A clinical study of the local injection of a freshly manufactured 35 kDa hyaluronan fragment for treating chronic wounds.

This study manufactured a 35 kDa hyaluronan fragment (HA35) by enzymatically degrading high-molecular-weight HA using hyaluronidase PH20 derived from bovine testis. The research then examined the therapeutic efficacy of locally administered, tissue-permeable HA35 in alleviating chronic wounds and their associated neuropathic pain. For 20 patients with nonhealing wounds and associated pain lasting over three months, 100 mg of HA35 was injected daily into the healthy skin surrounding the chronic wound for 10 days. Self-assessments before and after treatment indicated that HA35 significantly enhanced wound healing. This was evidenced by the formation of fresh granulation tissue on the wounds (p < 0.0001); reduced darkness, redness, dryness, and damage in the skin surrounding the wounds (p < 0.0001), and a decrease in wound size (p < 0.001). Remarkably, HA35 injections alleviated pain associated with chronic wounds within 24 hours (p < 0.0001). It can be concluded that the low-molecular-weight hyaluronan fragment HA35 potentially enhances the immune response and angiogenesis during wound healing.

Characterization, Bioactivity, and Biodistribution of 35 kDa Hyaluronan Fragment.

It has been reported that hyaluronic acid (HA) with a 35 kDa molecular weight (HA35) acts biologically to protect tissue from injury, but its biological properties are not yet fully characterized. This study aimed to evaluate the cellular effects and biodistribution of HA35 compared to HA with a 1600 kDa molecular weight (HA1600). We assessed the effects of HA35 and HA1600 on cell migration, NO and ROS generation, and gene expression in cultured macrophages, microglia, and lymphocytes. HA35 was separately radiolabeled with 99mTc and 125I and administered to C57BL/6J mice for in vivo biodistribution imaging. In vitro studies indicated that HA35 and HA1600 similarly enhanced cell migration through HA receptor binding mechanisms, reduced the generation of NO and ROS, and upregulated gene expression profiles related to cell signaling pathways in immune cells. HA35 showed a more pronounced effect in regulating a broader range of genes in macrophages and microglia than HA1600. Upon intradermal or intravenous administration, radiolabeled HA35 rapidly accumulated in the liver, spleen, and lymph nodes. In conclusion, HA35 not only exhibits effects on cellular bioactivity comparable to those of HA1600 but also exerts biological effects on a broader range of immune cell gene expression. The findings herein offer valuable insights for further research into the therapeutic potential of HA35 in inflammation-mediated tissue injury.

Protective Properties of Intestinal Alkaline Phosphatase Supplementation on the Intestinal Barrier: Interactions and Effects.

The intestinal barrier is critical for maintaining intestinal homeostasis, and its dysfunction is associated with various diseases. Recent findings have revealed the multifunctional role of intestinal alkaline phosphatase (IAP) in diverse biological processes, including gut health maintenance and function. This review summarizes the protective effects of IAP on intestinal barrier integrity, encompassing the physical, chemical, microbial, and immune barriers. We discuss the results and insights from in vitro, animal model, and clinical studies as well as the available evidence regarding the impact of diet on IAP activity and expression. IAP can also be used as an indicator to assess intestinal-barrier-related diseases. Further research into the mechanisms of action and long-term health effects of IAP in maintaining overall intestinal health is essential for its future use as a dietary supplement or functional component in medical foods.

Extrusion of casein and whey protein isolate enhances anti-hardening and performance in high-protein nutrition bars.

