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This work identified a class of cyanomethylquinolones (CQs) and their carboxyl analogues as potential multitargeting antibacterial candidates. Most of the prepared compounds showed high antibacterial activities against most of the tested bacteria, exhibiting lower MIC values (0.125-2 μg/mL) than those of clinical norfloxacin, ciprofloxacin, and clinafloxacin. The low hemolysis, drug resistance, and cytotoxicity, as well as good predictive pharmacokinetics of active CQs and carboxyl analogues revealed their development potential. Furthermore, they could eradicate the established biofilm, facilitating bacterial exposure to these antibacterial candidates. These active compounds could induce bacterial death through multitargeting effects, including intercalating into DNA, up-regulating reactive oxygen species, damaging membranes directly, and impeding metabolism. Moreover, the highly active cyclopropyl CQ 15 exhibited more effective in vivo anti-MRSA potency than ciprofloxacin. These findings highlight the potential of CQs and their carboxyl analogues as multitargeting broad-spectrum antibacterial candidates for treating intractable bacterial infections.

This study developed a class of novel structural antifungal hydrazylnaphthalimidols (HNs) with multitargeting broad-spectrum potential via multicomponent hybridization to confront increasingly severe fungal invasion. Some prepared HNs exhibited considerable antifungal potency; especially nitrofuryl HN 4a (MIC = 0.001 mM) exhibited a potent antifungal activity against Candida albicans , which is 13-fold higher than that of fluconazole. Furthermore, nitrofuryl HN 4a displayed low cytotoxicity, hemolysis and resistance, as well as a rapid fungicidal efficacy. Preliminary mechanistic investigations revealed that nitrofuryl HN 4a could inhibit lactate dehydrogenase to decrease metabolic activity and promote the accumulation of reactive oxygen species, leading to oxidative stress. Moreover, nitrofuryl HN 4a did not exhibit membrane-targeting ability; it could embed into DNA to block DNA replication but could not cleave DNA. These findings implied that HNs are promising as novel structural scaffolds of potential multitargeting broad-spectrum antifungal candidates for treating fungal infection.

Cannabis is the most prevalent abused substance after alcohol, and its consumption severely harms human health and thus adversely impacts society. The identification and quantification of cannabis in urine play important roles in practical forensics. Excitation-emission matrix (EEM) fluorescence spectroscopy coupled with parallel factor (PARAFAC) analysis was developed to identify and quantify the four main ingredients of cannabis in urine samples. The main ingredients of cannabis including Δ-9-tetrahydrocannabinol (THC), cannabidiol, cannabinol, and tetrahydrocannabinolic acid (THC-COOH) exhibited diverse fluorescence characteristics, and the concentrations of these compounds depicted a positive linear relationship with the fluorescence intensity at the ng/mL level. The EEM/PARAFAC method adequately characterized and discriminated the four ingredients in calibration and prediction samples with a low root-mean-square error of prediction (RMSEP; 0.03-0.07 μg/mL) and limit of quantitation (LOQ; 0.26-0.71 μg/mL). The prediction results of the EEM/PARAFAC method well correlated with that of GC-MS with a low RMSEP range (0.01-0.05 μg/mL) and LOQ range (0.07-0.44 μg/mL) in urine samples. The EEM spectroscopic investigation coupled with the PARAFAC algorithm results in an organic, solvent-less, fast, reliable tool to perform accurate and rapid screening of cannabis abusers., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Published by American Chemical Society.)

An unexpected Ugi cascade reaction was developed for the facile construction of γ-lactam-fused pyridone derivatives with high tolerance of substrates. A C(sp 3 )-N bond and a C(sp 2 )-C(sp 2 ) bond were formed together, accompanied by a chromone ring-opening in Ugi adducts, under the basic conditions without any metal catalyst for the whole process. Screening data of several difficult-to-inhibit cancer cell lines demonstrated that 7l displayed a high cytotoxicity against HCT116 cells (IC 50 = 5.59 ± 0.78 μM). Taken together, our findings revealed new insights into the molecular mechanisms underlying compound 7l and provided potential usage of this scaffold for cancer therapeutics.

A unique class of antibacterial azolylpyrimidinediols (APDs) and their analogues were developed. Some synthesized compounds showed excellent bacteriostatic potency; especially, triazolylpyrimidinediol (triazolyl PD) 7a exhibited good anti- Acinetobacter baumannii potential with a low MIC of 0.002 mmol/L. Triazolyl PD 7a with inconspicuous cytotoxicity and hemolytic activity could eradicate the established biofilm, showed low resistance, and exhibited favorable drug-likeness. Mechanistic explorations revealed that compound 7a without membrane-targeting ability could decrease metabolic activity, interact with DNA through groove binding action to block DNA replication rather than intercalate into and cleave DNA, and thus inhibit bacterial growth. Further computations displayed that the low E HOMO and large energy gap might help triazolyl PD 7a binding to biological targets more easily. Moreover, compound 7a gave appreciable in vivo pharmacokinetic properties and pharmacodynamics. These findings of azolylpyrimidinediols as novel structural scaffolds of DNA-groove binders might imply a large promise for the treatments of Acinetobacter baumannii infection.

ATP and reactive oxygen species (ROS) are considered significant indicators of cell apoptosis. However, visualizing the interplay between apoptosis-related ATP and ROS is challenging. Herein, we developed a metal-organic framework (MOF)-based nanoprobe for an apoptosis assay using duplex imaging of cellular ATP and ROS. The nanoprobe was fabricated through controlled encapsulation of gold nanorods with a thin zirconium-based MOF layer, followed by modification of the ROS-responsive molecules 2-mercaptohydroquinone and 6-carboxyfluorescein-labeled ATP aptamer. The nanoprobe enables ATP and ROS visualization via fluorescence and surface-enhanced Raman spectroscopy, respectively, avoiding the mutual interference that often occurs in single-mode methods. Moreover, the dual-modal assay effectively showed dynamic imaging of ATP and ROS in cancer cells treated with various drugs, revealing their apoptosis-related pathways and interactions that differ from those under normal conditions. This study provides a method for studying the relationship between energy metabolism and redox homeostasis in cell apoptosis processes.

New antibacterial 3-(aminothiazolyl)quinolones (ATQs) were designed and efficiently synthesized to counteract the growing multidrug resistance in animal husbandry. Bioactive assays manifested that N , N -dicyclohexylaminocarbonyl ATQ 10e and methyl ATQ 17a , respectively, showed better antibacterial behavior against Staphylococcus aureus ATCC 29213 and Pseudomonas aeruginosa than reference drug norfloxacin. Notably, highly active ATQ 17a with low hemolysis, negligible mammalian cytotoxicity, and good pharmacokinetic properties displayed low trends to induce resistance and synergistic combinations with norfloxacin. Preliminary mechanism exploration implied that representative ATQ 17a could inhibit the formation of biofilms and destroy bacterial membrane integrity, further binding to intracellular DNA and DNA gyrase to hinder bacterial DNA replication. ATQ 17a could also induce the production of excess reactive oxygen species and reduce bacterial metabolism to accelerate bacterial death. These results provided a promise for 3-(aminothiazolyl)quinolones as new potential multitargeting antibacterial agents to treat bacterial infection of animals.

