Effect of olive oil-based emulsion on human lymphocyte and neutrophil death

Background The incorporation of lipid emulsions in parenteral diets is a requirement for energy and essential fatty acid supply to critically ill patients. The most frequently used IV lipid emulsions (LE) are composed with long-chain triacylglycerols rich in omega-6 polyunsaturated fatty acids (PUFA) from soybean oil, but these LE promote lymphocyte and neutrophil death. A new emulsion containing 20% soybean oil and 80% olive oil rich in (omega-9 monounsaturated fatty acids (MUFA) has been hypothesized not to cause impairment of immune function. In this study, the toxicity of an olive oil-based emulsion (OOE) on lymphocytes and neutrophils from healthy volunteers was investigated. Methods: Twenty volunteers were recruited and blood was. collected before a 6-hour infusion of an OOE, immediately after infusion, and again 18 hours postinfusion. Lymphocytes and neutrophils were isolated by gradient density. The cells were studied immediately after isolation and after 24 hours or 48 hours in culture. The following determinations were carried out: triacylglycerol levels and fatty acid composition and levels in plasma, lymphocyte proliferation, production of reactive oxygen species, and parameters of lymphocyte and neutrophil death (viability, DNA fragmentation, phosphatidylserine externalization, mitochondrial depolarization, and neutral lipid accumulation). Results: OOE decreased lymphocyte proliferation, provoked lymphocyte necrosis, and had no effect on the proportion of viable neutrophils. The mechanism of cell death induced by OOE involved neutral lipid accumulation but had no effect on mitochondrial membrane depolarization. Conclusions: The OOE given as a single dose of 500 mL induced low toxicity to lymphocytes from healthy volunteers, probably by necrosis.

Effects of dietary lipid composition bioavailability and inulin-type fructans on mineral in growing rats

Objective: This study reports the effects of feeding with a combination of inulin-type fructans (ITF) and fish oil (FO) on mineral absorption and bioavailability as part of a semipurified diet offered to rats. Methods: Male Wistar rats (n = 24) were fed a 15% lipid diet (soybean oil [SO] or a 1:0.3 fish:soybean oil mixture [FSO]) and diets containing the same sources of lipids supplemented with 10% ITF (Raftilose Synergy 1) ad libitum for 15 d. Feces and urine were collected for mineral analyses during the last 5 d of the test period. Fatty acid composition was determined in liver and cecal mucosa homogenates. Liver and bone mineral analyses were performed by atomic absorption spectrophotometry. Bone biomechanical analyses were evaluated by a 3-point bending test. Results: Compared with the controls, ITF-fed rats had enlarged ceca and a significant decrease in cecal content pH (P < 0.001). The apparent mineral absorption was improved in these rats, and this effect was enhanced by dietary combination with FO for all minerals except for magnesium. Addition of ITF to the diet resulted in higher bone mineral content (calcium and zinc) and bone strength, but increased bone mineral content was only statistically significant in FO-fed animals. A decrease in liver iron stores (P = 0.015) was observed in rats fed FO, considering that ITF consumption returned to levels comparable to the SO control group. Conclusion: These findings confirm the positive influence of ITF on mineral bioavailability, which was potentiated by addition of FO to the diet. (C) 2009 Published by Elsevier Inc.

Cell activation state influences the modulation of HLA-DR surface expression on human monocytes/macrophages by parenteral fish oil lipid emulsion

