Prokaryotic Expression, Purification, and Antibacterial Activity of the Hepcidin Peptide of Crescent Sweetlips (Plectorhinchus cinctus).

The hepcidin peptide of crescent sweetlips (Plectorhinchus cinctus) is a cysteine-rich, cationic antimicrobial peptide that plays a crucial role in the innate immune system's defense against invading microbes. The aim of this study was to identify the optimal parameters for prokaryotic expression and purification of this hepcidin peptide and characterize its antibacterial activity. The recombinant hepcidin peptides were expressed in Escherichia coli strain Arctic Express (DE3), with culture and induction conditions optimized using response surface methodology (RSM). The obtained hepcidin peptides were then purified before tag cleavage, and their antibacterial activity was determined. The obtained results revealed that induction temperature had the most significant impact on the production of soluble recombinant peptides. The optimum induction conditions were determined to be an isopropylthio-ß-galactoside (IPTG) concentration of 0.21 mmol/L, induction temperature of 18.81 °C, and an induction time of 16.01 h. Subsequently, the recombinant hepcidin peptide was successfully purified using Ni-IDA affinity chromatography followed by SUMO protease cleavage. The obtained hepcidin peptide (without His-SUMO tag) demonstrated strong antimicrobial activity in vitro against V. parahaemolyticus, E. coli, and S. aureus. The results showed prokaryotic (E. coli) expression is a feasible way to produce the hepcidin peptide of crescent sweetlips in a cost-effective way, which has great potential to be used as an antimicrobial agent in aquaculture.

Polyoxometalate K6[P2Mo18O62] Inactivates Escherichia coli O157:H7 by Inducing recA Expression and Apoptosis-like Bacterial Death.

Polyoxometalates have emerged as promising bactericidal agents. In the current study, the bactericidal activity of polyoxometalate K6[P2Mo18O62] against Escherichia coli (E. coli) O157:H7 and its possible underlying mechanisms were explored. The obtained results demonstrated that K6[P2Mo18O62] could effectively kill E. coli O157:H7 at millimolar levels. Moreover, K6[P2Mo18O62] treatment also induced significant increases in recA protein expression and further triggered characteristic apoptosis-like bacterial death events such as DNA fragmentation and phosphatidylserine exposure. In conclusion, polyoxometalate K6[P2Mo18O62] possesses a desirable antibacterial activity, and induction of bacterial apoptosis-like death might be involved in its underlying bactericidal mechanisms.

High-Amylose Corn Starch Regulated Gut Microbiota and Serum Bile Acids in High-Fat Diet-Induced Obese Mice.

High-amylose corn starch is well known for its anti-obesity activity, which is mainly based on the regulatory effects on gut microbiota. Recently, the gut microbiota has been reported to improve metabolic health by altering circulating bile acids. Therefore, in this study, the influence of high-amylose corn starch (HACS) on intestinal microbiota composition and serum bile acids was explored in mice fed with a high fat diet (HFD). The results demonstrated HACS treatment reduced HFD-induced body weight gain, hepatic lipid accumulation, and adipocyte hypertrophy as well as improved blood lipid profiles. Moreover, HACS also greatly impacted the gut microbiota with increased Firmicutes and decreased Bacteroidetes relative abundance being observed. Furthermore, compared to ND-fed mice, the mice with HFD feeding exhibited more obvious changes in serum bile acids profiles than the HFD-fed mice with the HACS intervention, showing HACS might restore HFD-induced alterations to bile acid composition in blood. In summary, our results suggested that the underlying mechanisms of anti-obesity activity of HACS may involve its regulatory effects on gut microbiota and circulating bile acids.

Curcumin-Based Photodynamic Sterilization for Preservation of Fresh-Cut Hami Melon.