A new hybridization of berberine and benzimidazoles was performed to produce 13-(benzimidazolylmethyl)berberines (BMB) as potentially broad-spectrum antibacterial agents with the hope of confronting multidrug-resistant bacterial infections in the livestock industry. Some of the newly prepared hybrids showed obvious antibacterial effects against tested strains. Particularly, 13-((1-octyl-benzimidazolyl)methyl)berberine 6f (OBMB- 6f ) was found to be the most promising compound that not only exerted a strong activity (MIC = 0.25-2 μg/mL) and low cytotoxicity but also possessed a fast bactericidal capacity and low propensity to develop resistance toward Staphylococcus aureus and Escherichia coli even after 26 serial passages. Moreover, OBMB- 6f displayed the ability to prevent bacterial biofilm formation at low and high temperatures. The mechanistic exploration revealed that OBMB- 6f could significantly disintegrate bacterial membranes, markedly facilitate intracellular ROS generation, and efficiently intercalate into DNA. These results provided a profound insight into BMB against multidrug-resistant bacterial infections in the livestock industry.

An efficient one-pot reaction of propargylamides, isocyanides, and water catalyzed by zinc was developed for the rapid construction of 2-oxazolines with a wide functional group tolerance. The methylene-3-oxazoline was proven to play a vitally important role to start the tandem cascade transformation through unfunctionalized alkynes with sequential nucleophilic addition approaches of isocyanide and water. Notably, with a slight alteration of the reaction temperature and the addition of one molecule of water, various β-amino amide derivatives were synthesized in good to excellent yields.

Novel carbazole-oxadiazoles were developed as new potential antibacterial agents to combat dreadful resistance. Some target compounds displayed predominant inhibitory effects on the tested Gram-positive and -negative bacteria, and carbazole-oxadiazoles 5g , 5i - k , 16a - c , and tetrazole analogues 23b - c were found to be efficient in impeding the growth of MRSA and Pseudomonas aeruginosa ATCC 27853 (MICs = 0.25-4 μg/mL). Furthermore, compounds 5g and 23b - c not only possessed rapid bactericidal ability and low tendency to develop resistance but also exhibited low cytotoxic effects toward Hek 293T, HeLa, and red blood cells (RBCs), especially molecule 5g also showed low toxicity in vivo, which showed the therapeutic potential of these compounds. Further exploration indicated that compounds 5g , 5i , and 23b - c could disintegrate the integrity of bacterial cell membranes to leak the cytoplasmic contents, thus exerting excellent antibacterial effects. These facts mean that carbazole-based antibacterial agents might have bright prospects in confronting bacterial infections.

Natural berberine-derived azolyl ethanols as new structural antibacterial agents were designed and synthesized for fighting with dreadful bacterial resistance. Partial target molecules exhibited potent activity against the tested strains, particularly, nitroimidazole derivative 4d and benzothiazole-2-thoil compound 18b , with low cytotoxicity both exerted strong antibacterial activities against multidrug-resistant Escherichia coli at low concentrations as 0.007 and 0.006 mM, respectively. Meanwhile, the active compounds 4d and 18b possessed the ability to rapidly kill bacteria and observably eradicate the E. coli biofilm by reducing exopolysaccharide content to prevent bacterial adhesion, which was conducive to alleviating the development of E. coli resistance. Preliminary mechanistic explorations suggested that the excellent antibacterial potential of molecules 4d and 18b might be attributed to their ability to disintegrate membrane, accelerate ROS accumulation, reduce bacterial metabolism, and intercalate into DNA groove. These results provided powerful information for the further exploitation of natural berberine derivatives against bacterial pathogens.

Hydrogen peroxide (H 2 O 2 ) widely involves in intracellular and intercellular redox signaling pathways, playing a vital role in regulating various physiological events. Nevertheless, current analytical methods for the H 2 O 2 assay are often hindered by relatively long response time, low sensitivity, or self-interference. Herein, a zeolitic imidazolate framework-8 (ZIF-8)-based surface-enhanced Raman scattering (SERS) sensor has been developed to detect H 2 O 2 released from living cells by depositing ZIF-8 over SERS active gold nanoparticles (AuNPs) grafted with H 2 O 2 -responsive probe molecules, 2-mercaptohydroquinone. Combining the superior fingerprint identification of SERS and the highly efficient enrichment and selective response of H 2 O 2 by ZIF, the ZIF-8-based SERS sensor exhibits a high anti-interference ability for H 2 O 2 detection, with a limit of detection as low as 0.357 nM. Satisfyingly, owing to the enhanced catalytic activity derived from the successful integration of AuNPs and ZIF, the response time as short as 1 min can be obtained, demonstrating the effectiveness of the SERS sensor for rapid H 2 O 2 detection. Furthermore, the developed SERS sensor enables real-time detection of H 2 O 2 secreted from living cells under phorbol myristate acetate stimulation, as cells can be cultured on-chip. This study will pave the way toward the development of a metal-organic framework-based SERS platform for application in the fields of biosensing and early disease diagnosis associated with H 2 O 2 secretion, thus exhibiting promising potential for future therapies.

Natural berberine-hybridized benzimidazoles as potential antibacterial agents were constructed to treat Staphylococcus aureus infection in the livestock industry. Bioassay showed that some new berberine-benzimidazole hybrids exhibited potent antibacterial efficacies, especially, the 2,4-dichlorobenzyl derivative 7d not only showed strong activity against S. aureus ATCC 29213 with the MIC value of 0.006 mM but also effectively eradicated bacterial biofilm and exhibited low toxicity toward mammalian cells. The drug combination experiments showed that compound 7d together with norfloxacin could enhance the antibacterial efficacy. Moreover, the 2,4-dichlorobenzyl derivative 7d did not show obvious propensity to develop bacterial resistance. Preliminary mechanism studies revealed that the active molecule 7d could damage the membrane integrity, stimulate ROS generation, and bind with DNA as well as S. aureus sortase A, thus exerting powerful antibacterial ability. In light of these facts, berberine-benzimidazole hybrid 7d showed a large potentiality as a new bactericide for treating S. aureus in the livestock industry.

Heteroprotein complex coacervate (HPCC) is a liquid-like protein concentrate produced by liquid-liquid phase separation. We revealed the protein dynamic exchange and thermodynamic mechanism of β-conglycinin/lysozyme coacervate, and clarified the effect of HPCC on protein structure and activity. β-conglycinin and lysozyme assembled into coacervate at pH 5.75-6.5 and assembled into amorphous precipitates at higher pH. As the pH dropped from 8 to 6, the number of binding sites of the complex decreased in half, and the desolvation degree corresponding to the entropy gain was greatly reduced, conducing to the formation of coacervates rather than precipitates. The coacervates achieved the unique dynamic exchange by exchanging proteins with the diluted phase, making the uniform distribution of proteins in coacervates. The lysozyme activity was completely retained in β-conglycinin/lysozyme coacervates. These results proved that β-conglycinin-based heteroprotein complex coacervate is a feasible method to encapsulate and enrich active proteins in a purely aqueous environment.

