Effects of ultrasound and thermal treatment on the interaction between hyaluronic acid and lactoferrin: Preparation, structures and functionalities.

Complexes of polysaccharides and proteins have superior physicochemical and functional properties compared to single proteins or polysaccharides. In this study, lactoferrin-hyaluronic acid (LF-HA) complexes were prepared by both ultrasonic and thermal treatment. Appropriate preparation conditions, including ultrasonic and thermal treatment conditions, have been established. The complexes formed by different methods were structurally characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis, fourier transform infrared spectroscopy, and circular dichroism spectroscopy. Ultrasound formed non-covalent binding, while thermal treatment generated covalent bonding, altering the structure of LF. The LF-HA complexes showed improved heat stability, foaming stability, emulsifying activity and antioxidant capacity, but deceased foaming ability. Iron binding ability could only be improved by HA through thermal treatment. Moreover, the in vitro digestibility of LF-HA complexes decreased to below 80 % compared to LF.

Novel Human Milk Fat Substitutes Based on Medium- and Long-Chain Triacylglycerol Regulate Thermogenesis, Lipid Metabolism, and Gut Microbiota Diversity in C57BL/6J Mice.

Human milk is naturally rich in medium- and long-chain triacylglycerols (MLCT), accounting for approximately 30% of the total fat. However, infant formula fat is prepared using a physical blend of vegetable oils, which rarely contains MLCT, similar to human milk. The differences in MLCT between human milk and infant formulas may cause different lipid metabolisms and physiological effects on infants, which are unknown. This study aimed to analyze the metabolic characteristics of formula lipid containing novel human milk fat substitutes based on MLCT (FL-MLCT) and compare their effects with those of the physical blend of vegetable oils (FL-PB) on lipid metabolism and gut microbiota in mice. Compared with the FL-PB group, the FL-MLCT group showed increased energy expenditure, decreased serum triacylglycerol level, and significantly lower aspartate aminotransferase level, epididymal and perirenal fat weight, and adipocyte size. Moreover, the abundances of Firmicutes/Bacteroidota, Actinobacteriota, and Desulfovibrionaceae were significantly decreased in the FL-MLCT group. Novel human milk fat substitutes MLCT could inhibit visceral fat accumulation, improve liver function, and modulate the mice gut microbiota composition, which may contribute to controlling obesity.

Characterization of ultrasound-assisted covalent binding interaction between ß-lactoglobulin and dicaffeoylquinic acid: Great potential for the curcumin delivery.

The low bioavailability and poor gastrointestinal instability of curcumin hampers its application in pharmaceutical and food industries. Thus, it is essential to explore efficient carrier (e.g. a combination of polyphenols and proteins) for food systems. In this study, covalent ß-lactoglobulin (LG)-dicaffeoylquinic acids (DCQAs) complexes were prepared by combining ultrasound and free radical induction methods. Covalent interactions between LG and DCQAs were confirmed by analyzing reactive groups. Variations in secondary or tertiary structure and potential binding sites of covalent complexes were explored using Fourier transform infrared spectroscopy and circular dichroism. Results showed that the ß-sheet content decreased and the unordered content increased significantly (P < 0.05). The embedding rate of curcumin in prepared LG-DCQAs complexes using ultrasound could reach 49 % - 62 %, proving that complexes could embed curcumin effectively. This study highlights the benefit of ultrasound application in fabrication of protein-polyphenol complexes for delivering curcumin.

Analysis of non-covalent interaction between ß-lactoglobulin and hyaluronic acid under ultrasound-assisted treatment: Conformational structures and interfacial properties.

Hyaluronic acid (HA) used as a food ingredient is gaining acceptance and popularity. However, the studies available for the effect of HA concentrations on the properties of ß-lactoglobulin (ß-LG) were limited. In this study, we investigated that the molecular characterization and functional properties of the complex formed by the non-covalent binding of ß-LG and HA, as well as the ultrasound-assisted treatment at acidic pH. The optimal pH and ratio of ß-LG/HA were set as 7 and 4:1, respectively. The fluorescence spectroscopy, circular dichroism spectroscopy, and molecular docking results revealed that the addition of HA and ultrasound induced a decrease in random coil and α-helix and an increase in ß-sheet contents in ß-LG. By the complexation with HA, the thermal stability, freezing stability, and antioxidant properties of ß-LG were all improved under ultrasound treatment. The results of the present study can be useful for the modulation of HA based biopolymer complexes and the exploitation as encapsulating or structuring agents in food industry.

