Oral delivery of Eimeria acervulina transfected sequentially with two copies of the VP2 gene induces immunity against infectious bursal disease virus in chickens.

Chicken coccidiosis caused by Eimeria spp. can occur on almost all poultry farms, causing huge economic losses to the industry. Genetically manipulated Eimeria parasites as a vaccine vector to deliver viral antigens have been reported. In our preliminary study, transgenic E. acervulina expressing a VP2 gene (Ea-VP2) of the infectious bursal disease virus (IBDV) demonstrated partial protection against IBDV infection. To enhance immune responses, we aimed to increase the VP2 gene copy number in transgenic E. acervulina. In this study, we used a novel plasmid vector carrying a VP2 gene fused with three flag tags and a red fluorescent reporter gene (mCherry). The vector was introduced into Ea-VP2 sporozoites through nucleofection, leading to the generation of Ea-2VP2. Subsequent analysis revealed a notable escalation in the fluorescent rate, increasing from 0.11 to 95.1% following four consecutive passages facilitated by fluorescent-activated cell sorting. Verification via PCR, Western blot, and immunofluorescence confirmed the successful construction of the Ea-2VP2 population. Despite lower fecundity compared to wild-type E. acervulina, Ea-2VP2 maintained immunogenicity. Our research effectively created a transgenic E. acervulina strain transfected sequentially with two copies of the VP2 gene from IBDV. This modification resulted in an increased humoral immune response after primary immunization in chickens. Additionally, it demonstrated a degree of protection within the bursa against IBDV infection. Future studies will focus on further enhancing immune response levels.

Glucans from Armillaria luteo-virens: Structural Characterization and In Vivo Immunomodulatory Investigation under Different Administration Routes.

Polysaccharides fromArmillaria luteo-virens (ALP) were investigated for structural characterization and immunomodulatory activities. Three fractions (ALP-1, ALP-2, and ALP-3) were obtained with the yield of 2.4, 3.7, and 3.0 wt %, respectively. ALP-1 was proposed as a ß-(1 → 3)(1 → 6)-glucan with a triple-helix conformation; ALP-2 and ALP-3 were both identified as α-(1 → 4)(1 → 6)-glucan differing in their Mw and branching degree with a spherical conformation. The in vitro digestibility experiment and in vivo experiments using cyclophosphamide (CY)-treated mice demonstrated that intraperitoneal injection of α-glucan (1 mg·kg-1·day-1) and intragastric gavage of ß-glucan (10 mg·kg-1·day-1) both effectively restored the decrease in body weight, immune organ indexes, immune cell activities, serum immune marker levels, colonic short-chain fatty acids (SCFA) levels, and Bacteroidetes/Firmicutes ratio in immunosuppression mice. This study provides novel insights into the immunomodulatory activity of α- and ß-glucans under different administration routes, thereby promoting their application in both food and pharmaceutical areas.

Unveiling the breadmaking transformation: Structural and functional insights into Arabinoxylan.

To understand the changes in arabinoxylan (AX) during breadmaking, multi-step enzyme digestion was conducted to re-extract arabinoxylan (AX-B) from AX-fortified bread. Their structural changes were compared using HPSEC, HPAEC, FT-IR, methylation analysis, and 1H NMR analysis; their properties changes in terms of enzymatic inhibition activities and in vitro fermentability against gut microbiota were also compared. Results showed that AX-B contained a higher portion of covalently linked protein while the molecular weight was reduced significantly after breadmaking process (from 677.1 kDa to 15.6 kDa); the structural complexity of AX-B in terms of the degree of branching was increased; the inhibition activity against α-amylase (76.81 % vs 73.89 % at 4 mg/mL) and α-glucosidase (64.43 % vs 58.08 % at 4 mg/mL) was improved; the AX-B group produced a higher short-chain fatty acids concentration than AX (54.68 ± 7.86 mmol/L vs 44.03 ± 4.10 mmol/L). This study provides novel knowledge regarding the structural and properties changes of arabinoxylan throughout breadmaking, which help to predict the health benefits of fibre-fortified bread and achieve precision nutrition.

Disruption of UDP-galactopyranose mutase expression: A novel strategy for regulation of galactomannan biosynthesis and monascus pigments secretion in Monascus purpureus M9.