Fresh-cut fruits and vegetables are the main sources of foodborne illness outbreaks with implicated pathogens such as Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes. This study aimed at investigating the influence of two key parameters (concentration of curcumin and illumination time) on the effects of curcumin-based photodynamic sterilization on the preservation of fresh-cut Hami melons. The results indicated that illumination with 50 µmol/L curcumin for 60 min using a blue LED lamp reduced the total aerobic microorganism count by ~1.8 log CFU/g in fresh-cut Hami melons. Besides this, the effects of photodynamic sterilization on the soluble solids content, color, water content, firmness, and sensory indices of the fresh-cut Hami melons were also evaluated. Compared to the control group, photodynamic sterilization can effectively delay the browning rate and maintain the luminosity, firmness, water content, and soluble solids content of fresh-cut Hami melon. The sensory quality was indeed preserved well after 9 days of storage in a fridge. These results showed that photodynamic sterilization is an effective and promising technology to prolong the shelf life of fresh-cut Hami melons.

Short-chain fatty acids in control of energy metabolism.

Short-chain fatty acids (SCFAs), especially acetate, propionate and butyrate, are the end products from the intestinal microbial fermentation of dietary fibers and resistant starch. It has been well documented that plasma and colonic SCFAs are associated with metabolic syndromes. Recently, the involvement of SCFAs in energy homeostasis regulation has been extensively studied. The importance of SCFAs on energy metabolism has highlighted the potential of modulating SCFAs as a nutritional target to prevent and counteract metabolism disorders and its associated diseases such as obesity and type 2 diabetes. Here, we summarize the current knowledge about the biological properties of SCFAs with their impact on the energy homeostasis.

Steam Explosion-Assisted Extraction of Polysaccharides from Pleurotus eryngii and Its Influence on Structural Characteristics and Antioxidant Activity.

Pleurotus eryngii (PE) has been sought after for its various health benefits and high content of phenolic compounds. This study explored the feasibility of steam explosion (SE)-assisted extraction of polysaccharides with high antioxidant capacities from PE. An orthogonal experimental design (OED) was used to optimize the SE-assisted extraction of PE. The influence of the optimized SE-assisted extraction on the physicochemical properties of PE polysaccharides was determined by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), monosaccharide compositional analysis and antioxidant capacity assays. Under optimal SE conditions, SE-assisted extraction increased the polysaccharide yield by 138% compared to extraction without SE-assistance. In addition, SEM demonstrated that SE-assisted extraction markedly altered the spatial structure of Pleurotus eryngii polysaccharides (PEP), and monosaccharide compositional analysis revealed that this pretreatment significantly increased the proportions of some monosaccharides, such as glucose, rhamnose and arabinose, in the isolated PEP. FTIR spectra indicated no change in the major chemical functional groups of PEP. PEP extracted by SE-assisted extraction had significantly increased free radical scavenging and antioxidant capacities. In conclusion, SE-assisted extraction appears to be a novel polysaccharide extraction technology, which markedly increases extraction yields and efficiency and can increase the biological activity of polysaccharide extracts.

Palmatine alleviates cardiac fibrosis by inhibiting fibroblast activation through the STAT3 pathway.

Cardiac fibrosis, the hallmark of cardiovascular disease, is characterized by excessive deposition of extracellular matrix in the heart. Emerging evidence indicates that cardiac fibroblasts (CFs) play pivotal roles in driving cardiac fibrosis. However, due to incomplete insights into CFs, there are limited effective approaches to prevent or reverse cardiac fibrosis currently. Palmatine, a protoberberine alkaloid extracted from traditional Chinese botanical remedies, possesses diverse biological effects. This study investigated the potential therapeutic value and mechanism of palmatine against cardiac fibrosis. Adult male C57BL/6 mice were treated with vehicle, isoproterenol (ISO), or ISO plus palmatine for one week. After echocardiography assessment, mice hearts were collected for histopathology, real-time polymerase chain reaction, and Western blot analyses. Primary rat CFs were utilized in vitro. Compared to control, ISO-treated mice exhibited cardiac hypertrophy and structural abnormalities; however, treatment with palmatine ameliorated these effects of ISO. Moreover, palmatine treatment mitigated ISO-induced cardiac fibrosis. Network pharmacology and molecular docking analysis showed that palmatine strongly binds the regulators of cardiac fibrosis including signal transducer and activator of transcription 3 (STAT3) and mammalian target of rapamycin. Furthermore, palmatine reduced the elevated fibrotic factor expressions and overactivated STAT3 induced by ISO, Transformed growth factor ß1 (TGF-ß1), or interleukin-6 both in vivo and in vitro. Additionally, blocking STAT3 suppressed the TGF-ß1-induced CF activation. Collectively, these data demonstrated that palmatine attenuated cardiac fibrosis partly by inhibiting fibroblast activation through the STAT3 pathway. This provides an experimental basis for the clinical treatment of cardiac fibrosis with palmatine.