New Insights into Phospholipid Profile Alteration of Bigeye Tuna (Thunnus obesus) during Daily Cooking Processes Using Rapid Evaporative Ionization Mass Spectrometry.

Bigeye tuna (BET, Thunnus obesus) is one of the most nutritious and luxurious cosmopolitan fish. The cooked BET products are capturing the interests of consumers by enhancing flavor and ensuring microbiological safety; however, the lipidomic fingerprints during daily cooking processes have not been investigated. In this work, lipid phenotypic data variation in BET during air-frying, roasting, and boiling was studied comprehensively using iKnife rapid evaporative ionization mass spectrometry (REIMS). The outstanding lipid ions mainly including fatty acids (FAs) and phospholipids (PLs) were identified structurally. It was demonstrated that the rates of heat transfer and lipid oxidation in air-fried BET were slower than those in roasted and boiled BET by elucidating the lipid oxidation and PL hydrolysis mechanism. Furthermore, multivariate REIMS data analysis (e.g., discriminant analysis, support vector machine, neutral network, and machine learning models) was used to characterize the lipid profile change in different cooked BET samples, among which FAC22:6, PL18:3/22:6, PL18:1/22:6, and others were the salient contributing features for determining the cooked BET samples. These results may provide a potential strategy for a healthy diet by controlling and improving functional food quality in daily cooking.

The phenolics, antioxidant activity and in vitro digestion of pomegranate (Punica granatum L.) peels: an investigation of steam explosion pre-treatment.

Pomegranate peels, the main byproduct of pomegranate production, are rich in phenolic compounds that are known for their effective antioxidant properties and have vast application prospects. In this study, steam explosion, an environmentally friendly technique, was applied to pretreat pomegranate peels for phenol extraction. We investigated the effects of explosion pressure, duration, and particle size on the content of total and individual phenolics, and antioxidant activity of pomegranate peels before and after in vitro digestion. The optimal conditions for a steam explosion for pomegranate peels in terms of total phenol content were a pressure of 1.5 MPa, a maintenance time of 90 s, and a particle size of 40 mesh. Under these conditions, pomegranate peel extract presented a higher yield of total phenols, gallic acid, and ellagic acid. However, it also had a lower content of punicalin and punicalagin, compared to the unexploded peels. There was no improvement in the antioxidant activity of pomegranate peels after the steam explosion. Moreover, the content of total phenol, gallic acid, ellagic acid, punicalin, and punicalagin, as well as the antioxidant activity of pomegranate peels, all increased after gastric digestion. Nevertheless, there was a large variation in the pomegranate peel processed by different pressure, duration, and sieve fractions. Overall, this study demonstrated that steam explosion pre-treatment could be an efficient method for improving the release of phenolics, especially gallic acid, and ellagic acid, from pomegranate peels.

Degradation, isomerization and stabilization of three dicaffeoylquinic acids under ultrasonic treatment at different pH.

Dicaffeoylquinic acids (diCQAs) are found in a variety of edible and medicinal plants with various biological activities. An important issue is the low stability of diCQAs during extraction and food processing, resulting in the degradation and transformation. This work used 3,5-diCQA as a representative to study the influence of different parameters in ultrasonic treatment on the stability of diCQAs, including solvent, temperature, treatment time, ultrasonic power, duty cycle, and probe immersion depth. The generation of free radicals and its influence were investigated during the treatment. The stability of three diCQAs (3,5-diCQA, 4,5-diCQA and 3,4-diCQA) under the certain ultrasonic condition at different pH conditions was evaluated and found to decrease with the increase of pH, further weakened by ultrasonic treatment. Ultrasound was found to accelerate the degradation and isomerization of diCQAs. Different diCQAs showed different pattern of degradation and isomerization. The stability of diCQAs could be improved by adding epigallocatechin gallate and vitamin C.

Lipidomic fingerprinting of plasmalogen-loaded zein nanoparticles during in vitro multiple-stage digestion using rapid evaporative ionization mass spectrometry.