The impact of the cell wall structure of Monascus purpureus M9 on the secretion of extracellular monascus pigments (exMPs) was investigated. To modify the cell wall structure, UDP-galactopyranose mutase (GlfA) was knocked out using Agrobacterium-mediated transformation method, leading to a significant reduction in the Galf-based polysaccharide within the cell wall. Changes in mycelium morphology, sporogenesis, and the expression of relevant genes in M9 were also observed following the mutation. Regarding MPs secretion, a notable increase was observed in six types of exMPs (R1, R2, Y1, Y2, O1 and O2). Specifically, these exMPs exhibited enhancement of 1.33, 1.59, 0.8, 2.45, 2.89 and 4.03 times, respectively, compared to the wild-type strain. These findings suggest that the alteration of the cell wall structure could selectively influence the secretion of MPs in M9. The underlying mechanisms were also discussed. This research contributes new insights into the regulation of the synthesis and secretion of MPs in Monascus spp..

Hyperoside Prevents Aß42-Induced Neurotoxicity in PC12 Cells and Caenorhabditis elegans.

Traditional Chinese medicines such as hyperoside-rich Acanthopanax senticosus and Crataegus pinnatifida have been confirmed to exhibit anti-oxidative stress properties. Hyperoside, the main ingredient of numerous antioxidant herbs, may have the ability to postpone the onset of neurodegenerative diseases. This study investigates the possible therapeutic mechanism of hyperoside as a natural antioxidant against Alzheimer's disease (AD) in Caenorhabditis elegans and PC12 cells. Specifically, hyperoside reduced reactive oxygen species (ROS) level and Aß42-induced neurotoxicity in C. elegans worms. Meanwhile, hyperoside reduced ROS production and increased mitochondrial membrane potentialin Aß42-induced PC12 cells, which possibly due to the increase of antioxidant enzymes activity and the diminution of malondialdehyde levels. Hoechst 33,342 staining and flow cytometry analysis results suggested that hyperoside reverses cell apoptosis. Network pharmacology predicts potentially relevant hyperoside targets and pathways in AD therapy. As anticipated, hyperoside reversed Aß42-stimulated downregulation of the PI3K/Akt/Nrf2/HO-1. The PI3K inhibitor LY294002 partially abolished the protective capability of hyperoside. The results of molecular docking further indicated that the PI3K/Akt pathways may be involved in the protection of Aß42-induced PC12 cells by hyperoside treatment. The study provides theoretical information for research and development of hyperoside as an antioxidant dietary supplement.

Structural, rheological and functional properties of ultrasonic treated xanthan gums.

Xanthan gum can improve the freeze-thaw stability of frozen foods. However, the high viscosity and long hydration time of xanthan gum limits its application. In this study, ultrasound was employed to reduce the viscosity of xanthan gum, and the effect of ultrasound on its physicochemical, structural, and rheological properties was investigated using High-performance size-exclusion chromatography (HPSEC), ion chromatograph, methylation analysis, 1H NMR, rheometer, etc.. The application of ultrasonic-treated xanthan gum was evaluated in frozen dough bread. Results showed that the molecular weight of xanthan gum was reduced significantly by ultrasonication (from 3.0 × 107 Da to 1.4 × 106 Da), and the monosaccharide compositions and linkage patterns of sugar residues were altered. Results revealed that ultrasonication treatment mainly broke the molecular backbone at a lower intensity, then mainly broke the side chains with increasing intensity, which significantly reduced the apparent viscosity and viscoelastic properties of xanthan gum. The results of specific volume and hardness showed that the bread containing low molecular weight xanthan gum was of better quality. Overall, this work offers a theoretical foundation for broadening the application of xanthan gum and improving its performance in frozen dough.

(Epi)catechin damage effects on the development of mouse intestinal epithelial structure through the PERK-eIF2α-ATF4-CHOP pathway.