A class of quinazolone thiazoles was identified as new structural scaffolds for potential antibacterial conquerors to tackle dreadful resistance. Some prepared compounds exhibited favorable bacteriostatic efficiencies on tested bacteria, and the most representative 5j featuring the 4-trifluoromethylphenyl group possessed superior performances against Escherichia coli and Pseudomonas aeruginosa to norfloxacin. Further studies revealed that 5j with inappreciable hemolysis could hinder the formation of bacterial biofilms and trigger reactive oxygen species generation, which could take responsibility for emerging low resistance. Subsequent paralleled exploration discovered that 5j not only disintegrated outer and inner membranes to induce leakage of cytoplasmic contents but also broke the metabolism by suppressing dehydrogenase. Meanwhile, derivative 5j could intercalate into DNA to exert powerful antibacterial properties. Moreover, compound 5j gave synergistic effects against some Gram-negative bacteria in combination with norfloxacin. These findings indicated that this novel structural type of quinazolone thiazoles showed therapeutic foreground in struggling with Gram-negative bacterial infections.

Gas aggregation and formation of interfacial nanobubbles (INBs) provide challenges and opportunities in the operation of micro-/nanofluidic devices. In the current study, we used molecular dynamics(MD) simulations to investigate the effects of hydrophobicity and various homogeneous surface conditions on gas aggregation and INB stability with a series of 3D argon-water-solid and water-solid systems. Among various signatures of surface hydrophobicity, the potential of mean force (PMF) minima exhibited the strongest correlation with the water molecular orientation at the liquid-solid interface, compared to the depletion layer width and the droplet contact angle. Our results indicated that argon aggregation on the substrate was a function of hydrophobicity as well as competition between gas-solid and water-solid PMFs. Thus, one precondition for gas aggregation on a surface is that the free energy minima of gas induced by the surface be much lower than that induced by water. We found that although the presence of gas molecules had little effect on the measures of wettability, it enhanced density fluctuations near liquid-solid interfaces. The PMF of gas along the surface tangential plane exhibited a small energy barrier between the epitaxial gas layer (EGL) in the bubble and the gas enrichment layer (GEL) in the liquid, which may benefit nanobubble stability. Much lower PMF in the EGL compared to that in the GEL indicated that gas molecules could migrate from the GEL to the nanobubble basement. However, density fluctuations enhanced by the GEL could reduce the energy barrier, thus reducing the stability of INBs.

Porous materials derived from natural and biodegradable polymers have received growing interest. We demonstrate here an attractive method for the preparation of protein-based porous materials using emulsions stabilized by gliadin-chitosan hybrid particles (GCHPs) as the template, with the addition of gelatin and kosmotropic ions to improve the mechanical strength. The microstructure, mechanical properties, cytotoxicity, and fluid absorption behavior of porous materials were systematically investigated. This strategy facilitated the formation of porous materials with highly open and interconnected pore structure, which can be manipulated by altering the mass ratio of hexane or gelatin in the matrix. The Hofmeister effect resulted from kosmotropic ions greatly enhanced the Young's modulus and the compressive stress at 40% strain of porous materials from 0.56 to 6.84 MPa and 0.26 to 1.11 MPa, respectively. The developed all-natural porous materials were nontoxic to HaCaT cells; they also had excellent liquid (i.e., simulated body fluid and rabbit blood) absorption performance and advantages in resisting stress and maintaining geometry shape. The effects of different concentration amounts and type of salts in the Hofmeister series on the formation and performance of porous materials were also explored. Mechanical strength of porous materials was gradually enhanced when the (NH 4 ) 2 SO 4 concentration increased from 0 to 35 wt %, and the other four kosmotropic salts, including Na 2 S 2 O 3 , Na 2 CO 3 , NaH 2 PO 4 , and Na 2 SO 4 , also showed positive effects. This work opens a simple and feasible way to produce nontoxic and biodegradable porous materials with favorable mechanical strength and controllable pore structure. These materials have broad potential application in many fields involving biomedical and material science, such as cell culture, (bio)catalysis, and wound or bone defect healing.

Ternary chalcogenides, GeSb 2 Se 3 and Ge 3 Sb 4 Se 7 , were synthesized and characterized. These chalcogenides are the first ternary selenides in a ternary Ge-Sb-Se system that feature a layer structure related to black phosphorus and SnSe-type structures. Both compounds contain a ∞ 1 [Sb 2 Se 2 ] 2- unit with Sb + cations in a zigzag Sb-Sb chain structure, and Sb 3+ cations in a distorted NaCl 100 -type of ∞ 1 [Ge n -2 Sb 2 Se n ] 2+ unit ( n = 4, 5). These materials exhibit n-type semiconducting properties with thermal conductivity significantly lower than that of GeSe and Sb 2 Se 3 , which could be correlated to the 1D Sb + chain and disordered sites with different Ge/Sb compositions. It is anticipated that these newly discovered ternary chalcogenides may provide unique properties with enhanced thermoelectric properties.

A palladium-catalyzed alkenylation involving remote δ-position C(alkenyl)-H activation of cycloalkenes reacting with electron-deficient alkenes is described. This method features excellent site selectivity and stereoselectivity to efficiently afford only E -selective highly substituted 1,3-diene derivatives with extra-ligand-free and good functional group tolerance including estrone and free N-H tryptamine under weakly alkaline conditions. Mechanistic studies suggest that picolinamide as a bidentate directing group enables the formation of unique alkenyl palladacycle intermediates.

A transition-metal-free protocol for the difluoroalkylation of imidazopyridines with bromodifluoroaryl ketones promoted by visible light irradiation is presented. This protocol is distinguished by simple, mild, and catalyst-free reaction conditions with a wide reaction scope, which is complementary to existing difluoroalkylation strategies by photoredox scenarios. Additionally, this protocol potentially offers a new way for streamlining the synthesis of compounds containing the difluoro moiety.

The development of high-performance nanocarriers for nutraceuticals or drugs has become one of the topical research subjects in the functional food fields. In this work, we for the first time propose a novel and ecofriendly process to obtain a kind of nanostructured soy β-conglycinin (β-CG; a major soy storage globulin) as outstanding nanocarriers for poorly soluble bioactives (e.g., curcumin), by a urea-assisted disassembly and reassembly strategy. At urea concentrations > 4 M, the structure of β-CG gradually dissociated into its separate subunits (α, α', and β) and even denatured (depending on the type of subunits); after dialysis to remove urea, the dissociated subunits would reassemble into a kind of core-shell nanostructured particles, in which aggregated β-subunits acted as the core while the shell layer was mainly composed of α- and α'-subunits. The core-shell nanoparticles were favorably formed at protein concentrations of 1.0-2.0 wt %. Curcumin crystals were directly introduced into the β-CG solution at high urea concentrations (e.g., 8 M) and would preferentially interact with the denatured β-subunits. As a consequence, almost all of the curcumin molecules were encapsulated in the core part of the reassembled core-shell nanoparticles. The loading amount of curcumin in these nanoparticles could reach 18 g of curcumin per 100 g of protein, which far exceeds those reported previously. The encapsulated curcumin exhibited a high water solubility, extraordinary thermal stability, and improved bioaccessibility, as well as a sustained release behavior. The findings provide a novel strategy to fabricate a kind of high-encapsulation-performance, organic solvent-free, and biocompatible nanocarrier for hydrophobic nutraceuticals and drugs.