Plasmalogens (Pls) as the hydrophobic bioactive compound have shown potential in enhancing neurological disorders. However, the bioavailability of Pls is limited because of their poor water solubility during digestion. Herein, the hollow dextran sulfate/chitosan - coated zein nanoparticles (NPs) loaded with Pls was prepared. Subsequently, a novel in situ monitoring method utilizing rapid evaporative ionization mass spectrometry (REIMS) coupled with electric soldering iron ionization (ESII) was proposed to assess the lipidomic fingerprint alteration of Pls-loaded zein NPs during in vitro multiple-stage digestion in real time. A total of 22 Pls in NPs were structurally characterized and quantitatively analyzed, and the lipidomic phenotypes at each digestion stage were evaluated by multivariate data analysis. During multiple-stage digestion, Pls were hydrolyzed to lyso-Pls and free fatty acids by phospholipases A2, while the vinyl ether bond was retained at the sn-1 position. The result revealed that the contents of Pls groups were significantly reduced (p < 0.05). The multivariate data analysis results indicated that the ions at m/z 748.28, m/z 750.69, m/z 774.38, m/z 836.58, and etc. were the significant candidate contributors for monitoring the variation of Pls fingerprints during digestion. Results demonstrated that the proposed method exhibited potential for real-time tracking the lipidomic characteristics of nutritional lipid NPs digestion in the human gastrointestinal tract.

Sonochemical effects on fabrication, characterization and antioxidant activities of ß-lactoglobulin-chlorogenic acid conjugates.

The ß-lactoglobulin-chlorogenic acid (LG-CA) conjugate was explored to be formed through ultrasonication, redox-pair method and their combination, the ultrasonication used a probe ultrasonic machine with a 6 mm probe at 270 W, and the frequency was 20-25 kHz. The formation of the conjugate was confirmed by SDS-PAGE with a larger molecular weight. Besides, Fourier infrared spectroscopy (FTIR) and Circular dichroism (CD) indicated changes in the secondary structure of the LG-CA conjugate. The α-helix and ß-sheet contents of LG decreased and the unordered content increased significantly after the formation of covalent complexes. In addition, both the ultrasonic treatment and its combination with redox-pair method could significantly improve the antioxidant properties of LG. The former increased to 23.16 µmol Trolox/g sample, the latter 82-106 µmol Trolox/g sample. Therefore, ultrasonication could be used both individually and in combination with the redox-pair method to produce LG-CA conjugates with stronger antioxidant activities.

Aptamer-conjugated graphene oxide-based surface assisted laser desorption ionization mass spectrometry for selective extraction and detection of Aß1-42 in an Alzheimer's disease SH-SY5 cell model.

The generation and accumulation of amyloid-beta peptide (Aß1-42) in amyloid plaques are key characteristics of Alzheimer's disease (AD); thus, specific detection of Aß1-42 is essential for the diagnosis and treatment of AD. Herein, an aptamer-conjugated graphene oxide (Apt-GO) sensor was synthesized by π-π and hydrophobic interactions using thiol poly (ethylene glycol) amine (SH-PEG-NH2) as a spacer unit. Then, it was applied to selective capture of Aß1-42, and the resulting complex was directly analyzed by surface-assisted laser desorption ionization mass spectrometry (SALDI-MS). The results revealed that the Apt-GO could enhance the detection specificity and reduce non-specific adsorption. This method was validated to be sensitive in detecting Aß1-42 at a low level in human serum (ca. 0.1 µM) within a linear range from 0.1 to 10 µM. The immobilizing amount of aptamer on the GO was calculated to be 36.1 nmol/mg (RSD = 11.5%). In conclusion, this Apt-GO-based SALDI-MS method was sensitive and efficient in selective extraction and detection of Aß1-42, which proved to be a good option for early AD diagnosis.

Comprehensive lipidomic analysis of milk polar lipids using ultraperformance supercritical fluid chromatography-mass spectrometry.

Polar lipids in milk are receiving increasing interest due to their bioactivities. However, milk polar lipids present a wide range of physical-chemical properties at different concentrations, making their analysis challenging. In this study, we presented a comprehensive lipidomic method using ultraperformance supercritical fluid chromatography (UPSFC)-quadrupole-time of flight-mass spectrometry (Q-TOF-MS), which enabled the separation of 18 lipid classes (including nonpolar lipids, cholesterol, ceramide, glycerophospholipids, sphingomyelin, and gangliosides) within 10 min. The method was used to analyze the polar lipids in seven samples, including human milk, other mammalian milk and milk fat globule membrane ingredients, identifying 14 lipid classes containing 219 lipid molecular species. A mass spectrometry data processing strategy applicable for high-throughput studies was also developed and validated.

Role Medium-Chain Fatty Acids in the Lipid Metabolism of Infants.

Human breastmilk, the ideal food for healthy infants, naturally contains a high concentration of medium-chain fatty acids (MCFAs, about 15% of total fatty acids). MCFAs are an important energy source for infants due to their unique digestive and metabolic properties. MCFA-enriched oils are widely used in an infant formula, especially the formula produced for preterm infants. Recently, there has been a growing interest in the triglyceride structure of MCFAs in human milk, their metabolism, and their effects on infant health. This study summarized the MCFA composition and structure in both human milk and infant formula. Recent studies on the nutritional effects of MCFAs on infant gut microbiota have been reviewed. Special attention was given to the MCFAs digestion and metabolism in the infants. This paper aims to provide insights into the optimization of formulations to fulfill infant nutritional requirements.