As powerful bioactive compounds found in a variety of plant-based foods, (epi)catechins have been identified to be associated with an abundant array of health benefits. While their adverse impacts have also been gaining increasing attention, their intestinal impact is still unclear. In this study, intestinal organoids were used as an in vitro model to analyze the effects of four (epi)catechins on the development of the intestinal epithelial structure. Morphological characteristics, oxidative stress, and endoplasmic reticulum (ER) stress assays with (epi)catechins treatment showed that (epi)catechins promoted intestinal epithelial apoptosis and stress response. These effects had dose-dependent and structural differences (EGCG > EGC > ECG > EC). Furthermore, GSK2606414, a protein kinase RNA (PKR)-like ER kinase (PERK) pathway inhibitor, confirmed that the PERK-eukaryotic translation initiation factor 2α (eIF2α)-activating transcription factor 4 (ATF4)-C/EBP-homologous protein (CHOP) pathway is closely related to the damage. In addition, the results for the intestinal inflammatory mouse model further verified that (epi)catechins significantly delayed intestinal repair. Taken together, these findings revealed that overdosage of (epi)catechins has damage potential on the intestinal epithelium and may increase the risk of intestinal damage.

Can environmentally friendly technology help China to achieve a carbon neutrality target by 2060? An asymmetrical based study in China.

Climate change-related environmental challenges are prompting an increasing number of countries to set carbon-neutral targets. Since 2007, China has pursued numerous initiatives to attain carbon neutrality by 2060, including increasing the percentage of non-fossil energy, developing zero-emission and low-emission technologies, and taking actions that reduce CO2 emissions or boost carbon sinks. As a result, utilizing quarterly data from 2008/Q1 to 2021/Q4, and applying the nonlinear autoregressive distributed lag (NARDL) approach, this study evaluates the effectiveness of the measures taken by China to improve the ecological situation. The results of the study show that the measures enacted to reduce CO2 emissions did not accomplish their ultimate purpose. Specifically: (i) high-speed railways and new energy vehicles do not improve the environment in the long run; (ii) investments and patents in the energy sector, as well as low-carbon sources, will degrade the environment; (iii) only investments in the treatment of environmental pollution will improve the ecological situation. Various policy implications are suggested based on the empirical results in order to attain environmental sustainability.

Insight into the phylogeny and metabolic divergence of Monascus species (M. pilosus, M. ruber, and M. purpureus) at the genome level.

Background: Species of the genus Monascus are economically important and widely used in the production of food colorants and monacolin K. However, they have also been known to produce the mycotoxin citrinin. Currently, taxonomic knowledge of this species at the genome level is insufficient. Methods: This study presents genomic similarity analyses through the analysis of the average nucleic acid identity of the genomic sequence and the whole genome alignment. Subsequently, the study constructed a pangenome of Monascus by reannotating all the genomes and identifying a total of 9,539 orthologous gene families. Two phylogenetic trees were constructed based on 4,589 single copy orthologous protein sequences and all the 5,565 orthologous proteins, respectively. In addition, carbohydrate active enzymes, secretome, allergic proteins, as well as secondary metabolite gene clusters were compared among the included 15 Monascus strains. Results: The results clearly revealed a high homology between M. pilosus and M. ruber, and their distant relationship with M. purpureus. Accordingly, all the included 15 Monascus strains should be classified into two distinctly evolutionary clades, namely the M. purpureus clade and the M. pilosus-M. ruber clade. Moreover, gene ontology enrichment showed that the M. pilosus-M. ruber clade had more orthologous genes involved with environmental adaptation than the M. purpureus clade. Compared to Aspergillus oryzae, all the Monascus species had a substantial gene loss of carbohydrate active enzymes. Potential allergenic and fungal virulence factor proteins were also found in the secretome of Monascus. Furthermore, this study identified the pigment synthesis gene clusters present in all included genomes, but with multiple nonessential genes inserted in the gene cluster of M. pilosus and M. ruber compared to M. purpureus. The citrinin gene cluster was found to be intact and highly conserved only among M. purpureus genomes. The monacolin K gene cluster was found only in the genomes of M. pilosus and M. ruber, but the sequence was more conserved in M. ruber. Conclusion: This study provides a paradigm for phylogenetic analysis of the genus Monascus, and it is believed that this report will lead to a better understanding of these food microorganisms in terms of classification, metabolic differentiation, and safety.

Hyperoside inhibits pancreatic lipase activity in vitro and reduces fat accumulation in vivo.

