Effect of mechanical properties on in vitro dynamic digestion of starch contained in hydrogels.

BACKGROUND: This study evaluates the effect of mechanical properties on the in vitro dynamic gastrointestinal digestion of hydrogels containing starch (HCSs) as a model for studying the nutrient digestibility of solid foods. It provides a useful theoretical basis for the processing of specific foods. RESULT: Four types of HCSs with two levels of fracture stress (17.4-20.9 kPa and 55.5-57.6 kPa) and two levels of fracture strain (25.4-28.5% and 53.7-57.4%) were prepared. For these HCSs, the degree of gastric disintegration of hydrogels reduced significantly when fracture strain exceeded 30% (P < 0.05). The gastric emptying of HCS particles was also affected by mechanical properties. For example, even at the same level of fracture stress (ca. 20 kPa), the dry solids retention ratio decreased markedly from 0.90 to 0.43 with a decrease in fracture strain from 53.7% to 25.4% (P < 0.05). For the starch hydrolysis of HCSs after gastric digestion, more than 70% of starch in the particles of all types of HCSs emptied did not undergo digestion. The starch hydrolysis of HCSs during small intestinal digestion was also influenced by their mechanical properties. Fracture strains of HCSs, rather than their fracture stress, affected starch digestibility in hydrogels. CONCLUSION: The gastric disintegration, the gastric emptying, and the starch hydrolysis of HCSs are suppressed when fracture strain exceeded 30%. Even with the amount of nutritional components contained in hydrogels being the same, the in vitro gastrointestinal digestion behavior of HCSs depends on their mechanical properties. This behavior has the potential to be used in the design of processed foods with controlled bioaccessibility. © 2023 Society of Chemical Industry.

Facilitated encapsulation of a nonionic contrast agent for X-ray computed tomography into lipid vesicles by the multiple emulsification-solvent evaporation method.

We studied the encapsulation of iohexol (Ihex), a nonionic contrast agent used for X-ray computational tomography, into lipid vesicles using the multiple emulsification-solvent evaporation method to formulate a nanosized contrast agent. This lipid vesicle preparation method consists of three steps: (1) primary emulsification for producing water-in-oil (W/O) emulsions containing fine water droplets that will be converted to the internal water phase of the lipid vesicles, (2) secondary emulsification for formulating multiple water-in-oil-in-water (W/O/W) emulsions encapsulating the fine water droplets containing Ihex, and (3) solvent evaporation to remove the oil phase solvent (n-hexane) and to form lipid bilayers surrounding the fine inner droplets, resulting in the formation of lipid vesicles encapsulating Ihex. As the diameter and Ihex concentration of the primary W/O emulsion droplets decreased, a higher Ihex encapsulation yield was obtained for the final lipid vesicles. The entrapment yield of Ihex in the final lipid vesicles varied significantly with the emulsifier (Pluronic® F-68) concentration in the external water phase of W/O/W emulsion, and the highest yield (65%) was obtained when the emulsifier concentration was 0.1 wt%. We also investigated the powderization of lipid vesicles encapsulating Ihex via lyophilization. The powderized vesicles were dispersed in water after rehydration and maintained their controlled diameters. The entrapment yield of Ihex in powderized lipid vesicles was maintained for over 1 month at 25 ˚C, while significant leakage of Ihex was observed in the lipid vesicles suspended in the aqueous phase.

Improvements in Visual Aspects and Chemical, Techno-Functional and Rheological Characteristics of Cricket Powder (Gryllus bimaculatus) by Solvent Treatment for Food Utilization.