Ultrasound-assisted assembly of ß-lactoglobulin and chlorogenic acid for non covalent nanocomplex: fabrication, characterization and potential biological function.

It is essential to understand the ultrasound-induced changes in assembly of proteins and polyphenols into non covalent nanocomplex. ß-Lactoglobulin (LG) and chlorogenic acid (CA) with various biological activities can be combined to form food-grade nanocomplexes. This study systematically explored the role of high-intensity ultrasound pretreatment on the binding mechanisms of LG and CA, and the potential biological function for embedding curcumin (Cur). The scanning electron microscopy (SEM) revealed that ultrasound treatment could destroy the structure of LG, and the particle size of the protein was reduced to<50 nm. The change in secondary structure of the protein by ultrasound treatment could be revealed by the fourier transform infrared (FTIR) and fluorescence spectra. Besides, it was found that LG and CA were combined to form a complex under the hydrophobic interaction, and CA was bound in the internal cavity of LG with a relatively extended conformation. The result demonstrated that the ratio of Cur embedded in the ultrasonic sample could be effectively increased by 7% - 10%, the particle size in the emulsion was smaller, and the dispersion was more stable. This work contributes to the development of protein-polyphenol functional emulsion systems with the ability to deliver Cur.

Digestion of Medium- and Long-Chain Triacylglycerol and sn-2 Palmitate in Infant Formula: A Study Based on Dynamic In Vitro Simulation of Infant Gastrointestinal Lipolysis.

In this study, lipolysis of triacylglycerols (TAGs) in infant formula (IF) composed of different oils and supplied with different structured TAGs, including medium- and long-chain triacylglycerol (MLCT) and sn-2 palmitate, was studied using a dynamic digestion model simulating the infant gastrointestinal tract. The molecular species of digestion products released during digestion, including diacylglycerols, monoacylglycerols (MAGs), and free fatty acids, as well as undigested TAGs, were identified and quantified using liquid chromatography-mass spectrometry. We observed clearly different lipolysis degrees (LDs), with diversity in digestion products of different IFs. IFs supplied with MLCT showed moderate medium-chain fatty acid release during gastric digestion and higher LD after intestinal digestion. The presence of sn-2 palmitate in IF was associated with higher content of MAG-16:0 in digestion products. The species and contents of digestion products in IF were highly influenced by structured TAGs.

Effects of Chemical Composition and Microstructure in Human Milk and Infant Formulas on Lipid Digestion.

Infant formula (IF) is an important substitute for infants when human milk (HM) is unavailable; however, it was often observed with "insufficient fat" and showed different metabolic phenotypes, which may affect the growth and brain development of the infant. Considering that the milk fat digestion rate may affect the fat absorption and metabolism and further influence the metabolic phenotype, it is valuable to study the fat digestive behaviors of IF and HM. In the current study, we investigated the in vitro fat digestive properties of HM in comparison to four formulas (IF1, 2, 3, 4) including IFs enriched in OPO lipids (IF1 and IF3) and IFs with common mixed plant oils (IF2 and IF4). Results showed that the extent of eventual lipid hydrolysis of HM (98.9 ± 2.70%) was higher than those of IF1 and IF3 (90.4 ± 3.39 and 91.1 ± 1.67%, respectively) (p < 0.05) and IF2 and IF4 (81.9 ± 1.64 and 79.9 ± 1.05% respectively) (p < 0.01). Native fat globules and protein aggregation were observed at the end of HM gastric digestion, and the aggregates became smaller and then resolved from 60 to 120 min in intestinal digestion, while a large number of aggregates were observed in IF, which may slow the lipid digestion. The absorption differences between HM and IFs in lipid digestion need further study to elucidate the nutritional relevance to infant development and growth.

Triacylglycerol Containing Medium-Chain Fatty Acids: Comparison of Human Milk and Infant Formulas on Lipolysis during In Vitro Digestion.