Abnormal surface expression of HLA-DR by leukocytes is associated with a poor prognosis in critical care patients. Critical care patients often receive total parenteral nutrition with lipid emulsion (LE). In this study we evaluated the influence of fish oil LE (FO) on human monocyte/macrophage (M phi) expression of surface HLA-DR under distinct activation states. Mononuclear leukocytes from the peripheral blood of healthy volunteers (n = 18) were cultured for 24 hours without LE (control) or with 3 different concentrations (0.1, 0.25, and 0.5%) of the follow LE: a) pure FO b) FO in association (1:1 v/v) with LE composed of 50% medium-chain trygliceride and 50% soybean oil (MCTSO), and c) pure MCTSO. The leukocytes were also submitted to different cell activation states, as determinate by INF-gamma addition time: no INF-gamma addition, 18 hours before, or at the time of LE addition. HLA-DR expression on M phi surface was evaluated by flow cytometry using specific monoclonal antibodies. In relation to controls (for 0.1%, 0.25%, and 0.5%: 100) FO decreased the expression of HLA-DR when added alone [in simultaneously-activated M phi, for 0.1%: 70 (59 +/- 73); for 0.25%: 51 (48 +/- 56); and for 0.5%: 52.5(50 +/- 58)] or in association with MCTSO [in simultaneously-activated M phi, for 0.1%: 50.5 (47 +/- 61); for 25%: 49 (45 +/- 52); and for 05 %: 51 (44 +/- 54) and in previously-activated M phi, for 1.0 % : 63 (44 +/- 88); for 0.25%: 70 (41 +/- 88); and for 0.5%: 59.5 (39 +/- 79)] in culture medium (Friedman p<0.05). In relation to controls (for 0.1%, 0.25%, and 0.5%: 100), FO did not influence the expression of these molecules on non-activated M phi [for 0.1 % : 87.5 (75 +/- 93); for 0.25%: 111 (98 +/- 118); and for 0.5%: 101.5 (84 +/- 113)]. Results show that parenteral FO modulates the expression of HLA-DR on human M phi surface accordingly to leukocyte activation state. Further clinical studies evaluating the ideal moment of fish oil LE infusion to modulate leukocyte functions may contribute to a better understanding of its immune modulatory properties.

Anti-inflammatory effect of parenteral fish oil lipid emulsion on human activated mononuclear leukocytes

Background & aim: To compare the effect of fish oil-based (FO) lipid emulsions (LE) for parenteral administration with standard LE and a new FO containing LE composed of four different oils on the antigen presentation and inflammatory variables. Methods: Phytohemagglutinin (PHA) activated human mononuclear leukocytes were cultured with different LE - Control: without LE; SO: soybean oil; SO/FO: soybean and FO (4:1); MCT/SO: medium chain triglycerides and SO (1:1); MCT/SO/FO: MCT/SO and FO (4:1) and SMOF: a new LE containing FO. Cytokine production was evaluated by ELISA, the expression of antigen-presenting and co-stimulatory surface molecules were analyzed by flow cytometry and lymphocyte proliferation was assessed by H(3)-Thymidine incorporation, after tetanus toxoid-induced activation. Results: All LE decreased the HLA-DR and increased CD28 and CD152 expression on monocytes/macrophages and lymphocytes surface (p < 0.05). SO/FO and MCT/SO/FO decreased lymphocyte proliferation (p<0.05). All LE decreased IL-2 product ion, but this effect was enhanced with MCT/SO/FO and SMOF (p < 0.05). MCT/SOTO decreased IL-6 and increased IL-10, whereas SO had the opposite effect (p < 0.05). Conclusion: FO LE inhibited lymphocyte proliferation and had an anti-inflammatory effect. These effects seem to be enhanced when FO is mixed with MCT/SO. SMOF had a neutral impact on lymphocyte proliferation and IL-6 and IL-10 production.

Biosynthesis and characterization of biodegradable Poly(3-hydroxybutyrate) from renewable sources

Poly(3-hydroxybutyrate) was produced in fed-batch cultures of Ralstonia eutropha DSM 428 and Alcaligenes latus ATCC 29712 on a mineral medium with different carbon sources such as sucrose, sodium lactate, lactic acid, soybean oil and fatty acid. The bacteria converted the different carbon sources supplied into P3HB. The best results were obtained when lactate or soybean oil were supplied as the sole carbon source. The range of number average molar mass (Mn) for the polymers, analyzed by Gel Permeation Chromatography was 1.65 to 0.79 x 10(5) g mol(-1). FTIR spectroscopy revealed a characteristic absorbance associated with polyester structures. The crystallinity degree, determinate from X-ray diffractograms, was about 69% in all synthesized polymers. The thermal properties associated to semicrystalline polymers indicated a glass transition at 0.1 degrees C and a melting point at about 175 degrees C and enthalpy of 63-89 J g(-1). The (1)H-NMR and (13)C-NMR spectra of the polymers were in agreement with the calculated chemical shifts associated with P3HB structures.

