Comparative study on the treatment of swine wastewater by VFCW-MFC and VFCW: Pollutants removal, electricity generation, microorganism community.

Swine wastewater, characterized by high organic and nutrient content, poses significant environmental challenges. This study aims to compare the effectiveness of two treatment technologies, namely Vertical Flow Constructed Wetland-Microbial Fuel Cell (VFCW-MFC) and Vertical Flow Constructed Wetland (VFCW), in terms of pollutant removal, electricity generation, and microorganism community dynamics. The results showed that the average removal efficiencies of chemical oxygen demand (COD), ammonia nitrogen, total nitrogen (TN), total phosphorus (TP) and sulfadiazine antibiotics (SDZ) by VFCW-MFC were as high as 94.15%, 95.01%, 42.24%, 97.16% and 82.88%, respectively, which were all higher than that by VFCW. Both VFCW-MFC and VFCW have good tolerance to SDZ. In addition, VFCW-MFC has excellent electrical performance, with output voltage, power density, coulombic efficiency and net energy recovery up to 443.59 mV, 51.2 mW/m3, 52.91% and 2.04 W/(g·s), respectively, during stable operation. Moreover, the microbial community diversity of VFCW-MFC was more abundant, and the species abundance distribution in cathode region was more rich and even than in anode region. At phylum level, the dominant microorganisms in VFCW-MFC included Proteobacteria, Bacteroidota, Firmicutes and Actinobacteriota, which showed good degradation effect on SDZ. Proteobacteria and Firmicutes are also involved in electricity production. Chloroflexi, Proteobacteria and Bacteroidota play a major role in nitrogen reduction.

(-)-Epigallocatechin-3-gallate Directly Binds Cyclophilin D: A Potential Mechanism for Mitochondrial Protection.

(1) Background: (-)-Epigallocatechin-3-gallate (EGCG) has been reported to improve mitochondrial function in cell models, while the underlying mechanism is not clear. Cyclophilin D (CypD) is a key protein that regulates mitochondrial permeability transition pore (mPTP) opening. (2) Methods: In this study, we found that EGCG directly binds to CypD and this interaction was investigated by surface plasmon resonance (SPR), nuclear magnetic resonance (NMR) and molecular dynamic (MD) simulation. (3) Results: SPR showed an affinity of 2.7 × 10-5 M. The binding sites of EGCG on CypD were mapped to three regions by 2D NMR titration, which are Region 1 (E23-V29), Region 2 (T89-G104) and Region 3 (G124-I133). Molecular docking showed binding interface consistent with 2D NMR titration. MD simulations revealed that at least two conformations of EGCG-CypD complex exist, one with E23, D27, L90 and V93 as the most contributed residues and E23, L5 and I133 for the other. The major driven force for EGCG-CypD binding are Van der Waals and electrostatic interactions. (4) Conclusions: These results provide the structural basis for EGCG-CypD interaction, which might be a potential mechanism of how EGCG protects mitochondrial functions.

Glycan Determinants of Heparin-Tau Interaction.

Tau aggregates into paired helical filaments within neurons, a pathological hallmark of Alzheimer's disease. Heparin promotes tau aggregation and recently has been shown to be involved in the cellular uptake of tau aggregates. Although the tau-heparin interaction has been extensively studied, little is known about the glycan determinants of this interaction. Here, we used surface plasmon resonance (SPR) and NMR spectroscopy to characterize the interaction between two tau fragments, K18 and K19, and several polysaccharides, including heparin, heparin oligosaccharides, chemically modified heparin, and related glycans. Using a heparin-immobilized chip, SPR revealed that tau K18 and K19 bind heparin with a KD of 0.2 and 70 µM, respectively. In SPR competition experiments, N-desulfation and 2-O-desulfation had no effect on heparin binding to K18, whereas 6-O-desulfation severely reduced binding, suggesting a critical role for 6-O-sulfation in the tau-heparin interaction. The tau-heparin interaction became stronger with longer-chain heparin oligosaccharides. As expected for an electrostatics-driven interaction, a moderate amount of salt (0.3 M NaCl) abolished binding. NMR showed the largest chemical-shift perturbation (CSP) in R2 in tau K18, which was absent in K19, revealing differential binding sites in K18 and K19 to heparin. Dermatan sulfate binding produced minimal CSP, whereas dermatan disulfate, with the additional 6-O-sulfo group, induced much larger CSP. 2-O-desulfated heparin induced much larger CSP in K18 than 6-O-desulfated heparin. Our data demonstrate a crucial role for the 6-O-sulfo group in the tau-heparin interaction, which to our knowledge has not been reported before.

