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Cordycepin, an adenosine analog, exhibits diverse bioactivities and holds significant potential for applications in healthcare and agriculture. Fungi of the genus Cordyceps, such as Cordyceps militaris, can naturally produce cordycepin. Current sources of cordycepin primarily involve extraction from fruiting bodies or isolation from liquid fermentation using C. militaris, presenting challenges such as low production intensity, complex separation and purification systems, and high production costs, limiting industrial feasibility. Recent advancements have witnessed the utilization of various fungal chassis cells to successfully engineer heterologous biosynthetic platforms for cordycepin, such as Saccharomyces cerevisiae and unconventional yeasts, offering advantages of high yield, short fermentation cycles, and a broad substrate spectrum. This mini review summarizes the biosynthetic pathways of cordycepin and focused on the comparison of the characteristics, advantages, current performance and prospects for the microbial cell factories, analyzing potential targets for metabolic pathway modification and giving strategies in both genetic engineering and process engineering to enhance production intensity. The mini review particularly emphasizes the crucial role of chassis cell stress tolerance to the toxic product in determining cordycepin yield and highlights the urgent need for high-throughput screening methods for high-yield strains.

Abstract It is difficult to exhaustively screen all possible DNA binding sequences for a given transcription factor (TF). Here, we developed the KaScape method, in which TFs bind to all possible DNA sequences in the same DNA pool where DNA sequences are prepared by randomized oligo synthesis and the random length can be adjusted to a length such as 4, 5, 6, or 7. After separating bound from unbound double-stranded DNAs (dsDNAs), their sequences are determined by next-generation sequencing. To demonstrate the relative binding affinities of all possible DNA sequences determined by KaScape, we developed three-dimensional KaScape viewing software based on a K-mer graph. We applied KaScape to 12 plant TF family AtWRKY proteins and found that all AtWRKY proteins bound to the core sequence GAC with similar profiles. KaScape can detect not only binding sequences consistent with the consensus W-box “TTGAC(C/T)” but also other sequences with weak affinity. KaScape provides a high-throughput, easy-to-operate, sensitive, and exhaustive method for quantitatively characterizing the relative binding strength of a TF with all possible binding sequences, allowing us to comprehensively characterize the specificity and affinity landscape of transcription factors, particularly for moderate- and low-affinity binding sites.

In order to improve the stability and bioavailability, hawthorn polyphenols microcapsules were prepared by spray drying using β-cyclodextrin. Whey protein isolate and gum Arabic as concentrations wall materials, and hawthorn polyphenols as the core material. The particle size, microscopic appearance, stability, sustained release and antioxidant properties of the microcapsules were analyzed. The results showed that core-to-wall-material ratio was 1:3 (v/v), air inlet temperature was 165 ℃ and emulsification time was 11.4 min as the optimum preparation process. Under all conditions optimized, the encapsulation rate was 94.45%±1.03% and the drug loading was 17.99%±0.20% of hawthorn polyphenol microcapsules, which average particle size was 6.77 μm, the microcapsules gained complete particles and spherical structure. The microcapsules showed well stability in gastric juice and well release performance in intestinal juice. The release rates of hawthorn polyphenols and hawthorn polyphenol microcapsules after 2 h of simulated gastric juice digestion were 13.38% and 9.75%, respectively, and 6 h after intestinal juice digestion were 95.99% and 72.53%, respectively. The oxidations resistance and stabilities were improved obviously after encapsulation. The IC50 values of hawthorn polyphenols for DPPH, OH, O2− free radical scavenging rates were 12.669 μg/mL and 1.131, 0.581 mg/mL, respectively, and the hawthorn polyphenol microcapsules were 0.939 μg/mL and 0.129, 0.546 mg/mL, respectively. After 10 days of light exposure, the retention rates of hawthorn polyphenols and hawthorn polyphenols microcapsules were 64.99% and 86.74%, respectively. The retention rates at 80°C were 70.77% and 88.77%, respectively, and the retention rates of hawthorn polyphenols and hawthorn polyphenols microcapsules stored without packaging for 28 days were 73.48% and 91.15%, respectively. These above results indicated that microcapsule technology could improve the stability, sustained release and antioxidant activities of hawthorn polyphenols, and could be used as an effective method to improve the processing stability of hawthorn polyphenols.

