Influence of Polydispersity and Thermal Exchange on the Flocculation Rate of O/W Emulsions

According to recent simulations [ Langmuir 16 , 7975 (2000)], the flocculation rate ( k f ) of concentrated oil in water (O/W) emulsions interacting through van der Waals forces, can reach values considerably higher than the one expected for a very dilute system of non-interacting spheres (5.49 2 10 -18 m 3 /s). Similar calculations at a volume fraction J =0.001 using 64 particles only, already show a k f =5.83 2 10 -18 m 3 /s, reasonably close to the theoretical prediction. In this report Brownian Dynamics (BD) simulations are used to study the effect of the volume fraction and the drop size distribution (DSD) on the flocculation rate. First, the dependence of k f with the maximum value of the thermal interaction between the particles and the solvent is studied. Following, the variation of the flocculation rate is studied as a function of polydispersity for J =0.15. As expected, there is a strong dependence of k f on J . Faster and slower aggregation rates are observed depending on the characteristics of the DSD.     

Emulsions Based on the Interactions Between Lactoferrin and Chitosans

In this work, purification of lactoferrin from whey was performed with high recovery rate. Lactoferrin was then exploited in the preparation of food emulsions. Two tertiary emulsions, formed by olive oil, lecithin, chitosan, and lactoferrin, were compared: both the emulsions showed similar turbidity and stability. In the secondary emulsion formed by oil/lecithin/chitosan, the pH was increased to 9 before addition of lactoferrin. Then, lactoferrin was added, and the pH was stabilized above pH 9. Lactoferrin was found in amounts of 1 to 2.5 mg/ml in the multiple experiments. A fraction of the added lactoferrin was also present in a milky layer above the emulsion layer. This was, to our knowledge, the first study of emulsions made exploiting the interactions between lactoferrin and chitosan. It was noted that chitosan droplets remained soluble, although the hydrocolloid solubility occurs at pH lower than 5.9. These results showed the feasibility of manufacturing lactoferrin-based emulsions as functional foods.     

Analysis of Freeze-Thaw Behavior of Double (W1/O/W2) Emulsions by Differential Scanning Calorimetry: Effects of Inner Salt Concentration and Solid Fat Content

Food Biophysics - This work is focused on the study of freeze-thaw behavior of double (W1/O/W2) emulsions by the application of a cooling-heating-cooling cycle by differential scanning calorimetry....     

A Few Comments on Electrostatic Interactions in Cell Physiology

The role of fixed charges present at the surface of biological membranes is usually described by the Gouy-Chapman-Grahame theory of the electric double-layer where the Grahame equation is applied independently on each side of the membrane and where the capacitive charges (linked to the transmembrane ionic currents) are disregarded. In this article, we generalize the Gouy-Chapman-Grahame theory by taking into account both intrinsic charges (resulting from the dissociation of membrane constituents) and capacitive charges, in the density value of the membrane surface charges. In the first part, we show that capacitive charges couple electrostatic potentials present on both sides of the membrane. The intensity of this coupling depends both on the value of the membrane specific capacitance and the transmembrane electric potential difference. In the second part, we suggest some physiological implications of membrane electric double-layers.     

Interfacial Behavior and Antioxidant Efficiency of Olive Phenolic Compounds in O/W Olive oil Emulsions as Affected by Surface Active Agent Type

The aim of this study was to evaluate the interfacial behavior of syringic acid, tyrosol and oleuropein, phenolic antioxidant compounds naturally present in olives and olive oils, and their ability to influence the stability to lipid oxidation of olive oil O/W emulsions. To test also the interactions of these molecules with other components and the effects on their activity, two different surfactants were used to prepare the olive oil emulsions, Tween 20, and a whey protein concentrate (WPC). All the antioxidants affected the olive oil/water interfacial tension; among them, oleuropein showed the highest interfacial activity and, thus, is supposed to locate at the interface. In emulsified state, the presence of the phenolic compounds in WPC emulsions did not cause any significant effect on the dispersion degree if compared to the control whilst a general improvement was observed in Tween 20-emulsions, in particular when oleuropein was added systems. The antioxidants were thus proven not to impair the dispersed structure but rather to improve it. As regards the oxidative stability, the antioxidants under investigation caused the occurrence of similar induction phases in the hydroperoxides production not observed in the control emulsions. In the case of secondary oxidation products, the highest inhibition was achieved in both the emulsified systems by oleuropein. In general, however, a lower amount of both primary and secondary oxidation products were observed in WPC emulsions than in Tween 20-emulsions likely due to the antioxidative effect of whey proteins.     