Isolation of a lectin and a galactoxyloglucan from Mucuna sloanei seeds

A lectin and a galactoxyloglucan were characterized from Mucuna sloanei seed cotyledons. The galactoxyloglucan, isolated by water extraction and ethanol precipitation, had Glc:Xyl:Gal proportions in a molar ratio of 1.8:1.7:1.0 and a molar mass (M(w)) of 1.6 x 10(6) g mol(-1). The lectin (sloanin), isolated from the same seed by affinity chromatography on cross-linked Adenanthera pavonina galactomannan, gave two protein bands by SDS-PAGE (36 and 34 kDa) and one peak by gel filtration (63.6 kDa). Its N-terminal sequence indicated similar to 69% identity with soybean agglutinin to leguminous lectins. Circular dichroism (CD) spectra established that sloanin predominantly contains beta-sheet structures. Sloanin has similar to 5.5% carbohydrate and displayed hemagglutinating activity against rabbit and enzyme treated human erythrocytes, inhibited only by D-Gal containing sugars. The interaction between sloanin and storage cell-wall galactoxyloglucan was tested by affinity chromatography and fluorescence spectroscopy. (C) 2009 Elsevier Ltd. All rights reserved.

Camptosemin, a tetrameric lectin of Camptosema ellipticum: structural and functional analysis

Lectins have been classified into a structurally diverse group of proteins that bind carbohydrates and glycoconjugates with high specificity. They are extremely useful molecules in the characterization of saccharides, as drug delivery mediators, and even as cellular surface makers. In this study, we present camptosemin, a new lectin from Camptosema ellipticum. It was characterized as an N-acetyl-d-galactosamine-binding homo-tetrameric lectin, with a molecular weight around 26 kDa/monomers. The monomers were stable over a wide range of pH values and exhibited pH-dependent oligomerization. Camptosemin promoted adhesion of breast cancer cells and hemagglutination, and both activities were inhibited by its binding of sugar. The stability and unfolding/folding behavior of this lectin was characterized using fluorescence and far-UV circular dichroism spectroscopies. The results indicate that chemical unfolding of camptosemin proceeds as a two-state monomer-tetramer process. In addition, small-angle X-ray scattering shows that camptosemin behaves as a soluble and stable homo-tetramer molecule in solution.

Crystal structure and statistical coupling analysis of highly glycosylated peroxidase from royal palm tree (Roystonea regia)

Royal palm tree peroxidase (RPTP) is a very stable enzyme in regards to acidity, temperature, H(2)O(2), and organic solvents. Thus, RPTP is a promising candidate for developing H(2)O(2)-sensitive biosensors for diverse applications in industry and analytical chemistry. RPTP belongs to the family of class III secretory plant peroxidases, which include horseradish peroxidase isozyme C, soybean and peanut peroxidases. Here we report the X-ray structure of native RPTP isolated from royal palm tree (Roystonea regia) refined to a resolution of 1.85 angstrom. RPTP has the same overall folding pattern of the plant peroxidase superfamily, and it contains one heme group and two calcium-binding sites in similar locations. The three-dimensional structure of RPTP was solved for a hydroperoxide complex state, and it revealed a bound 2-(N-morpholino) ethanesulfonic acid molecule (MES) positioned at a putative substrate-binding secondary site. Nine N-glycosylation sites are clearly defined in the RPTP electron-density maps, revealing for the first time conformations of the glycan chains of this highly glycosylated enzyme. Furthermore, statistical coupling analysis (SCA) of the plant peroxidase superfamily was performed. This sequence-based method identified a set of evolutionarily conserved sites that mapped to regions surrounding the heme prosthetic group. The SCA matrix also predicted a set of energetically coupled residues that are involved in the maintenance of the structural folding of plant peroxidases. The combination of crystallographic data and SCA analysis provides information about the key structural elements that could contribute to explaining the unique stability of RPTP. (C) 2009 Elsevier Inc. All rights reserved.