The goal of the research was to find alternative protein sources for animal farming that are efficient and cost-effective. The researchers focused on distillers dried grains with solubles (DDGS), a co-product of bioethanol production that is rich in protein but limited in its use as a feed ingredient due to its high non-starch polysaccharides (NSPs) content, particularly for monogastric animals. The analysis of the Paenibacillus pabuli E1 genome revealed the presence of 372 genes related to Carbohydrate-Active enzymes (CAZymes), with 98 of them associated with NSPs degrading enzymes that target cellulose, hemicellulose, and pectin. Additionally, although lignin is not an NSP, two lignin-degrading enzymes were also examined because the presence of lignin alongside NSPs can hinder the catalytic effect of enzymes on NSPs. To confirm the catalytic ability of the degrading enzymes, an in vitro enzyme activity assay was conducted. The results demonstrated that the endoglucanase activity reached 5.37 U/mL, while beta-glucosidase activity was 4.60 U/mL. The filter paper experiments did not detect any reducing sugars. The xylanase and beta-xylosidase activities were measured at 11.05 and 4.16 U/mL, respectively. Furthermore, the pectate lyase and pectin lyase activities were found to be 8.19 and 2.43 U/mL, respectively. The activities of laccase and MnP were determined as 1.87 and 4.30 U/mL, respectively. The researchers also investigated the effect of P. pabuli E1 on the degradation of NSPs through the solid-state fermentation of DDGS. After 240 h of fermentation, the results showed degradation rates of 11.86% for hemicellulose, 11.53% for cellulose, and 8.78% for lignin. Moreover, the crude protein (CP) content of DDGS increased from 26.59% to 30.59%. In conclusion, this study demonstrated that P. pabuli E1 possesses various potential NSPs degrading enzymes that can effectively eliminate NSPs in feed. This process improves the quality and availability of the feed, which is important for animal farming as it seeks alternative protein sources to replace traditional nutrients.

A feeding trial was carried out for 30 days to investigate the impacts of dietary immunity induced A. pernyi pupae powder (IPP) and A. pernyi cecropin (AC) on the growth, immune-related enzyme activities and disease resistance in Yesso scallop, Patinopecten yessoensis. A total of 320 scallops, with an average weight of 54.75 ± 0.07 g were randomly divided into four treatment groups: control group (provided with a diet of spirulina powder and microalgae), groups with basic diet supplemented with A. pernyi pupae powder (NPP, 2 % of the final concentration), immunity induced A. pernyi pupae powder (IPP, 2 % of the final concentration), and A. pernyi cecropin (AC, 2 % of the final concentration), respectively. The results showed that the specific growth rate (SGR), lysozyme (LSZ), acid phosphatase (ACP) and superoxide dismutase (SOD) were significantly higher in the scallops fed with AC than in the other groups (P

Zearalenone (ZEA) is present worldwide as a serious contaminant of food and feed and causes male reproductive toxicity. The implication of paraptosis, which is a nonclassical paradigm of cell death, is unclear in ZEA-induced male reproductive disorders. In this study, the toxic effects of ZEA on the blood-testis barrier (BTB) and the related mechanisms of paraptosis were detected in goats. ZEA exposure, in vivo, caused a significant decrease in spermatozoon quality, the destruction of seminiferous tubules, and damage to the BTB integrity. Furthermore, ZEA exposure to Sertoli cells (SCs) in vitro showed similar dysfunction in structure and barrier function. Importantly, the formation of massive cytoplasmic vacuoles in ZEA-treated SCs corresponded to the highly swollen and dilative endoplasmic reticulum (ER), and paraptosis inhibition significantly alleviated ZEA-induced SC death and vacuolization, which indicated the important contribution of paraptosis in ZEA-induced BTB damage. Meanwhile, the expression of ER stress marker proteins was increased after ZEA treatment but decreased under the inhibition of paraptosis. The vacuole formation and SC death, induced by ZEA, were remarkably blocked by ER stress inhibition. In conclusion, these results facilitate the exploration of the mechanisms of the SC paraptosis involved in ZEA-induced BTB damage in goats.

Abstract Background ANGPTL8, an important regulator of lipid metabolism, was recently proven to have additional intracellular and receptor-mediated functions. This study aimed to investigate circulating levels of ANGPTL8 and its potential association with the risk of kidney function decline in a cohort study. Methods We analysed 2,311 participants aged 40 years old and older from the China Cardiometabolic Disease and Cancer Cohort (4C) Study. Kidney function decline was defined as an estimated glomerular filtration rate (eGFR) less than 60 mL per minute per 1.73 m2 of body surface area, a decrease in eGFR of ≥ 30% from baseline, chronic kidney disease (CKD)-related hospitalization or death, or end-stage renal disease. The association between baseline ANGPTL8 levels and kidney function decline was assessed using multivariable-adjusted Cox proportional hazards models, and inverse possibility of treatment weight (IPTW) was utilized to prevent overfitting. Results There were 136 (5.9%) cases of kidney function decline over a median of 3.8 years of follow-up. We found that serum ANGPTL8 levels at baseline were elevated in individuals with kidney function decline compared to those without kidney function decline during follow-up (718.42 ± 378.17 vs. 522.04 ± 283.07 pg/mL, p

