Molecular identification of Halomonas AZ07 and its multifunctional enzymatic activities to degrade Pyropia yezoensis under high-temperature condition.

BACKGROUND: Pyropia yezoensis a commercially important red seaweed species, is susceptible to various microorganisms infections, among which bacterial infections are the most prominent ones. Pyropia yezoensis is often affected by harmful bacterial communities under high temperatures that can lead to its degradation and economic losses. The current study aimed to explore Pyropia yezoensis-associated microbiota and further identify potential isolates, which can degrade Pyropia yezoensis under high-temperature conditions. METHODS AND RESULTS: The 16S rRNA gene sequencing was used to identify the agarolytic bacterial species. The results showed that Chromohalobacter sp. strain AZ6, Pseudoalteromonas sp. strain AZ, Psychrobacter sp. strain AZ3, Vibrio sp. strain AZ, and Halomonas sp. strain AZ07 exhibited algicidal properties as these strains were more abundant at high temperature (25 °C). Among the five isolated strains, the potential isolate Halomonas sp. strain AZ07 showed high production of agarolytic enzymes, including lipase, protease, cellulase, and amylase. This study confirmed that the isolated strain could produce these four different enzymes. The strain Halomonas AZ07 was co-treated with Pyropia yezoensis cells under two different temperature environments, including 13 °C and 25 °C. The degradation of Pyropia yezoensis occurred at the optimum temperature of 25 °C and effectively degraded their cell wall, proteins, lipids, and carbohydrates. CONCLUSION: The successful cultivation of Pyropia yezoensis in coastal farm environments is dependent on specific temperature and environmental factors, and lower temperatures have been observed to be particularly beneficial for the survival and growth of Pyropia yezoensis. The temperature below 13 °C was confirmed to be the best niche for the symbiotic relationship of microbiota associated with Pyropia yezoensis for its growth, development, and production.

Polysaccharides from Porphyra haitanensis: A Review of Their Extraction, Modification, Structures, and Bioactivities.

Porphyra haitanensis (P. haitanensis), an important food source for coastal residents in China, has a long history of medicinal and edible value. P. haitanensis polysaccharides are some of the main active ingredients in P. haitanensis. It is worth noting that P. haitanensis polysaccharides have a surprising and satisfactory biological activity, which explains the various benefits of P. haitanensis to human health, such as anti-oxidation, immune regulation, anti-allergy, and anticancer properties. Hence, a systematic review aimed at comprehensively summarizing the recent research advances in P. haitanensis polysaccharides is necessary for promoting their better understanding. In this review, we systematically and comprehensively summarize the research progress on the extraction, purification, structural characterization, modification, and biological activity of P. haitanensis polysaccharides and address the shortcomings of the published research and suggest area of focus for future research, providing a new reference for the exploitation of polysaccharides from P. haitanensis in the fields of medicine and functional foods.

Functional Characterization of the First Bona Fide Phytoene Synthase in Red Algae from Pyropia yezoensis.

The formation of phytoene by condensing two geranylgeranyl diphosphate molecules catalyzed by phytoene synthase (PSY) is the first committed and rate-limiting step in carotenoid biosynthesis, which has been extensively investigated in bacteria, land plants and microalgae. However, this step in macroalgae remains unknown. In the present study, a gene encoding putative phytoene synthase was cloned from the economic red alga Pyropia yezoensis-a species that has long been used in food and pharmaceuticals. The conservative motifs/domains and the tertiary structure predicted using bioinformatic tools suggested that the cloned PyPSY should encode a phytoene synthase; this was empirically confirmed by pigment complementation in E. coli. This phytoene synthase was encoded by a single copy gene, whose expression was presumably regulated by many factors. The phylogenetic relationship of PSYs from different organisms suggested that red algae are probably the progeny of primary endosymbiosis and plastid donors of secondary endosymbiosis.

Optimizing the Extraction of Bioactive Compounds from Porphyra linearis (Rhodophyta): Evaluating Alkaline and Enzymatic Hydrolysis for Nutraceutical Applications.