Novel method for measurement of heparin anticoagulant activity using SPR.

A novel method has been developed for the easy measurement of heparin's anticoagulant activity using surface plasmon resonance. The anticoagulant activity of target heparin was evaluated by measuring the competitive antithrombin III binding of analyte heparin in the solution phase and USP heparin immobilized on chip surface. Heparins, obtained from different animal sources, and low molecular weight heparins were analyzed. The results were reproducible and correlated well with the results of chromogenic assays (correlation coefficient r = 0.98 for anti-Xa and r = 0.94 for anti-IIa). This protocol provides many advantages, significantly minimizing time, cost and the complications of chromogenic assay methods.

Kinetic and Structural Studies of Interactions between Glycosaminoglycans and Langerin.

Langerin, a C-type lectin, is expressed in Langerhans cells. It was reported that langerin binds sulfated glycans, which is an important initial step for its role in blocking human immunodeficiency virus (HIV) transmission by capturing HIV pathogens and mediating their internalization into Birbeck granules for their elimination. It is fundamentally important to understand these interactions at the molecular level for the design of new highly specific therapeutic agents for HIV. Surface plasmon resonance (SPR), which allows for the real-time, direct, quantitative analysis of the label-free molecular interactions, has been used successfully for biophysical characterization of glycosaminoglycan (GAG)-protein interactions. In this study, we report kinetics, structural analysis, and the effects of physiological conditions (e.g., pH, salt concentration, and Ca(2+) and Zn(2+)concentrations) on the interactions between GAGs and langerin using SPR. SPR results revealed that langerin binds to heparin with high affinity (KD ∼ 2.4 nM) and the oligosaccharide length required for the interactions is larger than a tetrasaccharide. This heparin/heparan sulfate-binding protein also interacts with other GAGs, including dermatan sulfate, chondroitin sulfates C-E and KS. In addition, liquid chromatography-mass spectrometry analysis was used to characterize the structure of sulfated glycans that bound to langerin.

Epigallocatechin Gallate (EGCG) Decorating Soybean Seed Ferritin as a Rutin Nanocarrier with Prolonged Release Property in the Gastrointestinal Tract.

The instability and low bioavailability of polyphenols limit their applications in food industries. In this study, epigallocatechin gallate (EGCG) and soybean seed ferritin deprived of iron (apoSSF) were fabricated as a combined double shell material to encapsulate rutin flavonoid molecules. Firstly, due to the reversible assembly characteristics of phytoferritin, rutin was successfully encapsulated within apoSSF to form a ferritin-rutin complex (FR) with an average molar ratio of 28.2: 1 (rutin/ferritin). The encapsulation efficiency and loading capacity of rutin were 18.80 and 2.98 %, respectively. EGCG was then bound to FR to form FR-EGCG composites (FRE), and the binding number of EGCG was 27.30 ± 0.68 with a binding constant K of (2.65 ± 0.11) × 10(4) M(-1). Furthermore, FRE exhibited improved rutin stability, and displayed prolonged release of rutin in simulated gastrointestinal tract fluid, which may be attributed to the external attachment of EGCG to the ferritin cage potentially reducing enzymolysis in GI fluid. In summary, this work demonstrates a novel nanocarrier for stabilization and sustained release of bioactive polyphenols.

Screening and characterization of an anti-inflammatory pectic polysaccharide from Cucurbita moschata Duch.

