Solid phase microextraction, mass spectrometry and metabolomic approaches for detection of potential urinary cancer biomarkers: a powerful strategy for breast cancer diagnosis

A sensitive assay to identify volatile organic metabolites (VOMs) as biomarkers that can accurately diagnose the onset of breast cancer using non-invasively collected clinical specimens is ideal for early detection. Therefore the aim of this study was to establish the urinary metabolomic profile of breast cancer patients and healthy individuals (control group) and to explore the VOMs as potential biomarkers in breast cancer diagnosis at early stage. Solid-phase microextraction (SPME) using CAR/PDMS sorbent combined with gas chromatography–mass spectrometry was applied to obtain metabolomic information patterns of 26 breast cancer patients and 21 healthy individuals (controls). A total of seventy-nine VOMs, belonging to distinct chemical classes, were detected and identified in control and breast cancer groups. Ketones and sulfur compounds were the chemical classes with highest contribution for both groups. Results showed that excretion values of 6 VOMs among the total of 79 detected were found to be statistically different (p < 0.05). A significant increase in the peak area of (−)-4-carene, 3-heptanone, 1,2,4-trimethylbenzene, 2-methoxythiophene and phenol, in VOMs of cancer patients relatively to controls was observed. Statiscally significant lower abundances of dimethyl disulfide were found in cancer patients. Bioanalytical data were submitted to multivariate statistics [principal component analysis (PCA)], in order to visualize clusters of cases and to detect the VOMs that are able to differentiate cancer patients from healthy individuals. Very good discrimination within breast cancer and control groups was achieved. Nevertheless, a deep study using a larger number of patients must be carried out to confirm the results.

Effect of trypsin inhibitor from Crotalaria pallida seeds on Callosobruchus maculatus (cowpea weevil) and Ceratitis capitata (fruit fly)

GOMES, Carlos E. M. et al. Effect of trypsin inhibitor from Crotalaria pallida seeds on Callosobruchus maculatus (cowpea weevil) and Ceratitis capitata (fruit fly). Plant Physiology and Biochemistry (Paris), v. 43, n. 12, p. 1095-1102, 2005.ISSN 0981-9428. DOI:10.1016/j.plaphy.2005.11.004.

Flow-injection spectrophotometric determination of dipyrone in pharmaceutical formulations using ammonium molybdate as chromogenic reagent

A simple and rapid flow-injection spectrophotometric method is reported for the determination of dipyrone in pharmaceutical formulations. The method is based on the reaction of dipyrone with ammonium molybdate in acidic medium to produce blue molybdenum, which was detected spectrophotometrically at 620 nm. The analyte was determined in a single-line flow system. The calibration curve obtained was linear in the range of 5x10(-4) to 8x10(-3) mol L-1 for dipyrone concentration and the precision ( s r =1.7%) was satisfactory. The method proved to be selective and adequately sensitive. Application of the method to the analysis of pharmaceutical samples resulted in excellent accuracy; the percent mean recoveries were in the range 95.3%-101% and relative errors less than 5.0% for five pharmaceutical formulations were found.

Insights into the Specificity of Thioredoxin Reductase-Thioredoxin Interactions. A Structural and Functional Investigation of the Yeast Thioredoxin System

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Structural Aspects of the Distinct Biochemical Properties of Glutaredoxin 1 and Glutaredoxin 2 from Saccharomyces cerevisiae

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Structural and Biochemical Characterization of Peroxiredoxin Q beta from Xylella fastidiosa CATALYTIC MECHANISM and HIGH REACTIVITY

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Purification, characterization, crystallization and theoretical molecular modeling of gyroxin fraction from Crotalus durissus terrificus venom (Laurenti, 1768)