This study aimed to improve the visual aspects and chemical, techno-functional and rheological characteristics of Gryllus bimaculatus cricket powder through the use of different solvents, with the objective of using it as a protein source in food production. Four treatments (pH 5 aqueous solution, ethanol 20%, ethanol 99.5%, and hexane) were applied to the powder, and analyses were conducted to assess changes in the previously mentioned parameters. The results showed that the treatments led to an increase in protein concentration (from 55.4 to 72.5%) and a decrease in fat concentration (from 33.0 to 6.8%) in ethanol 99.5% treated powder, as well as a reduction in anti-nutritional compounds concentration, such as tannins (from 13.3 to 5.9 g/kg), in pH 5 treated powder, which is important for the nutritional value of the final product. The color of the powders was improved, being lighter after hexane and ethanol 99.5% treatments due to the removal of melanin with the defatting process. Flowability, water, and oil holding capacity were also improved in the defatted powders. All the results suggest that the main composition of the powder directly influences the analyzed parameters. These findings suggest that cricket powder treated with solvents can be used as a protein source in different food applications.

Analysis of Calcium Sulfate Scaling Phenomena on Reverse Osmosis Membranes by Scaling-Based Flux Model.

In this study, the behavior of permeate flux decline due to scale precipitation of calcium sulfate on reverse osmosis membranes was investigated. The proposed scaling-based flux model is able to explain that permeate fluxes attributed to three mechanisms of scale precipitation-cake formation, surface blockage, and mixed crystallization-converge to the same newly defined scaling-based critical flux. In addition, a scaling index is defined, which determines whether scale precipitates on the membrane. The experimental results were analyzed based on this index. The mass-transfer coefficients of flat membrane cells used in the experiments were measured and, although the coefficients differed, they could be summarized in the same form as the Leveque equation. Considering the results of the scale precipitation experiments, where the operating conditions of pressure, solute concentration, temperature, and Reynolds number were varied, the convergent values of the permeate fluxes are explained by the scaling-based critical fluxes and the scale precipitation zones by the scaling indexes.

Effect of dehydrocholic acid conjugated with a hydrocarbon on a lipid bilayer composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine.

The effects of bile acids, dehydrocholic acid (DHA) and DHA conjugated with a hydrocarbon (6-aminohexanoate; 6A-DHA) were evaluated using a lipid bilayer composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). DOPC formed a homogenous thin membrane in presence or absence of the DHA, while 20 mol% 6A-DHA induced phase separation on the DOPC thin membrane. It was observed formation of a stomatocyte-like liposomes when these membranes were suspended in a basic solvent. Generally, liposome formation can be prevented by some bile acids. It was found that DHA and 6A-DHA did not disrupt liposome formation, while DHA and 6A-DHA perturbed the liposomal membrane, resulting in increased local-fluidity due to the bent structure of DHA and 6A-DHA. DHA and 6A-DHA showed completely different effects on the hydrophobicity of the boundary surface of DOPC liposome membranes. The steroidal backbone of DHA was found to prevent the insertion of water molecules into the liposomal membrane, whereas 6A-DHA did not show the same behavior which was attributed to its conjugated hydrocarbon.

Freeze-dryable lipid vesicles with size tunability and high encapsulation efficiency prepared by the multiple emulsification-solvent evaporation method.

We investigated the extent of potential applicability of our recently developed method for preparing lipid vesicles [T. Kuroiwa et al., J. Am. Oil Chem. Soc., 93 (2016) 421], designated as the multiple emulsification-solvent evaporation method, with the intention of controlling the vesicle diameter and achieving high entrapment efficiency for water-soluble compounds. Using this method, the diameter of lipid vesicles could be varied by selecting the methods for preparing the primary water-in-oil emulsion, which contained water droplets as templates for the internal water phases of lipid vesicles. We obtained lipid vesicles with mean diameters of 0.2-4.4µm from water-in-oil-in-water multiple emulsions after solvent evaporation. A high entrapment yield of calcein, a water-soluble fluorescent dye, into the lipid vesicles was obtained for each vesicle sample, depending on their diameter and the type of emulsifier added to the external water phase. The use of polymeric emulsifier was more effective in achieving a high entrapment yield. The obtained lipid vesicles were powderized via freeze-drying. Vesicles could be powderized while maintaining their original diameter, as confirmed by scanning electron microscopy. Furthermore, the powderized vesicles could be rehydrated and resuspended without significant change in their diameter. However, the entrapment yield of calcein decreased after freeze-drying and rehydration. The calcein leakage during the freeze-drying followed by rehydration could be suppressed by adding an appropriate amount of trehalose as a lyoprotectant.