Medium-chain triacylglycerol (MCT) is widely used in infant formulas (IFs) to provide medium-chain fatty acids (MCFAs) for infants with special fat absorption requirements. However, MCFAs naturally present in human milk are medium-and long-chain triacylglycerols (MLCTs). This study investigated the effect of triacylglycerol containing MCFAs (MLCT vs MCT) on lipolysis by comparison of human milk and IFs containing 0, 20, 30, and 55% of MCT (IF 1 to IF 4) using an in vitro digestion model. Rabbit gastric lipase showed an extent of digestion within the expected range, and was selected as the alternative to human gastric lipase. All IFs showed a lower lipolysis degree compared with human milk. There was no significant difference (p = 0.175) among IFs supplemented with MCT at the end of intestinal digestion. In addition, the digestion of IFs with different MCT contents led to different free fatty acid profiles, which may have health effects on infants.

Short-chain fatty acid (SCFA) and medium-chain fatty acid (MCFA) concentrations in human milk consumed by infants born at different gestational ages and the variations in concentration during lactation stages.

This study aimed to quantify the short-chain fatty acids (SCFAs) and medium-chain fatty acids (MCFAs) in human milk triacylglycerols (TAGs) and investigate their concentrations in human milk consumed during lactation by infants born at different gestational ages. One hundred and eighty milk samples from the mothers of 30 full-term, 10 early-preterm (≤32 weeks), 10 mild-preterm (32-34 weeks), and 10 near-term (34-37 weeks) infants were collected from the colostrum, transitional, and mature milk. The human milk TAGs were transferred into fatty-acid methyl esters via potassium methoxide in methanol and determined using gas chromatography (GC). The total SCFA (4:0) and MCFA concentrations (6:0 and 8:0) were highest in the mature milk (1.47 ± 0.66 mg g-1 fat from full-term infant milk), approximately 42.18% higher than those in transitional milk. Significantly higher SCFA and MCFA concentrations were found in full-term milk than in preterm milk (p = 0.001). The milk TAGs were analyzed using ultra-high-performance supercritical fluid chromatography with quadrupole-time-of-flight mass spectrometry (UHPSFC-Q-TOF-MS), which showed that the SCFAs and MCFAs were mainly esterified with long-chain fatty-acid groups (16:0, 18:1 n-9, and 18:2 n-6) at the glycerol backbone. The infants' daily SCFA intake from human milk was estimated; this was highest from mature milk for full-term infants (∼14 mg d-1) which was significantly different from that of preterm infants from colostrum and transitional milk (p < 0.001). The correlation between dietary SCFAs and MCFAs in human milk and nutrition in newborns, especially in the gut microbiotas of preterm infants, requires further study.

Quantification of Nervonic Acid in Human Milk in the First 30 Days of Lactation: Influence of Lactation Stages and Comparison with Infant Formulae.

Nervonic acid (24:1 n-9, NA) plays a crucial role in the development of white matter, and it occurs naturally in human milk. This study aims to quantify NA in human milk at different lactation stages and compare it with the NA measured in infant formulae. With this information, optimal nutritional interventions for infants, especially newborns, can be determined. In this study, an absolute detection method that uses experimentally derived standard curves and methyl tricosanoate as the internal standard was developed to quantitively analyze NA concentration. The method was applied to the analysis of 224 human milk samples, which were collected over a period of 3-30 days postpartum from eight healthy Chinese mothers. The results show that the NA concentration was highest in colostrum (0.76 ± 0.23 mg/g fat) and significantly decreased (p < 0.001) in mature milk (0.20 ± 0.03 mg/g fat). During the first 10 days of lactation, the change in NA concentration was the most pronounced, decreasing by about 65%. Next, the NA contents in 181 commercial infant formulae from the Chinese market were compared. The NA content in most formulae was <16% of that found in colostrum and less than that found in mature human milk (p < 0.05). No significant difference (p > 0.05) was observed among NA content in formulae with different fat sources. Special attention was given to the variety of n-9 fatty acids in human milk during lactation, and the results indicated that interindividual variation in NA content may be primarily due to endogenous factors, with less influence from the maternal diet.

Triacylglycerol Composition of Breast Milk during Different Lactation Stages.

Triacylglycerol (TAG) composition of breast milk plays an important role in improving digestion, absorption, and metabolism when consumed by infants. This study characterized the TAG profile of human colostrum, transitional, and mature milk samples from 103 women. Significant differences in the TAGs composition of breast milk fat from three lactation stages were observed. The TAGs with high molecular weight and unsaturated fatty acid (such as 1,3-olein-2-palmitin (OPO) and 1(3)-olein-2-palmitin-3(1)-linolein (OPL)) were enriched in colostrum, while the TAGs containing medium-chain fatty acids were more abundant in transitional and mature milk than that in colostrum. Of note, OPL was the most common TAG in breast milk of Chinese women while the most common TAG in breast milk of Western women was OPO. This data will promote the development of infant formulas in terms of the TAG composition more suitable for infants.