The venom of Crotalus durissus terrificus snakes presents various substances, including a serine protease with thrombin-like activity, called gyroxin, that clots plasmatic fibrinogen and promote the fibrin formation. The aim of this study was to purify and structurally characterize the gyroxin enzyme from Crotalus durissus terrificus venom. For isolation and purification, the following methods were employed: gel filtration on Sephadex G75 column and affinity chromatography on benzamidine Sepharose 6B; 12% SDS-PAGE under reducing conditions; N-terminal sequence analysis; cDNA cloning and expression through RT-PCR and crystallization tests. Theoretical molecular modeling was performed using bioinformatics tools based on comparative analysis of other serine proteases deposited in the NCBI (National Center for Biotechnology Information) database. Protein N-terminal sequencing produced a single chain with a molecular mass of similar to 30 kDa while its full-length cDNA had 714 bp which encoded a mature protein containing 238 amino acids. Crystals were obtained from the solutions 2 and 5 of the Crystal Screen Kit (R), two and one respectively, that reveal the protein constitution of the sample. For multiple sequence alignments of gyroxin-like B2.1 with six other serine proteases obtained from snake venoms (SVSPs), the preservation of cysteine residues and their main structural elements (alpha-helices, beta-barrel and loops) was indicated. The localization of the catalytic triad in His57, Asp102 and Ser198 as well as S1 and S2 specific activity sites in Thr193 and Gli215 amino acids was pointed. The area of recognition and cleavage of fibrinogen in SVSPs for modeling gyroxin B2.1 sequence was located at Arg60, Arg72, Gln75, Arg81, Arg82, Lis85, Glu86 and Lis87 residues. Theoretical modeling of gyroxin fraction generated a classical structure consisting of two alpha-helices, two beta-barrel structures, five disulfide bridges and loops in positions 37, 60, 70, 99, 148, 174 and 218. These results provided information about the functional structure of gyroxin allowing its application in the design of new drugs.

Impacto de quatro substratos e parcelamento da fertirrigação na produção de tomate sob cultivo protegido

Analisou-se a produção do tomateiro, híbrido longa vida Carmen, cultivado em quatro substratos, com fertirrigação semanal e duas vezes por semana. O delineamento experimental utilizado foi blocos casualizados, em esquema fatorial 4 x 2 (quatro substratos e duas formas de fertirrigação), em quatro repetições. Os substratos testados foram: S1 = areia fina (0,250 - 0,105 mm); S2 = 1/2 areia fina + 1/2 bagaço de cana-de-açúcar; S3 = 1/2 areia fina + 1/2 casca de amendoim moída (passada em peneira com abertura de 7 x 18 mm) e S4 = 1/3 areia fina + 1/3 bagaço de cana-de-açúcar + 1/3 casca de amendoim moída. As fertirrigações avaliadas foram: F1 = fertirrigação realizada uma vez por semana, com aplicação da quantidade total de nutrientes e, F2 = fertirrigação realizada duas vezes por semana, com aplicação da metade da quantidade total de nutrientes, demandada por semana, em cada vez. A quantidade de nutrientes, em mg L-1, fornecida por meio de fertirrigação, foi: 200 de nitrogênio, 60 de fósforo, 350 de potássio, 206 de cálcio, 60 de magnésio, 150 de enxofre, 0,50 de boro, 0,10 de cobre, 2 de ferro, 0,75 de manganês, 0,10 de zinco e 0,01 de molibdênio. A dotação hídrica foi realizada em função dos dados obtidos em um tanque classe A, localizado no centro da casa de vegetação. Avaliou-se a altura das plantas, o número de cachos e de frutos por planta, o peso individual do fruto e a produção por planta. Os dados foram submetidos à análise de variância e as médias foram comparadas pelo teste de Tukey, ao nível de 5% de probabilidade. Verificou-se que a fertirrigação realizada duas vezes por semana, suprindo a demanda total de nutrientes da semana, resultou em consideráveis melhorias de produção. Os substratos utilizados, com exceção do substrato S2, apresentaram potencial de uso para o cultivo do tomateiro em ambiente protegido.

Calagem e aplicação de cobalto e molibdênio na cultura do amendoim

O experimento compreendeu dois cultivos consecutivos de amendoim das águas em um latossolo vermelho-escuro distrófico, textura média, com o objetivo de avaliar os efeitos da calagem e da aplicação de cobalto e molibdênio nas sementes sobre a produção de amendoim. Empregaram-se quatro doses de calcário dolomítico calcinado: 0, 4, 6 e 8 t/ha; dois cultivares de amendoim: Tatu e Tupã, e quatro tratamentos de sementes: não tratadas, tratadas com cobalto, com molibdênio e com cobalto mais molibdênio. Verificou-se que a calagem elevou a produção de amendoim em função do aumento proporcionado no número e massa de grãos por vagem. A máxima produção de vagens foi obtida para pH (CaCl2 0,01M) 5,0, teor de cálcio trocável de 23,5 mmol c/dm³ e saturação por bases do solo de 50%. A aplicação de cobalto, com ou sem molibdênio, nas sementes, não influenciou a produção de amendoim, independentemente das condições de acidez do solo. A maior produtividade de vagens do cultivar Tupã, em relação ao 'Tatu', quando ocorre, é devida à maior proporção de cascas e não de grãos.