Structural Investigation of MFe(2)O(4) (M = Fe, Co) Magnetic Fluids

Ferrites of the type M(II)Fe(2)O(4) (M = Fe and Co) have been prepared by the traditional coprecipitation method. These ferrites were modified by the adsorption of fatty acids derived from soybean and castor oil and were then dispersed in cyclohexane, providing very stable magnetic fluids, readily usable in nonpolar media. The structural properties of the ferrites and modified ferrites as well as the magnetic fluids were characterized by XRD (X-ray powder diffraction), TEM (transmission electron microscopy), DRIFTS (diffusion reflectance infrared Fourier transform spectroscopy), FTMR (Fourier transform near-infrared), UV-vis, normal Raman spectroscopy, and surface-enhanced Raman scattering (SERS). XRD and TEM analysis have shown that the magnetic nanoparticles (nonmodified and modified) present diameters in the range of 10-15 nm. DRIFTS measurements have shown that the carboxylate groups of soybean and castor oil fatty acids adsorb on the ferrite surface, forming three different structures: a bridging bidentate, a bridging monodentate, and a bidentate chelate structure. The FTIR and Raman spectra of nonmodified Fe(3)O(4) and CoFe(2)O(4) nanoparticles have shown that the number of observed phonons is not compatible with the expected O(h)(7) symmetry, since IR-only active phonons were observed. in the Raman spectra and vice versa. SERS measurements of a CoFe(2)O(4) thin film on a SERS-active gold electrode at different applied potentials made possible the assignment of the signals near 550 and 630 cm(-1) to Co-O motions and the signals near 470 and 680 cm(-1) to Fe-O motions.

Characterization and Identification of Proteolytic Bacteria From the Gut of the Velvetbean Caterpillar (Lepidoptera: Noctuidae)

The characterization and identification of proteolytic bacteria from the gut of the velvetbean caterpillar (Anticarsia gemmatalis) were the objectives of this study. Twelve aerobic and anaerobic isolates of proteolytic bacteria were obtained from the caterpillar gut in calcium caseinate agar. The number of colony forming units (CFUs) of proteolytic bacteria was higher when the bacteria were extracted from caterpillars reared on artificial diet rather than on soybean leaves (1.73 +/- 0.35 X 10(3) and 0.55 +/- 0.22 X 10(3) CFU/mg gut, respectively). The isolated bacteria were divided into five distinct groups, according to their polymerase chain reaction restriction fragment-length polymorphism profiles. After molecular analysis, biochemical tests and fatty acid profile determination, the bacteria were identified as Bacillus subtilis, Bacillus cereus, Enterococcus gallinarum, Enterococcus mundtii, and Staphylococcus xylosus. Bacterial proteolytic activity was assessed through in vitro colorimetric assays for (general) proteases, serine proteases, and cysteine proteases. The isolated bacteria were able of hydrolyzing all tested substrates, except Staphylococcus xylosus, which did not exhibit serine protease activity. This study provides support for the hypothesis that gut proteases from velvetbean caterpillar are not exclusively secreted by the insect cells but also by their symbiotic gut bacteria. The proteolytic activity from gut symbionts of the velvetbean caterpillar is suggestive of their potential role minimizing the potentially harmful consequences of protease inhibitors from some of this insect host plants, such as soybean, with implications for the management of this insect pest species.

Dietas de proteína reduzida e de diferentes digestabilidades suplementadas com aminoácidos sintéticos para frangos de corte

O conceito de proteína ideal adquiriu grande importância, devido à necessidade de maximizar o uso dos aminoácidos para a deposição de proteína corporal. O objetivo deste trabalho foi avaliar a formulação de rações com teor reduzido de PB suplementadas com aminoácidos sintéticos e de diferentes digestibilidades. Foram realizados dois experimentos (exp) utilizando frangos de corte machos das linhagens Ross (exp I) e Cobb (exp 11),no período de 3 a 6 semanas. No exp I foram testadas rações à base de milho e farelo de soja com quatro níveis de PB (20,8; 19,7; 18,6 e 17,5%). No exp 11,além dos tratamentos com os mesmos níveis de PB do exp I e digestibilidade padrão (DP), compostos principalmente por milho e farelo de soja, também foram testados dois tratamentos com dietas contendo 20,8 e 17,5% de PB e alta digestibilidade (AD), à base de milho, farelo de soja, amido de milho e proteína isolada de soja. Nos dois experimentos o ganho de peso, a conversão alimentar e o rendimento de cortes nobres foram negativamente afetados pela redução da PB dietética (P<0,05). Entretanto, as respostas de digestibilidade da matéria seca e digestibilidade da matéria orgânica, a retenção relativa de PB e a energia metabolizável aparente corrigida para nitrogênio foram superiores nas aves alimentadas com as dietas de baixos níveis de PB (P<0,05), mas não suficientes para compensar as perdas no desempenho. Não houve efeito da dieta AD sobre as respostas avaliadas, talvez em função das características físicas dos ingredientes utilizados.

