3D printing of responsive chiral photonic nanostructures.

Finely controlled flow forces in extrusion-based additive manufacturing can be exploited to program the self-assembly of malleable nanostructures in soft materials by integrating bottom-up design into a top-down processing approach. Here, we leverage the processing parameters offered by direct ink-writing (DIW) to reconfigure the photonic chiral nematic liquid crystalline phase in hydroxypropyl cellulose (HPC) solutions prior to deposition on the writing substrate to direct structural evolution from a particular initial condition. Moreover, we incorporate polyethylene glycol (PEG) into iridescent HPC inks to form a physically cross-linked network capable of inducing kinetic arrest of the cholesteric/chiral pitch at length scales that selectively reflect light throughout the visible spectrum. Based on thorough rheological measurements, we have found that printing the chiral inks at a shear rate where HPC molecules adopt pseudonematic state results in uniform chiral recovery following flow cessation and enhanced optical properties in the solid state. Printing chiral inks at high shear rates, on the other hand, shifts the monochromatic appearance of the extruded filaments to a highly angle-dependent state, suggesting a preferred orientation of the chiral domains. The optical response of these filaments when exposed to mechanical deformation can be used in the development of optical sensors.

Adaptive Laboratory Evolution of Probiotics toward Oxidative Stress Using a Microfluidic-Based Platform.

Adaptive laboratory evolution (ALE) can be used to make bacteria less susceptible to oxidative stress. An alternative to large batch scale ALE cultures is to use microfluidic platforms, which are often more economical and more efficient. Microfluidic ALE platforms have shown promise, but many have suffered from subpar cell passaging mechanisms and poor spatial definition. A new approach is presented using a microfluidic Evolution on a Chip (EVoc) design which progressively drives microbial cells from areas of lower H2O2 concentration to areas of higher concentration. Prolonged exposure, up to 72 h, revealed the survival of adaptive strains of Lacticaseibacillus rhamnosus GG, a beneficial probiotic often included in food products. After performing ALE on this microfluidic platform, the bacteria persisted under high H2O2 concentrations in repeated trials. After two progressive exposures, the ability of L. rhamnosus to grow in the presence of H2O2 increased from 1 mm H2O2 after a lag time of 31 h to 1 mm after 21 h, 2 mm after 28 h, and 3 mm after 42 h. The adaptive strains have different morphology, and gene expression compared to wild type, and genome sequencing revealed a potentially meaningful single nucleotide mutation in the protein omega-amidase.

Mammalian sperm hyperactivation regulates navigation via physical boundaries and promotes pseudo-chemotaxis.

Mammalian sperm migration within the complex and dynamic environment of the female reproductive tract toward the fertilization site requires navigational mechanisms, through which sperm respond to the tract environment and maintain the appropriate swimming behavior. In the oviduct (fallopian tube), sperm undergo a process called "hyperactivation," which involves switching from a nearly symmetrical, low-amplitude, and flagellar beating pattern to an asymmetrical, high-amplitude beating pattern that is required for fertilization in vivo. Here, exploring bovine sperm motion in high-aspect ratio microfluidic reservoirs as well as theoretical and computational modeling, we demonstrate that sperm hyperactivation, in response to pharmacological agonists, modulates sperm-sidewall interactions and thus navigation via physical boundaries. Prior to hyperactivation, sperm remained swimming along the sidewalls of the reservoirs; however, once hyperactivation caused the intrinsic curvature of sperm to exceed a critical value, swimming along the sidewalls was reduced. We further studied the effect of noise in the intrinsic curvature near the critical value and found that these nonthermal fluctuations yielded an interesting "Run-Stop" motion on the sidewall. Finally, we observed that hyperactivation produced a "pseudo-chemotaxis" behavior, in that sperm stayed longer within microfluidic chambers containing higher concentrations of hyperactivation agonists.

Biphasic Chemokinesis of Mammalian Sperm.