Estudo do aproveitamento do rejeito da mineração de esmeraldas na produção de refratários conformados isolantes

Emerald mining is an important area of the economy in Brazil, country which is in second place among the exporting nations of this gem. Due to the process of extraction, a great amount of reject is generated. Since there is no appropriate destination, the reject is abandoned around the mining industries, contributing to environment degradation. Nowadays, some of the most relevant things to an industry in general are: energy conservation, cost reduction, quality and productivity enhancement. The production of isolating, transformed refractory materials achieves the sustainability dimension when protection of the environment is incorporated to such process. This work investigates the use of emerald mining rejects in the ceramic body of refractory materials, aiming at obtaining a product whose characteristics are compatible with commercial products and, at the same time, allow the use of such rejects to solve the environmental issue caused by its disposal in nature. X-ray fluorescence analysis show that the emerald reject obtained after the flotation to extract molybdenum and mica has 70% of silica and alumina (SiO2+Al2O3) and 21% of a basic oxides and alkaline metals and earthy alkaline mixture (Na2O, K2O, CaO e MgO). Because of the significant amount of silica and alumina present in the reject, four refractory ceramic bodies were prepared. Samples with a rectangular shape and dimensions 100x50x10 mm were pressed in a steel mold at 27,5 MPa and sintered at 1200ºC for 40 min. under environment atmosphere in a resistive oven. The sintered samples were characterized in relation to the chemical composition (FRX), mineralogical composition (DRX), microstructure (MEV) and physical and mechanical properties. The results indicate that the mixture with 45% of reject, 45% of alumina and 10% of kaolin presents a refractory quality of 1420ºC, dimensional linear variation below 2.00%, apparent specific mass of 1,56 g/cm3 and porosity of 46,68%, which demonstrates the potential use of the reject as raw material for the industry of isolating transformed refractory materials

Aplicação foliar de molibdênio, produtividade e qualidade fisiológica de sementes de feijoeiro irrigado

O objetivo deste trabalho foi avaliar o efeito de doses e épocas de aplicação de molibdênio, por via foliar, na produtividade e qualidade fisiológica de sementes do feijoeiro Pérola irrigado, cultivado em Neossolo Quatizarênico. Os tratamentos foram constituídos pela aplicação de quatro doses de Mo (0, 40, 80 e 160 g ha-1), aplicadas em duas épocas: 15 ou 26 dias após a emergência das plântulas (DAE). A aplicação de Mo via foliar aumentou a produtividade de sementes e a matéria seca da parte aérea de plântulas do feijoeiro irrigado, cultivado em solo arenoso, independentemente da época de aplicação (15 ou 26 DAE). A germinação das sementes do feijoeiro foi aumentada pela aplicação de Mo aos 26 DAE. Na avaliação do vigor das sementes de feijão, pelo teste de primeira contagem, constatou-se efeito negativo da aplicação de Mo via foliar. A aplicação foliar de Mo aumentou a produtividade do feijoeiro e promoveu a obtenção de sementes que originaram plântulas com maior desenvolvimento inicial.

Qualidade fisiológica de sementes de milho tratadas com molibdênio

O tratamento de sementes com micronutrientes, como o molibdênio, garante uma maior uniformidade de aplicação, sendo que, a quantidade a ser aplicada desse elemento nas sementes deve ser suficiente para provir à exigência para o desenvolvimento e produção da cultura. Assim, objetivou-se com o presente trabalho avaliar a qualidade fisiológica de sementes de milho tratadas com molibdênio. A qualidade das sementes foi avaliada por meio da determinação do teor de água, do teste de germinação, da primeira contagem de germinação e da emissão de raízes primárias. Os tratamentos testados consistiram de cinco híbridos (DOW CO32; DOW 2B587; DOW 2B688; PIONEER 30F35 e PIONEER 30K73) e cinco doses de molibdênio aplicadas via semente (0; 7,5; 22,5; 67,5; 202,5 g ha-1 de molibdênio). A fonte de molibdênio utilizada foi o molibdato de sódio dihidratado (39% de molibdênio), sendo que a aplicação do molibdênio foi efetuada por meio da mistura com o fungicida líquido de suspensão concentrada carboxina+thiram sobre as sementes. A qualidade fisiológica das sementes de milho é influenciada negativamente por doses crescentes de molibdênio aplicadas. O híbrido de milho DOW 2B587 obteve melhor resposta à aplicação da maior dose de molibdênio em relação aos demais híbridos estudados.

