Enhancing functionality and bioactivity of walnut protein through limited enzyme digestion.

BACKGROUND: Walnut protein (WP) is recognized as a valuable plant protein. However, the poor solubility and functional properties limit its application in the food industry. It is a great requirement to improve the physicochemical properties of WP. RESULTS: Following a 90 min restricted enzymatic hydrolysis period, the solubility of WP significantly increased from 3.24% to 54.54%, with the majority of WP hydrolysates (WPHs) possessing a molecular weight exceeding 50 kDa. Circular dichroism spectra showed that post-hydrolysis, the structure of the protein became more flexible, while the hydrolysis time did not significantly alter the protein's secondary structure. After hydrolysis, WP's surface hydrophobicity significantly increased from 2279 to 6100. Furthermore, WPHs exhibited a strong capacity for icariin loading and micelle formation with critical micelle concentration values of 0.71, 0.99 and 1.09 mg mL-1, respectively. Moreover, similar immuno-enhancement activities were observed in WPHs. After exposure to WPHs, the pinocytosis of RAW264.7 macrophages was significantly improved. WPH treatment also increased the production of nitric oxide, interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in macrophages. Up-regulation of mRNA expressions of IL-6, inducible nitric oxide synthase (iNOS) and TNF-α was observed in a dose-dependent manner. CONCLUSION: The enhancement of functionality and bioactivity in WP can be achieved through the application of limited enzyme digestion with trypsin. This process effectively augments the nutritional value and utility of the protein, making it a valuable component in various dietary applications. © 2024 Society of Chemical Industry.

Diversified carbon intensity under global value chains: A measurement and decomposition analysis.

The rise and gradually developing of global value chains (GVC) have changed the essence of international trade, and further complicated the issue of carbon emission. It is of great significance to identify and study the practical carbon intensity in a country under the condition of GVC. Based on Inter-country Input-Output (ICIO) model, this paper establishes a framework for measuring the carbon intensities embodied in aggregated demand and distinguishing pure domestic demand, traditional trade, simple GVC route, complex GVC route at the global level and for 41 economies from 2010 to 2014 according to demand heterogeneity, and uses multiplicative structural decomposition analysis (SDA) method to decompose the driving factors affecting their changes. The results indicate that, in most developed countries, the carbon intensities embodied in international trade were significantly greater than pure domestic carbon intensity, whereas it was opposite in most developing countries. The carbon intensity embodied in traditional trade was generally lower than that in GVC routes, and in low carbon intensity (high carbon intensity) sub-group, the carbon intensity embodied in simple GVC route was lower (higher) than that in complex GVC route. Moreover, the emission coefficient effect was the main driver of the decline in almost all carbon intensities, especially embodied carbon intensity in GVC routes. On the contrary, the intermediate import effect was the main inhibitor, and this effect would cause more rise to embodied carbon intensity in simple GVC route, compared to embodied carbon intensity in complex GVC route.

Molecular dynamics simulation on the allosteric analysis of the c-di-GMP class I riboswitch induced by ligand binding.

Riboswitches are RNA molecules that regulate gene expression using conformation change, affected by binding of small molecule ligands. Although a number of ligand-bound aptamer complex structures have been solved, it is important to know ligand-free conformations of the aptamers in order to understand the mechanism of specific binding by ligands. In this paper, we use dynamics simulations on a series of models to characterize the ligand-free and ligand-bound aptamer domain of the c-di-GMP class I (GEMM-I) riboswitch. The results revealed that the ligand-free aptamer has a stable state with a folded P2 and P3 helix, an unfolded P1 helix and open binding pocket. The first Mg ions binding to the aptamer is structurally favorable for the successive c-di-GMP binding. The P1 helix forms when c-di-GMP is successive bound. Three key junctions J1/2, J2/3 and J1/3 in the GEMM-I riboswitch contributing to the formation of P1 helix have been found. The binding of the c-di-GMP ligand to the GEMM-I riboswitch induces the riboswitch's regulation through the direct allosteric communication network in GEMM-I riboswitch from the c-di-GMP binding sites in the J1/2 and J1/3 junctions to the P1 helix, the indirect ones from those in the J2/3 and P2 communicating to P1 helix via the J1/2 and J1/3 media.

Preparation, characterization, and release properties of Rosa roxburghii Tratt seed oil and ß-carotene-coloaded proliposomes.