The female reproductive tract (FRT) continuously modulates mammalian sperm motion by releasing various clues as sperm migrate toward the fertilization site. An existing gap in our understanding of sperm migration within the FRT is a quantitative picture of how sperm respond to and navigate the biochemical clues within the FRT. In this experimental study, we have found that in response to biochemical clues, mammalian sperm display two distinct chemokinetic behaviors which are dependent upon the rheological properties of the media: chiral, characterized by swimming in circles; and hyperactive, characterized by random reorientation events. We used minimal theoretical modeling, along with statistical characterization of the chiral and hyperactive trajectories, to show that the effective diffusivity of these motion phases decreases with increasing concentration of chemical stimulant. In the context of navigation this concentration dependent chemokinesis suggests that the chiral or hyperactive motion refines the sperm search area within different FRT functional regions. Further, the ability to switch between phases indicates that sperm may use various stochastic navigational strategies, such as run and tumble or intermittent search, within the fluctuating and spatially heterogeneous environment of the FRT.

Understanding animal-based flavor generation, mechanisms and characterization: Cheddar cheese and bacon flavors.

Natural animal-based flavors have great appeal to consumers and have broad applications in the food industry. In this review, we summarized findings related to bacon and Cheddar cheese flavors' components and their precursors, reaction mechanisms, influential factors, and characterization methods. The results show that free sugars, free amino acids, peptides, vitamins, lipids, and nitrites are precursors to bacon flavor. The conditions governing the formation of bacon flavor are thermally dependent, which facilitates the use of thermal food processing to generate such a flavor. For Cheddar cheese flavor, milk ingredients such as lactose, citrate, fat, and casein are reported as precursors. The optimum conditions to generate Cheddar cheese flavor from precursors are quite strict, which limits its application in food processing. As an alternative, it is more practical to generate Cheddar cheese flavor by combining key aroma compounds using thermal food processing. This review provides the food industry the comprehensive information about the generation of bacon and Cheddar cheese flavors using precursor molecules.

Encapsulation and stabilization of lactoferrin in polyelectrolyte ternary complexes.

Effective delivery of the bioactive protein, lactoferrin (LF), remains a challenge as it is sensitive to environmental changes and easily denatured during heating, restricting its application in functional food products. To overcome these challenges, we formulated novel polyelectrolyte ternary complexes of LF with gelatin (G) and negatively charged polysaccharides, to improve the thermal stability of LF with retained antibacterial activity. Linear, highly charged polysaccharides were able to form interpolymeric complexes with LF and G, while coacervates were formed with branched polysaccharides. A unique multiphase coacervate was observed in the gum Arabic GA-LF-G complex, where a special coacervate-in-coacervate structure was found. The ternary complexes made with GA, soy soluble polysaccharide (SSP), or high methoxyl pectin (HMP) preserved the protein structures and demonstrated enhanced thermal stability of LF. The GA-LF-G complex was especially stable with >90% retention of the native LF after treatment at 90 °C for 2 min in a water bath or at 145 °C for 30 s, while the LF control had only ~ 7% undenatured LF under both conditions. In comparison to untreated LF, LF in ternary complex retained significant antibacterial activity on both Gram-positive and Gram-negative bacteria, even after heat treatment. These ternary complexes of LF maintain the desired functionality of LF, thermal stability and antibacterial activity, in the final products. The ternary complex structure, particularly the multiphase coacervate, may serve as a template for the encapsulation and stabilization of other bioactives and peptides.

Thermal Degradation of Thaumatin at Low pH and Its Prevention Using Alkyl Gallates.