Síntese e caracterização do carbeto de molibdênio nanoestruturado para fins catalíticos na reação de oxidação parcial do metano

The nanostructures materials are characterized to have particle size smaller than 100 nm and could reach 1 nm. Due to the extremely reduced dimensions of the grains, the properties of these materials are significantly modified relatively when compared with the conventional materials. In the present work was accomplished a study and characterization of the molybdenum carbide, seeking obtain it with particles size in the nanometers order and evaluate its potential as catalyst in the reaction of partial methane oxidation. The method used for obtaining the molybdenum carbide was starting from the precursor ammonium heptamolybdate of that was developed in split into two oven, in reactor of fixed bed, with at a heating rate of 5ºC/min, in a flow of methane and hydrogen whose flow was of 15L/h with 5% of methane for all of the samples. The studied temperatures were 350, 500, 600, 650, 660, 675 and 700ºC and were conducted for 0, 60, 120 and 180 minutes, and the percent amount and the crystallite size of the intermediate phases were determined by the Rietveld refinement method. The carbide obtained at 660ºC for 3 hours of reaction showed the best results, 24 nm. Certain the best synthesis condition, a passivating study was accomplished, in these conditions, to verify the stability of the carbide when exposed to the air. The molybdenum carbide was characterized by SEM, TEM, elemental analysis, ICP-AES, TG in atmosphere of hydrogen and TPR. Through the elemental analysis and ICP-AES the presence carbon load was verified. TG in atmosphere of hydrogen proved that is necessary the passivating of the molybdenum carbide, because occur oxidation in room temperature. The catalytic test was accomplished in the plant of Fischer-Tropsch of CTGAS, that is composed of a reactor of fixed bed. Already the catalytic test showed that the carbide presents activity for partial oxidation, but the operational conditions should be adjusted to improve the conversion

Adubação nitrogenada em cobertura no feijoeiro com suplementação de molibdênio via foliar

The objective of this work was to evaluate the effect of leaf spray of molybdenum (Mo) and nitrogen (N) rates in common bean (Phaseolus vulgaris L.), cultivar Carioca Precoce, evaluating the yield components, yield and leaves nitrate levels. The research was carried out in Botucatu, State of São Paulo, Brazil. The sowing of the beans (06/03/2003) was made with the supply of 230 kg ha(-1) of NPK (4-30-10) granulated fertilizer at planting. The experiment was a complete randomized block, with four replications and a 5 x 2 factorial arrangement, involving five N rates (25; 50; 75 and 125 kg ha(-1)), applied 20 days after emergency (DAE), with or without 80 g ha(-1) of Mo, applied on leaves at 25 DAE. The Mo absence by leaf spray promoted the nitrate accumulation in the leaves in treatments with higher rates of N. The weight of 100 grains increased with the Mo by leaf spray, reducing the need for N to get the best results. The molybdenum fertilization increased bean yield regardless of N rate applied to the beans.

Síntese e caracterização de aluminofosfato impregnado com molibdênio/zinco visando a dessulfurização por adsorção

Discussions about pollution caused by vehicles emission are old and have been developed along the years. The search for cleaner technologies and frequent weather alterations have been inducing industries and government organizations to impose limits much more rigorous to the contaminant content in fuels, which have an direct impact in atmospheric emissions. Nowadays, the quality of fuels, in relation to the sulfur content, is carried out through the process of hydrodesulfurization. Adsorption processes also represent an interesting alternative route to the removal of sulfur content. Both processes are simpler and operate to atmospheric temperatures and pressures. This work studies the synthesis and characterization of aluminophosphate impregnate with zinc, molybdenum or both, and its application in the sulfur removal from the gasoline through the adsorption process, using a pattern gasoline containing isooctane and thiophene. The adsorbents were characterized by x-ray diffraction, differential thermal analysis (DTG), x-ray fluorescence and scanning electron microscopy (SEM). The specific area, volume and pore diameter were determined by BET (Brunauer- Emmet-Teller) and the t-plot method. The sulfur was quantified by elementary analysis using ANTEK 9000 NS. The adsorption process was evaluated as function of the temperature variation and initial sulfur content through the adsorption isotherm and its thermodynamic parameters. The parameters of entropy (ΔS), enthalpy variation (ΔH) and free Gibbs energy (ΔG) were calculated through the graph ln(Kd) versus 1/T. Langmuir, Freundlich and Langmuir-Freundlich models were adjusted to the experimental data, and the last one had presented better results. The thermodynamic tests were accomplished in different temperatures, such as 30, 40 and 50ºC, where it was concluded the adsorption process is spontaneous and exothermic. The kinetic of adsorption was studied by 24 h and it showed that the capability adsorption to the adsorbents studied respect the following order: MoZnPO > MoPO > ZnPO > AlPO. The maximum adsorption capacity was 4.91 mg/g for MoZnPO with an adsorption efficiency of 49%.