Model high-protein nutrition bars (HPNBs) were formulated by incorporating whey protein isolate (WPI) and casein (CN) at various extrusion temperatures (50, 75, 100, 125, and 150 °C) with a protein content of 45 g per 100 g. The free sulfhydryl groups, amino groups, hardness, and microstructures of HPNBs were analyzed periodically at 37 °C over a storage period of 45 days. Specifically, sulfhydryl group, amino group and surface hydrophobicity of extruded whey protein isolate (WPE) and extruded casein (CE) were significantly reduced (P < 0.05) compared to those of unextruded protein. HPNBs formulated with WPE (HWPE) and CE (HWCE) exhibited a slower hardening rate compared to those formulated with unmodified protein. Moreover, the color difference, hardness and sensory score of HPNBs after 45 days of storage were used as indicators, and the results of the TOPSIS multiple index analysis indicated that HPNB formulated with WPI extruded at 150 °C possessed the best quality characteristics.

Evaluating the role of glycyrrhizic acid on the dynamic stabilization mechanism of the emulsion prepared by α-Lactalbumin: Experimental and silico approaches.

The role of glycyrrhizic acid (GA) on the dynamic stabilization mechanism of the α-Lactalbumin (α-La) emulsion was evaluated in this study. Smaller particle size and higher zeta potential value were observed in the α-La/GA emulsion as compared to the α-La emulsion. Ultra-high-resolution microscopy revealed that the interfacial film formed around oil droplets by α-La/GA complex was thicker compared to that of either α-La or GA. The appearance of a new peak at 1679 cm-1 in FTIR of the α-La/GA emulsion attributed to the stretching vibration of CO, providing evidence of the formation of a stable emulsion system. The results from dynamic molecular simulation showed GA induced the formation of an interfacial adsorption layer at the oil-water interface, reducing the migration ability of GA. The findings indicate that the presence of GA in the α-La emulsion effectively enhances its stability, highlighting its potential as a valuable emulsifying agent for various industrial applications.

Sustainable emerging high-intensity sonication processing to enhance the protein bioactivity and bioavailability: An updated review.

High-intensity ultrasound (HIU) is considered one of the promising non-chemical eco-friendly techniques used in food processing. Recently (HIU) is known to enhance food quality, extraction of bioactive compounds and formulation of emulsions. Various foods are treated with ultrasound, including fats, bioactive compounds, and proteins. Regarding proteins, HIU induces acoustic cavitation and bubble formation, causing the unfolding and exposure of hydrophobic regions, resulting in functional, bioactive, and structural enhancement. This review briefly portrays the impact of HIU on the bioavailability and bioactive properties of proteins; the effect of HIU on protein allergenicity and anti-nutritional factors has also been discussed. HIU can enhance bioavailability and bioactive attributes in plants and animal-based proteins, such as antioxidant activity, antimicrobial activity, and peptide release. Moreover, numerous studies revealed that HIU treatment could enhance functional properties, increase the release of short-chain peptides, and decrease allergenicity. HIU could replace the chemical and heat treatments used to enhance protein bioactivity and digestibility; however, its applications are still on research and small scale, and its usage in industries is yet to be implemented.

Physical treatment synergized with natural surfactant for improving gas-water interfacial behavior and foam characteristics of α-lactalbumin.

The purpose of this study was to investigate effect of physical treatment (ultrasound, U/high pressure homogenization, H/combined treatment, UH or HU) and surfactant (Mogroside V, Mog) on air/water interface adsorption and foaming properties of α-lactalbumin (ALa). Firstly, the binding of Mog and all physical-treated ALa was a static quenching process. Mog had the greatest binding affinity for HU-ALa among all treated samples. U or H treatment could change surface hydrophobicity of ALa/Mog complex. Secondly, at the molar ratio (ALa:Mog) of 1:50, foaming ability (FA) of all ALa samples got the maximum. The sequence of FA in ALa and ALa/Mog complex was listed as follow: HU > U > H > UH. Moreover, foaming stability (FS) of HU-ALa was the highest, followed by H-ALa, U-ALa and UH-ALa. Meanwhile, low concentration Mog increased FS of ALa or UH-ALa, but it reduced FS of H-ALa, U-ALa and HU-ALa. Quartz crystal microbalance with dissipation monitoring (QCM-D) experiment indicated that ALa/Mog complex after U or H treatment was quickly absorbed at air/water interface, compared with the treated ALa, and HU-ALa/Mog had the largest frequency shift. In addition, HU-ALa had the thickest bubble membrane and the highest dissipation shift in all samples, indicating that the absorbed membrane thickness and viscoelasticity of samples was correlated with foam stability. Therefore, U and H treatment synergism with Mog was an effective approach to enhance foam properties of ALa, which indicated that HU-treated ALa/Mog complex could be viewed as the safe and efficient foaming agent applied in food processing.