Pectin polysaccharides have demonstrated diverse biological activities, however, the inflammatory potential of pectin polysaccharides extracted from Cucurbita moschata Duch remains unexplored. This study aims to extract, characterize and evaluate the effects of pumpkin pectin polysaccharide on lipopolysaccharide (LPS)-induced inflammatory response in RAW264.7 cells and dextran sulfate sodium (DSS)-induced colitis in mice, along with its underlying mechanism of action. Initially, we extracted three fractions of pectin polysaccharides from pumpkin and screened them for anti-inflammatory activity in LPS-induced macrophages, identifying CMDP-3a as the most potent anti-inflammatory fraction. Subsequently, CMDP-3a underwent comprehensive characterization through chromatography and spectroscopic analysis, revealing CMDP-3a as an RG-I-HG type pectin polysaccharide with →4)-α-D-GalpA-(1 â†’ and →4)-α-D-GalpA-(1 â†’ 2,4)-α-L-Rhap-(1 â†’ as the main chain. Further, in the LPS-induced RAW264.7 cells model, treatment with CMDP-3a significantly down-regulated the mRNA expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and pro-inflammatory cytokines (IL-1ß, TNF-α, and IL-6) by inhibiting the MAPK and NF-κB signaling pathways. Finally, in a mouse colitis model, CMDP-3a administration obviously inhibited DSS-induced pathological alterations and reduced inflammatory cytokine expressions in the colonic tissues by down-regulating the TLR4/NF-κB and MAPK pathways. These findings provide a molecular basis for the potential application of CMDP-3a in reducing inflammatory responses.

Advances in protein-based microcapsules and their applications: A review.

Due to their excellent emulsification, biocompatibility, and biological activity, proteins are widely used as microcapsule wall materials for encapsulating drugs, natural bioactive substances, essential oils, probiotics, etc. In this review, we summarize the protein-based microcapsules, discussing the types of proteins utilized in microcapsule wall materials, the preparation process, and the main factors that influence their properties. Additionally, we conclude with examples of the vital role of protein-based microcapsules in advancing the food industry from primary processing to deep processing and their potential applications in the biomedical, chemical, and textile industries. However, the low stability and controllability of protein wall materials lead to degraded performance and quality of microcapsules. Protein complexes with polysaccharides or modifications to proteins are often used to improve the thermal instability, pH sensitivity, encapsulation efficiency and antioxidant capacity of microcapsules. In addition, factors such as wall material composition, wall material ratio, the ratio of core to wall material, pH, and preparation method all play critical roles in the preparation and performance of microcapsules. The application area and scope of protein-based microcapsules can be further expanded by optimizing the preparation process and studying the microcapsule release mechanism and control strategy.

Structural characterization of squash polysaccharide and its effect on STZ-induced diabetes mellitus model in MIN6 cells.

A novel natural water-soluble acidic polysaccharide (PWESP-3) was isolated from squash with a molecular mass of 140.519 kDa, which was composed of arabinose (Ara, 35.30 mol%), galactose (Gal, 61.20 mol%), glucose (Glc, 1.80 mol%), and Mannuronic acid (ManA, 1.70 mol%) and contained Araf-(1→, →3)-Araf-(1→, →5)-Araf-(1→, Glcp-(1→, Galp-(1→, →3,5)-Araf-(1→, →2)-Glcp-(1→, →2)-Manp-(1→, →3)-Glcp-(1→, →4)-Galp-(1→, →3)-Galp-(1→, →6)-Galp-(1→, →3,4)-Galp-(1→, →4,6)-Galp-(1→ residues in the backbone. Moreover, the structure of PWESP-3 was identified by NMR spectra. The branch chain was connected to the main chain by the O-3 and O-4 atom of Gal. In addition, the effect of PWESP-3 on STZ-induced type I diabetes mellitus model in MIN6 cells was investigated. The results showed that PWESP-3 can increase the viability and insulin secretion of MIN6 cells and reduce the oxidative stress caused by ROS and NO. Meanwhile, PWESP-3 can also reduce the content of ATP, Ca2+, mitochondrial membrane potential and Caspase-3 activity in MIN6 cells. Furthermore, treatment with PWESP-3 can prevent single or double stranded DNA breaking to form DNA fragments and improve DNA damage in MIN6 cells, thereby avoiding apoptosis. Therefore, the above data highlight that PWESP-3 can improve the function of insulin secretion in STZ-induced MIN6 cells in vitro and can be used as an alternative food supplement to diabetes drugs.

Insight to starch retrogradation through fine structure models: A review.