W／O微乳液中糖化酶的研究

在十六烷基三甲基溴化铵/正戊醇/异辛烷/水体系中,选取W/O型微乳液作为糖化酶的微环境,研究了其催化活性.并与其在水溶液中进行了比较.结果表明,在微乳液中,糖化酶催化淀粉水解反应的最佳反应温度和pH比在水溶液中低,反应的最大速度V_m和米氏常数K_m比其在水溶液中分别提高了近6倍和3倍.     

Effect of Electrostatic Interactions on Phase Stability of Cubic Phases of Biomembranes

We investigated effect of electrostatic interactions due to surfacecharges on structures and stability of cubic phases of monoolein (MO)membrane using the small-angle X-ray scattering method. Firstly, wechanged the surface charge density of the membrane by usingdioleoylphosphatidic acid (DOPA). As increasing DOPA concentration in themembrane at 30 wt % lipid concentration, a Q²²⁴ to Q²²⁹ phasetransition occurred at 0.6 mol % DOPA, and at and above 25 mol %, DOPA/MOmembranes were in the L phase. NaCl in the bulk phase reduced theeffect of DOPA. These results indicate that as the electrostaticinteractions increase, the most stable phase changes as follows: Q²²⁴ Q²²⁹ L. The increase in DOPAconcentration reduced the absolute value of spontaneous curvature of themembrane, | H₀ |. Secondly, we changed the surface charge of themembrane by adding a de novo designed peptide, which has netpositive charges and a binding site on the electrically neutral membraneinterface. The peptide-1 (WLFLLKKK) induced a Q²²⁴ to Q²²⁹phase transition in the MO membrane at low peptide concentration. As NaClconcentration increases, the MO/peptide-1 membrane changed from Q²²⁹to Q²²⁴ phase. The increase in peptide-1 concentration reduced |H₀ |. Based on these results, the stability of the cubic phases and themechanism of phase transition between cubic phase and L phase arediscussed.     

Possible Specificity of Cellular Interactions Due to Electrostatic Forces

It is known that the cells from a mixed population in a culture medium will finally segregate. This “social behavior” of the cells is a direct consequence of both physical and chemical interactions between cells. The physical forces involved in cell-cell interactions are considered to be the electrostatic, van der Waals, and very-short-range hydration forces.

It has been believed until now that the electrostatic forces acting between identical cells were always repulsive and nonspecific. In the present work, we try to suggest that even the electrostatic forces could manifest a slightly specific character, so that a completely random Brownian motion of the cells could be influenced, corrected, and transformed into a “docking maneuver,” which could favor the specific interactions between cells suspended in a culture medium. The main point of our approach is that the peculiar patterns of electric charge distribution on the cell surfaces act like specific electrostatic fingerprints that could be responsible for the preferential interactions which precede cellular segregation (1).     

The Role of Electrostatic Interactions for Cytochrome c Oxidase Function

In recent years, the enormous increase in high-resolution three-dimensional structures of proteins together with the development of powerful theoretical techniques have provided the basis for a more detailed examination of the role of electrostatics in determining the midpoint potentials of redox-active metal centers and in influencing the protonation behavior of titratable groups in proteins. Based on the coordinates of the Paracoccus denitrificans cytochrome c oxidase, we have determined the electrostatic potential in and around the protein, calculated the titration curves for all ionizable residues in the protein, and analyzed the response of the protein environment to redox changes at the metal centers. The results of this study provide insight into how charged groups can be stabilized within a low-dielectric environment and how the range of their electrostatic effects can be modulated by the protein. A cluster of 18 titratable groups around the heme a ₃–Cu_B binuclear center, including a hydroxide ion bound to the copper, was identified that accounts for most of the proton uptake associated with redox changes at the binuclear site. Predicted changes in net protonation were in reasonable agreement with experimentally determined values. The relevance of these findings in the light of possible mechanisms of redox-coupled proton movement is discussed.     

Uncovering Specific Electrostatic Interactions in the Denatured States of Proteins

The stability and folding of proteins are modulated by energetically significant interactions in the denatured state that is in equilibrium with the native state. These interactions remain largely invisible to current experimental techniques, however, due to the sparse population and conformational heterogeneity of the denatured-state ensemble under folding conditions. Molecular dynamics simulations using physics-based force fields can in principle offer atomistic details of the denatured state. However, practical applications are plagued with the lack of rigorous means to validate microscopic information and deficiencies in force fields and solvent models. This study presents a method based on coupled titration and molecular dynamics sampling of the denatured state starting from the extended sequence under native conditions. The resulting denatured-state pK_as allow for the prediction of experimental observables such as pH- and mutation-induced stability changes. I show the capability and use of the method by investigating the electrostatic interactions in the denatured states of wild-type and K12M mutant of NTL9 protein. This study shows that the major errors in electrostatics can be identified by validating the titration properties of the fragment peptides derived from the sequence of the intact protein. Consistent with experimental evidence, our simulations show a significantly depressed pK_a for Asp⁸ in the denatured state of wild-type, which is due to a nonnative interaction between Asp⁸ and Lys¹². Interestingly, the simulation also shows a nonnative interaction between Asp⁸ and Glu⁴⁸ in the denatured state of the mutant. I believe the presented method is general and can be applied to extract and validate microscopic electrostatics of the entire folding energy landscape.     

