Labeling of Salmonella Typhymurium with iodine-131 to study phagocytic function in rats

The present study describes a method for labeling Salmonella typhymurium with iodine-131 to evaluate both the morphological and the functional characteristics of the reticulo-endothelial system. A suspension containing 2 x 10(9) bacteria per ml was labeled with carrier-free Na131I without reductor, with a labeling yield of 46.5 ± 3% and 3.5 ± 1.3% of free Iodine-131. The biodistribution of the labeled bacteria in rats was studied with a large field-of-view scintillation camera equiped with a pinhole collimator. Whole body images were obtained 15 and 30 minutes after intravenous injection of the labeled microorganisms. Images showed accumulation of bacteria in the liver and both normal and transplanted spleens of the animals. Autoradiographs of liver and spleen demonstrated labeled bacteria within the cells of the reticulo-endothelial system. The method described is easy to perform, has a good labeling yield and allows the functional evaluation of the reticulo-monophagocytic system, including transplanted spleens.O presente trabalho descreve um método para marcação de Salmonella typhymurium com iodo 131, útil para avaliar tanto os aspectos morfológicos como funcionais do sistema retícu lo endotelial. Uma suspensão contendo 2 x 10" bactérias por ml foi marcada com Na mI livre de carre gador e de redutor, resultando em um rendimen to de marcação de 46.5 : 3% e 3.5 ± 1.3% de iodelo 131 livre. Estudou se a biodistribuiçáo das bactérias marcadas em ratos, obtendo se imagens de cor po total aos 15 e 30 minutos após injeção em uma câmara de cintilação de campo de visão amplo, equipada com um colimador "pinhole" As imagens mostraram captação das bactérias pelo fígado e pelo baço normal ou transplantado, dos animais estudados. Os cortes autoradiográ ficos de baço e fígado mostraram bactérias marcadas dentro das células do sistema retículo-endotelial. O método de marcação da Salmonella typhymurium com iodo-131 se mostrou de fácil execução com rendimento adequado, possibilitando a avaliação funcional de órgãos do sistema retículo-monofagocitário como o baço transplantado

Crystallization and Melting Behavior of Cocoa Butter in Lipid Bodies of Fresh Cacao Beans

The present study aims at observing the crystallization and polymorphic behavior of cocoa butter (CB) in fresh cacao beans with DSC and X-ray diffraction techniques. Underlying idea of this study was to relate the necessary conditions of germination of cacao beans to the crystallization of CB, which are present in oil-in-water emulsion droplets (lipid bodies) with diameters of 1-2 μm. Different cooling and heating conditions, with rates of 15, 2, 0.5, and 0.1 °C/min, were applied to fresh cacao beans and compared to bulk CB. The results showed that the crystallization temperatures (Tc) of CB in fresh cacao beans were lower than those of bulk CB at all the experimental conditions. In addition, polymorphic behavior of the occurrence of metastable and stable forms of CB was also different between the fresh cacao beans and the bulk state. The thermal behavior of fresh and dried cacao beans that have different geographical origins was also analyzed. This study indicates that the germination conditions of cacao beans are extended to low-temperature environmental areas by reducing the Tc value of CB in fresh beans than of the bulk oil. The results were consistent with the germination experiments of fresh cacao beans at different temperatures

External factors affecting polymorphic crystallization of lipids

Many physical properties (e.g., hardness, texture, rheology, and spreadability) of lipid‐based products largely depend on the extent of crystallization and transformation of lipids, and their network formation. Therefore, many studies have focused on controlling the crystallization of lipids in order to determine the functionality of lipid crystals. Both internal and external factors greatly affect the processes of lipid crystallization. The most important internal factors are polymorphism, which depends on variation in fatty acid moieties, and the composition or blending of different lipid materials. Important external factors are thermal treatment, additives, application of shear, sonication, and pressure. This paper briefly reviews recent advances in research on these external factors. We discuss the results by considering the relationships between external factors and thermodynamics, as well as kinetic properties of the crystallization and transformation of polymorphic forms of lipid crystals

Polymorphic crystallization and kinetic pathways of triacylglycerols

The kinetic properties of crystallization and structural transformation are important, particularly for the application of polymorphic crystal systems in pharmaceutical, biomedical, food technology, and other applications. Triacylglycerols (TAGs) are the main components of alimentary fats and oils. Fat structures and compositions determine their physical properties (e.g., rheology, morphology, and texture), where polymorphism exerts a strong influence. There are different polymorphic forms of TAGs: the most general ones (in increasing stability) are: α form, which crystallizes in the hexagonal system, β' form (orthorhombic system) and β form (triclinic system). In this study, several techniques, like Synchrotron Radiation X-Ray Diffraction, Thermal Analysis and Thermo-optical Microscopy, were used in order to characterize the polymorphism under the kinetic point of view of some TAGs present in edible oils and fats. Therefore, the role of some kinetic parameters (e.g. cooling and heating rates) is highlighted