Rosa roxburghii Tratt seed oil (RSO) and ß-carotene (ßC) were chosen to prepare proliposomes by the thin-film dispersion method. The characteristics of unloaded proliposome, RSO proliposome (L-R), ßC proliposome (L-ß), and RSO/ßC proliposome (L-R-ß) were analyzed, and their antioxidant activity, storage stability, and release properties were investigated. The proliposomes had an encapsulation efficiency (RSO, ßC) higher than 83.10%, nanometer size, smooth surface, and irregular structure. L-R-ß showed better dispersibility, stability, and antioxidant activity than L-R and L-ß. Simultaneous encapsulation of RSO and ßC reduced the phospholipid oxidation of proliposomes and improved the retention rate of RSO in storage environments of 4, 25, and 40°C. Moreover, the RSO and ßC release kinetics of proliposomes in the simulated saliva fluid and gastric fluid phases can be described by the first-order model, and the Korsmeyr-Peppas method was applied to describe their release mechanism in the simulated intestinal fluid phase.

Molecular dynamics simulation study of DNA conformation changes caused by the dinuclear platinum(II) complexes with the bisphosphonate group.

Bisphosphonate (BP) has been widely used as a bone-targeting group, and the BP-modified platinum(II) complexes have shown potential to as anticancer drugs against bone-related diseases, such as osteosarcoma. DNA conformation changes induced by the BP-modified dinuclear platinum(II) complexes have been investigated using molecular dynamics simulations. The results indicated that the BP-modified dinuclear platinum(II) complexes coordinated to DNA results in DNA structural distortions, including twisting, unwinding and bending. Furthermore, the rigidity of the bridging linkers in the BP-modified platinum(II) complex may induce more significant DNA structural distortions with same spans. The results provide the detail information of DNA conformational changes induced by the BP-modified platinum(II) complexes with different flexibility of bridging linkers, and are helpful for exploring novel platinum-based antitumor drugs.

Effect of drying methods on Lactobacillus Rhamnosus GG microcapsules prepared using the complex coacervation method.

Lactobacillus rhamnosus GG (LGG) microcapsules prepared using the complex coacervation method were dried through spray and vacuum freeze drying. Physical properties, resistance to simulated gastrointestinal and bile salt conditions, and the stability of the LGG microcapsules during storage and heat treatment were estimated. Spray and vacuum freeze drying are suitable for LGG microcapsule preparation because of lower than 2.80% of the water content, and higher than 93.21% of the encapsulation efficiency. Under the simulated gastrointestinal condition, the survival rates of two kinds of microencapsulated LGG were over 96%. At 0.6% bile salt concentration, the survival rate of microencapsulated LGG subjected to spray drying was 110.89% higher than that of LGG subjected to vacuum freeze drying. In addition, the stability of the former was better than that of the latter during storage under low water activity condition (0.11). However, under high water activity condition (0.75), the two showed opposite results. The two kinds of microencapsulated LGG were completely inactivated after 4 weeks in 0.75 of water activity. The survival counts of microencapsulated LGG had no significant difference at -18°C after 8 weeks. After 8 weeks at 25°C and 4°C, the survival loss of microencapsulated LGG through drying and vacuum freeze drying was approximately 2 and 1 log CFU/g, respectively. PRACTICAL APPLICATION: LGG is a kind of microorganism beneficial to the human body, but it is easy to inactivate and lose its physiological value. LGG microcapsules can improve the stability of LGG, prolong the release time, and expand its application range. The drying method will affect the properties of LGG microcapsules. So, the characteristics analysis of LGG microcapsules with different drying methods can provide a reference for the preparation and application of LGG microcapsules.

Structure characterization of soybean peptides and their protective activity against intestinal inflammation.

Soybean peptides serve as functional foods with impressive health benefits. The structure characteristics of peptides are highly related to the health benefits. The structure-activity relationship and mechanism underlined are important scientific questions in this field. To answer these questions, soybean peptides were produced by combinatory enzymatic hydrolysis in this work. Fifty-two peptide sequences were identified by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The anti-inflammatory activities of these peptides were investigated by using a lipopolysaccharide (LPS)-induced inflammation cell model. Soybean peptides could significantly promote cell proliferation. Additionally, soybean peptides could alleviate LPS-induced inflammation by reducing the production and expression of nitric oxide (NO), tumor necrosis factor alpha (TNF-α), interleukin-1ß (IL-1ß) and interleukin-6 (IL-6). Moreover, soybean peptides could promote the mRNA expression of proteins related to inflammation inhibition (IL-10) and tight junction modulation. The structure-activity relationship was addressed. The results documented the potential of soybean peptides as functional foods.