Vicenistatin induces early endosome-derived vacuole formation in mammalian cells.

Homotypic fusion of early endosomes is important for efficient protein trafficking and sorting. The key controller of this process is Rab5 which regulates several effectors and PtdInsPs levels, but whose mechanisms are largely unknown. Here, we report that vicenistatin, a natural product, enhanced homotypic fusion of early endosomes and induced the formation of large vacuole-like structures in mammalian cells. Unlike YM201636, another early endosome vacuolating compound, vicenistatin did not inhibit PIKfyve activity in vitro but activated Rab5-PAS pathway in cells. Furthermore, vicenistatin increased the membrane surface fluidity of cholesterol-containing liposomes in vitro, and cholesterol deprivation from the plasma membrane stimulated vicenistatin-induced vacuolation in cells. These results suggest that vicenistatin is a novel compound that induces the formation of vacuole-like structures by activating Rab5-PAS pathway and increasing membrane fluidity.

Formulation and stabilization of nano-/microdispersion systems using naturally occurring edible polyelectrolytes by electrostatic deposition and complexation.

This review paper presents an overview of the formulation and functionalization of nano-/microdispersion systems composed of edible materials. We first summarized general aspects on the stability of colloidal systems and the roles of natural polyelectrolytes such as proteins and ionic polysaccharides for the formation and stabilization of colloidal systems. Then we introduced our research topics on (1) stabilization of emulsions by the electrostatic deposition using natural polyelectrolytes and (2) formulation of stable nanodispersion systems by complexation of natural polyelectrolytes. In both cases, the preparation procedures were relatively simple, without high energy input or harmful chemical addition. The properties of the nano-/microdispersion systems, such as particle size, surface charge and dispersion stability were significantly affected by the concerned materials and preparation conditions, including the type and concentration of used natural polyelectrolytes. These dispersion systems would be useful for developing novel foods having high functionality and good stability.

PIV and CFD studies on analyzing intragastric flow phenomena induced by peristalsis using a human gastric flow simulator.

This study quantitatively analyzed the flow phenomena in model gastric contents induced by peristalsis using a human gastric flow simulator (GFS). Major functions of the GFS include gastric peristalsis simulation by controlled deformation of rubber walls and direct observation of inner flow through parallel transparent windows. For liquid gastric contents (water and starch syrup solutions), retropulsive flow against the direction of peristalsis was observed using both particle image velocimetry (PIV) and computational fluid dynamics (CFD). The maximum flow velocity was obtained in the region occluded by peristalsis. The maximum value was 9 mm s(-1) when the standard value of peristalsis speed in healthy adults (UACW = 2.5 mm s(-1)) was applied. The intragastric flow-field was laminar with the maximum Reynolds number (Re = 125). The viscosity of liquid gastric contents hardly affected the maximum flow velocity in the applied range of this study (1 to 100 mPa s). These PIV results agreed well with the CFD results. The maximum shear rate in the liquid gastric contents was below 20 s(-1) at UACW = 2.5 mm s(-1). We also measured the flow-field in solid-liquid gastric contents containing model solid food particles (plastic beads). The direction of velocity vectors was influenced by the presence of the model solid food particle surface. The maximum flow velocity near the model solid food particles ranged from 8 to 10 mm s(-1) at UACW = 2.5 mm s(-1). The maximum shear rate around the model solid food particles was low, with a value of up to 20 s(-1).

Microcompartmentalized cell-free protein synthesis in semipermeable microcapsules composed of polyethylenimine-coated alginate.