Effective Preservation of Chilled Pork Using Photodynamic Antibacterial Film Based on Curcumin-ß-Cyclodextrin Complex.

A biodegradable photodynamic antibacterial film (PS-CF) was prepared using the casting method, with κ-Carrageenan (κ-Car) as the film-forming substrate and curcumin-ß-cyclodextrin (Cur-ß-CD) complex as photosensitizer. Chilled pork samples were coated with PS-CF and stored at 4 °C to investigate the effects of PS-CF combined with LED light irradiation (425 nm, 45 min) (PS+L+) on pork preservation during 10 days of storage. The total viable count (TVC) of bacteria, total volatile basic nitrogen value (TVB-N) and the pH of pork treated with PS+L+ were all lower than the control, and the water-holding capacity (WHC) was higher. Ten days later, the TVB-N value was 12.35 ± 0.57 mg/100 g and the TVC value was 5.78 ± 0.17 log CFU/g, which was within the acceptable range. Sensory evaluation determined that the color, odor, and overall acceptability of pork treated with PS+L+ were significantly better than the control. These findings suggest that PS+L+ treatment effectively extended the shelf life of chilled pork from ~4-5 to 10 days. Correlation analysis showed that the sensory quality of the chilled pork significantly correlated with total bacterial counts, TVB-N and thiobarbituric acid reactive substances (TBARS) (p < 0.05), suggesting that these biomarkers could be used as standard indicators for evaluating the freshness of chilled pork. These findings demonstrate the effectiveness of Cur-ß-CD photodynamic antibacterial film for the preservation of chilled pork and provide a theoretical basis for the application of the film for the preservation of fresh food in general.

Current Perspectives on Viable but Non-Culturable Foodborne Pathogenic Bacteria: A Review.

Foodborne diseases caused by foodborne pathogens pose risks to food safety. Effective detection and efficient inactivation of pathogenic bacteria has always been a research hotspot in the field of food safety. Complicating these goals, bacteria can be induced to adopt a viable but non-culturable (VBNC) state under adverse external environmental stresses. When in the VBNC state, pathogens cannot form visible colonies during traditional culture but remain metabolically active and toxic. The resulting false negative results in growth-related assays can jeopardize food safety. This review summarizes the latest research on VBNC foodborne pathogens, including induction conditions, detection methods, mechanism of VBNC formation, and possible control strategies. It is hoped that this review can provide ideas and methods for future research on VBNC foodborne pathogenic bacteria.

RNA-seq Analysis Reveals Potential Synergic Effects of Acetate and Cold Exposure on Interscapular Brown Adipose Tissue in Mice.

Brown adipose tissue (BAT) exhibits remarkable morphological and functional plasticity in response to environmental (e.g., cold exposure) and nutrient (e.g., high-fat diet) stimuli. Notably, a number of studies have showed that acetate, the main fermentation product of dietary fiber in gut, profoundly influences the differentiation and activity of BAT. However, the potential synergic or antagonistic effects of acetate and cold exposure on BAT have not been well examined. In the present study, the C57BL/6J mice were treated with acetate at the systemic level before a short period of cold exposure. Physiological parameters including body weight, blood glucose, and Respiratory Exchange Ratio (RER) were monitored, and thermal imaging of body surface temperature was captured. Moreover, the transcriptome profiles of interscapular BAT were also determined and analyzed afterwards. The obtained results showed that acetate treatment prior to cold exposure could alter the gene expression profile, as evidenced by significant differential clusters between the two groups. GO analysis and KEGG analysis further identified differentially expressed genes being mainly enriched for a number of biological terms and pathways related to lipid metabolism and brown adipose activity such as "G-protein-coupled receptor activity", "cAMP metabolic process", "PPAR signaling pathway", and "FoxO signaling pathway". GSEA analysis further suggested that activation status of key pathways including "PPAR signaling pathway" and "TCA cycle" were altered upon acetate treatment. Taken together, our study identified the potential synergistic effect of acetic acid with cold exposure on BAT, which highlighted the positive dietary and therapeutic aspects of acetate.