复乳法制备红细胞代用品

以单甲氧基聚乙二醇聚乳酸共聚物 [methoxypoly(ethyleneglycol) poly DL lactide,PELA]为膜材 ,用W /O/W的复乳 溶剂扩散法制备了包埋牛血红蛋白 (bovinehemoglobin ,BHb)的微胶囊作为人造红细胞。研究发现复乳过程搅拌分散速率、有机溶剂种类和固化方法对BHb活性有明显影响。当搅拌分散速率小于 90 0 0r/min、以乙酸乙酯为有机相时 ,采用复乳 溶剂扩散法包埋BHb的过程对BHb活性无明显影响。当搅拌分散速率高于 12 0 0 0r/min时 ,BHb活性降低。复乳 溶剂扩散法制备微胶囊过程中固化液体积与微胶囊中BHb活性密切相关 ,通过增大固化液和复乳液的体积比可较好地保持BHb的活性。最后制得了粒径 10 μm左右、包埋率 93%、P50 和Hill系数均接近于天然BHb的微胶囊。     

Impact of Lauric Arginate Application Form on its Antimicrobial Activity in Meat Emulsions

Lauric arginate (LAE) is a food-grade cationic surfactant that is highly active against a wide range of food pathogens (Listeria monocytogenes, Salmonella, and Escherichia coli) and food spoilers (Lactobacilli, yeast, and molds). The antimicrobial efficacy of LAE in compositionally complex environments is likely to be negatively impacted by its interactions with food ingredients. Therefore, we investigated different application systems of LAE and their impact on its antimicrobial efficacy when added to “Lyoner style” sausages. LAE was applied as a powder, aqueous solution, in oil-in-water emulsions with different droplet sizes, and as solid lipid particles (SLP) with different droplet sizes. Structures of the systems were identified by optical microscopy, differential scanning calorimetry (DSC) and static light scattering. A recontamination on the surface of sliced sausages was simulated using Listeria innocua as the target organism (2 log colony forming units (CFU)/slice), and the antimicrobial impact of 1,000, 1,500 and 2,000 μg/g applied LAE in the sausage was examined by growth curves. A modeling of the CFU-time relationship was carried out to provide a better evaluation of the antimicrobial activity of LAE. Finally, we carried out an isothermal titration calorimetry (ITC) analysis to simulate the interactions between LAE and proteins in the sausage matrix. Results revealed that the application systems differed in their surface area and, therefore, showed different antimicrobial activities when incorporated into sausage. The study demonstrated that the SLP and emulsions as LAE application systems increased the antimicrobial activity against microbial growth on the surface of sliced “Lyoner style” sausages.     

Assessing the Impact of Electrostatic Drag on Processive Molecular Motor Transport

The bidirectional movement of intracellular cargo is usually described as a tug-of-war among opposite-directed families of molecular motors. While tug-of-war models have enjoyed some success, recent evidence suggests underlying motor interactions are more complex than previously understood. For example, these tug-of-war models fail to predict the counterintuitive phenomenon that inhibiting one family of motors can decrease the functionality of opposite-directed transport. In this paper, we use a stochastic differential equations modeling framework to explore one proposed physical mechanism, called microtubule tethering, that could play a role in this “co-dependence” among antagonistic motors. This hypothesis includes the possibility of a trade-off: weakly bound trailing molecular motors can serve as tethers for cargoes and processing motors, thereby enhancing motor–cargo run lengths along microtubules; however, this introduces a cost of processing at a lower mean velocity. By computing the small- and large-time mean-squared displacement of our theoretical model and comparing our results to experimental observations of dynein and its “helper protein” dynactin, we find some supporting evidence for microtubule tethering interactions. We extrapolate these findings to predict how dynein–dynactin might interact with the opposite-directed kinesin motors and introduce a criterion for when the trade-off is beneficial in simple systems.     

