Eletroforese em Gel Bidimensional/métodos , Metabolismo Energético , Fígado/metabolismo , Masculino , Espectrometria de Massas/métodos , Fenótipo , Carne Vermelha/análiseRESUMO
Mercury is a potentially toxic element that is present in the environment of the Brazilian Amazon and is responsible for adverse health effects in humans. This study sought to assess possible protein biomarkers of mercury exposure in breast milk samples from lactating women in the Madeira and Negro Rivers in the Brazilian Amazon. The mercury content of hair samples of lactating women was determined, and the proteome of breast milk samples was obtained using two-dimensional electrophoresis after protein precipitation with acetone. Mercury measurements of protein spots obtained via protein fractionation were performed by graphite furnace atomic absorption spectrometry (GFAAS), and it was observed that mercury is linked to proteins with molecular masses in the range of 14-26 kDa. The total mercury concentration was also determined by GFAAS in unprocessed milk, lyophilized milk, and protein pellets, with the purpose of determining the mercury mass balance in relation to the concentration of this element in milk and pellets. Approximately 85 to 97% of mercury present in the lyophilized milk from samples of lactating women of the Madeira River is bound in the protein fraction. From lactating women of the Negro River, approximately 49% of the total mercury is bound in the protein fraction, and a difference of 51% is bound in the lipid fraction.
Assuntos
Cabelo/química , Mercúrio/análise , Metaloproteínas/análise , Leite Humano/química , Brasil , Feminino , HumanosRESUMO
This paper presents the results of mercury fractionation in muscle samples of dourada (Brachyplatystoma rousseauxii) from the JIRAU Hydroelectric Power Plant in the Madeira River Basin in the Amazon region of Brazil. The proteome of the dourada muscle was separated by two-dimensional polyacrylamide gel electrophoresis (2D PAGE). The mercury present in the protein spots was determined by graphite furnace atomic absorption spectrometry (GFAAS) after acid mineralisation in an ultrasound bath. The protein spots in which the presence of mercury was detected were characterised by electrospray ionisation tandem mass spectrometry (ESI-MS/MS) after tryptic digestion. The GFAAS determinations indicated that 65% of the mercury was linked to the protein fraction with a molar mass (Mm) of less than 90 kDa. The mercury concentrations in the seven spots in which this protein fraction was present were in the range of 11.40-35.10 µg kg(-1). Based on the mercury concentrations, it was possible to estimate that the protein spots contained approximately 1-3 mercury atoms per protein molecule. The ESI-MS/MS analysis allowed characterisation of the seven protein spots as the following proteins: protein NLRC5 (molar mass=18.10, pI=6.30); 39S ribosomal protein L36 mitochondrial (molar mass=15.40, pI=8.23); N-alpha-acetyltransferase 20 (Mm=15.95, pI=8.80); Mth938 domain-containing protein (Mm=15.01, pI=9.60); ubiquitin-40S ribosomal protein S27a (Mm=9.80, pI=7.60); parvalbumin alpha (Mm=12.40, pI=3.80) and parvalbumin beta (Mm=13.10, pI=3.45).
Assuntos
Proteínas de Peixes/isolamento & purificação , Mercúrio/isolamento & purificação , Músculos/química , Proteoma/isolamento & purificação , Poluentes Químicos da Água/isolamento & purificação , Animais , Brasil , Peixes-Gato/metabolismo , Eletroforese em Gel Bidimensional , Proteínas de Peixes/química , Contaminação de Alimentos/análise , Proteoma/química , Rios , Espectrometria de Massas por Ionização por Electrospray , Espectrofotometria AtômicaRESUMO
A method was developed to attach 4-amino-2-mercaptopyrimidine (AMP) onto silica gel surface and to determine trace metals. The surface functionalization reaction was performed with a silylant agent, chloropropyltrimethoxysilane (Si-CPTS), and the product, Si-AMP, was characterized by FT-IR and elemental analysis to evaluate the surface modification. The functionalized silica was applied in the sorption of Cu(II) ions from an aqueous medium. The series of adsorption isotherms were adjusted to a modified Langmuir equation and the maximum number of moles of adsorbed copper was 0.447 mmol g(-1). The modified material was placed in a preconcentration system, where it reached an approximately 20-fold enrichment factor using 5mg of Si-AMP. The proposed method was applied in the preconcentration and determination of Cu(II) in a fresh water sample from the Paraná river and was validated through a comparative analysis of a standard reference material (1643e).