Molecular cloning of the hepcidin gene from crescent sweetlips (Plectorhinchus cinctus) and characterization of its encoded antimicrobial peptide.

Hepcidin has been identified as an important antimicrobial peptide exerting important innate immunomodulatory activities in many fish species. In the present study, reverse transcription PCR coupled with the rapid amplification of cDNA ends was used to obtain the full-length cDNA of the crescent sweetlips hepcidin gene, which is 829 bp in length and includes an 273 bp ORF encoding a peptide with 90 amino acid residues. Sequence alignment showed a typical RXKR motif and eight conserved cysteine residues in the deduced amino acid sequences. Four disulfide bonds were predicted to form between these eight cysteines, which may stabilize the hairpin structure in hepcidin molecule. Furthermore, phylogenetic analysis showed that the deduced amino acid sequences of crescent sweetlips hepcidin had high sequence homology to hepcidins from fish species of Eupercaria. In addition, the crescent sweetlips hepcidin peptide demonstrated a strong antimicrobial activity in vitro against several types of pathogenic bacteria in fish. In conclusion, the obtained results suggested that crescent sweetlips hepcidin possessed the typical structure similar to other fish hepcidins and had strong antibacterial activity, which showed great potential in the prevention of fish diseases in aquaculture.

K6[P2Mo18O62] as DNase-Mimetic Artificial Nucleases to Promote Extracellular Deoxyribonucleic Acid Degradation in Bacterial Biofilms.

In the current study, the DNase-like activity of the Dawson-type polyoxometalate K6[P2Mo18O62] was explored. The obtained findings demonstrated that K6[P2Mo18O62] could effectively cleave phosphoester bonds in the DNA model substrate (4-nitrophenyl phosphate) and result in the degradation of plasmid DNA. Moreover, the application potential of this Dawson-type polyoxometalate as a DNase-mimetic artificial enzyme to degrade extracellular DNA (eDNA) in Escherichia coli (E. coli) bacterial biofilm was explored. The results demonstrated that K6[P2Mo18O62] exhibited high cleavage ability toward eDNA secreted by E. coli and thus eradicated the bacterial biofilm. In conclusion, Dawson-type polyoxometalate K6[P2Mo18O62] possessed desirable DNase-like activity, which could serve as a bacterial biofilm eradication agent by cleaving and degrading eDNA molecules.

Lesson learned from COVID-19 pandemic for the future of food industry.

With WHO announcing COVID-19 no longer as a public health emergency of international concern (PHEIC) on May 5, 2023, coupled with the fact that the majority of the countries of the world have dropped strict city lockdown or border closure, this perhaps signals the end of the COVID-19 crisis caused by the SARS-CoV-2 virus. However, the COVID-19 pandemic has resulted in far-reaching effects affecting nearly every aspect of our lives and society. Notably, the food industry including agriculture, food manufacturers, food logistics, distributors and retailers have all felt the profound impact and had experienced significant stress during the pandemic. Therefore, it is essential to retrospect the lessons that can be learned from this pandemic for the food industry. This short review aims to address the food safety issues related to the COVID-19 pandemic by focusing on its foodborne transmission potential, innovations of virus detection strategies suitable for food industry; development of phathogenicaidal methods and devices to inactivate SARS-CoV-2 virus (particularly in industrial scale); and the set-up of related food regulations and guidelines as preventive and control measures for preventing the spread of SARS-CoV-2 virus through the food supply chain during the pandemic. This article may provide useful references for the food industry to minimize the food safety impact of COVID-19 (as well as other respiratory virus) and allows them to better prepare for similar future challenges.

Preparation and Characterization of Biodegradable κ-Carrageenan Based Anti-Bacterial Film Functionalized with Wells-Dawson Polyoxometalate.