The retrogradation of starch is crucial for the texture and nutritional value of starchy foods products. There is mounting evidence highlighting the significant impact of starch's fine structures on starch retrogradation. Because of the complexity of starch fine structure, it is a formidable challenge to study the structure-property relationship of starch retrogradation. Several models have been proposed over the years to facilitate understanding of starch structure. In this review, from the perspective of starch models, the intricate structure-property relationship is sorted into the correlation between different types of structural parameters and starch retrogradation performance. Amylopectin B chains with DP 24-36 and DP ≥36 exhibit a higher tendency to form ordered crystalline structures, which promotes starch retrogradation. The chains with DP 6-12 mainly inhibit starch retrogradation. Based on the building block backbone model, a longer inter-block chain length (IB-CL) enhances the realignment and reordering of starch. The mathematical parameterization model reveals a positive correlation between amylopectin medium chains, amylose short chains, and amylose long chains with starch retrogradation. The review is structured according to starch models; this contributes to a clear and comprehensive elucidation of the structure-property relationship, thereby providing valuable references for the selection and utilization of starch.

Comprehensive Assessment of Polysaccharides Extracted from Squash by Subcritical Water under Different Conditions.

The effects of subcritical water microenvironment on the physiochemical properties, antioxidant activity and in vitro digestion of polysaccharides (SWESPs) from squash were investigated. After single-factor experiments, twenty samples were successfully prepared at different extraction temperatures (110, 130, 150, 170 and 190 °C) and extraction times (4, 8, 12 and 16 min). Under a low temperature environment, the whole process was mainly based on the extraction of SWESP. At this time, the color of SWESP was white or light gray and the molecular mass was high. When the temperature was 150 °C, since the extraction and degradation of SWESP reached equilibrium, the maximum extraction rate (18.67%) was reached at 150 °C (12 min). Compared with traditional methods, the yield of squash SWESP extracted by subcritical water was 3-4 times higher and less time consuming. Under high temperature conditions, SWESPs were degraded and their antioxidant capacity and viscosity were reduced. Meanwhile, Maillard and caramelization reactions turned the SWESPs yellow-brown and produced harmful substances. In addition, different SWESPs had different effects on in vitro digestion. In brief, SWESPs prepared under different conditions have different structures and physicochemical properties, allowing the obtainment of the required polysaccharide. Our results show that squash polysaccharides prepared in different subcritical water states had good development potential and application in the food industry.

Structural diversity and physicochemical properties of polysaccharides isolated from pumpkin (Cucurbita moschata) by different methods.

Natural polysaccharides were isolated and purified from Cucurbita moschata by hot water extraction and mild acid-base sequential extraction. Chemical and instrumental studies revealed that hot water-extracted and mild acid-extracted polysaccharides with molecular masses of 48 kDa and 85 kDa were both pectic polysaccharides with homogalacturonan (HG) and rhamnogalacturonan-I (RG-I) domains, while mild acid-extracted polysaccharide was more dominated by branched RG-I with higher contents of galactose (10.59 %) and arabinose (8.08 %). Furthermore, mild acid-extracted polysaccharide exhibited better thickening and emulsifying properties, likely due to its larger molecular mass and higher branching degree. Mild base-extracted polysaccharide with a molecular mass of 18 kDa was a glucan-like polysaccharide. It showed the strongest thermostability and gel behavior among these pumpkin polysaccharides, likely attributed to its unique network structure stabilized by substantial intra/intermolecular hydrogen bonds. This study aimed to establish the structure-property relationships between these structurally diverse pumpkin polysaccharides from different extraction methods and provided theoretical foundations for their targeted application in foods.

The Preparation and Potential Bioactivities of Modified Pectins: A Review.

Pectins are complex polysaccharides that are widely found in plant cells and have a variety of bioactivities. However, the high molecular weights (Mw) and complex structures of natural pectins mean that they are difficult for organisms to absorb and utilize, limiting their beneficial effects. The modification of pectins is considered to be an effective method for improving the structural characteristics and promoting the bioactivities of pectins, and even adding new bioactivities to natural pectins. This article reviews the modification methods, including chemical, physical, and enzymatic methods, for natural pectins from the perspective of their basic information, influencing factors, and product identification. Furthermore, the changes caused by modifications to the bioactivities of pectins are elucidated, including their anti-coagulant, anti-oxidant, anti-tumor, immunomodulatory, anti-inflammatory, hypoglycemic, and anti-bacterial activities and the ability to regulate the intestinal environment. Finally, suggestions and perspectives regarding the development of pectin modification are provided.