Pickering high internal-phase emulsions (HIPEs) and porous materials derived from the Pickering HIPEs have received increased attention in various research fields. Nevertheless, nondegradable inorganic and synthetic stabilizers present toxicity risks, thus greatly limiting their wider applications. In this work, we successfully developed nontoxic porous materials through the Pickering HIPE-templating process without chemical reactions. The obtained porous materials exhibited appreciable absorption capacity to corn oil and reached the state of saturated absorption within 3 min. The Pickering HIPE templates were stabilized by gliadin-chitosan complex particles (GCCPs), in which the volume fraction of the dispersed phase (90%) was the highest of all reported food-grade-particle-stabilized Pickering HIPEs so far, further contributing to the interconnected pore structure and high porosity (>90%) of porous materials. The interfacial particle barrier (Pickering mechanism) and three-dimensional network formed by the GCCPs in the continuous phase play crucial roles in stabilization of HIPEs with viscoelastic and self-supporting attributes and also facilitate the development of porous materials with designed pore structure. These materials, with favorable biocompatibility and biodegradability, possess excellent application prospects in foods, pharmaceuticals, materials, environmental applications, and so on.

Novel sulfonamide-based four-component molecular hybrids as potential DNA-targeting antimicrobial agents were developed from marketed acetanilide through convenient procedures. Biological assays indicated that a few of the target compounds showed significant inhibitory efficiencies toward the tested bacteria and fungi. Noticeably, metronidazole hybrid 6a exhibited a lower minimum inhibitory concentration (MIC) value of 0.019 mM against clinical drug-resistant Escherichia coli ( E. coli), which showed to be 84-fold more active than clinical norfloxacin and had no obvious toxicity toward human breast cancer MCF-7 cells. Synergistic combinations of compound 6a with clinical antibacterial or antifungal drugs could improve the antimicrobial efficiency. Further molecular modeling indicated that the active molecule 6a could bind with THR-199, HIS-64, and GLN-92 residues of human carbonic anhydrase isozyme II through hydrogen bonds and was also able to insert into base-pairs of the DNA hexamer duplex by forming hydrogen bonds. The preliminary exploration of the antibacterial mechanism suggested that compound 6a was capable of disturbing the E. coli membrane effectively and intercalating into clinical resistant E. coli bacterial DNA through noncovalent bonds to form a supramolecular complex, thus exerting its powerful antimicrobial activity. This might suggest a great possibility for hybrid 6a to be a DNA-targeting membrane active potentiator against clinical drug-resistant E. coli.

In this work, three Cu metal-organic framework samples with tunable rhombic, squama, and trucated bipyramid morphologies have been synthesized at 0, 25, and 60 °C, respectively, and further employed as precursors to initially prepare Cu@C composites by the calcination-thermolysis procedure. Then Cu@C composites have been etched with HCl and subsequently activated with KOH to obtain activated porous carbon (APC-0, -25, and -60). Interestingly, APC-25 presents a loose multilevel morphology of cabbage and possesses the largest specific surface area (1880.4 m 2 g -1 ) and pore volume (0.81 cm 3 g -1 ) among these APC materials. Consequently, APC-25 also exhibits the highest specific capacitance of 196 F g -1 at 0.5 A g -1 , and the corresponding symmetric supercapacitor cell (SSC) achieves a remarkable energy density of 11.8 Wh kg -1 at a power density of 350 W kg -1 . Furthermore, APC-25 shows excellent cycling stability, and the loss of capacitance is only 7.7% even after 10000 cycles at 1 A g -1 . Significantly, five light-emitting diodes can be lit by six SSCs, which proves that APC-25 can be used in energy storage devices.

In this work, zein/chitosan nanoparticles (ZCPs-Q) were developed for encapsulating quercetin to overcome its lower water solubility and instability, and to concomitantly enhance its cellular uptake and intracellular antioxidant activity. This strategy enhanced quercetin solubility 753.6 and 9.95 times in water and PBS (7.4), respectively, and quercetin encapsulated in ZCPs remained stable after UV irradiation and heat treatment. ZCPs-Q could significantly attenuate AAPH induced erythrocyte hemolysis through the inhibition of ROS generation. It restored intracellular antioxidant enzyme (SOD and GSH-Px) activities to normal levels and inhibited intracellular malondialdehyde (MDA) formation. Simultaneously, ZCPs-Q showed a strong antioxidant activity in HepG2 cells with an EC 50 value of 31.18 μg/mL, which was lower than free quercetin's 41.02 μg/mL. ZCPs enhanced the uptake efficiency of quercetin in Caco-2 cells, which contributed to the improvement of cellular antioxidant activities (CAA) evaluated with the CAA assay and AAPH-induced erythrocyte hemolysis assay. The designed route is particularly suitable for the encapsulation of water-insoluble nutraceuticals and for enhancing cell uptake and CAA.

Glycation with carbohydrates has been considered to be an effective strategy to improve the emulsifying properties of plant storage globulins, but the knowledge is inconsistent and even contradictory. This work reported that the glycation with soy soluble polysaccharide (SSPS) progressively improved the emulsification efficiency of soy glycinin (SG) in a degree-of-glycation (DG)-dependent manner. The glycation occurred in both the acidic (A) and basic (B) polypeptides to a similar extent. The physicochemical and structural properties of glycated SG samples with different DG values of 0-35% were characterized. The emulsifying properties of unglycated and glycated SG were performed on the emulsions at an oil fraction of 0.3 and a protein concentration in the aqueous phase, produced using microfluidization as the emusification process. The glycation with increasing the DG led to a progressive decrease in solubility and surface hydrophobicity but remarkably increased the magnitude of ζ-potential. Dynamic latter scattering and spectroscopic results showed that the glycation resulted in a gradual dissociation of the 11S-form SG at the quaternary level (into different [AB] subunits), in a DG-dependent way, while their tertiary ([AB] subunits) and secondary structure were slightly affected. Besides the emulsification efficiency, the glycation progressively accelerated the droplet flocculation and facilitated the adsorption of the proteins at the interface and formation of bridged emulsions. The results demonstrated that the improvement of the emulsification efficiency of SG by the glycation with SSPS was largely attributed to the enhanced conformation flexibility at the [AB] subunit level as well as facilitated formation of bridged emulsions. It was also confirmed that once the glycated SG adsorbed at the interface, it would readily dissociated into subunits; the dissociated [AB] subunits exhibited an outstanding Pickering stabilization. The findings would be of importance for providing new knowledge about the molecular mechanism for the modification of emulsifying properties of oligomeric globulins by the glycation with polysaccharides.