Modelo de volatilidade estocástica com saltos aplicado a commodities agrícolas

Mensalmente são publicados relatórios pelo Departamento de Agricultura dos Estados Unidos (USDA) onde são divulgados dados de condições das safras, oferta e demanda globais, nível dos estoques, que servem como referência para todos os participantes do mercado de commodities agrícolas. Esse mercado apresenta uma volatilidade acentuada no período de divulgação dos relatórios. Um modelo de volatilidade estocástica com saltos é utilizado para a dinâmica de preços de milho e de soja. Não existe um modelo ‘ideal’ para tal fim, cada um dos existentes têm suas vantagens e desvantagens. O modelo escolhido foi o de Oztukel e Wilmott (1998), que é um modelo de volatilidade estocástica empírica, incrementado com saltos determinísticos. Empiricamente foi demonstrado que um modelo de volatilidade estocástica pode ser bem ajustado ao mercado de commodities, e o processo de jump-diffusion pode representar bem os saltos que o mercado apresenta durante a divulgação dos relatórios. As opções de commodities agrícolas que são negociadas em bolsa são do tipo americanas, então alguns métodos disponíveis poderiam ser utilizados para precificar opções seguindo a dinâmica do modelo proposto. Dado que o modelo escolhido é um modelo multi-fatores, então o método apropriado para a precificação é o proposto por Longstaff e Schwartz (2001) chamado de Monte Carlo por mínimos quadrados (LSM). As opções precificadas pelo modelo são utilizadas em uma estratégia de hedge de uma posição física de milho e de soja, e a eficiência dessa estratégia é comparada com estratégias utilizando-se instrumentos disponíveis no mercado.

Avaliação da eficiência de co-transformação de soja (Glycine max (L.) Merrill) via bombardeamento de partículas utilizando um gene de seleção e um gene de interesse em plamídeos diferentes

Visando aumentar a resistência a moléstias fúngicas, o presente trabalho teve como objetivo introduzir um gene (chit1) que codifica uma quitinase do fungo Metarhizium anisopliae em cultivares de soja [Glycine max (L.) Merrill]. A co-transformação foi a estratégia escolhida, visando a obtenção de plantas livres de transgenes marcadores na progênie das plantas transformadas. A co-transformação foi realizada via biolística, tendo como tecido-alvo conjuntos de embriões somáticos globulares das cultivares MG/BR46 Conquista e IAS-5. O plasmídeo pGusHyg, que contém o gene repórter gusA e o gene marcador hpt, foi bombardeado concomitantemente com o plasmídeo pMOG463chit1, que porta o gene chit1. Os conjuntos de embriões bombardeados foram transferidos para meio seletivo contendo higromicina, visando a obtenção de material estavelmente transformado. Os conjuntos embriogênicos higromicina-resistentes foram transferidos seqüencialmente para meios de proliferação D-20 (sem higromicina), maturação e regeneração. No total, foram obtidos 387 e 380 embriões histodiferenciados das cultivares MG/BR46 Conquista e IAS-5, respectivamente. Plantas transgênicas adultas e férteis foram regeneradas. Para avaliar a eficiência da estratégia de cotransformação, foram realizadas análises moleculares de embriões histodiferenciados e de plantas regeneradas. Os resultados obtidos neste trabalho permitiram o cálculo da taxa de co-transformação de 44% para os embriões histodiferenciados da cultivar MG/BR46 Conquista e de 50% para plantas de IAS-5. Não existem, até o momento, relatos de trabalhos em soja utilizando embriões somáticos globulares em proliferação como alvo para estudos de co-transformação.