Anti-Inflammatory Effects of the 35kDa Hyaluronic Acid Fragment (B-HA/HA35).

Background: Hyaluronic acid (HA) and HA fragments interact with a variety of human body receptors and are involved in the regulation of various physiological functions and leukocyte trafficking in the body. Accordingly, the development of an injectable HA fragment with good tissue permeability, the identification of its indications, and molecular mechanisms are of great significance for its clinical application. The previous studies showed that the clinical effects of injectable 35kDa B-HA result from B-HA binding to multiple receptors in different cells, tissues, and organs. This study lays the foundation for further studies on the comprehensive clinical effects of injectable B-HA. Methods: We elaborated on the production process, bioactivity assay, efficacy analyses, and safety evaluation of an injectable novel HA fragment with an average molecular weight of 35 kDa (35 kDa B-HA), produced by recombinant human hyaluronidase PH20 digestion. Results: The results showed that 35 kDa B-HA induced human erythrocyte aggregation (rouleaux formation) and accelerated erythrocyte sedimentation rates through the CD44 receptor. B-HA application and injection treatment significantly promoted the removal of mononuclear cells from the site of inflammation and into the lymphatic circulation. At a low concentration, 35 kDa B-HA inhibited production of reactive oxygen species and tumor necrosis factor by neutrophils; at a higher concentration, 35 kDa B-HA promoted the migration of monocytes. Furthermore, 35 kDa B-HA significantly inhibited the migration of neutrophils with or without lipopolysaccharide treatment, suggesting that in local tissues, higher concentrations of 35 kDa B-HA have antiinflammatory effects. After 99mTc radiolabeled 35 kDa B-HA was intravenously injected into mice, it quickly entered into the spleen, liver, lungs, kidneys and other organs through the blood circulation. Conclusion: This study demonstrated that the HA fragment B-HA has good tissue permeability and antiinflammatory effects, laying a theoretical foundation for further clinical studies.

Modification of fermented whey protein concentrates: Impact of sequential ultrasound and TGase cross-linking.

This study aimed to examine the impact of fermentation process on whey protein and improve the general properties of fermented whey protein concentrate (FWPC) recovered by a combined ultrafiltration-diafiltration (UF-DF) operation. Impacts of sequential ultrasound (US) pretreatment and transglutaminase (TGase) crosslinking on structural, functional, and physicochemical properties of FWPCs were investigated. Partially denatured and hydrolyzed fermented whey protein could replace heat denaturation prior to the TGase addition to a whey protein system. Sequential treatment increased the molecular weight of FWPCs as exhibited by both SEM and SDS-PAGE, which demonstrates that modification can lead to the polymers and oligomers production. The zeta potential value increased significantly after US treatment and enzyme catalysis, and all the modified FWPCs were strongly negatively charged. Compared with the secondary structure of untreated FWPCs, the percentage of α-helix and random coil in modified FWPCs significantly increased, while the percentage of ß-sheet and ß-turns reduced. Solubility, free sulfhydryl groups, and surface hydrophobicity of all FWPCs were significantly improved compared to non-fermented WPC (P < 0.05). Sequential treatment induced a substantial impact on the emulsifying activity and stability of modified samples in comparison with untreated FWPCs. Scanning electron microscope pictures confirmed the positive effects of sequential treatments on texture and void size reduction. Therefore, the application of recovering modified FWPCs is fully recommended as a commercially viable approach for enhanced protein production at the industrial scale.

