Search Results (33)

Purpose: Whey protein (WP) consumption prior to a meal curbs appetite and reduces postprandial glucose (PPG) through stimulating endogenous GLP-1 secretion and insulin. Methods: We assessed the metabolic effects of a concentrated WP, using a new micelle-technology (WPM), in people with type 2 diabetes (T2D) and overweight or obesity (NCT04639726). In a randomized-crossover design, participants performed two 240 min lunch meal (622 kcal) tests 7 ± 4 days apart. After an overnight fast and a standardized breakfast, 10 g (125 mL) WPM (40 kcal) or placebo (125 mL water, 0 kcal) was consumed 15 min ahead of the mixed-nutrient meal. Effects on PPG (primary endpoint), insulin, GLP-1, and branched-chain amino acids (BCAAs) were evaluated with frequent blood sampling. Changes in incremental areas under the concentration curve (iAUC) were compared using a mixed model. Results: Twenty-six individuals (14 females, mean ± SD age 62.0 ± 8.3 years, HbA1c 58 ± 12 mmol/mol/7.5 ± 1.1%, BMI 29.2 ± 4.8 kg/m²) completed both tests. WPM significantly reduced PPG iAUC_0–2h by 22% (p = 0.028), and iAUC_0–3h numerically by −18% (p = 0.090) vs. placebo. WPM also increased insulin iAUC_0–1h by 61% (p < 0.001), and iAUC_0–3h by 30% (p = 0.004), respectively. Total GLP-1 iAUC_0–2h was enhanced by 66% (p < 0.001). Postprandial plasma BCAA patterns were characterized by a rapid increase and larger iAUC_0–2h (all p < 0.001) after WPM. No adverse events were ascribed to consuming WPM. Conclusions: A 125 mL pre-meal drink containing just 10 g WPM before a mixed meal reduced PPG and increased insulin, GLP-1, and BCAAs. WPM may therefore serve as a metabolic modulator in people with T2D living with overweight or obesity. Full article

Directed evolution is a powerful technique for creating biomolecules such as proteins and nucleic acids with tailor-made properties for therapeutic and industrial applications by mimicking the natural evolution processes in the laboratory. Droplet microfluidics improved classical directed evolution by enabling time-consuming and laborious steps in this iterative process to be performed within monodispersed droplets in a highly controlled and automated manner. Droplet microfluidic chips can generate, manipulate, and sort individual droplets at kilohertz rates in a user-defined microchannel geometry, allowing new strategies for high-throughput screening and evolution of biomolecules. In this review, we discuss directed evolution studies in which droplet-based microfluidic systems were used to screen and improve the functional properties of biomolecules. We provide a systematic overview of basic on-chip fluidic operations, including reagent mixing by merging continuous fluid streams and droplet pairs, reagent addition by picoinjection, droplet generation, droplet incubation in delay lines, chambers and hydrodynamic traps, and droplet sorting techniques. Various microfluidic strategies for directed evolution using single and multiple emulsions and biomimetic materials (giant lipid vesicles, microgels, and microcapsules) are highlighted. Completely cell-free microfluidic-assisted in vitro compartmentalization methods that eliminate the need to clone DNA into cells after each round of mutagenesis are also presented. Full article

The behavior of polyzwitterions, constituted by dipole-like zwitterionic monomers, is significantly different from that of uniformly charged polyelectrolytes. The origin of this difference lies in the intrinsic capacity of polyzwitterions to self-associate intramolecularly and associate with interpenetrating chains driven by dominant dipolar interactions. Earlier attempts to treat polyzwitterions implicitly assume that the dipoles of zwitterion monomers are randomly oriented. At ambient temperatures, the dipolar zwitterion monomers can readily align with each other generating quadrupoles and other multipoles and thus generating heterogeneous structures even in homogeneous solutions. Towards an attempt to understand the role of such dipolar associations, we present a mean field theory of solutions of polyzwitterions. Generally, we delineate a high-temperature regime where the zwitterion dipoles are randomly oriented from a low-temperature regime where quadrupole formation is significantly prevalent. We present closed-form formulas for: (1) Coil-globule transition in the low-temperature regime, the anti-polyelectrolyte effect of chain expansion upon addition of low molar mass salt, and chain relaxation times in dilute solutions. (2) Spontaneous formation of a mesomorphic state at the borderline between the high-temperature and low-temperature regimes and its characteristics. A universal law is presented for the radius of gyration of the microgel, as a proportionality to one-sixth power of the polymer concentration. (3) Swelling equilibrium of chemically cross-linked polyzwitterion gels in both the high temperature and low-temperature regimes. Addressing the hierarchical internal dynamics of polyzwitterion gels, we present a general stretched exponential law for the time-correlation function of gel displacement vector, that can be measured in dynamic light scattering experiments. The present theory is of direct experimental relevance and additional theoretical developments to all polyzwitterion systems, and generally to biological macromolecular systems such as intrinsically disordered proteins. Full article