We describe microcompartmentalized cell-free protein synthesis in semipermeable microcapsules prepared from water-in-oil-in-water droplets by a rupture-induced encapsulation method. An aqueous solution of template DNA coding for green fluorescent protein and enzymes for the cell-free protein synthesis was aliquoted into water-in-oil droplets using a microfluidic device, and the droplets were transformed into semipermeable microcapsules. Substrates for protein synthesis diffused into the microcapsules through their semipermeable polyion complex membranes composed of polyethylenimine-coated alginate. Cell-free protein synthesis was confirmed by detection of the fluorescence of the synthesized green fluorescence protein in the microcapsules. We also used this microcompartmentalized system to synthesize protein from a single molecule of template DNA encapsulated by limiting dilution.

Industrial lab-on-a-chip: design, applications and scale-up for drug discovery and delivery.

Microfluidics is an emerging and promising interdisciplinary technology which offers powerful platforms for precise production of novel functional materials (e.g., emulsion droplets, microcapsules, and nanoparticles as drug delivery vehicles- and drug molecules) as well as high-throughput analyses (e.g., bioassays, detection, and diagnostics). In particular, multiphase microfluidics is a rapidly growing technology and has beneficial applications in various fields including biomedicals, chemicals, and foods. In this review, we first describe the fundamentals and latest developments in multiphase microfluidics for producing biocompatible materials that are precisely controlled in size, shape, internal morphology and composition. We next describe some microfluidic applications that synthesize drug molecules, handle biological substances and biological units, and imitate biological organs. We also highlight and discuss design, applications and scale up of droplet- and flow-based microfluidic devices used for drug discovery and delivery.

Efficient preparation of giant vesicles as biomimetic compartment systems with high entrapment yields for biomacromolecules.

The 'lipid-coated ice-droplet hydration method' was applied for the preparation of milliliter volumes of a suspension of giant phospholipid vesicles containing in the inner aqueous vesicle pool in high yield either calcein, α-chymotrypsin, fluorescently labeled bovine serum albumin or dextran (FITC-BSA and FITC-dextran; FITC=fluorescein isothiocyanate). The vesicles had an average diameter of ca. 7-11 µm and contained 20-50% of the desired molecules to be entrapped, the entrapment yield being dependent on the chemical structure of the entrapped molecules and on the details of the vesicle-formation procedure. The 'lipid-coated ice droplet hydration method' is a multistep process, based on i) the initial formation of a monodisperse water-in-oil emulsion by microchannel emulsification, followed by ii) emulsion droplet freezing, and iii) surfactant and oil removal, and replacement with bilayer-forming lipids and an aqueous solution. If one aims at applying the method for the entrapment of enzymes, retention of catalytic activity is important to consider. With α-chymotrypsin as first model enzyme to be used with the method, it was shown that high retention of enzymatic activity is possible, and that the entrapped enzyme molecules were able to catalyze the hydrolysis of a membrane-permeable substrate which was added to the vesicles after their formation. Furthermore, one of the critical steps of the method that leads to significant release of the molecules from the water droplets was investigated and optimized by using calcein as fluorescent probe.

Characterization of phospholipids in membrane vesicles derived from Pseudomonas aeruginosa.

Many Gram-negative bacteria release membrane vesicles (MVs), but their phospholipid properties are poorly understood. Phosphatidylglycerol was present at high levels in MVs derived from Pseudomonas aeruginosa, but not in the cellular outer membrane. The ratio of stearic acid in MVs was high compared to that in the cellular outer membrane. These findings suggest that membrane rigidity is associated with MV biogenesis.

Formation of monodisperse calcium alginate microbeads by rupture of water-in-oil-in-water droplets with an ultra-thin oil phase layer.