Phase behavior of binary mixture systems of saturated-unsaturated mixed-acid triacylglycerols: effects of glycerol structures and chain−chain interactions

We systematically examined the phase behavior of binary mixtures of mixed-acid triacylglycerols (TAGs) containing palmitic and oleic acid moieties 1,3-dioleoyl-2-palmitoyl-glycerol (OPO), 1,2-dipalmitoyl-3-oleoyl-rac-glycerol (PPO), and 1,2-dioleoyl-3-palmitoyl-rac-glycerol (OOP), which are widely present in natural fats and are employed in the food, pharmaceutical, and cosmetic industries. Differential scanning calorimetry and X-ray diffraction methods were applied to observe the mixing behavior of PPO/OPO, OOP/OPO, and PPO/OOP under metastable and stable conditions. The results led to three conclusions: (1) Eutectic behavior was observed in PPO/OPO. (2) Molecular compound (MC) crystals were formed in the mixtures of OOP/OPO and PPO/OOP. (3) However, the MC crystals occurred only under metastable conditions and tended to separate into component TAGs to form eutectic mixture systems after 17 months of incubation. These results were contrary to those of previous studies on 1,3-dipalmitoyl-2-oleoyl glycerol (POP)/OPO and POP/PPO in which the MC crystals were thermodynamically stable. We determined that specific molecular interactions may cause this different phase behavior (stability of POP/OPO and POP/PPO MC crystals and metastability of OOP/OPO and PPO/OOP MC crystals). All results confirm the significant effects of molecular structures of glycerol groups, interactions of fatty acid chains, and polymorphism of the component TAGs on the mixing behavior of mixed-acid TAGs

Heterogeneous microstructures of spherulites of lipid mixtures characterized with synchrotron radiation microbeam X-ray diffraction

We observed microstructures of spherulites made of two triacylglycerols (TAGs) (POP (1,3-dipalmitoyl-2-oleoyl-sn-glycerol) and OPO (1,3-dioleoyl-2-palmitoyl-sn-glycerol)), which exhibit the mixing behavior of a molecular compound (MC) formation at a ratio of POP : OPO = 50 : 50, using synchrotron radiation microbeam (beam area of 5 × 5 μm2) X-ray diffraction (SR-μ-XRD). The spherulites were grown from neat liquid, and from solution containing 50% n-dodecane and 50% POP + OPO. SR-μ-XRD analysis revealed heterogeneous distributions of MCPOP:OPO and component TAGs in every spherulite; TAG compositions in the inner and outer areas differed when the relative ratios of POP and OPO were changed. In the 75POP : 25OPO spherulites, MCPOP:OPO always occupied the inner areas and POP dominated the outer areas as a result of different rates of crystallization of MCPOP:OPO and POP in the spherulite. In contrast, in the 25POP : 75OPO spherulites, the entire area was homogeneously occupied either with almost all OPO or with coexisting MCPOP:OPO and OPO. The microstructures of the spherulites grown from n-dodecane solutions demonstrated basically the same characteristics as those grown from the neat liquid both for 75POP : 25OPO and 25POP : 75OPO. SR-μ-XRD also clarified that the lamellar planes of the crystals are directed parallel to the radial direction from the central to the outer regions of the spherulites

Crystallization and transformation of polymorphic forms of trioleoyl glycerol (OOO) and 1,2-dioleoyl-3-rac-linoleoyl glycerol (OOL)

This study examined the influence of different thermal treatments on the crystallization and transformation of trioleoyl glycerol (OOO) and 1,2-dioleoyl-3-rac-linoleoyl glycerol (OOL). Two triacylglycerol (TAG) samples were cooled at 0.5-15 °C·min-1 and heated at 2 and 15 °C·min-1. The polymorphic characteristics of the two TAGs were analyzed in situ using differential scanning calorimetry, Raman spectroscopy, and synchrotron radiation X-ray diffraction. Multiple polymorphic forms were identified in OOO (α, β′2, β′1, β2, and β1) and OOL (α, β′2, and β′1). Larger quantities of more stable forms (e.g., β2 and β1 of OOO and β′1 of OOL) were obtained when the samples were slowly cooled and heated. In contrast, less stable polymorphs were obtained with increased cooling and heating rates. Polymorphic transformations occurred in either solid-state or melt-mediation and were influenced by heating rates. The results were analyzed by considering the activation energies for crystallization and transformation of stable and less stable polymorphic forms in comparison with previous studies on 1,3-dipalmitoyl-2-oleoyl-glycerol and 1, 3-dioleoyl-2-palmitoyl-glycerol