Hyperoside, the main component of many anti-obesity plants, might exhibit a lipase inhibition effect to reduce fat accumulation. The anti-obesity effect of hyperoside was investigated by studying its inhibitory effect and mechanism on pancreatic lipase in vitro and evaluating its ability to reduce lipid accumulation in vivo. Hyperoside is a mixed-type inhibitor of lipase with an IC50 of 0.67 ± 0.02 mmol L-in vitro. Hyperoside changed the secondary conformation of lipase, increased the α-helix content, and changed the microenvironment of lipase through static quenching. The interaction between hyperoside and lipase results from a strong binding spontaneous exothermic reaction, mainly through hydrogen bonding, van der Waals force and electrostatic force. Hyperoside protected hepatic lipid accumulation and adipose tissue hypertrophy and reduced the expression of inflammatory factors in high-fat diet-induced rats. Moreover, hyperoside had a good inhibitory effect on lipase activity in serum and increased fecal fat excretion, thereby reducing lipid absorption in vivo. This study provides theoretical support for the research and development of hyperoside in fat-reducing functional foods.

Binding of ankaflavin with bovine serum albumin (BSA) in the presence of carrageenan and protective effects of Monascus yellow pigments against oxidative damage to BSA after forming a complex with carrageenan.

Ankaflavin (AK) is a typical yellow pigment extracted from Monascus-fermented rice with several biological effects; however, its solubility is poor. Thus, research studies of the delivery systems of AK, especially those constructed from protein-polysaccharide complexes, have attracted considerable attention. However, the interactions that exist in the system have rarely been investigated. This work focused on the interactions between AK and bovine serum albumin (BSA) as well as the influence of carrageenan (Car) on the binding of AK to BSA. Results revealed that the quenching of BSA by AK involved the static quenching mechanism. The formed BSA-AK complexes were mainly maintained by hydrophobic forces and AK was located within the hydrophobic cavity of BSA. Compared to free AK or AK only complexed with BSA, a higher absorption intensity of AK was observed for the formed BSA-AK-Car complexes, indicating changes in the microenvironment of AK. This was confirmed by the increase in the α-helix content of BSA after the formation of BSA-AK-Car complexes. Hydrogen bond, van der Waals, and electrostatic interactions were verified to be the primary forces preserving the BSA-AK-Car complexes. Moreover, the antioxidant potential of Monascus-fermented products rich in AK (denoted as Mps), namely BSA-Mps and BSA-Mps-Car was evaluated. The antioxidant activity of Mps was negatively impacted by BSA, while the addition of Car could enhance the antioxidant capacity of BSA-Mps-Car complexes. Meanwhile, Mps showed a protective effect against free radical-induced oxidation damage to BSA, and Car could further improve this effect.

Novel magnetic graphoxide/biochar composite derived from tea for multiple SAs and QNs antibiotics removal in water.

Antibiotics pollution is an urgent public health issue. Biochar is a kind of promising composite for removal antibiotic in aqueous environment. In this study, a novel magnetic graphoxide/biochar composite (mGO/TBC) was synthesized by simple impregnation method and used as an efficient and recyclable persulfate (PS) activator for degradation and removal of sulfonamides (SAs) and quinolones (QNs) antibiotics. Based on the synergism pre-adsorption and degradation between graphoxide and biochar, the removal rates of mGO/TBC on sarafloxacin hydrochloride, sulfadimethoxine, sulfapyridine, sulfadoxine, sulfamonomethoxine, sulfachloropyridazine, enrofloxacin, and ciprofloxacin were increased above 95%. Moreover, the mGO/TBC could be reused at least seven times after degradation-recovery cycles. Quenching experiment and ESR analysis proved that 1O2, •OH, and SO4•- from mGO/TBC/PS system were the primary oxidation active species to degrade SAs and QNs. It is a promising substrate for antibiotic bioremediation with good application prospects.

Black rice anthocyanin extract enhances the antioxidant capacity in PC12 cells and improves the lifespan by activating IIS pathway in Caenorhabditis elegans.

Black rice is rich in anthocyanins, and the antioxidant effect of anthocyanins is recognized by consumers. The aim of this study was to identify the molecular mechanisms underlying the antioxidant activity of black rice anthocyanin extract (BRAE) in PC12 cells and C. elegans. Results showed that BRAE increased antioxidant enzyme activities and decreased the accumulation of reactive oxygen species (ROS) and malondialdehyde in PC12 cells induced by H2O2. Meanwhile, BRAE extended the lifespan, enhanced resistance to stress, increased antioxidant enzyme activities, and reduced lipofuscin, ROS, and MDA accumulation in wild-type C. elegans. The polyQ40 aggregation in AM141, paralysis in CL4176, and chemotaxis deficit in CL2355 were alleviated by BRAE administration. BRAE downregulated the mRNA expression of age-1 and daf-2, while upregulated the daf-16 mRNA level and SOD-3, CTL-1, and GST-4 protein expression. Mutational lifespan tests and molecular docking showed that insulin pathway might be involved in the mechanism of lifespan extension.