This paper reports a novel formation method of monodisperse calcium alginate microbeads from water-in-oil-in-water (W/O/W) droplets with an ultra-thin oil phase layer. W/O/W droplets containing sodium alginate in an internal aqueous phase were formed as a template of calcium alginate microbeads using a microfluidic device. The ultra-thin oil phase layer of the W/O/W droplets was ruptured by an osmotic pressure difference between the internal and external aqueous phase. Immediately after the rupture, polyanionic alginate in the internal aqueous phase was cross-linked with calcium ion diffused from the external aqueous phase, and monodisperse and spherical calcium alginate microbeads were formed.

Variation of physiochemical properties and cell association activity of membrane vesicles with growth phase in Pseudomonas aeruginosa.

Pseudomonas aeruginosa and other Gram-negative bacteria release membrane vesicles (MVs) from their surfaces, and MVs have an ability to interact with bacterial cells. Although it has been known that many bacteria have mechanisms that control their phenotypes with the transition from exponential phase to stationary phase, changes of properties in released MVs have been poorly understood. Here, we demonstrate that MVs released by P. aeruginosa during the exponential and stationary phases possess different physiochemical properties. MVs purified from the stationary phase had higher buoyant densities than did those purified from the exponential phase. Surface charge, characterized by zeta potential, of MVs tended to be more negative as the growth shifted to the stationary phase, although the charges of PAO1 cells were not altered. Pseudomonas quinolone signal (PQS), one of the regulators related to MV production in P. aeruginosa, was lower in MVs purified from the exponential phase than in those from the stationary phase. MVs from the stationary phase more strongly associated with P. aeruginosa cells than did those from the exponential phase. Our findings suggest that properties of MVs are altered to readily interact with bacterial cells along with the growth transition in P. aeruginosa.

Microfluidic preparation of water-in-oil-in-water emulsions with an ultra-thin oil phase layer.

We developed a novel microfluidic device to prepare monodisperse water-in-oil-in-water (W/O/W) emulsions with an ultra-thin (<1 microm) oil phase layer. This microfluidic device was composed of two microchannel junctions, one of which had a step structure, and a uniformly hydrophobic surface for effective oil removal from W/O/W droplets. At the first junction, an internal aqueous phase was transformed into slug-shaped water-in-oil (W/O) droplets by a flow-focusing mechanism. At the second junction equipped with the step structure, the preformed slug-shaped W/O droplets were introduced into an external aqueous phase and were transformed into spherical W/O droplets. In the downstream area of the second junction, the W/O droplets were released from the hydrophobic surface of the microchannel into the external aqueous phase by inertial lift force and were transformed into W/O/W droplets. During this process, most of the oil phase was effectively removed from the W/O droplets: the bulk of the oil phase flowed along the hydrophobic surface of the microchannel. The thickness of the oil phase layer of the resulting W/O/W droplets was ultra-thin, less than 1 microm. The volume of the internal aqueous phase of the W/O/W droplets reflected that of the W/O droplets and was controlled by the flow rates of the internal aqueous phase and oil phase during W/O droplet formation. We successfully demonstrated encapsulation of water-soluble molecules and polymer particles into the prepared W/O/W emulsion.

Characterization of a soybean oil-based biosurfactant and evaluation of its ability to form microbubbles.

This paper characterizes the physico-chemical properties of the soybean oil-based polymeric surfactant, Palozengs R-004 (hereafter referred to as R-004). The surface activity of R-004 is comparable to the reported activities of biosurfactants produced by microorganisms and higher than some of the conventional synthetic surfactants. The surface tension of Milli-Q water was reduced to a minimum value of roughly 30mN/m at a concentration of about 0.07wt.%. R-004 exhibited a unique aggregation behavior: small aggregates (pre-micelles) were formed at very low concentrations. Zeta-potential measurements showed that the micelles of R-004 are negatively charged due to the presence of carboxylic groups. The ability of R-004 to form and stabilize microbubbles was evaluated and was found to be greatly affected by filtration while remaining independent of R-004 concentration over the concentration range studied (0.05-0.5wt.%). These results suggest that a very low level of surfactant can be used to produce microbubbles without affecting their properties. Our results suggest the possibility of using soybean oil-based surfactants to food, pharmaceutical, and cosmetic applications.