In situ synchrotron radiation X-ray diffraction study of crystallization kinetics of polymorphs of 1,3-dioleoyl-2-palmitoyl glycerol (OPO)

We examined the influence of kinetics on the polymorphic behavior of 1,3-dioleoyl-2-palmitoyl glycerol (OPO), a triacylglycerol (TAG) present in natural oils such as olive oil and human breast milk. Pure OPO was heated at 15 °C min−1 in all cases. The polymorphic crystallization of OPO was studied at different cooling rates (15, 2, 1, and 0.5 °C min−1), and the dynamic polymorphic transformations were characterized on heating by simultaneously using Differential Scanning Calorimetry (DSC) and Synchrotron Radiation X-Ray Diffraction (SR-XRD) with small-angle X-ray diffraction (SAXD) and wide-angle X-ray diffraction (WAXD). Thermo-Optical Microscopy (TOM) was also used in order to observe the polymorphic transitions. Polymorphic forms of OPO were identified and characterized. As the cooling rate decreased, more stable forms crystallized, not following the Ostwald step rule, although in most cases concurrent crystallization occurred and the whole process was highly complex

Detecção radiométrica da atividade metabólica do Paracoccidioides brasiliensis e da sua sensibilidade à Anfotericina B e ao Dietilestilbestrol

Paracoccidioidomycosis (South American blastomycosis) is a systemic disease, strikingly more frequent in males, caused by the dimorphic fungus Paracoccidioides brasiliensis. A radiometric assay system has been applied to study the metabolic activity and the effect of drugs on this fungus "in vitro". The Y form of the yeast, grown in liquid Sabouraud medium was inoculated into sterile reaction vials containing the 6B aerobic medium along with 2.0 μCi of 14C-substrates. Control vials, prepared in the same way, contained autoclaved fungi. To study the effects of amphotericin B (AB) (0.1 and 10 μg/ml) and diethylstilbestrol (DSB) (1.0, 5.0 and 10 μg/ml) extra controls with live fungi and no drug were used. All vials were incubated at 35°C and metabolism measured daily with a Bactec instrument. 14CO2 production by P. brasiliensis was slow and could be followed for as long as 50 days. AB at 10mg/ml and DSB at 5 μg/ml inhibited the metabolism and had a cidal effect on this fungus. The results with DSB might explain the low incidence of the disease in females. This technique shows promise for studying metabolic pathways, investi gating more convenient 14C-substrates to expedite radiometric detection and for monitoring the effects of other drugs and factors on the metabolism of P. brasiliensis "in vitro".A paracoccidiodomicose (blastomicose sul-americana) é uma doença sistêmica muito mais freqüente no sexo masculino, causada pelo fungo dimórfico Paracoccidioides brasiliensis. Um sistema radiométrico foi utilizado para estudar a atividade metabólica e o efeito deftrogas sobre este fungo "in vitro". A forma Y do fungo, cultivada em Sabouraud líquido, foi inoculada em frascos estéreis contendo o meio aeróbio 6B, juntamente com 2,0 uCi de substâncias marcadas com carbono-14. Frascos-controle, preparados da mesma forma, foram inoculados com fungos autoclavados. Para estudar os efeitos da anfotericina B (AB) (0,1 e 10 μg/ml) e do dietilestilbestrol (DEB) (1, 5 e 10 μg/ml), controles adicionais foram preparados, contendo fungos viáveis mas não a droga. Todos os frascos foram incubados a 35°C e o metabolismo medido diariamente com uma máquina Bactec. A produção de 14CO2 pelo P. brasiliensis foi lenta e pôde ser acompanhada por 50 dias. Concentrações de 10 μg/ml de AB e 5 μg/ml de DEB inibiram o metabolismo e tiveram efeito fungicida. Os resultados com DEB poderiam explicar a baixa incidência da doença em mulheres. Esta técnica é promissora para estudar as vias metabólicas, investigar substâncias marcadas mais adequadas para tornar mais rápida a detecção radiométrica do fungo e para acompanhar os efeitos de outras drogas e fatores sobre o metabolismo do P. brasiliensis "in vitro"