Diets containing partially hydrogenated oils (PHOs) expose the human body to trans fatty acids, thus endangering cardiovascular health. Pickering high internal phase emulsions (HIPEs) is a promising alternative of PHOs. This work attempted to construct stable Pickering HIPEs by engineering interface architecture through manipulating the interfacial, self-assembly, and packing behavior of zein particles using the interaction between protein and pectin. Partially wettable zein/pectin hybrid particles (ZPHPs) with three-phase contact angles ranging from 84° to 87° were developed successfully. ZPHPs were irreversibly anchored at the oil-water interface, resulting in robust and ordered interfacial structure, evidenced by the combination of LB-SEM and CLSM. This situation helped to hold a percolating 3D oil droplet network, which facilitated the formation of Pickering HIPEs with viscoelasticity, excellent thixotropy (>91.0%), and storage stability. Curcumin in HIPEs was well protected from UV-induced degradation and endowed HIPEs with ideal oxidant stability. Fabricated Pickering HIPEs possess a charming application prospect in foods and the pharmaceutical industry.

There is still a debate about the effectiveness of native globular proteins to perform as Pickering-like stabilizers for oil-in-water high internal phase emulsions (HIPEs). In the work, we report one native globular protein (ovalbumin) with strong structural integrity and high refolding ability, exhibits an outstanding Pickering stabilization for HIPEs. Ultrastable gel-like HIPEs can be formed through a facile one-pot homogenization even at a concentration as low as 0.2 wt %. The HIPEs formed in the protein-poor regime are a kind of self-supporting and remoldable hydrogel consisting of bridging droplets. The formed HIPEs also exhibit other unique characteristics, such as extraordinary coalescence stability (against prolonged storage or heating), susceptibility to freeze-thawing, enhanced oxidation stability (to encapsulated bioactives), and inhibited vaporization of volatile oils. The findings would be of importance for extending the HIPEs to be applied in food, cosmetic, and petroleum industries.

This work attempted to engineer emulsions' interface using the special affinity between proline-rich gliadin and proanthocyanidins (PA), to develop surfactant-free antioxidant Pickering emulsions with digestive-resistant properties. This binding interaction between gliadin and PA benefited the interfacial adsorption of the particles to corn oil droplets. Pickering droplets as building units assembled into an interconnected three-dimensional network structure, giving the emulsions viscoelasticity and ultrastability. Oxidative markers in Pickering emulsions were periodically monitored under thermally accelerated storage. Lipid digestion and oxidation fates were characterized using in vitro gastrointestinal (GI) models. The interfacial membrane constructed by antioxidant particles served as a valid barrier against lipid oxidation and digestion, in a PA dose-dependent manner. Briefly, lipid oxidation under storage and simulated GI tract was retarded. Free fatty acid (FFA) fraction released decreased by 55% from 87.9% (bulk oil) to 39.5% (Pickering emulsion), implying engineering interfacial architecture potentially benefited to fight obesity. This study opens a facile strategy to tune lipid oxidation and digestion profiles through the cooperation of the Pickering principle and the interfacial delivery of antioxidants.

A series of isopropanol-bridged carbazole azoles as potential antimicrobial agents were designed and synthesized from commercial carbazoles. Bioassay revealed that 3,6-dichlorocarbazolyl triazole 3f could effectively inhibit the growth of E. faecalis with minimal inhibitory concentration of 2 μg/mL. The active molecule 3f showed lower propensity to trigger the development of resistance in bacteria than norfloxacin and exerted rapidly bactericidal ability. Compound 3f also exhibited low cytotoxicity to normal mammalian RAW264.7 cells. Further mechanism exploration indicated that conjugate 3f was membrane active against E. faecalis and could form 3f -DNA complex by intercalating into DNA of resistant E. faecalis , which might be responsible for its antimicrobial action. Molecular docking showed an efficient binding of triazole derivative 3f with DNA gyrase enzyme through noncovalent interactions., Competing Interests: The authors declare no competing financial interest.

A series of novel naphthalimide aminothiazoles were developed for the first time and evaluated for their antimicrobial activity. Some prepared compounds possessed good inhibitory activity against the tested bacteria and fungi. Noticeably, the piperazine derivative 4d displayed superior antibacterial activity against MRSA and Escherichia coli with MIC values of 4 and 8 μg/mL, respectively, to reference drugs. The most active compound 4d showed low toxicity against mammalian cells with no obvious triggering of the development of bacterial resistance, and it also possessed rapid bactericidal efficacy and efficient membrane permeability. Preliminarily investigations revealed that compound 4d could not only bind with gyrase-DNA complex through hydrogen bonds but could effectively intercalate into MRSA DNA to form 4d -DNA supramolecular complex, which might be responsible for the powerful bioactivity. Further transportation behavior evaluation indicated that molecule 4d could be effectively stored and carried by human serum albumin, and the hydrophobic interactions and hydrogen bonds played important roles in the binding process., Competing Interests: The authors declare no competing financial interest.

The complexation between soy soluble polysaccharide (SSPS) and curcumin at pH 7.0 and 4.0, as well as some physicochemical characteristics of the resultant complexes, was investigated. The encapsulation efficiency and loading amount of curcumin in the complexes at pH 4.0 reached 67.3% and 4.49 μg/mg SSPS, respectively. Ethanol-induced denaturation and structural unfolding of the protein fraction in SSPS was essential for complex formation. The complexation with curcumin resulted in aggregation of SSPS and the subsequent formation of compacted nanoparticles with curcumin as the core. The complexation greatly improved the heat stability and in vitro bioaccessibility of curcumin. In general, the encapsulation efficiency, heat stability, and bioaccessibility of curcumin in the complexes at pH 4.0 were better than those at pH 7.0. The findings are of importance for the development of food grade nanovehicles for enhanced water solubility, stability, and bioaccessibility of hydrophobic bioactives.

Inter/intramolecular approaches to sp 2 C-N bond formation of N-alkenyl benzimidazoles have been accomplished in the presence of an iodide anion associated with a copper catalyst. Both intermolecular and intramolecular reactions included tandem processes, in which selective iodination of sp 3 C-H bond at the α-position of ester under mild conditions was demonstrated for the first time. Tandem reactions involving sp 3 C-H activation via α-iodo ester intermediate under copper catalysis efficiently provided more than 20 novel azole compounds, and free radicals were not involved in this transformation.

Using soy protein isolate (SPI) and soy-soluble polysaccharides (SSPS) as polymer matrixes, this study reported a novel process to fabricate unique core-shell complex (nano)particles to perform as carriers for curcumin (a typical poorly soluble bioactive). In the process, curcumin-SPI nanocomplexes were first formed at pH 7.0 and then coated by SSPS. At this pH, the core-shell complex was formed in a way the SPI nanoparticles might be incorporated into the interior of SSPS molecules without distinctly affecting the size and morphology of particles. The core-shell structure was distinctly changed by adjusting pH from 7.0 to 4.0. At pH 4.0, SSPS was strongly bound to the surface of highly aggregated SPI nanoparticles, and as a consequence, much larger complexes were formed. The bioaccessibility of curcumin in the SPI-curcumin complexes was unaffected by the SSPS coating. However, the core-shell complex formation greatly improved the thermal stability and controlled release properties of encapsulated curcumin. The improvement was much better at pH 4.0 than that at pH 7.0. All of the freeze-dried core-shell complex preparations exhibited good redispersion behavior. The findings provide a simple approach to fabricate food-grade delivery systems for improved water dispersion, heat stability, and even controlled release of poorly soluble bioactives.