Acetyl-glucomannan from Dendrobium officinale: Structural modification and immunomodulatory activities.

To understand the mechanisms of immunomodulatory effect, Dendrobium Officinale polysaccharides (DOP) were treated by ultrasound and mild base separately to generate fractions of various weight-average molecular weight (Mw) and degrees of acetylation (DA). The structural features, conformational properties, functional properties and immunomodulatory activities of original and modified DOPs were investigated. Ultrasonic treatment decreased the Mw and apparent viscosity and improved the water solubility of DOP. Mild base treatment remarkably reduced the DA and the water solubility, while the overall apparent viscosity was increased. Conformational analysis by triple-detector high performance size-exclusion chromatography showed that the molecular chain of DOP turned more compact coil conformation with decreased DA. Results from the macrophages RAW 264.7 analysis showed that samples sonicated for 200 min (Mw 34.2 kDa) showed the highest immune-regulation effects. However, the immunomodulatory effects of the samples after de-acetylation were all compromised compared to the original DOP. This study inspires further research to establish the structural-immunomodulatory relationships, which promote the application of DOP in both the food and medicine fields.

Therapeutic potential of non-starch polysaccharides on type 2 diabetes: from hypoglycemic mechanism to clinical trials.

Non-starch polysaccharides (NSPs) have been reported to exert therapeutic potential on managing type 2 diabetes mellitus (T2DM). Various mechanisms have been proposed; however, several studies have not considered the correlations between the anti-T2DM activity of NSPs and their molecular structure. Moreover, the current understanding of the role of NSPs in T2DM treatment is mainly based on in vitro and in vivo data, and more human clinical trials are required to verify the actual efficacy in treating T2DM. The related anti-T2DM mechanisms of NSPs, including regulating insulin action, promoting glucose metabolism and regulating postprandial blood glucose level, anti-inflammatory and regulating gut microbiota (GM), are reviewed. The structure-function relationships are summarized, and the relationships between NSPs structure and anti-T2DM activity from clinical trials are highlighted. The development of anti-T2DM medication or dietary supplements of NSPs could be promoted with an in-depth understanding of the multiple regulatory effects in the treatment/intervention of T2DM.

The impact of renewable and non-renewable energy consumption on carbon emission intensity in China: Fresh evidence from novel dynamic ARDL simulations.

Given the dire state of global warming, it is critical to investigate the elements that influence carbon emissions intensity and to precisely monitor progress in carbon emissions intensity growth in order to meet the aim of lowering CO2 emissions. This research explores the association among renewable energy and non-renewable energy consumption, the urban population, research and development expenditure, technological innovation, and carbon emissions intensity in China using annual time series data over the period 1990 to 2019. The Dynamic ARDL simulation technique was utilized to investigate the long-run and short-run correlations between renewable and non-renewable energy consumption and CEI. The results suggest that there is strong evidence of a long-run correlation between the variables. The findings indicate that in the long-run, renewable energy and non-renewable energy consumption, and research and development expenditure have a positive influence on CEI by 0.27%, 0.75%, and 0.21%, whereas the urban population has a negative influence by 2.31%, respectively. However, the urban population and technological innovation have positively affected the short-run CEI by 12.17% and 0.23%, respectively. Policies should focus on continuous investment in renewable energy sources, clean energy innovation, improving energy efficiency, forest restoration, and carbon neutrality initiatives to lessen the environmental extreme pressure associated with CO2 emissions.

Conformational Properties of Flaxseed Rhamnogalacturonan-I and Correlation between Primary Structure and Conformation.