In the present study, an anti-bacterial film (Carr/POM film) was prepared through the incorporation of Wells-Dawson polyoxometalate K6[Mo18O62P2] into κ-carrageenan-based polymers using the tape-casting method. The mechanical properties, thermal stability, and morphology of the prepared film were characterized. The obtained results showed that incorporation of K6[Mo18O62P2] significantly affected the morphology and structure of the films. Moreover, the polyoxometalate-based film demonstrated desirable bactericidal activity against Escherichia coli (Gram-negative) and Staphylococcus aureus (Gram-positive). Carr/POM (@8 mg/mL) film resulted in an obvious inhibition zone around the film in Kirby-Bauer disk diffusion test, which could also remove 99% of S. aureus and E. coli on plastic, glass, and stainless steel. In addition, this anti-bacterial film also demonstrated good biodegradability, which could be decomposed in soil in around 1 week. In conclusion, the polyoxometalate-based film showed good anti-bacterial property against food-borne pathogenic microbes, suggesting the prepared film has great potential to be developed as promising food packaging.

Comprehensive properties of photodynamic antibacterial film based on κ-Carrageenan and curcumin-ß-cyclodextrin complex.

In this study, a biodegradable photodynamic antibacterial film (Car-Cur) was prepared using casting method with κ-Carrageenan (κ-Car) as film-forming substrate and curcumin-ß-cyclodextrin (Cur-ß-CD) complex as photosensitizer. The comprehensive performance of this Car-Cur film was investigated. The obtained results showed that the concentration of Cur-ß-CD was an important factor determining the properties of film including tensile strength (TS) elongation at break (EB), water vapor permeability (WVP), water content (WC) and thermal stability. When the concentration of Cur-ß-CD is 1%, the film demonstrated the maximum TS and EB, increased thermal stability, with desirable WVP and WC. Furthermore, this film also showed good photodynamic antibacterial potential against Staphylococcus aureus and Escherichia coli upon irradiation of blue LED light. Moreover, the film can be degraded in the soil in one week. In conclusion, our results suggested Car-Cur photodynamic film could be developed as biodegradable antimicrobial packaging material for food preservation.

Curcumin-Mediated Sono-Photodynamic Treatment Inactivates Listeria monocytogenes via ROS-Induced Physical Disruption and Oxidative Damage.

Sono-photodynamic sterilization technology (SPDT) has become a promising non-thermal food sterilization technique because of its high penetrating power and outstanding microbicidal effects. In this study, Listeria monocytogenes (LMO) was effectively inactivated using curcumin as the sono-photosensitizer activated by ultrasound and blue LED light. The SPDT treatment at optimized conditions yielded a 4-log reduction in LMO CFU. The reactive oxygen species (ROS) production in LMO upon SPDT treatment was subsequently investigated. The results demonstrated SPDT treatment-induced excessive ROS generation led to bacterial cell deformation and membrane rupture, as revealed by the scanning electron microscope (SEM) and cytoplasmic material leakage. Moreover, agarose gel electrophoresis and SDS-PAGE further revealed that SPDT also triggered bacterial genomic DNA cleavage and protein degradation in LMO, thus inducing bacterial apoptosis-like events, such as membrane depolarization.

Enhancement of photodynamic bactericidal activity of curcumin against Pseudomonas Aeruginosa using polymyxin B.

BACKGROUND: Pseudomonas aeruginosa (P. aeruginosa) is an emerging opportunistic pathogen, which can cause bacterial skin diseases such as green nail syndrome, interdigital infections and folliculitis. Curcumin-mediated antimicrobial photodynamic therapy (aPDT) has been demonstrated as a promising therapeutic option for the treatment of skin infection though its inactivation of gram-negative bacteria such as P. aeruginosa. MATERIALS AND METHODS: In the present study, we examined the adjuvant effect of polymyxin B on the antibacterial activity of curcumin-mediated aPDT against P. aeruginosa. P. aeruginosa was treated with curcumin in the presence of 0.1-0.5 mg/L polymyxin B and irradiated by blue LED light (10 J/cm2). Bacterial cultures treated with curcumin alone served as controls. Colony forming units (CFU) were counted and the viability of P. aeruginosa was calculated after aPDT treatment. The possible underlying mechanisms for the enhanced killing effects were also explored. RESULTS: The killing effects of curcumin-mediated aPDT against P. aeruginosa was significantly enhanced by polymyxin B (over 2-log reductions). Moreover, it was also observed that addition of polymyxin B in the curcumin-mediated aPDT led to the apparent bacterial membrane damage with increased leakage of cytoplasmic contents and extensive DNA and protein degradation. DISCUSSION: The photodynamic action of curcumin against P. aeruginosa could be significantly enhanced by the FDA-approved drug polymyxin B. Our results highlight the potential of introducing polymyxin B to enhance the effects of aPDT treatment against gram-negative skin infections, in particular, P. aeruginosa.