A series of novel 3-aminothiazolquinolones as analogues of quinolone antibacterial agents were designed and synthesized in an effort to circumvent quinolone resistance. Among these 3-aminothiazolquinolones, 3-(2-aminothiazol-4-yl)-7-chloro-6-(pyrrolidin-1-yl) quinolone 12b exhibited potent antibacterial activity, low cytotoxicity to hepatocyte cells, strong inhibitory potency to DNA gyrase, and a broad antimicrobial spectrum including against multidrug-resistant strains. This active molecule 12b also induced bacterial resistance more slowly than norfloxacin. Analysis of structure-activity relationships (SARs) disclosed that the 2-aminothiazole fragment at the 3-position of quinolone plays an important role in exerting antibacterial activity. Molecular modeling and experimental investigation of aminothiazolquinolone 12b with DNA from a sensitive methicillin-resistant Staphylococcus aureus (MRSA) strain revealed that the possible antibacterial mechanism might be related to the formation of a compound 12b-Cu(2+)-DNA ternary complex in which the Cu(2+) ion acts as a bridge between the backbone of 3-aminothiazolquinolone and the phosphate group of the nucleic acid.

The complexation of nanoparticles in unheated and heated (at 75-95°) soy protein isolate (SPI) with curcumin and the effects on curcumin stability/bioaccessibility and in vitro protein digestibility were investigated. The nanoparticles did not display noticeable changes in size and morphology upon nanocomplexation with curcumin, except their surface hydrophobicity. The encapsulation efficiency of curcumin progressively decreased with increasing initial curcumin concentration in the dispersion, while the load amount linearly increased. The solubility of curcumin in water was enhanced by the complexation above 98000-fold (vs free curcumin in water). The formation of the nanocomplexes considerably improved the storage stability of curcumin. In vitro simulated digestion experiments indicated that the complexation also improved the bioaccessibility of curcumin; the bioaccessibility was greatly impaired by hydrolysis-induced protein aggregation. Addtionally, the nanocomplexation significantly improved the in vitro protein digestibility of both unheated and heated SPI.

Water-insoluble phytosterols were developed into a kind of colloidal particle as Pickering stabilizers for emulsions by a classic anti-solvent method using 100% ethanol as the organic phase to solubilize the phytosterols and whey protein concentrate (WPC) as the emulsifier. The colloidal particles in the dispersion, with morphology of stacked platelet-like sheets, had a mean diameter of 44.7 and 24.7 μm for the volume- and surface-averaged sizes, respectively. The properties and stability of the emulsions stabilized by these colloidal particles were highly dependent upon the applied total solid concentration (c; in the dispersion) and oil fraction (ø). The results indicated that (1) at a low c value (<1.0%, w/v) the emulsions were susceptible to phase separation, even at a low ø of 0.2, (2) at low ø values (e.g., 0.2 or 0.3) and a relatively high c value (1.0%, w/v, or above), a severe droplet flocculation occurred for the emulsions, and (3) when both c and ø were appropriately high, a kind of self-supporting gel-like emulsions could be formed. More interestingly, a phase inversion of the emulsions from the oil-in-water to water-in-oil type was observed, upon the ø increasing from 0.2 to 0.6 (especially at high c values, e.g., 3.0%, w/v). The elaborated Pickering emulsions stabilized by the phytosterol colloidal particles with a gel-like behavior would provide a candidate to act as a novel delivery system for active ingredients.

The interfacial and emulsifying properties of soy protein isolate nanoparticles formed by combined treatments of heating and electrostatic screening, as affected by variation of the protein concentration (c) and emulsification process, were investigated. These nanoparticles (with a z-average diameter of 103 nm at c = 0.1%, w/v) tended to aggregate at higher c values, and their internal structure was mainly maintained by hydrophobic interactions and disulfide bondings. In general, increasing c progressively favored diffusion and/or adsorption at the interface and formation of finer emulsions; increasing the energy input level of emulsification improved the emulsification efficiency and extent of droplet flocculation, as well as the emulsion coalescence and creaming stability. The rheological and creaming behavior of these emulsions was predominately determined by the amount of proteins adsorbed at the interface. The results confirmed that these nanoparticles can formulate Pickering emulsions with properties tailored by selecting c and the emulsification process.

An efficient FeCl3 catalyzed protocol for the synthesis of amino functionalized indolylnitroalkanes from easily available precursor indolylnitroalkenes and substituted amines has been developed. Regioselective C-alkylation in the presence of free amino substituted arenes occurred. The scope of this methodology shows good functional group tolerance, and further, this protocol was used to prepare indolylquinoline derivatives.

This work attempted to develop novel high barrier zein/SC nanoparticle (ZP)-stabilized emulsion films through microfluidic emulsification (ZPE films) or in combination with solvent (ethyl acetate) evaporation techniques (ZPE-EA films). Some physical properties, including tensile and optical properties, water vapor permeability (WVP), and surface hydrophobicity, as well as the microstructure of ZP-stabilized emulsion films were evaluated and compared with SC emulsion (SCE) films. The emulsion/solvent evaporation approach reduced lipid droplets of ZP-stabilized emulsions, and lipid droplets of ZP-stabilized emulsions were similar to or slightly lower than that of SC emulsions. However, ZP- and SC-stabilized emulsion films exhibited a completely different microstructure, nanoscalar lipid droplets were homogeneously distributed in the ZPE film matrix and interpenetrating protein-oil complex networks occurred within ZPE-EA films, whereas SCE films presented a heterogeneous microstructure. The different stabilization mechanisms against creaming or coalescence during film formation accounted for the preceding discrepancy of the microstructures between ZP-and SC-stabilized emulsion films. Interestingly, ZP-stabilized emulsion films exhibited a better water barrier efficiency, and the WVP values were only 40-50% of SCE films. A schematic representation for the formation of ZP-stabilized emulsion films was proposed to relate the physical performance of the films with their microstructure and to elucidate the possible forming mechanism of the films.

Although the functionality of plant proteins (and soy proteins in particular) has been widely investigated in the last decades, the importance of conformational characteristics to their functionalities is still far away from being understood. The aim of the present work was to unravel the role of conformational flexibility at the quaternary and/or tertiary levels in the emulsifying and interfacial properties of phaseolin, an ideal vicilin (or 7S globulin) from red kidney bean. The conformational flexibility at quaternary and tertiary levels of phaseolin was modulated by urea with increasing concentrations from 0 to 8 M, as characterized by using dynamic light scattering (DLS), intrinsic fluorescence and derivative UV spectroscopy, and differential scanning calorimetry (DSC). The emulsifying and interfacial properties, including emulsifying ability, flocculated state of oil droplets (in fresh emulsions), emulsion stability against creaming, and adsorption dynamics at the oil-water interface, were characterized at a specific protein concentration of 0.5% (w/v). The results indicated that increasing the urea concentration resulted in a progressive dissociation of trimeric phaseolin molecules into monomeric subunits, and even a structural unfolding of dissociated subunits; the urea-induced conformational changes at quaternary and/or tertiary levels were reversible, and the molecules at high urea concentrations shared similar structural features to the "molten globule state". On the other hand, increasing the urea concentration progressively improved the emulsifying ability of the protein, and flocculated extent of oil droplets in the fresh emulsions, but led to a progressive decrease in interfacial protein concentration. The improvement of the emulsifying ability was not related to diffusion (during initial adsorption) and penetration at the interface, but highly dependent on ease of structural rearrangement of the adsorbed proteins. These observations clearly confirmed that the flexibility of phaseolin at quaternary and/or tertiary levels plays a vital role in its emulsifying ability, mainly through the way of affecting the ease of structural rearrangement of adsorbed proteins at the interface. The findings could provide an in-depth understanding of the importance of conformational flexibility for the emulsifying properties of oligomeric storage globulins, and thus are of great help to guide the modifications of the proteins for better emulsifying properties.