Structure and rheological properties of extruded whey protein isolate: Impact of inulin.

Impacts of inulin addition (0, 5, 10, 15 %) on structure, functional and rheological properties of whey protein isolate (WPI) after extrusion pretreatment (E-WPI) were investigated. The results proved that after adding 15 % inulin, water holding capacity of gels, emulsifying activity, emulsion stability, foaming ability and foaming stability of E-WPI were the best and increased by 24.38 %, 7.43 %, 12.35 %, 162.97 % and 41.31 %, compared with those of unextruded WPI, respectively. Rheology analysis showed that apparent viscosity and consistency index of all the samples after inulin addition were enhanced and exhibited pseudoplastic fluids. FTIR spectroscopy indicated that E-WPI/WPI and inulin was linked together due to hydrogen bonds and addition of inulin increased the proportion of their ß-turn structure. These findings demonstrated that the addition of inulin in combination with extrusion pretreatment could jointly improve the functional properties of WPI. Therefore, E-WPI with the addition of inulin shows potential commercial applications in the production of novel food foaming agents and emulsifiers.

Comparison of Oxidative and Physical Stabilities of Conjugated Linoleic Acid Emulsions Stabilized by Glycosylated Whey Protein Hydrolysates via Two Pathways.

The objective of the research was to analyze and compare the oxidative and physical stabilities of conjugated linoleic acid (CLA) emulsions stabilized by two glycosylated hydrolysates (GPP-A and GPP-B) that were formed via two different pathways. This study showed that GPP-A exhibited higher browning intensity and DPPH radical scavenging ability in comparison with GPP-B. Moreover, the CLA emulsion formed by GPP-A exhibited a lower creaming index, average particle size, primary and secondary oxidative products, in comparison with GPP-B-loaded emulsion. However, the GPP-A-loaded emulsion showed a higher absolute potential and fraction of interfacial adsorption than that of the CLA emulsion formed by GPP-B. Therefore, the CLA emulsion formed by GPP-A exhibited stronger stabilities in comparison with the GPP-B-loaded emulsion. These results suggested that GPP-A showed an emulsification-based delivery system for embedding CLA to avoid the loss of biological activities. Additionally, the development of CLA emulsions could exert its physiological functions and prevent its oxidation.

Characterization of major volatile compounds in whey spirits produced by different distillation stages of fermented lactose-supplemented whey.

This research aimed to advance the understanding of acceptable sensory qualities of potable whey-based spirit from nonsupplemented, mid-supplemented, and high-supplemented whey samples by analyzing major volatile compounds during different stages of distillation (head, heart, and tail). The results demonstrated that commercial Saccharomyces cerevisiae strain in lactase-hydrolyzed whey showed rapid and complete sugar hydrolysis and efficient ethanol production in 24, 30, and 36 h on average, producing up to 29.5, 42.1, and 56.4 g/L of ethanol, respectively. The variations in titratable acidity, specific gravity, pH value, residual protein, sugar content, and alcohol yield were investigated during the fermentation. The total amount of volatile compound concentrations significantly decreased from the head (2,087-2,549 mg/L) to the tail whey spirits (890-1,407 mg/L). In the whey spirit, 2-methyl-1-butanol, 3-methyl-1-butanol, 2-methyl-1-propanol, 1-propanol, acetaldehyde, and ethyl acetate were the most prevalent dominant compounds, accounting for the largest proportion of total volatile compounds. The volatile compounds detected were far below the acceptable legal limit. The results suggest that high sensory qualities of potable whey-based spirits can be produced by fermentation of lactose-supplemented whey with S. cerevisiae cells.

Laccase cross-linking of sonicated α-Lactalbumin improves physical and oxidative stability of CLA oil in water emulsion.