Impact of Phytic Acid on the Physical and Oxidative Stability of Protein-Stabilized Oil-in-Water Emulsions

The effects of phytic acid on the physical and oxidative stability of flaxseed oil-in-water emulsions containing whey protein-coated lipid droplets were investigated. The surface potential, particle size, microstructure, appearance, and oxidation of these emulsions were monitored when they were stored at pH 3.5 and 7.0 for 25 days in the dark (37 °C). The phytic acid and protein-coated lipid droplets had similar charges (both negative) at pH 7.0, but had opposite charges (negative and positive) at pH 3.5. At pH 7.0, the addition of phytic acid had no impact on the physical stability of the emulsions but significantly improved their oxidative stability, which was attributed to its ability to sequester pro-oxidant transition metals (iron ions). At pH 3.5, extensive droplet aggregation and creaming occurred in the emulsions containing phytic acid, which was ascribed to charge neutralization and ion bridging. The oxidative stability of the acidified emulsions, however, still increased after addition of phytic acid, which was again attributed to its ability to chelate iron ions. Interestingly, the antioxidant activity of phytic acid decreased as its level was increased. Our results suggest that phytic acid may be used as a natural antioxidant to improve the oxidative stability of food emulsions containing polyunsaturated fatty acids, but its level must be carefully controlled.

     

Development of W/O Microemulsion for Transdermal Delivery of Iodide Ions

The objective of this study was to develop a water-in-oil (w/o) microemulsion which can be utilized as a transdermal delivery for iodide ions. Several w/o microemulsion formulations were prepared utilizing Span 20, ethanol, Capryol 90®, and water. The selected formulations had 5%, 10%, 15%, 20%, and a maximum of 23% w/w water content. Potassium iodide (KI) was incorporated in all formulations at 5% w/v. Physicochemical characterizations were conducted to evaluate the structure and stability. These studies included: mean droplet size, pH, viscosity, conductivity, and chemical stability tests. In vitro human skin permeation studies were conducted to evaluate the diffusion of the iodide ion through human skin. The w/o microemulsion formulations were stable and compatible with iodide ions with water content ranging from 5% to 23% w/w. The addition of KI influenced the physicochemical properties of microemulsion as compared to blank microemulsion formulations. In vitro human skin permeation studies indicated that selected formulations improved iodide ion diffusion significantly as compared to control (KI solution; P value < 0.05). Iodide ions were entrapped within the aqueous core of w/o microemulsion. Span 20, ethanol and Capryol 90 protected the iodide ions against oxidation and formed a stable microemulsion. It is worth to note that according to Hofmeister series, iodide ions tend to lower the interfacial tension between water and oil and consequently enhance overall stability. This work illustrates that microemulsion system can be utilized as a vehicle for the transdermal administration of iodide.KEY WORDS: iodide, microemulsion, skin permeation, transdermal     

Novel preparation method for surfactant-coated enzymes using W/O emulsion

Summary A novel preparation method for surfactant-coated enzymes has been developed using a W/O emulsion. The enzymatic activity of chymotrypsin in isooctane significantly increased with the coating of surfactants. The surfactant-coated chymotrypsin showed a high enzymatic activity for amidation, although powdered chymotrypsin did not show the activity. Further, the coated enzyme showed a remarkably high storage stability.     

Electrostatic immobilization of pectinase on negatively charged AOT-Fe3O4 nanoparticles

Enzyme immobilization on magnetic nanoparticles (MNPs) has been a field of intense studies in biotechnology during the past decade. The present study suggests MNPs negatively charged by docusate sodium salt (AOT) as a support for pectinase immobilization. AOT is a biocompatible anionic surfactant which can stabilize MNPs. Electrostatic adsorption can occur between enzyme with positive charge and oppositely charged surface of MNPs (ca. 100 nm). The effect of three factors, i.e. initial enzyme concentration, aqueous pH and AOT concentration in different levels was investigated on pectinase immobilization. Maximum specific activity (1.98 U/mg enzyme) of immobilized pectinase and maximum enzyme loading of 610.5 mg enzyme/g support was attained through the experiments. Initial enzyme concentration is significantly important on both loading and activity of immobilized enzyme, while pH and AOT concentration only affect the amount of immobilized enzyme. Immobilized enzyme on MNPs was recovered easily through magnetic separation. At near pH of immobilization, protein leakage in reusability of immobilized enzyme was low and activity loss was only 10–20% after six cycles. Since pH is associated with immobilization by electrostatic adsorption, the medium pH was changed to improve the release of protein from the support, as well. MNPs properties were investigated using Scanning Electron Microscopy (SEM), Fourier Transform Infrared (FT-IR) spectroscopy, and Dynamic Light Scattering (DLS) analysis.