Short-Chain Fatty-Acid-Producing Micro-Organisms Regulate the Pancreatic FFA2-Akt/PI3K Signaling Pathway in a Diabetic Rat Model Affected by Pumpkin Oligosaccharides.

Herein, we applied the Illumina MiSeq pyrosequencing platform to amplify the V3-V4 hypervariable regions of the 16 S rRNA gene of the gut microbiota (GM) and a gas chromatograph-mass spectrometer to detect the metabolites after supplementation with pumpkin oligosaccharides (POSs) to determine the metabolic markers and mechanisms in rats with type 2 diabetes (T2D). The POSs alleviated glucolipid metabolism by decreasing the serum low-density lipoprotein (LDL), total cholesterol (TC), and glucose levels. These responses were supported by a shift in the gut microbiota, especially in the butyric-acid-producing communities. Meanwhile, elevated total short-chain fatty acid (SCFA), isovaleric acid, and butyric acid levels were observed after supplementation with POSs. Additionally, this work demonstrated that supplementation with POSs could reduce TNF-α and IL-6 secretion via the FFA2-Akt/PI3K pathway in the pancreas. These results suggested that POSs alleviated T2D by changing the SCFA-producing gut microbiota and SCFA receptor pathways.

Analyzing the Effect of Baking on the Flavor of Defatted Tiger Nut Flour by E-Tongue, E-Nose and HS-SPME-GC-MS.

In order to screen for a proper baking condition to improve flavor, in this experiment, we analyzed the effect of baking on the flavor of defatted tiger nut flour by electronic tongue (E-tongue), electronic nose (E-nose) and headspace solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS). According to E-tongue and E-nose radar plots and principal component analysis (PCA), baking can effectively change the taste and odor of defatted tiger nut flour, and the odors of samples with a baking time of >8 min were significantly different from the original odor of unbaked flour. Moreover, bitterness and astringency increased with longer baking times, and sweetness decreased. HS-SPME-GC-MS detected a total of 68 volatile organic compounds (VOCs) in defatted tiger nut flour at different baking levels, and most VOCs were detected at 8 min of baking. Combined with the relative odor activity value (ROAV) and heat map analysis, the types and contents of key flavor compounds were determined to be most abundant at 8 min of baking; 3-methyl butyraldehyde (fruity and sweet), valeraldehyde (almond), hexanal (grassy and fatty), and 1-dodecanol, were the key flavor compounds. 2,5-dimethyl pyrazine, and pyrazine, 2-ethylalkyl-3,5-dimethyl- added nutty aromas, and 1-nonanal, 2-heptanone, octanoic acid, bicyclo [3.1.1]hept-3-en-2-ol,4,6,6-trimethyl-, and 2-pentylfuran added special floral and fruity aromas.

In vivo pharmacokinetic study of a Cucurbita moschata polysaccharide after oral administration.

Orally administrated is the primary way of pumpkin polysaccharides intake, and this is the way that they elicit their multiple bioactivities. However, little is known about how those orally ingested polysaccharides work, and the pharmacokinetics of pumpkin polysaccharides after orally administrated has not been described. This study aimed to elucidate the pharmacokinetic information (the plasma concentration-time curve, tissue distribution, and excretion profiles) of the pumpkin polysaccharide (PPc) in vivo after oral administration. Results revealed that PPc could enter into the blood and exhibited a relatively long circulation in the blood with a mean residence time (MRT) of 7.45 h and presented a higher aggregation of PPc in the liver and kidneys. To obtain the visualization of the systemic circulation of PPc, in vivo imaging was used with near-infrared fluorescence (NIRF) imaging, which distributed into various tissues with different region of interest (ROI) values after oral administration. The in-depth understanding of oral delivery of PPc in vivo was provided, which will provide the instruction clinical medication and deepen the understanding of bioactivities mechanisms of oral pumpkin polysaccharides.

Antitumor mechanisms of an exopolysaccharide from Lactobacillus fermentum on HT-29 cells and HT-29 tumor-bearing mice.