Structure identification of soybean peptides and their immunomodulatory activity.

Soybean peptides are functional food with good health benefits. The health benefits presented are highly dependent on the peptide structure. In this work, soybean peptides were prepared by alkaline protease hydrolysis of soybean proteins. The peptide structure was identified by UPLC-MS/MS. The full peptide composition was revealed. The sequences of 51 peptides were identified and 46 peptides were assigned as immunomodulatory peptides. By evaluating the immumonodulatory activity and mechanism, soybean peptides could facilitate the proliferation of macrophages. The pinocytotic activity and NO level were increased. Induction of iNOS mRNA expression by soybean peptides was responsible for the increased NO production. The release of cytokines IL-6 and TNF-α was elevated and their levels were equal to positive control. The mRNA expression levels of IL-6 and TNF-α were also improved by soybean peptides, but much lower than positive control. The results were helpful for application of soybean peptides in functional foods.

Extraction of flavonoids from Rhodiola sachlinesis A. Bor by UPE and the antioxidant activity of its extract.

Rhodiola sachlinesis, A. Bor is one of the most popular traditional Chinese medicines and has been proved to have many effective compounds, such as flavonoids, anthraglycosides, essential oil with cinnamic aldehyde and citral and organic acids. In this article, a new method of ultrahigh pressure extraction (UPE) was used to extract flavonoids from R. sachlinesis. Uniform Design was employed to get the optimum extraction conditions. The optimum UPE conditions were as follows: pressure was 500 MPa; ethanol concentration was 40%; solid/liquid ratio (g mL(-1)) was 1 : 70. The optimum extraction yield of flavonoids was 52.33 mg g(-1). The antioxidant activity of the crude extract was tested with DPPH assay. It showed a higher activity, compared with tertiary butyhydroquinone (TBHQ) at the same concentration (1 mg mL(-1)).

Clinical effect and action mechanism of Weicao Capsule in treating gout.

OBJECTIVE: To study the therapeutic effect of Weicao Capsule (WCC) on gout. METHODS: Two hundred gout patients were assigned to two groups. The treated group was treated with WCC and the control group was treated with Tongfengding Capsule. Both groups were given the respective treatments orally 3 times a day, 2 capsules each time with 2 weeks as one course and all patients received 2 successive courses of treatment. Changes of blood beta(2)-microglobulin (beta(2)-M), hemoglobin (Hb), 24 h urinary protein (24 h UP), pH value of urine and blood uric acid (BUA) as well as kidney function were observed. RESULTS: After treatment, level of beta(2)-M got lowered significantly, Hb and 24 h UP, blood urea nitrogen, serum creatinine and the clearance rate of creatinine, as well as blood lipids all improved obviously in the treated group (all P<0.01), while these parameters remained unchanged in the control group (P>0.05). The pH value of urine was improved in both groups showing an insignificant difference between them (P>0.05). BUA was decreased in both groups with a decrease to a larger extent in the treated group (P<0.01). The total effective rate was 87% in the treated group, which was superior to that in the control group (62%, P<0.05). CONCLUSION: WCC has a favorable therapeutic effect on gout and its mechanism of action for improving renal function and reducing urinary protein could be related with the lowering of blood beta(2)-M, BUA and lipids.

Single-Molecule Detection Reveals Different Roles of Skp and SurA as Chaperones.

Skp and SurA are both periplasmic chaperones involved in the biogenesis of Escherichia coli ß-barrel outer membrane proteins (OMPs). It is commonly assumed that SurA plays a major role whereas Skp is a minor factor. However, there is no molecular evidence for whether their roles are redundant. Here, by using different dilution methods, we obtained monodisperse and aggregated forms of OmpC and studied their interactions with Skp and SurA by single-molecule fluorescence resonance energy transfer and fluorescence correlation spectroscopy. We found that Skp can dissolve aggregated OmpC while SurA cannot convert aggregated OmpC into the monodisperse form and the conformations of OmpC recognized by the two chaperones as well as their stoichiometries of binding are different. Our study demonstrates the functional distinctions between Skp and SurA. In particular, the role of Skp is not redundant and is probably more significant under stress conditions.