BackgroundPreterm premature rupture of membranes (PPROM) is a common pregnancy complication. Yet, the main cause of PPROM remains poorly understood. In this study, we used 16S rRNA gene sequencing technology to identify the differences in vaginal microbiota between pregnant women with PPROM and those who delivered at term.MethodsVaginal samples were collected from 48 patients with PPROM and 54 age- and gestational age-matched pregnant women who delivered at term (controls). The vaginal microbiota of the two groups was compared using 16S rRNA gene sequencing of the V3-V4 regions.ResultsThe vaginal microbial composition of the PPROM group was significantly different from that of the control group. Our results showed that the diversity of vaginal microbiota in patients with PPROM increased compared with controls. The relative abundance of Lactobacillus iners, Gardnerella vaginalis, Prevotella bivia, Ochrobactrum sp., Prevotella timonensis, and Ureaplasma parvum were more abundant in patients with PPROM, while Lactobacillus crispatus and Lactobacillus gasseri were more abundant in controls. Ochrobactrum sp., Prevotella timonensis, and Gardnerella vaginalis, could serve as biomarkers for PPROM. Finally, we proposed several metabolic pathways, including PWY-6339, PWY-6992, and PWY-7295.ConclusionPPROM is characterized by vaginal microbial dysbiosis. The dysbiotic vaginal microbiota signatures in patients with PPROM include a higher bacterial diversity, decreased autochthonous bacteria, and increased pathogenic bacteria. These results may be beneficial for developing biomarkers for screening and early diagnosis of PPROM and may provide effective preventative treatments.

Summary The emergence and rapid spread of multidrug‐resistant bacteria has induced intense research for novel therapeutic approaches. In this study, the Acinetobacter baumannii bacteriophage D2 (vB_AbaP_D2) was isolated, characterized and sequenced. The endolysin of bacteriophage D2, namely Abtn‐4, contains an amphipathic helix and was found to have activity against multidrug‐resistant Gram‐negative strains. By more than 3 log units, A. baumannii were killed by Abtn‐4 (5 µM) in 2 h. In absence of outer membrane permeabilizers, Abtn‐4 exhibited broad antimicrobial activity against several Gram‐positive and Gram‐negative bacteria, such as Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumonia, Enterococcus and Salmonella. Furthermore, Abtn‐4 had the ability to reduce biofilm formation. Interestingly, Abtn‐4 showed antimicrobial activity against phage‐resistant bacterial mutants. Based on these results, endolysin Abtn‐4 may be a promising candidate therapeutic agent for multidrug‐resistant bacterial infections.

Abstract Background Angiopoietin-like protein 8 (ANGPTL8), an important regulator of lipid metabolism, is increased in diabetes and is associated with insulin resistance. However, the role of ANGPTL8 in the outcomes of diabetic patients remains unclear. This study aimed to investigate circulating levels of ANGPTL8 in participants with and without diabetes and its potential associations with clinical outcomes in a 5 year cohort study. Methods Propensity-matched cohorts of subjects with and without diabetes from the Risk Evaluation of Cancers in Chinese Diabetic Individuals: A longitudinal (REACTION) study were generated on the basis of age, sex and body mass index at baseline. The primary outcome was all-cause mortality. The secondary outcomes were a composite of new-onset major adverse cardiovascular events, hospitalization for heart failure, and renal dysfunction (eGFR

Objective. This study explored the colorectal cancer exosome lncRNA prostate cancer associated transcript 1– (PCAT1) mediated circulating tumors and the mechanism of cell colorectal cancer liver metastasis. Methods. Exosomes were extracted from the primary colorectal cancer (CRC) cell lines HCT116 and SW480 and cultured with T84 and human umbilical vein endothelial (HUVE) cells. The expression of PCAT1 and miR-329-3p was detected by real-time quantitative polymerase chain reaction (RT-qPCR), the expression of Netrin-1, CD146, and epithelial mesenchymal transition (EMT) related proteins was detected by Western blot, the proliferation activity of T84 cells was detected by cell counting kit 8 (CCK-8), and cell migration was detected by Transwell. The expression of the F-actin signal was detected by immunofluorescence after coculture of exosomes with human umbilical vein endothelial cells (HUVECs). Changes in subcutaneous tumor and liver nodule size after PCAT1 deletion were observed in a mouse model of liver metastasis from rectal cancer. Results. PCAT1 expression was upregulated in primary cell lines and their exosomes. After exosomes were cocultured with colorectal cancer tumor circulating T84 cells, the expression of Netrin-1 and CD146 was upregulated, the expression of miR-329-3p was downregulated, the proliferation and migration ability of T84 cells were enhanced, and EMT occurred. After knocking down PCAT1, the above phenomenon was reversed. Similarly, after exosomes were cocultured with HUVECs, the expression of the F-actin signal increased, and after PCAT1 was knocked down, the F-actin signal also decreased. PCAT1 regulates miR-329-3p/Netrin-1 and affects the biological behavior of T84 and F-actin signal expression in HUVECs. In a mouse model of colorectal cancer liver metastasis, knocking down PCAT1 significantly reduced the nodules formed by liver metastasis in mice. Conclusions. LncRNA PCAT1 derived from colorectal cancer exosomes regulates the activity of the Netrin-1-CD146 complex in circulating tumor cells (CTCs) to promote the occurrence of colorectal cancer EMT and liver metastasis and provides new molecular targets for the treatment of colorectal cancer liver metastasis.