Thaumatin, a potent sweet tasting protein extracted from the Katemfe Plant, is emerging as a natural alternative to synthetic non-nutritive sweeteners and flavor enhancer. As a food additive, its stability within the food matrix during thermal processing is of great interest to the food industry. When heated under neutral or basic conditions, thaumatin was found to lose its sweetness due to protein aggregation caused by sulfhydryl catalyzed disulfide bond interchange. At lower pH, while thaumatin was also found to lose sweetness after heating, it does so at a slower rate and shows more resistance to sweetness loss. SDS-PAGE indicated that thaumatin fragmented into multiple smaller pieces under heating in acidic pH. Using BEMPO-3, a lipophilic spin trap, we were able to detect the presence of a free-radical within the hydrophobic region of the protein during heating. Protein carbonyl content, a byproduct of protein oxidation, also increased upon heating, providing additional evidence for protein cleavage by a radical pathway. Hexyl gallate successfully inhibited the radical generation as well as protein carbonyl formation of thaumatin during heating.

Flavor components, precursors, formation mechanisms, production and characterization methods: garlic, onion, and chili pepper flavors.

There is an enormous demand in the food industry to shift toward natural flavors. However, most flavor molecules are significantly unstable outside their original sources. Moreover, limited studies are focused on the flavor formation mechanisms, regeneration methods, and stability, which could help facilitate this replacement by establishing a link between food processing conditions and flavor generation.This scoping review summarizes major findings related to the identification of garlic, onion, and chili pepper flavors and their precursor molecules, formation mechanisms, generation of flavors and precursors, characterization methods, and precursor stability under thermal food processing conditions. The findings confirmed that the allium flavors could be generated by alliin and isoalliin precursors through thermal processing. Also, the literature lacks detailed knowledge about chili pepper flavor's precursors, and only capsaicinoids have been reported as a thermally stable chili pepper flavor.Although numerous studies have focused on this area, there is still a lack of detailed applicable knowledge. Future investigations can be framed into (1) Development of efficient methods to generate flavors during food processing; (2) Improvement of flavors' stability; (3) Understanding the interactions of flavors and their precursors with other food ingredients and additives; and (4) Characterization of the organoleptic properties of flavors.

The influence of the female reproductive tract and sperm features on the design of microfluidic sperm-sorting devices.

Although medical advancements have successfully helped a lot of couples with their infertility by assisted reproductive technologies (ART), sperm selection, a crucial stage in ART, has remained challenging. Therefore, we aimed to investigate novel sperm separation methods, specifically microfluidic systems, as they do sperm selection based on sperm and/or the female reproductive tract (FRT) features without inflicting any damage to the selected sperm during the process. In this review, after an exhaustive studying of FRT features, which can implement by microfluidics devices, the focus was centered on sperm selection and investigation devices. During this study, we tried not to only point to the deficiencies of these systems, but to put forth suggestions for their improvement as well.

Heat- and shear-reversible networks in food: A review.

While nature behaves like an irreversible network with respect to entropy and time, certain systems in nature exist that are, to some extent, reversible. The property of reversibility imparts unique benefits to systems that possess them, making them suitable for designing self-healing, stimuli-responsive, and smart materials that can be used in widely divergent fields. Reversible networks are currently being exploited for applications in tissue engineering, drug delivery, and soft robotics. They are also being utilized as low-calorie fat mimetics with melt-in-your-mouth textures, as well as being explored as potential scaffolds for three-dimensional (3D) printable food, among other applications. This review aims to gather representative examples of heat- and shear-reversible networks in the food science literature from the last 30 or so years, in other words, reversible food gels made either from linear biopolymers or from colloidal, particulate dispersions, including those that have been modified specifically to induce reversibility. An overview of the network mechanisms involved that impart reversibility, including a discussion of the strength and range of forces involved, will be highlighted. A model that explains why certain networks are thermoreversible while others are shear-reversible, and why others are both, will also be proposed. A fundamental understanding of these mechanisms will prove invaluable when designing reversible networks in the future, making possible the precise control of their properties, thus fostering innovative applications within the food industry and beyond.

Diffusion-Convection Hybrid Microfluidic Platform for Rapid Antibiotic Susceptibility Testing.