Determination of Equilibrium Constant and Relative Brightness in Fluorescence Correlation Spectroscopy by Considering Third-Order Correlations.

Fluorescence correlation spectroscopy (FCS) is a powerful tool to investigate molecular diffusion and relaxations, which may be utilized to study many problems such as molecular size and aggregation, chemical reaction, molecular transportation and motion, and various kinds of physical and chemical relaxations. This article focuses on a problem related to using the relaxation term to study a reaction. If two species with different fluorescence photon emission efficiencies are connected by a reaction, the kinetic and equilibrium properties will be manifested in the relaxation term of the FCS curve. However, the conventional FCS alone cannot simultaneously determine the equilibrium constant (K) and the relative fluorescence brightness (Q), both of which are indispensable in the extraction of thermodynamic and kinetic information from the experimental data. To circumvent the problem, an assumption of Q = 0 is often made for the weak fluorescent species, which may lead to numerous errors when the actual situation is not the case. We propose to combine the third-order FCS with the conventional second-order FCS to determine K and Q without invoking other resources. The strategy and formalism are verified by computer simulations and demonstrated in a classical example of the hairpin DNA-folding process.

Scanning Single-Molecule Fluorescence Correlation Spectroscopy Enables Kinetics Study of DNA Hairpin Folding with a Time Window from Microseconds to Seconds.

Single-molecule fluorescence measurements have been widely used to explore kinetics and dynamics of biological systems. Among them, single-molecule imaging (SMI) is good at tracking processes slower than tens of milliseconds, whereas fluorescence correlation spectroscopy (FCS) is good at probing processes faster than submilliseconds. However, there is still shortage of simple yet effective single-molecule fluorescence method to cover the time-scale between submilliseconds and tens of milliseconds. To effectively bridge this millisecond gap, we developed a single-molecule fluorescence correlation spectroscopy (smFCS) method that works on surface-immobilized single molecules through surface scanning. We validated it by monitoring the classical DNA hairpin folding process. With a wide time window from microseconds to seconds, the experimental data are well fitted to the two-state folding model. All relevant molecular parameters, including the relative fluorescence brightness, equilibrium constant, and reaction rate constants, were uniquely determined.

[Effects of puerarin on plasma membrane GLUT4 content in skeletal muscle from insulin-resistant Sprague-Dawley rats under insulin stimulation].

OBJECTIVE: To explore the effect of puerarin injection on the amount of GLUT4 protein at the plasma membrane in insulin-resistant rat skeletal muscle. METHOD: The rat model of insulin resistance (IR) was made by being fed with high-fat diet. The animals were divided into three groups (ten in each group): group I: controls; group II: Insulin-resistant rats; group III: Insulin-resistant rats + Puerarin treatment. Insulin-resistant rats were injected with 100 mg puerarin injection per kg body weight through abdominal cavity once a day for 4 weeks. Fasting blood glucose and fasting serum insulin levels were measured before and after Puerarin treatment, respectively. Insulin treatment was achieved by intraperitoneal injection of insulin (1 unit insulin per kg body weight.) 15 minute before killing the animals. The right hindlimb skeletal muscle was rapidly dissected. Then the expression of GLUT4 protein at the plasma membrane in all the animals was assessed with Western blot method. RESULT: The GLUT4 content at the plasma membrane in insulin-resistant rats skeletal muscle was significantly lower (about 31%) than that of controls (P < 0.01). Puerarin Injection partly corrected fasting blood glucose (from 6.17 +/- 0.67 mmol x L(-1) to 5.54 +/- 0.35 mmol x L(-1)) and fasting serum insulin levels (from 17.09 +/- 2.02 mU x L(-1) to 11.86 +/- 1.35 mU x L(-1)) and increased the GLUT4 content at the plasma membrane by 1.18-fold in insulin-resistant rats skeletal muscle. CONCLUSION: Puerarin Injection can ameliorate IR, and the mechanism may be involved in increasing cell-surface level of GLUT4 through decreasing fasting blood glucose and fasting serum insulin levels, improving GLUT4 trafficking and intracellular insulin signaling.