Comparative metallomics for transgenic and non-transgenic soybeans

In this work, a comparative metallomics of transgenic and non- transgenic soybeans [ Glycine max ( L.) Merrill] was performed. Soybean proteins were extracted with a proper buffer and separated by two- dimensional polyacrylamide gel electrophoresis. Metal ions bound to a set of eight proteins randomly selected ( ranging from 13.98 to 54.87 kDa), were characterized by matrixassisted laser desorption- ionization quadrupole- time of flight mass spectrometry and mapped using synchrotron radiation X- ray spectrometry. The metal ions detected were: Ca( II), Cu( II), Fe( II), Mn( II), Ni( II) and Zn( II). Transgenic and non- transgenic soybeans proteins were found to display typical and random profiles for metal ions binding. To test the reliability of the qualitative metal ions profiles, quantification of Ca( II), Cu( II) and Fe( II) was performed via microwave-assisted decomposition in mini- vials followed by atomic absorption spectrometry determination. Qualitative and quantitative metallomics was found to be coherent and to match profiles expected from the known protein functions. The protein of spot 5, with molar mass of 37.62 kDa ( amino acid sequence presented), was found to display the most characteristic change in metal ions content, with higher Ca( II), Cu( II) and Fe( II) concentrations for transgenic soybeans.22121501150

Combination of PAGE and LA-ICP-MS as an analytical workflow in metallomics: state of the art, new quantification strategies, advantages ans limitations

Metallomics (more specifically, metalloproteomics) is an emerging field that encompasses the role, uptake, transport and storage of trace metals, which are essential to preserve the functions of proteins within a biological system. The current strategies for metal-binding and metalloprotein analysis based on the combination of polyacrylamide gel electrophoresis (PAGE) and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) are discussed in this review. The advantages, limitations and the most recently developed and applied quantification approaches for this methodology are also described

Evaluation of metabonomic, proteomic and metallomic profiles for bipolar disorder and its treatment with lithium in blood serum samples