Assuntos
Cobre/química , Dióxido de Silício/química , Extração em Fase Sólida/métodos , Poluentes Químicos da Água/isolamento & purificação , Purificação da Água/métodos , Adsorção , Géis , Íons , Cinética , Ligantes , Pirimidinas/química , Espectrofotometria Atômica/métodos , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Compostos de Sulfidrila/química , Poluentes Químicos da Água/químicaRESUMO
In this work the copper(II) complexation parameters of aquatic organic matter, aquatic and soil humic substances from Brazilian were determined using a new versatile approach based on a single-stage tangential-flow ultrafiltration (TF-UF) technique (cut-off 1 kDa) and sensitive atomic spectrometry methods. The results regarding the copper(II) complexation capacity and conditional stability constants obtained for humic materials were compared with those obtained using direct potentiometry with a copper-ion-selective electrode. The analytical procedure based on ultrafiltration is a good alternative to determine the complexation parameters in natural organic material from aquatic and soil systems. This approach presents additional advantages such as better sensibility, applicability for multi-element capability, and its possible to be used under natural conditions when compared with the traditional ion-selective electrode.
Assuntos
Solo/análise , Ultrafiltração/métodos , Água/química , Brasil , Cobre/química , ÍonsRESUMO
This work describes the synthesis and characterization of 2-aminothiazole-modified silica gel (SiAT), as well as its application for preconcentration (in batch and column technique) of Cu(II), Ni(II) and Zn(II) in ethanol medium. The adsorption capacities of SiAT determined for each metal ion were (mmol g(-1)): Cu(II)=1.20, Ni(II)=1.10 and Zn(II)=0.90. In addition, results obtained in flow experiments, showed a recovery of ca. 100% of the metal ions adsorbed in a column packed with 500 mg of SiAT. The eluent was 2.0 mol L(-1) HCl. The sorption-desorption of the studied metal ions made possible the development of a preconcentration method for metal ions at trace level in fuel ethanol using flame AAS for their quantification.
Assuntos
Cromatografia/métodos , Etanol/análise , Metais Pesados/análise , Automóveis , Quelantes , Cobre/análise , Fontes Geradoras de Energia , Níquel/análise , Sílica Gel , Dióxido de Silício , Tiazóis , Zinco/análiseRESUMO
Cadmium uptake and its effects on the growth of beans (Phaseolus vulgaris, L. cv carioca) was studied in a complete Hoagland &Arnon nutritive solution, containing cadmium levels ranging from 10 to 200 mmol L-1. The concentration of cadmium in leaves and roots increased as the concentration of cadmium in the solution increased. The cadmium level in roots was greater than in leaves. The inhibition of leave and root growths by cadmium was reversible.
A absorção de cádmio e seus efeitos no crescimento de feijoeiro (Phaseolus vulgaris, L. cv carioca) foi estudada em solução nutritiva completa de Hoagland & Arnon acrescida de doses de cádmio com concentrações entre 10 e 200 mmol L-1. Os teores de cádmio em folhas e raízes aumentaram à medida que aumentou a concentração de cádmio na solução nutritiva. Os teores de cádmio nas raízes foram maiores que nas folhas. Os resultados mostraram que a inibição do crescimento de folhas e raízes provocadas pela presença de cádmio são reversíveis.
RESUMO
Cadmium uptake and its effects on the growth of beans (Phaseolus vulgaris, L. cv carioca) was studied in a complete Hoagland &Arnon nutritive solution, containing cadmium levels ranging from 10 to 200 mmol L-1. The concentration of cadmium in leaves and roots increased as the concentration of cadmium in the solution increased. The cadmium level in roots was greater than in leaves. The inhibition of leave and root growths by cadmium was reversible.
A absorção de cádmio e seus efeitos no crescimento de feijoeiro (Phaseolus vulgaris, L. cv carioca) foi estudada em solução nutritiva completa de Hoagland & Arnon acrescida de doses de cádmio com concentrações entre 10 e 200 mmol L-1. Os teores de cádmio em folhas e raízes aumentaram à medida que aumentou a concentração de cádmio na solução nutritiva. Os teores de cádmio nas raízes foram maiores que nas folhas. Os resultados mostraram que a inibição do crescimento de folhas e raízes provocadas pela presença de cádmio são reversíveis.