Conventional antibiotic susceptibility testing (AST) assays such as broth microdilution and Kirby-Bauer disk diffusion are time-consuming (e.g., 24-72 h) and labor-intensive. Here, we present a microfluidic platform to perform AST assays with a broad range of antibiotic concentrations and controls. A culture medium stream was serially enriched with antibiotics along the length of the platform via diffusion and flow-directing mass convection mechanisms, generating a concentration gradient captured in a series of microchamber duplicates. We observed an agreement between the simulated and experimental concentration gradients and applicability to a variety of different molecules by changing the loading time according to a simple linear equation. The AST assay in our platform is based on bacterial metabolism, indicated by resazurin fluorescence. The small reaction volume enabled a minimum inhibitory concentration (MIC) to be determined in 4-5 h. Proof-of-concept functionality testing, using human isolates and clinically important antibiotics from different classes, indicated a high rate of agreement (94%: MIC within ±1 two-fold dilution of the reference method) of on-chip MICs and conventional broth microdilution. Overall, our results showed that this microfluidic platform is capable of determining antibiotic susceptibility in a rapid and reliable manner.

Tuning C-Phycocyanin Photoactivity via pH-Mediated Assembly-Disassembly.

Environment-triggered protein conformational changes have garnered wide interest in both fundamental research, for deciphering in vivo acclimatory responses, and practical applications, for designing stimuli-responsive probes. Here, we propose a protein-chromophore regulatory mechanism that allows for manipulation of C-phycocyanin (C-PC) from Spirulina platensis by environmental pH and UV irradiation. Using small-angle X-ray scattering, a pH-mediated C-PC assembly-disassembly pathway, from monomers to nonamers, was unraveled. Such flexible protein matrices impart tunability to the embedded tetrapyrroles, whose photochemical behaviors were found to be modulated by protein assembly states. UV irradiation on C-PC triggers pH-dependent singlet oxygen (1O2) generation and conformational changes. Intermolecular photo-crosslinking occurs at pH 5.0 via dityrosine species, which bridges solution-based C-PC oligomers into unprecedented dodecamers and 24-mers. These supramolecular assemblies impart C-PC at pH 5.0, which significantly enhanced 1O2 yield, fluorescence, and photostability relative to those at other pH values, a finding that makes C-PC appealing for tumor-targeted photodynamic therapy.

Rheotaxis-based separation of sperm with progressive motility using a microfluidic corral system.

The separation of motile sperm from semen samples is sought after for medical infertility treatments. In this work, we demonstrate a high-throughput microfluidic device that can passively isolate motile sperm within corrals inside a fluid channel, separating them from the rest of the diluted sample. Using finite element method simulations and proposing a model for sperm motion, we investigated how flow rate can provide a rheotaxis zone in front of the corral for sperm to move upstream/downstream depending on their motility. Using three different flow rates that provided shear rates above the minimum value within the rheotaxis zone, we experimentally tested the device with human and bovine semen. By taking advantage of the rheotactic behavior of sperm, this microfluidic device is able to corral motile sperm with progressive velocities in the range of 48-93 µm⋅s-1 and 51-82 µm⋅s-1 for bovine and human samples, respectively. More importantly, we demonstrate that the separated fractions of both human and bovine samples feature 100% normal progressive motility. Furthermore, by extracting the sperm swimming distribution within the rheotaxis zone and sperm velocity distribution inside the corral, we show that the minimum velocity of the corralled sperm can be adjusted by changing the flow rate; that is, we are able to control the motility of the separated sample. This microfluidic device is simple to use, is robust, and has a high throughput compared with traditional methods of motile sperm separation, fulfilling the needs for sperm sample preparation for medical treatments, clinical applications, and fundamental studies.

Combination of copigmentation and encapsulation strategies for the synergistic stabilization of anthocyanins.