Orientadores: Marco Aurelio Zezzi Arruda, Claudio Eduardo Muller BanzatoTese (doutorado) - Universidade Estadual de Campinas, Instituto de QuimicaResumo: O transtorno afetivo bipolar é uma doença psiquiátrica caracterizada por alterações de humor marcantes, oscilando entre episódios de mania e depressão, que afeta entre 1 a 3% da população mundial. Os mecanismos em nível molecular deste transtorno, assim como de seu tratamento com lítio, que é o medicamento mais utilizado, ainda não são estabelecidos. Assim sendo, o objetivo deste trabalho de Tese consistiu em explorar biomarcadores potenciais (metabólitos, proteínas e íons metálicos livres ou ligados a proteínas) para o transtorno afetivo bipolar e seu tratamento com lítio. Para isso, foi realizada a comparação dos perfis metabonômicos (utilizando espectroscopia de ressonância magnética nuclear de hidrogênio e análise quimiométrica), proteômicos (utilizando eletroforese bidimensional em gel de poliacrilamida e diferentes técnicas de espectrometria de massas molecular) e metalômicos (utilizando espectrometria de massas com fonte de plasma indutivamente acoplado) de amostras de soro sangüíneo de pacientes com transtorno afetivo bipolar utilizando o lítio (n = 15) ou outras drogas excluindo o lítio (n = 10) e de indivíduos saudáveis (n = 25). A análise metabonômica indicou os lipídeos como sendo os metabólitos mais afetados na presença do transtorno afetivo bipolar e do tratamento com lítio, o que corroborou com os resultados da análise proteômica, onde a apolipoproteína A-I foi uma das proteínas que sofreu maior alteração em seus níveis, sendo subexpressa em pacientes bipolares, independentemente do tratamento, porém apresentando uma restauração ao nível do grupo controle após o tratamento com lítio. As análises ionômicas apontaram As, B, Cl, Cr, Fe, K, Li, Mg, P, S, Se, Si, Sr e Zn como os íons livres diferenciais e as análises metaloproteômicas apontaram, principalmente, Ca, Co, Fe, K, Mg, Mn, Na, Ti e Zn ligados a proteínas como sendo os metais que sofreram as maiores alterações na presença do transtorno afetivo bipolar e de seu tratamento com lítio. Dentre as proteínas ligadas a metais que apresentaram diferenças entre os grupos estudados em termos de metais ligados, destacam-se a apolipoproteína A-I, a transtiretina e a vitronectina, que haviam sido identificadas previamente nas análises proteômicas por apresentarem alterações em suas expressões. A partir destes estudos, foi possível identificar, de maneira exploratória, moléculas diferenciais que podem orientar futuros estudos envolvendo os mecanismos patofisiológicos do transtorno afetivo bipolar e de ação terapêutica de drogas como o lítio, bem como na descoberta de biomarcadores para a doença e/ou seu tratamento com lítioAbstract: Bipolar disorder is a psychiatric illness characterized by marked mood changes, oscillating between mania and depression episodes, which affects 1-3% of the worldwide population. Molecular level mechanisms of this disorder, as well as of its treatment with lithium, which is the most widely used medication, are not yet known. Thus, the aim of this work consisted in explore potential biomarkers (metabolites, proteins, metal ions free or bounded to proteins) for bipolar disorder and its treatment with lithium. For this purpose, it was performed the comparison of metabonomic (using hydrogen nuclear magnetic resonance spectroscopy and chemometric analysis), proteomic (using 2-D gel electrophoresis and different molecular mass spectrometry techniques), and metallomic (using inductively coupled plasma mass spectrometry) profiles for blood serum samples of bipolar disorder patients treated with lithium (n = 15) or other drugs than lithium (n = 10) and healthy individuals (n = 25). Metabonomic analyses indicated lipids as the most affected metabolites in the presence of bipolar disorder and of lithium treatment, which corroborated with the results of proteomic analyses, where apolipoprotein A-I was one of the proteins that showed highest alterations in its levels. It was downregulated in bipolar disorder patients, independently of the treatment, but showed a level restored to that of the control group after lithium treatment. Ionomic analyses detected As, B, Cl, Cr, Fe, K, Li, Mg, P, S, Se, Si, Sr e Zn as differential free ions, and metalloproteomic analyses detected mainly Ca, Co, Fe, K, Mg, Mn, Na, Ti and Zn bound to proteins as being the metals that presented highest alterations in the presence of bipolar disorder and lithium treatment. Among the metal-binding proteins that indicated differences between the studied groups, apolipoprotein A-I, transthyretin and vitronectin, which were previously identified in the proteomic analyses, are highlighted. With the studies described in this research work, it was possible to identify, in an exploratory way, differential molecules that can guide future studies on the patophysiological mechanisms of bipolar disorder or terapeutic action pathways of drugs like lithium, as well as on the discovery of biomarkers for the illness and/or its treatment with lithiumDoutoradoQuimica AnaliticaDoutor em Ciência

Determination Of Chromium (vi) By Flame Atomic Absorption Spectrometry After Cloud Point Extraction And Preconcentration [determinação De Cromo (vi) Por Espectrometria De Absorção Atômica Com Chama Após A Extraçã O E Pré-concentração No Ponto Nuvem]