Porphyra sensu lato is one of the most economically significant and widely cultured and consumed algae in the world. Porphyra species present excellent nutraceutic properties due to their bioactive compounds (BACs). This research aimed to find the most efficient aqueous extraction method for BACs by examining alkaline and enzymatic hydrolysis. Alkaline hydrolysis with 2.5% sodium carbonate (SC) and at 80 °C proved optimal for extracting all BACs (phycobiliproteins, soluble proteins, polyphenols, and carbohydrates) except mycosporine-like amino acids (MAAs), which were best extracted with water only, and at 80 °C. Enzymatic hydrolysis, particularly with the 'Miura' enzymatic cocktail (cellulase, xylanase, glycoside hydrolase, and ß-glucanase), showed superior results in extracting phycoerythrin (PE), phycocyanin (PC), soluble proteins, and carbohydrates, with increases of approximately 195%, 510%, 890%, and 65%, respectively, compared to the best alkaline hydrolysis extraction (2.5% SC and 80 °C). Phenolic content analysis showed no significant difference between the 'Miura' cocktail and 2.5% SC treatments. Antioxidant activity was higher in samples from alkaline hydrolysis, while extraction of MAAs showed no significant difference between water-only and 'Miura' treatments. The study concludes that enzymatic hydrolysis improves the efficiency of BACs extraction in P. linearis, highlighting its potential for the nutraceutical industry, and especially with respect to MAAs for topical and oral UV-photoprotectors.

Porphyra haitanensis glycoprotein regulates glucose homeostasis: targeting the liver.

In this study, we investigated the effects of glycoprotein (PG)-mediated regulation of Porphyra haitanensis on liver glucose metabolism in hyperglycemic mouse models, and sought to establish the underlying mechanism, as determined by the changes in liver gene expression and metabolic profiles. The results showed that 30-300 mg kg-1 PG upregulated the expression of the liver genes Ins1, Ins2, Insr, Gys2, Gpi1, Gck, and downregulated the expression of G6pc, G6pc2, and G6pc3, in a concentration-dependent manner. 300 mg kg-1 PG downregulated the concentrations of glucose-related metabolites in the liver, but upregulated lactic acid, 2-aminoacetic acid, and glucose-1-phosphate concentrations. It was assumed that PG regulated liver glucose metabolism by enriching insulin secretion, glycolysis/gluconeogenesis, and the AMPK signaling pathway, and promoting insulin secretion, glycogen synthesis, and glycolysis. Our findings supported the development of P. haitanensis and its glycoproteins as novel natural antidiabetic compounds that regulated blood glucose homeostasis.

Peptidomics- inspired discovery and activity evaluation of antioxidant peptides in multiple strains mixed fermentation of Porphyra yezoensis.

This study investigated the production of antioxidant peptides from Porphyra yezoensis through fermentation with three strains of microorganisms: Lactiplantibacillus plantarum L13, Bacillus amyloliquefaciens MMB-02, and Saccharomyces cerevisiae A8. The crude peptides were extracted by aqueous acid precipitation and purified by Sephadex G-25 gel column to produce highly active antioxidant components with molecular weight of <4000 Da. The LC-MS/MS result revealed that the fermentation group contained more hydrophobic amino acids and oligopeptides, which were mainly originated from phycobiliproteins and algal blue proteins. Finally, the antioxidant activity of Porphyra yezoensis was determined with DPPH· and ABTS· scavenging rates of 54.87% and 57.39%, respectively. The ferric ion-reducing power (FRAP) and enzyme activities of SOD and CAT were significantly higher than those of the control group. This study provides a scientific foundation for the deep processing of striped seaweed and contributes to the theoretical understanding of synthetic antioxidant substitutes.

Purified Pyropia yezoensis Pigment Extract-Based Tandem Dye Synthesis.