In recent years, there have been increasing interests in developing food-grade Pickering stabilizers, due to their potential applications in formulations of novel functional foods. The present work was to investigate the potential of soy proteins to be developed into a kind of Pickering-like stabilizer for oil-in-water emulsions. The nanoparticle aggregates of soy protein isolate (SPI) were formed by sequential treatments of heating at 95 °C for 15 min and then electrostatic screening with NaCl addition. The particle size and microstructure of these aggregates were characterized using dynamic light scattering and atomic force microscopy, indicating that the fabricated nanoparticle aggregates were ∼100 nm in size with more surface hydrophobic nature (relative to unheated SPI). The influence of particle concentration (c; 0.5-6.0%, w/w) and increasing oil fraction (ϕ; in the range 0.2-0.6) on the droplet size and coalescence and/or creaming stability of the emulsions stabilized by these nanoparticle aggregates was investigated. The results showed that, at ϕ = 0.2, increasing the c resulted in a progressive but slight decrease in droplet size, and improved the stability against coalescence and creaming; at a specific c, the creaming stability was progressively increased by increasing the ϕ, with better improvement observed at a higher c (e.g., 6.0% vs 2.0%). The improvement of creaming stability was largely associated with the formation of a gel-like network that could entrap the oil droplets within the network. The observations are generally consistent with those observed for the conventional Pickering emulsions, confirming that soy proteins could be applied as a kind of effective Pickering-like stabilizer. The finding may have important implications for the design and fabrication of protein-based emulsion formulations, and even for the development of soy protein products with some unique functions. To the authors' knowledge, this is the first work to report that heat-induced soy protein aggregates exhibit a good potential to act as Pickering-type stabilizers.

A protocol for the N-bromosuccinimide (NBS)- and trichloroisocyanuric acid (TCCA)-mediated synthesis of novel 2-halomethylene-3-oxoketoximes via one-pot halogenation/oxidation of isoxazoline N-oxide derivatives is described here. The keto functionality of 3-ketoximes was selectively reduced by lithiumaluminum hydride to synthesize an unprecedented type of Baylis-Hillman oxime, which underwent N-O coupling to produce new isoxazoline N-oxide derivative.

Although it is well recognized that the conformation state of a protein affects its surface properties, the importance of conformation flexibility to its functionality is still not well understood. This study systemically investigated the influence of protein concentration (c) and disulfide bond (S-S) cleavage with a reducing agent, β-mercaptoethanol (2-ME), on the conformation and emulsifying properties of an ideal globular protein, bovine serum albumin (BSA), with the aim to unravel the role of conformational flexibility in the functionality. The conformations were evaluated using size exclusion chromatography, dynamic light scattering (DLS), extrinsic fluorescence, and derivative UV spectroscopy. The emulsifying properties, including emulsifying ability, extent of droplet flocculation at a specific period of storage, and stability against flocculation and/or coalescence as well as creaming, were characterized using droplet size and creaming index analyses. The results indicated that the tertiary conformation of native BSA was closely dependent on its c (in the range of 0.05-1.0%), and increasing c resulted in a more compacted and rigid conformation. The c dependence largely determined the susceptibility of S-S bridges to reduction and even refolding of reduced BSA molecules. Interestingly, there was approximately a critical c (e.g., 0.25-0.5%) below which the S-S cleavage resulted in a gradual structural unfolding of the molecules and above which the situation was the reverse. On the other hand, the alteration with protein and 2-ME concentrations led to a variety of changes in emulsion size (d4,3; in water or 1% SDS) at 4 and 24 h and creaming index (up to 2 weeks). In general, at a low c value (e.g., 0.25%) increasing the S-S cleavage progressively improved the emulsifying ability and emulsion stability (especially against coalescence and creaming), whereas at c = 0.5 or 0.75%, the S-S cleavage, on the contrary, impaired the emulsifying properties, especially emulsion stability against flocculation and/or coalescence. These results suggest that the conformational flexibility of the protein (in solution) plays a vital role in different aspects of its emulsifying properties, for example, ease of structural unfolding at the interface, lateral interactions between adsorbed proteins, and formation of a viscoelastic interfacial layer (or multilayers). This knowledge could provide an in-depth understanding of the relationships between tertiary conformational flexibility and emulsifying properties of globular proteins.

The objective of this research was to fabricate novel antimicrobial films based on zein colloidal nanoparticles coated with sodium caseinate (SC), an emulsifier/stabilizer. Thymol-loaded zein-SC nanoparticles were prepared using an antisolvent technique, with the average particle size and zeta potential about 200 ± 20 nm and -40 mV, respectively. Zein-SC nanoparticle-based films exhibited higher mechanical resistance and water barrier capacity than the SC films and concomitant good extensibility as compared with zein films. Thymol loadings endowed zein-SC nanoparticle-based films with antimicrobial activity against Escherichia coli and Salmonella as well as DPPH radical scavenging activity. Water vapor permeability, microstructure, mechanical, and controlled release properties of the films were evaluated. The possible relationship between some selected physical properties and microstructure were also discussed. Atomic force microscopy (AFM) analysis indicated that thymol loadings resulted in the emergence phenomena of the nanoparticles to form large particles or packed structure, consisting of clusters of nanoparticles, within the film matrix, in a thymol loading dependent manner. The appearance of large particles or an agglomerate of particles may weaken the compactness of protein network of films and thus impair the water barrier capacity, mechanical resistance, and extensibility of the films. The release kinetics of thymol from nanoparticle-based films can be described as a two-step biphasic process, that is, an initial burst effect followed by subsequent slower release, and zein-SC nanoparticles within the films matrices gave them the ability to sustain the release of thymol. In addition, a schematic illustration of the formation pathway of zein-SC nanoparticle-based films with or without thymol was proposed to illuminate the possible relationship between some selected physical properties and the microstructure of the films.