The present work reports a method for chromium (VI) determination by flame atomic absorption spectrometry (FAAS) after cloud point extraction and preconcentration. Chromium (VI) is complexed with 1,5-diphenylcarbazide (DFC) in acidic medium (pH 2.0) and it is extracted into 25 μL of surfactant-rich phase containing Triton X-114. The variables that affect the cloud point formation, such as the surfactant (0.1-1.0% m/v) and complexant (0.01-0.80% m/v) concentrations, time of complexation (0-60 min), and effect of electrolyte NaCl (0-20% m/v) addition are evaluated. Under optimized conditions, 0.3% m/v Triton X-114, 0.05% m/v DFC and 10% m/v NaCl are used for chromium (VI) extraction. This method allows detection and quantification limits of 0.4 μg L -1 and 1.5 μg L-1, respectively, and a linear calibration range from 5 to 500 μg L-1. The preconcentration factor obtained is 27 and the extraction efficiency achieved varies from 87 to 99.3%.3117380Manzoori, J.L., Sorouraddin, M.H., Shemiran, F., (1996) Anal. Lett., 29 (11), p. 2007Wang, C., Martin, D.F., Martin, B.B., (1999) J. Environ. Sci. Health, A34 (3), p. 705Padarauskas, A., Judzentiene, A., Naujalis, E., Paliulionyte, V., (1998) J. Chromatogr. A, 808 (1-2), p. 193Shemirani, F., Abkenar, S.D., Mirroshandel, A.A., Niasari, M.S., Kozania, R.R., (2003) Anal. Sci., 19 (10), p. 1453Paleologos, E.K., Vlessidis, A.G., Karayannis, M.I., Evmiridis, N.P., (2003) Anal. Chim. Acta, 477 (2), p. 223Paleologos, E.K., Stalikas, C.D., Tzouwara-Karayanni, S.M., Karayannis, M.I., (2001) Anal. Chim. Acta, 436 (1), p. 49Giokas, D.L., Paleologos, E.K., Tzouwara-Karayanni, S.M., Karayannis, M.I., (2001) J. Anal. At. Spectrom., 16 (5), p. 521Nascentes, C.C., Arruda, M.A.Z., (2003) Talanta, 61, p. 759Paleologos, E.K., Stalikas, C.D., Tzouwara-Karayanni, S.M., Pilidis, G.A., Karayannis, M.I., (2000) J. Anal. At. Spectrom., 15 (3), p. 287Coelho, L.M., Arruda, M.A.Z., (2005) Spectrochim. Acta B, 60 (5), p. 743Cordero, B.M., Pavón, J.L.P., Pinto, C.G., Laespada, M.E.F., (1993) Talanta, 40 (11), p. 1703Maniasso, N., (2001) Quim. Nova, 24 (1), p. 87Hinze, W.L., Pramauro, E., (1993) Crit. Rev. Anal. Chem., 24 (2), p. 133Stalikas, C.D., (2002) Trends Anal. Chem., 21 (5), p. 343Bezerra, M.A., Arruda, M.A.Z., Ferreira, S.L.C., (2005) Appl. Spectrosc. Rev., 40 (4), p. 269Sanz-Medel, A., De La Campa, M.R.F., Gonzalez, E.B., Fernandez-Sanchez, M.L., (1999) Spectrochim. Acta B, 54 (2), p. 251Willems, G.J., Blaton, N.M., Peeters, O.M., De Ranter, C.J., (1977) Anal. Chim. Acta, 88 (2), p. 345Nascentes, C.C., (2002) Emprego de Diferentes Estratégias para Análises Em Larga Escala: Screening, Extração Ultra-sônica e Pré-concentração Por Ponto Nuvem, p. 140. , Tese de Doutorado, Unicamp, CampinasKomaromy-Hiller, G., Calkins, N., Wandruszka, R., (1996) Langmuir, 12, p. 916Tarley, C.R.T., Arruda, M.A.Z., (2004) Anal. Sci., 20, p. 961Ferreira, S.L.C., Lemos, V.A., Moreira, B.C., Costa, A.C.S., Santelli, R.E., (2000) Anal. Chim. Acta, 403, p. 259(1987) Analyst, 11, p. 119http://www.mma.gov.br/port/conama/res/res86/res2086.htm

Laser Ablation Icp-ms: Application In Biomedical Research

In the last decade, the development of diverse bioanalytical methodologies based on mass spectrometry imaging has increased, as has their application in biomedical questions. The distribution analysis of elements (metals, semimetals, and nonmetals) in biological samples is a point of interest in life sciences, especially within the context of metallomics, which is the scientific field that encompasses the global analysis of the entirety of elemental species inside a cell or tissue. Laser ablation inductively coupled plasma mass spectrometry (LAICP-MS) has been efficiently employed to generate qualitative and quantitative maps of elemental distribution in thin tissue sections of a variety of biological samples, for example, brain, cartilage, spinal cord, etc. The combination of elemental with molecular mass spectrometry allows obtaining information about the elements bound to proteins, when they are previously separated by gel electrophoresis (metalloproteomics), and also adding a new dimension to molecular mass spectrometry imaging by the correlation of molecular and elemental distribution maps in definite regions in a biological tissue. In the present review, recent biomedical applications in LA-ICP-MS imaging as a stand-alone technique and in combination with molecular mass spectrometry imaging techniques are discussed. Applications of LA-ICP-MS in the study of neurodegenerative diseases, distribution of contrast agents and metallodrugs, and metalloproteomics will be focused in this review. (C) 2015 Wiley Periodicals, Inc.3614757Alexander von Humboldt Foundation (Bonn, Germany)Deutsche Forschungsgemeinschaft (DFG) [BE 2649/5-1