α-lactalbumin was modified by ultrasound (US, 20 kHz, 43 ± 3.4 W/cm-2) pre-treatments (0, 15, 30 and 60 min) and laccase cross-linking of sonicated α-lactalbumin was used to evaluate the physical and oxidative stability of conjugated linoleic acid (CLA) emulsions. The emulsions prepared with laccase cross-linking US-α-lactalbumin (α-lactalbumin treated with US pre-treatment) and US-α-lactalbumin were scrutinized for oxidative and physical stability at room temperature for two weeks of storage. Laccase cross-linking US-α-lactalbumin (Lac-US-α-lactalbumin) revealed improved physical stability in comparison with US-α-lactalbumin, specified by droplet size, structural morphology, adsorbed protein, emulsifying properties and creaming index. SDS-PAGE analysis showed that there was formation of polymers in Lac-US-α-lactalbumin emulsion. Surface hydrophobicity of Lac-US-α-lactalbumin was higher than that of US-α-lactalbumin, and gradually enhanced with the increase of ultrasound time. More importantly, the measurements of peroxide values and conjugated dienes were used to study the oxidative stability of the CLA emulsions. The Lac-US-α-lactalbumin emulsion proved to be reducing the synthesis of fatty acid hydroperoxides and less conjugated dienes compared to the native and US-α-lactalbumin emulsions. This study revealed that the combination of US pre-treatment and laccase cross-linking might be an effective technique for the modification of CLA emulsions.

Citric acid promotes disulfide bond formation of whey protein isolate in non-acidic aqueous system.

Impacts of citric acid (CA) treatment under non-acidic conditions (pH 7.0, 8.0 and 9.0) on whey protein isolate (WPI) were examined in this study. Size exclusion chromatography and SDS-PAGE indicated that molecular size and weight of WPI-CA became larger at pH 7.0, 8.0 and 9.0 with CA ranged from 0 to 15 mg/mL, but the protein aggregates disappeared after ß-mercaptoethanol was added. The free SH groups of WPI-CA gradually decreased. This could be deduced that CA could promote disulfide bond formation of WPI at the non-acidic pH values. Furthermore, fourier transform infra-red (FTIR) spectroscopy and fluorescence spectroscopy data confirmed the conformational changes of secondary and tertiary structures of CA-modified WPI, respectively. Therefore, these results suggested that disulfide bond formation of WPI occurred at citric acid treatment under non-acidic conditions, being contributed to production of its larger molecular size substances and alteration of its structural characteristics.

Comparison in bioactivity and characteristics of Ginkgo biloba seed polysaccharides from four extract pathways.

The effects of hot-water extraction (HWE), ultrasound-treated extraction (UTE), enzyme-treated extraction (ETE) and ultrasound-enzyme treated extraction (UETE) on the α-glucosidase inhibitory activity, antioxidant activities and characteristics of Ginkgo biloba seed polysaccharides were investigated and compared in this study. Among the four extracted polysaccharides, the UETE-polysaccharide initially exhibited the highest α-glucosidase inhibitory activity and antioxidant activities. The HWE-polysaccharide showed a large number of small compact spherical structures, and the UTE-polysaccharide exhibited an irregular pleated porous shape; meanwhile, the ETE-polysaccharide and UETE-polysaccharide were spongy with smooth surface topography, as observed by scanning electron microscopy (SEM). The four polysaccharides varied in monosaccharide composition. The HWE-polysaccharide mainly consisted of homogeneous mannose; the UETE-polysaccharide was primarily composed of mannose, rhamnose, and glucose in a molar ratio of 8.25:1.00:1.53. The HWE-polysaccharide had the largest molecular weight (4.2 × 105 Da), reduced by the order of the UETE-polysaccharide (2.02 × 104 Da), ETE-polysaccharide (1.72 × 104 Da), and UTE-polysaccharide (1.34 × 104 Da). Thus, the four extract methods exerted significant effects on the bioactivity and characteristics of the polysaccharides. The UETE-polysaccharide from G. biloba seeds showed the highest bioactive activities and distinctive structural characteristics.