Gestão de marketing nas associações e cooperativas de agronegócio : o caso da Associação dos Produtores de Soja e Milho do Estado de Mato Grosso

Esta dissertação analisar gestão de marketing no agronegócio, especificamente no contexto da Associação dos Produtores de Soja e Milho do Estado de Mato Grosso (Aprosoja/MT); trata-se de uma iniciativa que pode ser considerada um novo perfil de liderança no sistema cooperativista. O problema de pesquisa é identificar quais são os fundamentos da gestão de marketing da Aprosoja/MT. Para isso, foi realizado um estudo de caso único, com finalidade exploratória e enfoque qualitativo. Durante a coleta de dados, além da análise documental, foram realizadas vinte entrevistas pessoais, primeiramente utilizando o método da teoria fundamentada, para posterior elaboração de um roteiro semiestruturado. Os relatos das entrevistas e o estudo de caso foram interpretados tomando por base o referencial teórico proporcionado por autores como Kotler, Levy, Vargo, Lusch, Zylbersztajn, Bialoskorski e Tejon, dentre outros estudiosos das áreas da lógica dominante de serviços (LDS), do marketing de relacionamento e do marketing do agronegócio. Os resultados da pesquisa indicam que a Aprosoja/MT tem uma visão e entendimento bastante avançados sobre os conceitos mais modernos do campo do marketing, não se restringindo ao marketing tático. Dessa forma, pode-se concluir que a Aprosoja/MT opera no nível estratégico do marketing, com forte orientação para o marketing de relacionamento. É possível afirmar que a gestão de marketing da Aprosoja/MT está fundamentada no relacionamento com seus stakeholders, em função do modelo da LDS.

Viabilidade do confinamento de bovinos com feno de resíduos culturais de soja no sistema triângulo

O presente estudo buscou analisar a viabilidade econômica, agronômica e ambiental de um sistema que está em execução em uma propriedade no norte do estado de Goiás. Esse sistema, denominado Sistema Triângulo, vai de encontro com a Integração Lavoura-Pecuária e busca formas alternativas para o uso dos restos culturais (fardos) de soja fornecendo esse material como volumoso para o gado no sistema de confinamento. Para o estudo da viabilidade, foram usadas as informações da própria fazenda num estudo de caso. Para tal, realizou-se uma revisão quanto à adoção de sistema de produção na área agrícola, quanto às interações dos macronutrientes que seriam retirados das áreas por intermédio dos fardos, o papel da matéria orgânica quanto à sustentabilidade do sistema, o cenário atual dos confinamentos no Brasil e os custos que compõem e determinam a lucratividade dessa atividade. Por ser um estudo de caráter incremental, a viabilidade econômica ficou focada à redução dos custos alimentares. Foram utilizadas informações produzidas pela Fazenda Triângulo que é produtora de soja desde 2002. Sendo assim, foi possível acompanhar a evolução dos teores de macronutrientes (nitrogênio, fósforo, potássio, cálcio, magnésio e enxofre) e do carbono em uma área total de 351 ha. Os resultados mostraram uma estabilidade nos níveis dos macroelementos após o início da retirada dos restos culturais mas, apresentou-se inconclusivo no caso do carbono. Foi estimada também a exportação dos macronutrientes por intermédio da retirada dos fardos das áreas de soja. Levantada a composição química dos fardos, foram estabelecidos seis cenários de reposição via adubação específica e esses custos foram acrescidos de custos de processamento dos fardos e o custo dos outros componentes da dieta no confinamento. Calculado o valor presente líquido para cada um dos cenários e comparando com a alternativa tradicional de dieta de confinamento, onde se usa a silagem de milho como 100% do volumoso, mostrou-se uma redução de até 5,02% nos custos alimentares. Com as práticas agronômicas da produção de soja vigentes hoje, recomenda-se a adoção do Sistema Triângulo com a ressalva de uma necessidade de maiores estudos quando seu impacto nos teores de carbono orgânico no solo.