We have obtained an exopolysaccharide (YL-11 EPS) produced by Lactobacillus fermentum YL-11 isolated from fermented milk and confirmed that it can effectively inhibit colon cancer HT-29 cells proliferation in vitro. The aim of this study is to study anti-colon cancer effect in vivo and its possible mechanisms. Animal assays indicated YL-11 EPS treatment significantly suppressed the growth of HT-29 tumor xenograft without exhibiting obvious negative effects on normal cells. Cell experiments demonstrated YL-11 EPS treatment up regulated the ratio of Bax/Bcl-2 and induced the decrease in mitochondrial membrane potential and improved the expression of cleaved caspases-3 and cleaved PARP proteins, and finally induced HT-29 cells apoptosis, suggesting the involvement of mitochondrial pathway. Moreover, YL-11 EPS can block the PI3K/AKT signaling pathway and arrest the cell cycle in G1-phase to exert its anti-colon cancer activity. Overall, YL-11 EPS can be explored as a potential nutraceutical to prevent colorectal cancer.

Nutritional factors for anemia in pregnancy: A systematic review with meta-analysis.

Background: Anemia in pregnancy is a serious threat to maternal and child health and is a major public health problem. However, the risk factors associated with its incidence are unclear and controversial. Methods: PubMed, Ovid Embase, Web of Science, and Cochrane databases were systematically searched (inception to June 27, 2022). The screening of search results, extraction of relevant data, and evaluation of study quality were performed independently by two reviewers. Results: A total of 51 studies of high quality (NOS score ≥ 7) were included, including 42 cross-sectional studies, six case-control studies, and three cohort studies. Meta-analysis showed that infected parasite, history of malarial attack, tea/coffee after meals, meal frequency ≤ 2 times per day, frequency of eating meat ≤ 1 time per week, frequency of eating vegetables ≤ 3 times per week, multiple pregnancies, multiparous, low household income, no antenatal care, rural residence, diet diversity score ≤ 3, have more than 3 children, history of menorrhagia, underweight, family size ≥ 5, middle upper arm circumference < 23, second trimester, third trimester, birth interval ≤ 2 year were all risk factors for anemia in pregnancy. Conclusions: Prevention of anemia in pregnancy is essential to promote maternal and child health. Sufficient attention should be paid to the above risk factors from the social level and pregnant women's own aspects to reduce the occurrence of anemia in pregnancy. Systematic review registration: https://www.crd.york.ac.uk/prospero/, identifier: CRD42022344937.

Optimization of germination and ultrasonic-assisted extraction for the enhancement of γ-aminobutyric acid in pumpkin seed.

Germination and ultrasonic-assisted extraction (UAE) are economical and effective methods to enhance bioactive compounds in plant seeds. We optimized the germination parameters and UAE parameters by using response surface methodology to maximize the recovery of γ-aminobutyric acid (GABA) in pumpkin seeds. The optimal germination conditions were as follows: soaking the seeds at 28°C for 6 h with 0.2% CaCl2, 3.8 mg/ml monosodium glutamate, and 4.0 mg/ml vitamin B6, then germination at 30°C for 61.6 h. The optimal conditions for UAE were as follows: 1:75 (w/v) material-to-solvent ratio, 220 W ultrasonic power, and ultrasonic treatment at 50°C for 14.4 min, which afforded an extraction yield of 2679 ± 10 mg/100 g. Moreover, the GABA-enhanced extract showed the potential of hypolipidemic effect in type 2 diabetes rats. These results confirmed that a combination of germination and UAE increased the GABA yield from pumpkin seeds and provided a basis for GABA-enhanced production to improve lifestyle-associated diseases.

Homogalacturonan from squash: Characterization and tau-binding pattern of a sulfated derivative.

A pectic polysaccharide (WAP) was isolated from squash and identified as a homogalacturonan with a molecular mass of 83.2 kDa by GPC, monosaccharide composition analysis, FT-IR and NMR spectra. Sulfation modification of WAP was carried out and a sulfated derivative (SWAP) was obtained with a substitution degree of 1.81. The NMR spectrum indicated that the sulfation modification mainly occurred at the C-2 and C-3 positions of galacturonan residues. The binding pattern of SWAP to tau K18 protein was observed in 2D 1H15N HSQC spectra of tau, which resembled the tau-heparin interaction, with R2 domain as the major binding region. These results suggest that SWAP has the potential to act as a heparin mimic to inhibit the transcellular spread of tau; thus natural polysaccharide from squash may be developed into therapies for AD and related tauopathies.