Red phycoerythrin (R-PE) is a highly valuable protein found in an edible seaweed, Pyropia yezoensis. It is used extensively in biotechnological applications due to its strong fluorescence and stability in diverse environments. However, the current methods for extracting and purifying R-PE are costly and unsustainable. The aim of the present study was to enhance the financial viability of the process by improving the extraction and purification of R-PE from dried P. yezoensis and to further enhance R-PE value by incorporating it into a tandem dye for molecular biology applications. A combination of ultrafiltration, ion exchange chromatography, and gel filtration yielded concentrated (1 mg·mL-1) R-PE at 99% purity. Using purified PE and Cyanine5 (Cy5), an organic tandem dye, phycoerythrin-Cy5 (PE-Cy5), was subsequently established. In comparison to a commercially available tandem dye, PE-Cy5 exhibited 202.3% stronger fluorescence, rendering it suitable for imaging and analyzes that require high sensitivity, enhanced signal-to-noise ratio, broad dynamic range, or shorter exposure times to minimize potential damage to samples. The techno-economic analysis confirmed the financial feasibility of the innovative technique for the extraction and purification of R-PE and PE-Cy5 production.

The Role of Nitrate Supply in Bioactive Compound Synthesis and Antioxidant Activity in the Cultivation of Porphyra linearis (Rhodophyta, Bangiales) for Future Cosmeceutical and Bioremediation Applications.

Porphyra sensu lato has economic importance for food and pharmaceutical industries due to its significant physiological activities resulting from its bioactive compounds (BACs). This study aimed to determine the optimal nitrate dosage required in short-term cultivation to achieve substantial BAC production. A nitrate experiment using varied concentrations (0 to 6.5 mM) revealed optimal nitrate uptake at 0.5 mM in the first two days and at 3 and 5 mM in the last five days. Polyphenols and carbohydrates showed no differences between treatments, while soluble proteins peaked at 1.5 and 3 mM. Total mycosporine-like amino acids (MAAs) were highest in algae incubated at 5 and 6.5 mM, and the highest antioxidant activity was observed in the 5 mM, potentially related to the MAAs amount. Total carbon and sulfur did not differ between treatments, while nitrogen decreased at higher nitrate. This discovery highlights the nuanced role of nitrate in algal physiology, suggesting that biological and chemical responses to nitrate supplementation can optimize an organism's health and its commercially significant bioactive potential. Furthermore, given its ability to absorb high doses of nitrate, this alga can be cultivated in eutrophic zones or even in out-/indoor tanks, becoming an excellent option for integrated multi-trophic aquaculture (IMTA) and bioremediation.

Exploring the therapeutic potential of porphyran extracted from Porphyra haitanensis in the attenuation of DSS-induced intestinal inflammation.

Ulcerative colitis is a chronic, spontaneous inflammatory bowel disease that primarily affects the colon. This study aimed to explore how Porphyra haitanensis porphyran (PHP) modulates the immune response and the associated mechanisms that alleviate dextran sulphate sodium-induced colitis in mice. Histological assessments via H&E staining and AB-PAS staining revealed that PHP intervention partially restored the number of goblet cells and improved intestinal mucosal function. Immunohistochemical and Western blot analyses of claudin-1, occludin, and MUC-2 demonstrated that PHP could repair the intestinal barrier and reduce colon damage by upregulating the expression of these proteins. PHP intervention was associated with a decrease in pro-inflammatory cytokine expression and an increase in anti-inflammatory cytokine expression. Moreover, the expression of proteins involved in intestinal immune homing, such as CCR-9, CCL-25, MAdCAM-1, and α4ß7, was significantly suppressed in response to PHP treatment. Conversely, PHP upregulates the expression of CD40 and TGF-ß1, both of these can promote healing and reduce inflammation in the gut lining. This study demonstrates that PHP can ameliorate ulcerative colitis by enhancing the intestinal barrier and modulating immune responses. These findings offer valuable insights into the potential utility of P. haitanensis as a promising natural product for managing ulcerative colitis.

Mixed acid treatment for removal of green macroalgae from Neopyropia aquaculture nets: Field experiment in the Subei Shoal, China.