The structure-function relationships of plant oligomeric globulins are still not fully recognized. The present work investigated the influence of glycation with glucose (at 1:50 and 1:100 protein/sugar molar ratios; incubation periods of 2.5, 5.0, and 10.0 h) on the physicochemical and conformational properties of kidney bean vicilin (phaseolin), with the aim of understanding the structure-function relationships of legume vicilins. Protein solubility (PS), surface charge (isoelectric point) and hydrophobicity (H0), and secondary, tertiary, and/or quaternary conformations, as well as the emulsifying activities (emulsifying activity and emulsion stability indices, EAI and ESI) were evaluated. The 2.5 h incubation period of glycation led to least PS and highest H0, and after that, the PS and H0, on the contrary, gradually changed with increasing incubation period. The glycation increased the α-helix content and highly ordered secondary structures (α-helix+β-strand), as evidenced by far-UV circular dichroism (CD) spectroscopy. Combined analyses of differential scanning calorimetry, intrinsic emission fluorescence, and near-UV CD spectroscopy indicated that phaseolin underwent a tertiary conformation unfolding and subsequent rearrangement process (to form a new tertiary conformation), whereas the quaternary conformational flexibility progressively increased upon increasing degree of glycation. The conformation rearrangement was more distinct at the 1:100 molar ratio than at the 1:50 counterpart. The glycation at 5.0 and 10.0 h periods considerably increased the EAI, but only at the 1:50 molar ratio was the ESI progressively increased with the incubation period. These results confirmed that besides surface properties (e.g., PS and H0), the flexibility in tertiary and/or quaternary conformations played a major role in the emulsifying properties of glycated vicilins. The findings would have important implications for understanding the structure-function relationships of legume oligomeric globulins, thus providing a direction to further improve the surface-related functional properties of these proteins.

The effects of high hydrostatic pressure (HHP) treatment (100-500 MPa) on solubility and structural properties of ethanol (EtOH)-denatured soy β-conglycinin and glycinin were investigated using differential scanning calorimetry, Fourier transform infrared and ultraviolet spectroscopy. HHP treatment above 200 MPa, especially at neutral and alkaline pH as well as low ionic strength, significantly improved the solubility of denatured soy proteins. Structural rearrangements of denatured β-conglycinin subjected to high pressure were confirmed, as evidenced by the increase in enthalpy value (ΔH) and the formation of the ordered supramolecular structure with stronger intramolecular hydrogen bond. HHP treatment (200-400 MPa) caused an increase in surface hydrophobicity (F(max)) of β-conglycinin, partially attributable to the exposure of the Tyr and Phe residues, whereas higher pressure (500 MPa) induced the decrease in F(max) due to hydrophobic rearrangements. The Trp residues in β-conglycinin gradually transferred into a hydrophobic environment, which might further support the finding of structural rearrangements. In contrast, increasing pressure induced the progressive unfolding of denatured glycinin, accompanied by the movement of the Tyr and Phe residues to the molecular surface of protein. These results suggested that EtOH-denatured β-conglycinin and glycinin were involved in different pathways of structural changes during HHP treatment.

The mechanical properties, water-holding capacities (WHC), and microstructures of emulsion gels, induced by glucono-δ-lactone (GDL), CaCl(2), and microbial transglutaminase (MTGase) from unheated and heated soy protein isolate (SPI)-stabilized emulsions (at protein concentration 5%, w/v; oil volume fraction, 20%, w/v), were investigated and compared. The influence of thermal pretreatments (at 90 °C for 5 min) before and/or after emulsification was evaluated. Considerable differences in mechanical, water-holding, and microstructural properties were observed among various emulsion gels. The thermal pretreatment after emulsification increased the strength of the emulsion gels induced by GDL and CaCl(2), whereas in the case of MTGase, thermal pretreatments before and/or after emulsification on the contrary greatly inhibited gel network formation. The application of the enzyme coagulant exhibited much higher potential to form SPI-stabilized emulsion gels with higher mechanical strength than that of the other two coagulants. The WHC of the emulsion gels seemed to be not directly related to their gel network strength. Confocal laser scanning microscope analyses indicated that the network microstructure of the formed emulsion gels, mainly composed of aggregated protein-stabilized oil droplets and protein aggregate clumps, varied with the type of applied coagulants and emulsions. The differences in microstructure were basically consistent with the differences in mechanical properties of the gels. These results could provide valuable information for the formation of cold-set soy protein-stabilized emulsion gels.

Fluorescence and differential scanning calorimetry (DSC) were used to study changes in the conformation of red kidney bean (Phaseolus vulgaris L.) protein isolate (KPI) under various environmental conditions. The possible relationship between fluorescence data and DSC characteristics was also discussed. Tryptophan fluorescence and fluorescence quenching analyses indicated that the tryptophan residues in KPI, exhibiting multiple fluorophores with different accessibilities to acrylamide, are largely buried in the hydrophobic core of the protein matrix, with positively charged side chains close to at least some of the tryptophan residues. GdnHCl was more effective than urea and SDS in denaturing KPI. SDS and urea caused variable red shifts, 2-5 nm, in the emission λ(max), suggesting the conformational compactness of KPI. The result was further supported by DSC characteristics that a discernible endothermic peak was still detected up to 8 M urea or 30 mM SDS, also evidenced by the absence of any shift in emission maximum (λ(max)) at different pH conditions. Marked decreases in T(d) and enthalpy (ΔH) were observed at extreme alkaline and/or acidic pH, whereas the presence of NaCl resulted in higher T(d) and ΔH, along with greater cooperativity of the transition. Decreases in T(d) and ΔH were observed in the presence of protein perturbants, for example, SDS and urea, indicating partial denaturation and decrease in thermal stability. Dithiothreitol and N-ethylmaleimide have a slight effect on the thermal properties of KPI. Interestingly, a close linear relationship between the T(d) (or ΔH) and the λ(max) was observed for KPI in the presence of 0-6 M urea.

The fibrillar aggregation at pH 2.0 of soy β-conglycinin, glycinin, and the 1:1 mixture thereof, induced by heating at 80 °C for various periods of time, was investigated using Th T and Congo Red spectroscopic techniques. The morphology of the formed fibrillar aggregates was characterized using atomic force microscopy (AFM), whereas the conformational changes and the polypeptide hydrolysis of the proteins upon heating were also evaluated. Th T fluorescence analysis indicated that β-conglycinin had a much higher potential to form heat-induced amyloid-like aggregates than glycinin. AFM analyses showed that all of the soy globulins could form twisted screw-structure fibrils with heights of 1.4-2.2 nm, but the morphology of the amyloid-like fibrils considerably varied among various soy proteins. Significantly lower width at half-heights and higher coil periodicity values were observed for the β-conglycinin fibrils than the glycinin counterpart. Far-UV CD spectral analysis indicated that upon heating, the secondary conformations of the proteins changed considerably, especially during initial heating (e.g., <4 h), and the changes were much more distinct in the β-conglycinin case than in the glycinin case. Furthermore, reducing electrophoresis analyses indicated that progressive polypeptide hydrolysis occurred upon heating, but the polypeptide hydrolysis for the β-conglycinin was much more severe than that of glycinin. The data suggest that soy β-conglycinin exhibited a much higher potential to form thermally fibrillar aggregates than glycinin, and the differences seem to be mainly associated with the differences in their conformational changes and extent of polypeptide hydrolysis by the heating. The results would be of vital importance for the utilization of soy proteins to produce thermally induced fibrillar gels with excellent properties.