Unsaturated fat replacement should be used to reduce the use of saturated fat and trans fatty acids in the diet. In this study, pea protein micro-gels (PPMs) with different structures were prepared by microparticulation at pH 4.0–7.0 and named as PPM (pH 4.0), PPM (pH 4.5), PPM (pH 5.0), PPM (pH 5.5), PPM (pH 6.0), PPM (pH 6.5), and PPM (pH 7.0). Pea protein was used as a control to evaluate the structure and interfacial properties of PPMs by particle size distribution, Fourier transform infrared spectroscopy (FTIR), free sulfhydryl group content, and emulsifying property. PPM (pH 7.0) was suitable for application in O/W emulsion stabilization because of its proper particle size, more flexible structure, high emulsifying activity index (EAI) and emulsifying stability index (ESI). The Pickering emulsion stabilized by PPM (pH 7.0) had a uniform oil droplet distribution and similar rheological properties to cream, so it can be used as a saturated fat replacement in the manufacture of ice cream. Saturated fat was partially replaced at different levels of 0%, 20%, 40%, 60%, 80%, and 100%, which were respectively named as PR0, PR20, PR40, PR60, PR80, and PR100. The rheological properties, physicochemical indexes, and sensory properties of low-saturated fat ice cream show that PPM (pH 7.0)-stabilized emulsion can be used to substitute 60% cream to manufacture low-saturated fat ice cream that has high structural stability and similar melting properties, overrun, and sensory properties to PR0. The article shows that it is feasible to prepare low-saturated fat ice cream with PPM (pH 7.0)-stabilized Pickering emulsion, which can not only maintain the fatty acid profile of the corn oil used, but also possess a solid-like structure. Its application is of positive significance for the development of nutritious and healthy foods and the reduction of chronic disease incidence. Full article

Many efforts are continuously undertaken to develop glucose-sensitive biomaterials able of controlling glucose levels in the body and self-regulating insulin delivery. Hydrogels that swell or shrink as a function of the environmental free glucose content are suitable systems for monitoring blood glucose, delivering insulin doses adapted to the glucose concentration. In this context, the development of sensors based on reversible binding to glucose molecules represents a continuous challenge. Concanavalin A (Con A) is a bioactive protein isolated from sword bean plants (Canavalia ensiformis) and contains four sugar-binding sites. The high affinity for reversibly and specifically binding glucose and mannose makes Con A as a suitable natural receptor for the development of smart glucose-responsive materials. During the last few years, Con A was used to develop smart materials, such as hydrogels, microgels, nanoparticles and films, for producing glucose biosensors or drug delivery devices. This review is focused on Con A-based materials suitable in the diagnosis and therapeutics of diabetes. A brief outlook on glucose-derived theranostics of cancer is also presented. Full article

The rheological properties, e.g., viscosity and yield stress, of fermented concentrated milk products (protein content > 8%) are strongly dependent on their volume fraction. Post-treatment with high-power ultrasound can reduce the volume fraction of these products and, hence, lead to reduced crowding effects and thus lower viscosities and yield stress. Besides that, the particle size distribution (span) should stay unaltered. Increasing the energy input during the sonication of fat-free fresh cheese with a protein content of 8.9 ± 0.4% decreased the volume fraction below the limit for concentrated products (ϕ = 0.4), while the particle size also decreased. This led to a narrowed span and, hence, the viscosity should have increased; however, the results showed that viscosity and yield stress were decreasing. Consequently, the influence of the span was neglectable for concentrated fermented milk products with volume fractions below the concentrated area. Furthermore, the sonicated samples showed no syneresis over a storage time of two weeks. The sonicated samples reached similar rheological properties to commercial stirred yogurt, which demonstrated the suitability of high-power ultrasound as a post-treatment to tailor the rheological properties of high-protein fermented milk products. Full article