Abstract DNA–protein interaction plays an essential role in the storage, expression, and regulation of genetic information. A 1D/3D facilitated diffusion mechanism has been proposed to explain the extraordinarily rapid rate of DNA‐binding protein (DBP) searching for cognate sequence along DNA and further studied by single‐molecule experiments. However, direct observation of the detailed chronological protein searching image is still a formidable challenge. Here, for the first time, a single‐molecule electrical monitoring technique is utilized to realize label‐free detection of the DBP–DNA interaction process based on high‐gain silicon nanowire field‐effect transistors (SiNW FETs). The whole binding process of WRKY domain and DNA has been visualized with high sensitivity and single‐base resolution. Impressively, the swinging of hydrogen bonds between amino acid residues and bases in DNA induce the dynamic collective motion of DBP–DNA. This in situ, label‐free electrical detection platform provides a practical experimental methodology for dynamic studies of various biomolecules.

Objective: To investigate the effects of sulfured polysaccharide from Undaria pinnatifida (SPUP) on the biological behaviors of ovarian cancer (OC) cells and its potential mechanism.Methods: Sulfated polysaccharide from Undaria pinnatifida (SPUP) was extracted and characterized through a combination of chemical analysis, IR spectra, UV-Vis, gas chromatography, and high-performance gel permeation chromatography. OC and human ovarian surface epithelial cells were used as working model in vitro for evaluation of SPUP’s therapeutic effects. A combination of CCK-8, Transwell, and flow cytometry assay was used to measure the proliferation, migration, invasion, and apoptosis of OC cells, respectively. In addition, the protein expression levels of cells were also measured by Western blot.Results: SPUP suppressed OC development from three different perspectives: 1) SPUP treatment significantly inhibited the proliferation of OC in a dosage-dependent manner (p < 0.05); 2) SPUP inhibited the migration and invasion of OC cells confirmed by scratch and Transwell experiments (p < 0.05); 3) SPUP induced apoptosis in OC cells and thus further inhibited the growth of OC cells evaluated using flow cytometry (p < 0.05). The underlying mechanism of the suppressing effects of SPUP might be related to the inhibition of the hedgehog (Hh) signaling pathway in OC cells after SPUP treatment. With additional suppression of the Hh signaling pathway, the anticancer effects of SPUP were enhanced (p < 0.05).Conclusion: Taken together, SPUP could inhibit the proliferation, migration, and invasion and induce apoptosis of OC cells by inhibiting the activation of the Hh signaling pathway, which proposes SPUP as a novel drug to treat OC clinically.

The effects of different doses of a multispecies probiotic (MSP) mixture on growth performance, the incidence of diarrhea rate and immune function, and fecal microbial diversity and structure were evaluated in pre-weaning Holstein dairy calves at WK2, WK4, WK6, and WK8. Forty Chinese Holstein female newborn calves were randomly assigned to four treatments with 10 calves in each group, C (control group), T1 (0.5 g MSP/calf/day, T2 (1 g MSP/calf/day), and T3 (2 g MSP/calf/day) groups. The experimental period was 56 days. Feed intake and health scoring were recorded every day until the end of the experiment. Fecal contents and blood samples were sampled at WK2, WK4, WK6, and WK8. Growth performance, incidence of diarrhea, and total serum concentrations (IgA, IgG, and IgM) were analyzed. Bacterial 16S rRNA and fungal ITS genes were high-throughput sequenced for fecal microbiota. The relationships among the populations of the principal fecal microbiota at WK2 and the growth performance or serum immunoglobulin concentrations were analyzed using Pearson’s rank correlation coefficients. The MSP supplementation reduced the incidence of diarrhea in the first 4 weeks of life, and serum IgA, IgG, and IgM concentrations increased between WK2 and WK8 in the T3 group. There was an increase in growth performance and reduction in the incidence of diarrhea until WK4 after birth in T3 group, compared with the control, T1, and T2 groups. The results of fecal microbiota analysis showed that Firmicutes and Bacteroides were the predominant phyla, with Blautia, Ruminococcaceae_UCG-005, norank_f__Muribaculaceae, Bacteroides, Subdoligranulum, and Bifidobacterium being the dominant genera in calf feces. Aspergillus, Thermomyces, and Saccharomyces were the predominant fungal phyla. Compared with the control, in T1 and T2 groups, the MSP supplementation reduced the relative abundance of Bacteroidetes and increased the relative abundance of Bifidobacterium, Lactobacillus, Collinsella, and Saccharomyces at WK2 in group T3. Thus, the fecal microbial composition and diversity was significantly affected by the MSP mixture during the first 2 weeks of the calves’ life. MSP mixtures reduced the incidence of diarrhea in pre-weaning calves (during the first 4 weeks of life). There was a significant improvement in growth performance, reduction in calf diarrhea, balance in the fecal microbiota, and an overall improvement in serum immunity, compared with the control group. We, therefore, recommend adding 2 g/day of multispecies probiotic mixture supplementation in diets of dairy calves during their first 4 weeks of life before weaning.