Copigmentation and encapsulation are the two most commonly used techniques for anthocyanin stabilization. However, each of these techniques by itself suffers from many challenges associated with the simultaneous achievement of color intensification and high stability of anthocyanins. Integrating copigmentation and encapsulation may overcome the limitation of usage of a single technique. This review summarizes the most recent studies and their challenges aiming at combining copigmentation and encapsulation techniques. The effective approaches for encapsulating copigmented anthocyanins are described, including spray/freeze-drying, emulsification, gelation, polyelectrolyte complexation, and their combinations. Other emerging approaches, such as layer-by-layer deposition and ultrasonication, are also reviewed. The physicochemical principles underlying the combined strategies for the fabrication of various delivery systems are discussed. Particular emphasis is directed toward the synergistic effects of copigmentation and encapsulation, for example, modulating roles of copigments in the processes of gelation and complexation. Finally, some of the major challenges and opportunities for future studies are highlighted. The trend of integrating copigmentation and encapsulation has been just started to develop. The information in this review should facilitate the exploration of the combination of multistrategy and the fabrication of robust delivery systems for copigmented anthocyanins.

A Spiderweb-Like Metal-Organic Framework Multifunctional Foam.

Processing metal-organic frameworks (MOFs) into hierarchical macroscopic materials can greatly extend their practical applications. However, current strategies suffer from severe aggregation of MOFs and limited tuning of the hierarchical porous network. Now, a strategy is presented that can simultaneously tune the MOF loading, composition, spatial distribution, and confinement within various bio-originated macroscopic supports, as well as control the accessibility, robustness, and formability of the support itself. This method enables the good dispersion of individual MOF nanoparticles on a spiderweb-like network within each macrovoid even at high loadings (up to 86 wt %), ensuring the foam pores are highly accessible for excellent adsorption and catalytic capacity. Additionally, this approach allows the direct pre-incorporation of other functional components into the framework. This strategy provides precise control over the properties of both the hierarchical support and MOF.

Nanoliter-Sized Microchamber/Microarray Microfluidic Platform for Antibiotic Susceptibility Testing.

The rise of antimicrobial resistance is challenging for physicians in clinical practice to prescribe antibiotics that are effective against bacterial infections. Conventional antibiotic susceptibility testing (AST) is labor-intensive and time-consuming (18-24 h). Newly emerging technologies such as microfluidics may enable more rapid AST assay time. In this study, we utilize a nanoliter-sized microchamber/microarray-based microfluidic (N-3M) platform to reduce the AST assay time and rapidly determine the minimum inhibitory concentrations of different antibiotics. Bacterial suspensions, with or without antibiotics, are loaded into small nanoliter-sized chambers, and the change in fluorescent intensity emitted from resazurin reduction, which correlated with bacterial growth, is measured. We demonstrate the reproducibility, functionality, and efficiency of our N-3M platform for numerous wild-type clinical bacterial isolates including Escherichia coli, Klebsiella pneumoniae, and Enterococcus faecalis. The time-to-result of our N-3M platform varies between ∼1-3 h, depending on growth rates of different bacterial species. We believe that our proposed N-3M platform is robust, is easy-to-implement, has a short time-to-result, and can be applicable for microbial AST in clinical applications.

Polyelectrolyte Complex Inclusive Biohybrid Microgels for Tailoring Delivery of Copigmented Anthocyanins.

This study fabricated a novel biohybrid microgel containing polysaccharide-based polyelectrolyte complexes (PECs) for anthocyanins. Herein, anthocyanins were encapsulated into PECs composed of chondroitin sulfate and chitosan, followed by incorporation into alginate microgels using emulsification/internal gelation method. We demonstrated that PECs incorporation strongly affected the properties of microgels, dependent on the polysaccharide concentration and pH in which they were fabricated. The dense internal network surrounded by an alginate shell was clearly visualized in cross-sectioned PECs-microgels. Stability studies carried out under varying ionic strength and pH conditions demonstrated the stimuli-responsiveness of the PECs-microgels. Additionally, the presence of PECs conferred microgels with high rigidity during freeze-drying and excellent reconstitution capacity upon rehydration. These observations were attributed to the modulation of electrostatic and hydrogen-bonding cross-linking between PECs and the alginate gel matrix and suggest the PECs inclusive microgels hold promise as delivery vehicles for the controlled release of hydrophilic bioactive compounds.