The field of soft matter teems with molecules and aggregates of molecules that have internal size-modulating degrees of freedom. Proteins, peptides, microgels, polymers, micelles, and even some colloids can exist in multiple—often just two dominating—states with different effective sizes, where size can refer to the volume or to the cross-sectional area for particles residing on surfaces. The size-dependence of their accessible states renders the behavior of these particles pressure-sensitive. The Bragg–Williams model is among the most simple mean-field methods to translate the presence of inter-particle interactions into an approximate phase diagram. Here, we extend the Bragg–Williams model to account for the presence of particles that are immersed in a solvent and exist in two distinct states, one occupying a smaller and the other one a larger size. The basis of the extension is a lattice–sublattice approximation that we use to host the two size-differing states. Our model includes particle–solvent interactions that act as an effective surface tension between particles and solvent and are ignorant of the state in which the particles reside. We analyze how the energetic preference of the particles for one or the other state affects the phase diagrams. The possibility of a single phase-two phases-single phase sequence of phase transitions as a function of increasing temperature is demonstrated. Full article

Soy-based yoghurt alternatives were highly requested by consumers over the last few years. However, their texture does not always fulfil consumers’ demands as such yoghurt alternatives are often perceived as too firm or too soft, sandy, or fibrous. In order to improve the texture, fibres, for example, in the form of microgel particles (MGP), can be added to the soy matrix. MGP are expected to interact with soy proteins, creating different microstructures and, thus, different gel properties after fermentation. In this study, pectin-based MGP were added in different sizes and concentrations, and the soy gel properties after fermentation were characterised. It was found that the addition of 1 wt.% MGP influenced neither the flow behaviour nor the tribological/lubrication properties of the soy matrix, regardless of the MGP size. However, at higher MGP concentrations (3 and 5 wt.%), the viscosity and yield stress were reduced, the gel strength and cross-linking density decreased, and the water-holding capacity was reduced. At 5 wt.%, strong and visible phase separation occurred. Thus, it can be concluded that apple pectin-based MGP serve as inactive fillers in fermented soy protein matrices. They can, therefore, be used to weaken the gel matrix purposely to create novel microstructures. Full article

In this study, oleogels were prepared by the emulsion-template method using egg-white protein microgel as a gelator and xanthan gum (XG) as thickener. The physicochemical properties of the emulsion and oleogels were investigated. The adsorption of protein on the surface of the oil droplet reached saturation when the protein microgel concentration reached 2%. The excess protein combined with XG and accumulated on the outer layer of the oleogel, which prevented the emulsion from flocculation, enhanced the oil-holding capacity of the oleogel, and had a positive effect on preventing the oxidation of oil. When the concentration of XG was less than 0.4%, the EWP microgel, combined with the XG, stabilized the emulsion. As the concentration of XG was greater than 0.4%, excessive XG in the emulsion improved the viscosity and mechanical properties of the emulsion to prevent the aggregation of oil droplets. However, the change in XG concentration had no significant effect on the oxidation of the oil. Full article

The increasing occurrence of obesity and other non-communicable diseases has shifted the human diet towards reduced calorie intake. This drives the market to develop low-fat/non-fat food products with limited deterioration of textural properties. Thus, developing high-quality fat replacers which can replicate the role of fat in the food matrix is essential. Among all the established types of fat replacers, protein-based ones have shown a higher compatibility with a wide range of foods with limited contribution to the total calories, including protein isolate/concentrate, microparticles, and microgels. The approach to fabricating fat replacers varies with their types, such as thermal–mechanical treatment, anti-solvent precipitation, enzymatic hydrolysis, complexation, and emulsification. Their detailed process is summarized in the present review with a focus on the latest findings. The fat-mimic mechanisms of fat replacers have received little attention compared to the fabricating methods; attempts are also made to explain the underlying principles of fat replacers from the physicochemical prospect. Finally, a future direction on the development of desirable fat replacers in a more sustainable way was also pointed out. Full article