To evaluate the potential health-promoting benefits of Berberis nummularia and B. atrocarpa fruits, the biochemical properties (nutrition component, mineral substance, organic acids), total phenolic and flavonoid content and antioxidant (DPPH, FRAP, ABTS and ORAC) capacity of ethanol extracts of B. nummularia and B. atrocarpa fruits wild-grown in Xinjiang were analyzed. The results indicated that there were no meaningful differences (p > 0.05) between the ash (1 ± 0.1 and 1 ± 0.0 g/100 g), fiber (16 ± 1.0 and 18 ± 1.4) and carbohydrate (57 ± 1.8 and 56 ± 1.8 g/100 g) content, respectively, in the dry fruits of B. nummularia and B. atrocarpa. The total fat (7 ± 0.4 and 5 ± 0.1 mg/100 g), soluble sugar (23 ± 0.6 and 12 ± 1.4 g/100 g), titratable acidity (18 ± 2.5% and 14 ± 1.3%) content, and energy value (330.86 and 314.41 kcal/100 g) of B. nummularia was significantly higher than that of B. atrocarpa fruits. Both species contain malic acid, acetic acid, tartaric acid, citric acid and fumaric acid, in which, malic acid is the dominant organic acid. The organic acid and mineral components of B. nummularia fruits were significantly higher than that of B. atrocarpa (p < 0.05). The total phenolic and flavonoid content of B. nummularia were 2 ± 0.0 mg GA/g DW and 2 ± 0.0 mg RE/g DW, respectively, which were significantly lower than the total phenolic and flavonoid content of B. atrocarpa (12 ± 0.1 mg GA/g DW and 9 ± 0.0 mg RE/g DW). The antioxidant capacity of B. nummularia (4 ± 0.1 mg Ascorbic acid/g DW for DPPH, 32 ± 0.1 mg Trolox/g DW for FRAP, 80 ± 3.0 mg Trolox/g DW for ABTS and 60 ± 3.6 mg Trolox/g for ORAC was significantly lower than that of B. atrocarpa (12 ± 0.0 mg Ascorbic acid/g DW for DPPH, 645 ± 1.1 mg Trolox/g DW for FRAP, 304 ± 3.0 mg Trolox/g DW for ABTS and 155 ± 2.8 mg Trolox/g for ORAC). B. atrocarpa fruits showed significantly higher antioxidant capacity than that of B. nummularia. The fruits of the two species can be used in food coloring and nutritional supplements, and consumption of the fruits can aid in weight control and reduce blood glucose or cholesterol.

The low fiber digestibility and unbalanced amino acids restricted the use of DDGS in swine diets. Key nutrition components dynamic monitoring and key regulatory pathways analysis were performed to find the rules of nutrition changes for DDGS fermented by Aspergillus niger. Cellulose and hemicellulose were reduced to 15.3% and 15.2%. 1,4-D-Xylobiose was decreased from 16.8 μg/mL to 0.2 μg/mL. Lys, Arg, and Thr were increased to 3.00%, 2.89%, and 4.40%, and met the requirements of pigs. The whole fermentation process was divided into three stages. Cellulose degradation and Lys and Arg synthesis occurred in the early stage, while Asp synthesis occurred in the last stage. α-Ketoglutarate was the key factor for Aspergillus niger degrading cellulose to synthesize Lys and Arg. The key active metabolic pathways that respond to the changes in nutrition were identified which preliminarily revealed the rules of nutrition adjustment of DDGS during fermentation with Aspergillus niger.

In this study, humidified air dielectric barrier discharge (DBD) plasma was used to inactivate Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), and bacteriophages in biofilms containing DNA, NaCl, carbohydrates, and proteins. The humidified DBD plasma was very effective in the inactivation of microbes in the (≤1.0 μm) biofilms. The number of surviving E. coli, S. aureus, and bacteriophages in the biofilms was strongly dependent on the constituent and thickness of the biofilms and was greatly reduced when the plasma treatment time increased from 5 s to 150 s. Our analysis shows that the UV irradiation was not responsible for the inactivation of microbes in biofilms. The short-lived RONS generated in the humidified air DBD plasma were not directly involved in the inactivation process; however, they recombined or reacted with other species to generate the long-lived RONS. Long-lived RONS diffused into the biofilms to generate very active species, such as ONOOH and OH. This study indicates that the geminated NO2 and OH pair formed due to the homolysis of ONOOH can cause the synergistic oxidation of various organic molecules in the aqueous solution. Proteins in the biofilm were highly resistant to the inactivation of microbes in biofilms, which is presumably due to the existence of the unstable functional groups in the proteins. The unsaturated fatty acids, cysteine-rich proteins, and sulfur–methyl thioether groups in the proteins were easily oxidized by the geminated NO2 and OH pair.