Bioactive whey peptide particles: An emerging class of nutraceutical carriers.

Whey-based diets have been linked with prolonged life expectancy and improved physical performance. These observations based on numerous clinical and simulated studies are attributed to diverse biological activities of whey peptides. Recently, bioactive whey peptides were exploited for enveloping nutraceuticals and drugs in view of fabricating capsules that the carrier matrix is also bioactive. Some of the most considered bioactivities of whey peptides including antihypertension, antioxidant, anti-obesity, anti-diabetes, and hypocholesterolemic properties with corresponding underlying mechanisms are briefly discussed. Then, we overview the supramolecular and gelation-prompted encapsulation of nutraceuticals with whey proteins, followed by summarizing recent developments in utilization of synthetic peptides for gene and drug delivery. Finally, particulation of bioactive whey peptides are communicated. Whey peptides may exert both biologically beneficial and technologically appreciated activities. Two procedures including desolvation and internal gelation have been so far employed for bioactive peptides particulation. Crosslinking is a prerequisite to confer acid-induced cold-set gelation to bioactive peptides. It also increases peptides Fe3+-reducing power. Surface activity of a population of peptides in whey protein hydrolysate may result in co-adsorption of the peptides together with small molecule surfactants onto oil-water interface, leading to modulated interfacial architecture and particle morphology.

Microfluidic Fabrication of Colloidal Nanomaterials-Encapsulated Microcapsules for Biomolecular Sensing.

Implantable sensors that detect biomarkers in vivo are critical for early disease diagnostics. Although many colloidal nanomaterials have been developed into optical sensors to detect biomolecules in vitro, their application in vivo as implantable sensors is hindered by potential migration or clearance from the implantation site. One potential solution is incorporating colloidal nanosensors in hydrogel scaffold prior to implantation. However, direct contact between the nanosensors and hydrogel matrix has the potential to disrupt sensor performance. Here, we develop a hollow-microcapsule-based sensing platform that protects colloidal nanosensors from direct contact with hydrogel matrix. Using microfluidics, colloidal nanosensors were encapsulated in polyethylene glycol microcapsules with liquid cores. The microcapsules selectively trap the nanosensors within the core while allowing free diffusion of smaller molecules such as glucose and heparin. Glucose-responsive quantum dots or gold nanorods or heparin-responsive gold nanorods were each encapsulated. Microcapsules loaded with these sensors showed responsive optical signals in the presence of target biomolecules (glucose or heparin). Furthermore, these microcapsules can be immobilized into biocompatible hydrogel as implantable devices for biomolecular sensing. This technique offers new opportunities to extend the utility of colloidal nanosensors from solution-based detection to implantable device-based detection.

Microcapsules for Enhanced Cargo Retention and Diversity.

Prevention of undesired leakage of encapsulated materials prior to triggered release presents a technological challenge for the practical application of microcapsule technologies in agriculture, drug delivery, and cosmetics. A microfluidic approach is reported to fabricate perfluoropolyether (PFPE)-based microcapsules with a high core-shell ratio that show enhanced retention of encapsulated actives. For the PFPE capsules, less than 2% leakage of encapsulated model compounds, including Allura Red and CaCl2 , over a four week trial period is observed. In addition, PFPE capsules allow cargo diversity by the fabrication of capsules with either a water-in-oil emulsion or an organic solvent as core. Capsules with a toluene-based core begin a sustained release of hydrophobic model encapsulants immediately upon immersion in an organic continuous phase. The major contribution on the release kinetics stems from the toluene in the core. Furthermore, degradable silica particles are incorporated to confer porosity and functionality to the otherwise chemically inert PFPE-based polymer shell. These results demonstrate the capability of PFPE capsules with large core-shell ratios to retain diverse sets of cargo for extended periods and make them valuable for controlled release applications that require a low residual footprint of the shell material.