Pseudomonas quinolone signal affects membrane vesicle production in not only gram-negative but also gram-positive bacteria.

Many Gram-negative bacteria naturally produce membrane vesicles (MVs) to the extracellular milieu. The Pseudomonas quinolone signal (PQS), a quorum-sensing signal of Pseudomonas aeruginosa, is a positive regulator of MV production. In this study, we investigated its effects on MV production in other Gram-negative and -positive bacterial species. The addition of PQS to an Escherichia coli K12 culture resulted in increased MV production and enlarged MVs. An excessive amount of MgCl(2) repressed E. coli MV production either with or without PQS, suggesting that an anionic repulsion of cellular surfaces increases MV production. PQS was found in the cellular membrane and MVs in E. coli. The enhancement of MV production by PQS occurred in other Gram-negative bacteria, including Burkholderia and Pseudomonas species. Moreover, PQS induced MV production in a Gram-positive bacterium, Bacillus subtilis 168, which does not normally produce MV under laboratory conditions. An excessive amount of MgCl(2) did not repress B. subtilis MV production in the presence of PQS, suggesting the production mechanism to be different from that in Gram-negative bacteria. Together, these results indicated that PQS enhances MV production in Gram-negative bacteria and induces it in Gram-positive bacteria.

Effect of a local anesthetic lidocaine hydrochloride on the bilayer structure of phospholipids.

The effect of a local anesthetic, lidocaine hydrochloride (LC x HCl), on the bilayer systems of purified egg phosphatidylcholine (EPC), dipalmitoylphosphatidylcholine (DPPC) and dioleoylphosphatidylcholine (DOPC) was studied by means of small-angle X-ray scattering (SAXS), Prodan fluorescence and electrophoretic light scattering. In the liquid crystalline phase of EPC and DOPC bilayers, the contraction of lamellar distance by ca. 0.8-1.0 nm and the decrease of average vesicle size were observed in the presence of LC x HCl. The adsorption of LC x HCl on the vesicle interface brought about the lateral expansion of bilayers and the decrease in the radius of curvature of vesicles. The contraction in the lamellar distance of EPC bilayer caused by high concentration of LC x HCl is attributable to the chain folding in the liquid crystalline state. In the gel phase of DPPC bilayer, the contraction of the lamellar distance in the presence of 0.37 M LC x HCl amounts to 1.6 nm, and the emission maximum of Prodan fluorescence was red-shifted from 440 nm to 518 nm. These phenomena are attributed to the formation of LC x HCl-induced interdigitated gel phase.

Effects of surfactant and electrolyte concentrations on bubble formation and stabilization.

As interest in the application of microbubbles grows, it is becoming increasingly important to understand the factors affecting their formation and properties in order to effectively generate microbubbles. This paper investigates the effect of surfactant concentration and electrolyte addition on the size distribution and stability of microbubbles. The anionic surfactant sodium dodecyl sulfate (SDS) was used as the surfactant. Minimum bubble diameter and maximum stability were achieved at surfactant concentrations above the CMC. The effect of the electrolyte addition was studied by adding sodium chloride (NaCl) at an SDS concentration below the critical micelle concentration (CMC). Addition of NaCl decreased bubble size and improved bubble preparation to a certain extent. The addition of salt at low concentrations did not affect the surface tension; however, the surface tension was reduced as salt concentration was increased and reached a constant value for NaCl concentrations above 0.25%. The presence of NaCl resulted in a significant decrease in zeta-potential, implying a reduction in the surface charge of SDS micelles. This result suggests that the presence of NaCl may improve the generation and stability of bubbles by enhancing the structures of the adsorption monolayer and interfacial film.