Particle size is not only important for the sensory perception of fat-free fermented concentrated milk products, but also for processing operations because of the direct relationship with apparent viscosity. The aim of this study was to apply inline particle size analysis using focused beam reflectance measurement (FBRM) to obtain real-time information regarding the particle size of a fat-free fermented concentrated milk product, namely, fresh cheese. By comparing inline particle size data to offline particle size, apparent viscosity, protein content and processing information, the potential to use inline particle size analysis as a process monitoring and control option during fresh cheese production was assessed. Evaluation of inline particle size after fermentation and before further processing, e.g., after a buffering tank, shows promise as a means to control variance of product entering downstream processing and, thus, improve final product consistency over time. Measurement of inline particle size directly before filling could allow for precise control of final product characteristics by the use of mechanical or mixing devices placed before the inline measurement. However, attention should be given to the requirements of the inline measurement technology for accurate measurement, such as product flow rate and pressure. Full article

In the last decade, alginate-based microgels have gained relevant interest as three-dimensional analogues of extracellular matrix, being able to support cell growth and functions. In this study, core-shell microgels were fabricated by self-polymerization of dopamine (DA) molecules under mild oxidation and in situ precipitation of polydopamine (PDA) onto alginate microbeads, processed by electro fluid dynamic atomization. Morphological (optical, SEM) and chemical analyses (ATR-FTIR, XPS) confirmed the presence of PDA macromolecules, distributed onto the microgel surface. Nanoindentation tests also indicated that the PDA coating can influence the biomechanical properties of the microgel surfaces—i.e., σmaxALG = 0.45 mN vs. σmaxALG@PDA = 0.30 mN—thus improving the interface with hMSCs as confirmed by in vitro tests; in particular, protein adsorption and viability tests show a significant increase in adhesion and cell proliferation, strictly related to the presence of PDA. Hence, we concluded that PDA coating contributes to the formation of a friendly interface able to efficiently support cells’ activities. In this perspective, core-shell microgels may be suggested as a novel symmetric 3D model to study in vitro cell interactions. Full article

In this study, the mechanism of ultrasound combined with corn starch granules (CSG) treatment improved the foam properties of whey protein isolates (WPI) and was systematically investigated. The results showed that ultrasound combined with corn starch granules treatment increased foam capacity and stability by 15.38% and 41.40%, respectively. Compared with the control group, corn starch granules enhanced the surface charge (52.38%) and system turbidity (51.43%), which certainly provided the necessary conditions for the improvement of foam stabilization stability. In addition, corn starch granules as microgel particles increased the mechanical properties of the interfacial protein film, thus delaying the instability of foam. This research would provide new insights into the design of new protein-based foam foods in the future food industry. Full article

Multiple myeloma is a hematologic neoplasm caused by an uncontrolled clonal proliferation of neoplastic plasma cells (nPCs) in the bone marrow. The development and survival of this disease is tightly related to the bone marrow environment. Proliferation and viability of nPCs depend on their interaction with the stromal cells and the extracellular matrix components, which also influences the appearance of drug resistance. Recapitulating these interactions in an in vitro culture requires 3D environments that incorporate the biomolecules of interest. In this work, we studied the proliferation and viability of three multiple myeloma cell lines in a microgel consisting of biostable microspheres with fibronectin (FN) on their surfaces. We also showed that the interaction of the RPMI8226 cell line with FN induced cell arrest in the G0/G1 cell cycle phase. RPMI8226 cells developed a significant resistance to dexamethasone, which was reduced when they were treated with dexamethasone and bortezomib in combination. Full article

In the last decade, different technological approaches have been proposed for the fabrication of 3D models suitable to evaluate in vitro cell response. Among them, electro fluid dynamic atomization (EFDA) belonging to the family of electro-assisted technologies allows for the dropping of polysaccharides and/or proteins solutions to produce micro-scaled hydrogels or microgels with the peculiar features of hydrogel-like materials (i.e., biocompatibility, wettability, swelling). In this work, a method to fabricate 3D scaffolds by the assembly of bicomponent microgels made of sodium alginate and gelatin was proposed. As first step, optical and scanning electron microscopy with the support of image analysis enabled to explore the basic properties of single blocks in terms of correlation between particle morphology and process parameters (i.e., voltage, flow rate, electrode gap, and needle diameter). Chemical analysis via ninhydrin essays and FTIR analysis confirmed the presence of gelatin, mostly retained by physical interactions into the alginate network mediated by electrostatic forces. In vitro tests confirmed the effect of biochemical signals exerted by the protein on the biological response of hMSCs cultured onto the microgels surface. Hence, it is concluded that alginate/gelatin microgels assemblies can efficiently work as 3D scaffolds able to support in vitro cells functions, thus providing a friendly microenvironment to investigate in vitro cell interactions. Full article