Botanical oils are staple consumer goods globally, but as a by-product of oil crops, meal is of low utilization value and prone to causing environmental problems. The development of proteins in meal into bioactive peptides, such as Perilla peptide, through biotechnology can not only solve environmental problems, but also create more valuable nutritional additives. In the present work, the hydrolysis process of Perilla meal protein suitable for industrial application was optimized with the response surface methodology (RSM) on the basis of single-factor experiments. Alcalase was firstly selected as the best-performing among four proteases. Then, based on Alcalase, the optimal hydrolysis conditions were as follows: enzyme concentration of 7%, hydrolysis temperature of 61.4 °C, liquid-solid ratio of 22.33:1 (mL/g) and hydrolysis time of 4 h. Under these conditions, the degree of hydrolysis (DH) of Perilla meal protein was 26.23 ± 0.83% and the DPPH scavenging capacity of hydrolysate was 94.15 ± 1.12%. The soluble peptide or protein concentration of Perilla meal protein hydrolysate rose up to 5.24 ± 0.05 mg/mL, the ideal yield of which was estimated to be 17.9%. SDS-PAGE indicated that a large proportion of new bands in hydrolysate with small molecular weights appeared, which was different from the original Perilla meal protein. The present data contributed to further, more specific research on the separation, purification and identification of antioxidant peptide from the hydrolysate of Perilla meal protein. The results showed that the hydrolysis of Perilla meal protein could yield peptides with high antioxidant activity and potential applications as natural antioxidants in the food industry.

In the present work, the extraction process of total flavonoids (TFs) from X. sorbifolia flowers by ultrasound-assisted extraction was optimized under the response surface methodology (RSM) on the basis of single-factor experiments. The optimal extraction conditions were as follows: ethanol concentration of 80%, solid–liquid ratio of 1:37 (g/mL), temperature of 84 °C, and extraction time of 1 h. Under the optimized conditions, the extraction yield of the TFs was 3.956 ± 0.04%. The radical scavenging capacities of TFs against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) were much greater than that of rutin. The results of antibacterial experiments indicated that the TFs displayed strong inhibitory activities on E. coli, S. aureus and Bacillus subtilis. Therefore, X. sorbifolia flowers can be used as a novel source of natural flavonoids, and the TFs have potential applications as natural antioxidants or antibacterial agents in the food and pharmaceutical industries.

Due to increasing safety concerns regarding human consumption of fish products, an increasing number of medicinal chemicals are prohibited from use in aquaculture. As a result, Chinese herbal medicines are being increasingly used, coining the use of the term “green medicine.” Research shows that Chinese herbal medicines have many beneficial effects on fish, including growth promotion, enhancement of disease resistance, and improvement in meat quality. Many effective ingredients have been discovered in Chinese herbal medicines, which function to promote feed intake, improve meat flavor, and increase digestive enzyme activity. They also regulate and participate in processes that improve the specific and non-specific immunity of fish; however, the composition of Chinese herbal medicines is very complex and it is often difficult to identify the effective ingredients. This article reviews the latest research and application progress in Chinese herbal medicines regarding growth and feed utilization, immunity and disease resistance, and the meat quality of cultured fish. It also discusses research on the chemical constituents of classical Chinese medicinal herbs and problems with the application of Chinese herbal medicines in fish culture. This article concludes by proposing that future studies on Chinese herbal medicines should focus on how to cheaply refine and extract the effective ingredients in classical Chinese medicinal herbs, as well as how to use them efficiently in aquaculture.

Accumulating evidence has shown that preeclampsia (PE) was associated with an aberrant maternal-fetal inflammatory response. In the present study, we first found that in human PE placentas levels of toll-like receptor 4 (TLR4), phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK) and inflammatory cytokines IL-6 and MCP-1 were significantly upregulated. Next, we demonstrated a notable increase in systolic blood pressure (SBP) and proteinuria in lipopolysaccharide (LPS)-treated pregnant rats and concomitant high levels of TLR4 and p-p38 in these PE-like rat placentas, which led to aberrant overexpression of both IL-6 and MCP-1, as well as deficient trophoblast invasion and spiral artery (SA) remodeling, and these abnormalities were ameliorated by SB203580, a reported inhibitor of p38. In vitro we further confirmed that LPS triggered the activation of TLR4/p38 signaling pathway, which promoted trophoblast apoptosis and damaged trophoblastic invasion via downstream effectors IL-6 and MCP-1; these mutations were rectified by silencing this signaling pathway. These findings elaborated potential mechanisms that aberrant TLR4/p38 signaling might contribute to PE and LPS-induced PE-like symptom by damaging trophoblast invasion and SA remodeling via activating inflammatory cytokines including IL-6 and MCP-1.

Background: The alga Laminaria japonica is the most economically important brown seaweed cultured in China, which is used as food and aquatic animal feedstuff. However, the use of L. japonica as a feedstuff in Apostichopus japonicas farming is not ideal because A. japonicas does not produce enough enzyme activity for degrading the large amount of algin present in L. japonica. In this study, semi solid fermentation of the L. japonica feedstuff employing a Bacillus strain as the microbe was used to as a mean to degrade the algin content in L. japonica feedstuff. Results: The Bacillus strain, Bacillus amyloliquefaciens WB1, was isolated by virtue of its ability to utilize sodium alginate as the sole carbon source. Eight factors affecting growth and algin-degrading capacity of WB1 were investigated. The results of Plackett–Burman design indicated that fermentation time, beef extract, and solvent to solid ratio were the significant parameters. Furthermore, the mutual interaction between the solvent to solid ratio and beef extract concentration was more significant than the other pairs of parameters on algin degradation. Optimal values obtained from Central-Composite Design were 113.94 h for fermentation time, 0.3% (w/v) beef extract and 44.87 (v/w) ratio of solvent to feedstuff. Under optimal conditions, 56.88% of the algin was degraded when a 50-fold scale-up fermentation was carried out, using a 5-L fermenter. Conclusions: This study provides an alternative and economical way to reduce the algin content in L. japonica through degradation by WB1, making it a promising potential source of feed for cultured L. japonica.