The pectic polysaccharides extracted from flaxseed (Linum usitatissiumum L.) mucilage and kernel were characterized as rhamnogalacturonan-I (RG-I). In this study, the conformational characteristics of RG-I fractions from flaxseed mucilage and kernel were investigated, using a Brookhaven multi-angle light scattering instrument (batch mode) and a high-performance size exclusion chromatography (HPSEC) system coupled with Viscotek tetra-detectors (flow mode). The Mw of flaxseed mucilage RG-I (FM-R) was 285 kDa, and the structure-sensitive parameter (ρ) value of FM-R was calculated as 1.3, suggesting that the FM-R molecule had a star-like conformation. The Mw of flaxseed kernel RG-I (FK-R) was 550 kDa, and the structure-sensitive parameter (ρ) values ranged from 0.90 to 1.21, suggesting a sphere to star-like conformation with relatively higher segment density. The correlation between the primary structure and conformation of RG-I was further discussed to better understand the structure-function relationship, which helps the scale-up applications of pectins in food, pharmaceutical, or cosmetic industries.

In Vitro Fermentability of Soybean Oligosaccharides from Wastewater of Tofu Production.

Soybean oligosaccharides (SBOS) isolated from wastewater of tofu production were studied in terms of their structural characteristics and in vitro fermentation by human fecal inocula. Three sub-fractions named Z1 (14%), Z2 (13%), and Z3 (17%) were obtained by Sephadex G-15 column separation. Z1 contained mainly stachyose; Z2 and Z3 contained stachyose, raffinose, and sucrose with different relative percentages. The in vitro batch fermentation model of human intestinal bacteria including 0, 12, 24, and 48 h was used to investigate the fermentation characteristics of SBOS. According to the results, during the fermentation process, the molecular weight of oligosaccharides decreased significantly with increasing fermentation time, indicating that oligosaccharides could be utilized and degraded by the colonic microbiota. Furthermore, SBOS could significantly promote the production of short-chain fatty acids (SCFAs), especially acetic, propionic, and butyric acids. SBOS increased the abundance of Firmicutes, while that of Proteobacteria was decreased. Additionally, SBOS could promote the proliferation of Dialister, Bacteroides, and Akkermansia at the genus level. Therefore, SBOS can be potentially used as prebiotic promoting gut health.

Polyphenol-Polysaccharide Complex: Preparation, Characterization, and Potential Utilization in Food and Health.

Polysaccharides and polyphenols coexist in many plant-based food products. Polyphenol-polysaccharide interactions may affect the physicochemical, functional, and physiological properties, such as digestibility, bioavailability, and stability, of plant-based foods. In this review, the interactions (physically or covalently linked) between the selected polysaccharides and polyphenols are summarized. The preparation and structural characterization of the polyphenol-polysaccharide conjugates, their structural-interaction relationships, and the effects of the interactions on functional and physiological properties of the polyphenol and polysaccharide molecules are reviewed. Moreover, potential applications of polyphenol-polysaccharide conjugates are discussed. This review aids in a comprehensive understanding of the synthetic strategy, beneficial bioactivity, and potential application of polyphenol-polysaccharide complexes.

Isotherm, kinetics, and adsorption mechanism studies of diethylenetriaminepentaacetic acid-modified banana/pomegranate peels as efficient adsorbents for removing Cd(II) and Ni(II) from aqueous solution.

Two novel absorbents were synthesized for the first time by banana and pomegranate peels using diethylenetriaminepentaacetic acid (DTPA) modification to eliminate Cd(II) and Ni(II) of sewage. The DTPA-modified peels performed significantly higher adsorption capacity than unmodified materials. The maximum adsorption capacities of DTPA-modified banana/pomegranate peel were 46.729/46.296 mg/g for Cd(II), and 29.240/16.611 mg/g for Ni(II). Adsorption isotherm and kinetics models were simulated to determine their removal efficiency and potential for recovery of these two heavy metals. As the results, the adsorption reached equilibrium within 5 min and was well described by the pseudo-second order model and Langmuir isotherm. The surface morphology analysis of the synthetic materials by Scanning Electron Microscopy-Energy Dispersive X-ray spectroscopy, Fourier Transform Infrared spectroscopy, and X-ray Photoelectron Spectroscopy, implied that ion exchange, complexation, and physical adsorption may together contribute to Cd(II) and Ni(II) loading on DTPA-modified peels. This study demonstrates the feasibility of waste peels as cost-efficient bio-absorbents to remove Cd(II) and Ni(II) in sewage systems, and discovers potential adsorption mechanism of efficiency improvements after DTPA modification.