Chlorella unsaturated fatty acids suppress high-fat diet-induced obesity in C57/BL6J mice.

Chlorella has been identified as a rich source of unsaturated fatty acids. Since the antiobesity effects of unsaturated fatty acids have been well documented; therefore, we explored the antiobesity actions of chlorella unsaturated fatty acids (C.UFAs) in the current study. The obtained results demonstrated C.UFAs, which contain abundant linoleic acid, could retard body weight gain (reducing body weigh by 13.93% after 16 weeks of treatment), improve blood glucose (19.29% lower) and lipid profile (23.45% lower in TG, 8.76% lower in TC) compared to high-fat diet-fed C57BL/6J mice. The possible underlying mechanisms might involve reducing hepatic lipid accumulation via down-regulation of lipogenic genes (PPARγ, C/EBPα, LPL, aP2, FAS, and SREBP-1c) and up-regulation of lipolytic gene (adiponectin). We also demonstrate C.UFAs could reduce HFD-induced adipocyte hypertrophy via activation of AMPK signaling pathway in adipose tissue and liver. In summary, our study highlights the potential of C.UFAs as a functional food for obesity management. PRACTICAL APPLICATION: Chlorella has already been commercialized as a functional food antiobesity function. In the current study, the unsaturated fatty acids isolated from chlorella were found to exert beneficial effects on hyperglycemia, hyperlipidemia, hepatic steatosis, and adipocyte hypertrophy in high-fat diet-fed mice. This may provide theoretical foundation for developing novel chlorella-based functional foods.

Control Measurements of Escherichia coli Biofilm: A Review.

Escherichia coli (E. coli) is a common pathogen that causes diarrhea in humans and animals. In particular, E. coli can easily form biofilm on the surface of living or non-living carriers, which can lead to the cross-contamination of food. This review mainly summarizes the formation process of E. coli biofilm, the prevalence of biofilm in the food industry, and inhibition methods of E. coli biofilm, including chemical and physical methods, and inhibition by bioactive extracts from plants and animals. This review aims to provide a basis for the prevention and control of E. coli biofilm in the food industry.

Hsian-Tsao (Mesona chinensis Benth.) Extract Improves the Thermal Tolerance of Drosophila melanogaster.

Global warming has prompted scientific communities to consider how to alleviate thermal stress in humans and animals. The present study assessed the supplementation of hsian-tsao extract (HTE) on thermal stress in Drosophila melanogaster and preliminarily explicated its possible physiological and molecular mechanisms. Our results indicated that the lethal time for 50% of female flies fed on HTE was significantly longer than that of male flies at the same heat stress temperature. Under thermal stress, the survival time of females was remarkably increased in the HTE addition groups compared to the non-addition group. Thermal hardening by acute exposure to 36°C for 30 min (9:00 to 9:30 a.m.) every day could significantly prolong the longevity of females. Without thermal hardening, HTE increased the antioxidant capacity of females under heat stress, accompanied by an increment of catalase (CAT) activity, and the inhibition for hydroxyl radicals (OH⋅) and superoxide anions (⋅O2 -). Superoxide dismutase (SOD) activity and the inhibition for ⋅O2 - was significantly affected by thermal hardening in the non-HTE addition groups, and significant differences were shown in CAT and SOD activities, and the inhibition for ⋅O2 - among groups with thermal hardening. After heat exposure, heat shock protein 70 (Hsp70) was only up-regulated in the group with high levels of added HTE compared with the group without and this was similar in the thermal hardening group. It was concluded that the heat stress-relieving ability of HTE might be partly due to the enhancement of enzymatic activities of SOD and CAT, and the inhibition for OH⋅ and ⋅O2 -. However, the expression levels of Hsp70 were not well related to thermal tolerance or heat survival.