Background/Aims: Preeclampsia is an idiopathic and serious complication during gestation in which placental trophoblast cells differentiate into several functional subtypes, including highly invasive extravillous trophoblasts (EVTs). Although the cause and pathogenesis of preeclampsia have remained unclear, numerous studies have suggested that the inadequacy of EVT invasion leads to imperfect uterine spiral artery remodelling, which plays a crucial role in the development of preeclampsia. Rac1, or Ras-related C3 botulinum toxin substrate 1, was found to be a key regulator of the migration, invasion uand apoptosis of various tumour cells. Because EVTs share similar invasive and migratory biological behaviours with malignant cells, this study aimed to determine whether the Rac1 signalling pathway affects trophoblast invasion and is thus involved in the pathogenesis of preeclampsia. Methods: We measured the activity of Rac1 and its downstream targets, β-catenin, Snail and MMP9 in placental tissues from patients experiencing a normal pregnancy and those with preeclampsia. Furthermore, we treated HTR-8/SVneo cells with a shRNA Rac1 vector and the β-catenin inhibitor IWP-2 and explored Rac1 signalling pathway activation as well as the effects of Snail and β-catenin on trophoblast invasion. Results: In placental samples from patients experiencing a normal pregnancy and those with preeclampsia, active Rac1 levels and MMP9 protein and mRNA levels were significantly decreased in term pregnancy samples compared to early pregnancy samples. Lower levels were found in preeclampsia samples than in normal term pregnancy samples, and these levels significantly declined in severe preeclampsia samples compared with mild preeclampsia samples. Further analyses demonstrated that both Rac1 shRNA and the β-catenin inhibitor significantly suppressed MMP9 and Snail activation in trophoblasts, thus impairing trophoblast invasion. Notably, silencing Rac1 down-regulated the expression of β-catenin in HTR-8/SVneo cells, demonstrating that β-catenin is a downstream effector of Rac1 in trophoblast invasion. Conclusion: Our data suggest that Rac1-mediated activation of β-catenin might regulate Snail and MMP9 expression subsequently promoting trophoblast invasion in pregnancy.

In order to find an efficient way for broccoli to increase the phenolic content, this study intended primarily to elucidate the effect of methyl jasmonate (MeJA) treatment on the phenolic accumulation in broccoli. The optimum concentration of MeJA was studied first, and 10 μM MeJA was chosen as the most effective concentration to improve the phenolic content in wounded broccoli. Furthermore, in order to elucidate the effect of methyl jasmonate (MeJA) treatment on phenolic biosynthesis in broccoli, the key enzyme activities of phenylpropanoid metabolism, the total phenolic content (TPC), individual phenolic compounds (PC), antioxidant activity (AOX) and antioxidant metabolism-associated enzyme activities were investigated. Results show that MeJA treatment stimulated phenylalanine ammonia-lyase (PAL), cinnamate 4-hydroxylase (C4H), and 4-coumarin coenzyme A ligase (4CL) enzymes activities in phenylpropanoid metabolism, and inhibited the activity of polyphenol oxidase (PPO), and further accelerated the accumulation of the wound-induced rutin, caffeic acid, and cinnamic acid accumulation, which contributed to the result of the total phenolic content increasing by 34.8% and ferric reducing antioxidant power increasing by 154.9% in broccoli. These results demonstrate that MeJA in combination with wounding stress can induce phenylpropanoid metabolism for the wound-induced phenolic accumulation in broccoli.

Fresh-cut cantaloupe is particularly susceptible to contamination with pathogenic bacteria, such as Escherichia coli O157:H7, Listeria monocytogenes, and Staphylococcus aureus. Therefore, development of rapid, yet accurate detection techniques is necessary to ensure food safety. In this study, a multiplex PCR system and propidium monoazide (PMA) concentration were optimized to detect all viable pathogens in a single tube. A dual filtration system utilized a filtration membrane with different pore sizes to enrich pathogens found on fresh-cut cantaloupe. The results revealed that an optimized multiplex PCR system has the ability to effectively detect three pathogens in the same tube. The viable pathogens were simultaneously detected for PMA concentrations above 10 μg/ml. The combination of a nylon membrane (15 μm) and a micro pore filtration membrane (0.22 μm) formed the dual filtration system used to enrich pathogens. The achieved sensitivity of PMA-mPCR based on this dual filtration system was 2.6 × 103 cfu/g for L. monocytogenes, 4.3 × 10 cfu/g for E. coli O157:H7, and 3.1 × 102 cfu/g for S. aureus. Fresh-cut cantaloupe was inoculated with the three target pathogens using concentrations of 103, 102, 10, and 1 cfu/g. After 6-h of enrichment culture, assay sensitivity increased to 1 cfu/g for each of these pathogens. Thus, this technique represents an efficient and rapid detection tool for implementation on fresh-cut cantaloupe.

Two experiments were conducted to study the effects of red pepper (Capsicum frutescens) powder or red pepper pigment on the performance and egg yolk color of laying hens. In Exp. 1, 210, thirty-wk old, Hy-line Brown laying hens were fed one of seven diets containing 0.3, 0.6, 1.2, 2.0, 4.8 or 9.6 ppm red pepper pigment or 0.3 ppm carophyll red. Each diet was fed to three replicate batteries of hens with each battery consisting of a row of five cages of hens with two hens per cage (n = 3). In Exp. 2, 180, thirty-wk old, Hyline Brown laying hens, housed similarly to those in Exp. 1, were fed an unsupplemented basal diet as well as treatments in which the basal diet was supplemented with 0.8% red pepper powder processed in a laboratory blender to an average particle size of 300 μm, 0.8% red pepper powder processed as a super fine powder with a vibrational mill (44 μm) and finally 0.8% red pepper powder processed as a super fine powder with a vibrational mill but mixed with 5% Na2CO3 either before or after grinding. A diet supplemented with 0.3 ppm carophyll red pigment was also included (n = 3). In both experiments, hens were fed the red pepper powder or pigment for 14 days. After feeding of the powder or pigment was terminated, all hens were fed the basal diet for eight more days to determine if the dietary treatments had any residual effects. In Exp. 1, there were no differences in egg-laying performance, feed consumption or feed conversion ratio due to inclusion of red pepper pigment in the diet. Average egg weight was higher (p0.05). However, compared with the control group, supplementation with all of the red pepper powder treatments increased egg weight (p

The objective of this study was to investigate the potential of using phages as a therapy against hemorrhagic pneumonia in mink both in vitro and in vivo. Five Pseudomonas aeruginosa (P. aeruginosa) strains were isolated from lungs of mink with suspected hemorrhagic pneumonia and their identity was confirmed by morphological observation and 16S rDNA sequence analysis. Compared to P. aeruginosa strains isolated from mink with hemorrhagic pneumonia in 2002, these isolates were more resistant to antibiotics selected. A lytic phage vB_PaeP_PPA-ABTNL (PPA-ABTNL) of the Podoviridae family was isolated from hospital sewage using a P. aeruginosa isolate as host, showing broad host range against P. aeruginosa. A one-step growth curve analysis of PPA-ABTNL revealed eclipse and latent periods of 20 and 35 min, respectively, with a burst size of about 110 PFU per infected cell. Phage PPA-ABTNL significantly reduced the growth of P. aeruginosa isolates in vitro. The genome of PPA-ABTNL was 43,227 bp (62.4% G+C) containing 54 open reading frames and lacked regions encoding known virulence factors, integration-related proteins and antibiotic resistance determinants. Genome architecture analysis showed that PPA-ABTNL belonged to the "phiKMV-like Viruses" group. A repeated dose inhalational toxicity study using PPA-ABTNL crude preparation was conducted in mice and no significantly abnormal histological changes, morbidity or mortality were observed. There was no indication of any potential risk associated with using PPA-ABTNL as a therapeutic agent. The results of a curative treatment experiment demonstrated that atomization by ultrasonic treatment could efficiently deliver phage to the lungs of mink and a dose of 10 multiplicity of infection was optimal for treating mink hemorrhagic pneumonia. Our work demonstrated the potential for phage to fight P. aeruginosa involved in mink lung infections when administered by means of ultrasonic nebulization.

Mycotoxins, ubiquitous contaminants in food, present a global threat to human health and well-being. Mitigation efforts, such as the implementation of sound agricultural practices, thorough food processing, and the advancement of mycotoxin control technologies, have been instrumental in reducing mycotoxin exposure and associated toxicity. To comprehensively assess mycotoxins and their toxicodynamic implications, the deployment of effective and predictive strategies is imperative. Understanding the manner of action, transformation, and cumulative toxic effects of mycotoxins, moreover, their interactions with food matrices can be gleaned through gene expression and transcriptome analyses at cellular and molecular levels. MicroRNAs (miRNAs) govern the expression of target genes and enzymes that play pivotal roles in physiological, pathological, and toxicological responses, whereas acute phase proteins (APPs) exert regulatory control over the metabolism of therapeutic agents, both endogenously and posttranscriptionally. Consequently, this review aims to consolidate current knowledge concerning the regulatory role of miRNAs in the initiation of toxicological pathways by mycotoxins and explores the potential of APPs as biomarkers following mycotoxin exposure. The findings of this research highlight the potential utility of miRNAs and APPs as indicators for the detection and management ofmycotoxins in food through biological processes. Thesemarkers offer promising avenues for enhancing the safety and quality of food products. [ABSTRACT FROM AUTHOR]