To develop an effective method to eliminate green macroalgae attached to Neopyropia aquaculture nets, we explored the influence of mixed acid solution on the photosynthetic fluorescence characteristics of Ulva spp. (green macroalgae) and Neopyropia yezoensis (red macroalgae) from Dafeng and Rudong aquaculture areas in Jiangsu Province, China. Treatment with mixed acid solution (0.0475 % hydrochloric acid:citric acid (pH 2.0) at a ratio of 4:3) for 60 s caused death of Ulva spp., but did not affect N. yezoensis. Additionally, a mixed acid solution effectively eliminated green macroalgae from Neopyropia aquaculture rafts and the marine environment remained unaffected. Hence, the application of mixed acid solution treatment has demonstrated significant efficacy in eradicating green macroalgae adhered to Neopyropia aquaculture rafts, thus presenting a promising strategy for mitigating green macroalgae proliferation in Neopyropia aquaculture areas and curbing their contribution to green tides.

Dehydration, Rehydration and Thermal Treatment: Effect on Bioactive Compounds of Red Seaweeds Porphyra umbilicalis and Porphyra linearis.

The nutritional and bioactive value of seaweeds is widely recognized, making them a valuable food source. To use seaweeds as food, drying and thermal treatments are required, but these treatments may have a negative impact on valuable bioactive compounds. In this study, the effects of dehydration, rehydration, and thermal treatment on the bioactive compounds (carotenoids, phycobiliproteins, total phenolic content (TPC), total flavonoids content (TFC)), antioxidant (ABTS and DPPH radical scavenging activities) and anti-Alzheimer's (Acetylcholinesterase (AchE) inhibitory activities, and color properties of Porphyra umbilicalis and Porphyra linearis seaweeds were evaluated. The results revealed significant reductions in carotenoids, TPC, TFC, and antioxidant activities after the seaweeds' processing, with differences observed between species. Thermal treatment led to the most pronounced reductions in bioactive compound contents and antioxidant activity. AchE inhibitory activity remained relatively high in all samples, with P. umbilicalis showing higher activity than P. linearis. Changes in color (ΔE) were significant after seaweeds' dehydration, rehydration and thermal treatment, especially in P. umbilicalis. Overall, optimizing processing methods is crucial for preserving the bioactive compounds and biological activities of seaweeds, thus maximizing their potential as sustainable and nutritious food sources or as nutraceutical ingredients.

Mechanism of Porphyra Yezoensis Polysaccharides in Inhibiting Hyperoxalate-Induced Renal Injury and Crystal Deposition.

Oxidative damage to the kidneys is a primary factor in the occurrence of kidney stones. This study explores the inhibitory effect of Porphyra yezoensis polysaccharides (PYP) on oxalate-induced renal injury by detecting levels of oxidative damage, expression of adhesion molecules, and damage to intracellular organelles and revealed the molecular mechanism by molecular biology methods. Additionally, we validated the role of PYP in vivo using a crystallization model of hyperoxalate-induced rats. PYP effectively scavenged the overproduction of reactive oxygen species (ROS) in HK-2 cells, inhibited the adhesion of calcium oxalate (CaOx) crystals on the cell surface, unblocked the cell cycle, restored the depolarization of the mitochondrial membrane potential, and inhibited cell death. PYP upregulated the expression of antioxidant proteins, including Nrf2, HO-1, SOD, and CAT, while decreasing the expression of Keap-1, thereby activating the Keap1/Nrf2 signaling pathway. PYP inhibited CaOx deposition in renal tubules in the rat crystallization model, significantly reduced high oxalate-induced renal injury, decreased the levels of the cell surface adhesion proteins, improved renal function in rats, and ultimately inhibited the formation of kidney stones. Therefore, PYP, which has crystallization inhibition and antioxidant properties, may be a therapeutic option for the treatment of kidney stones.

Fluorescent imaging and toxicology study of alga-derived carbon dots in zebrafish.

With the widespread application of carbon dots (CDs) in fluorescence imaging, their toxicity has become a focal point of concern. The potential toxicity of CDs synthesized from different raw materials remains an unresolved issue. Laver and wakame, which are commonly popular sea vegetable foods rich in nutrients, were utilized to investigate whether synthetic CDs derived from these alga sources retain medicinal value. Herein, two types of fluorescent alga-derived CDs were prepared through hydrothermal synthesis using laver and wakame respectively. Zebrafish were immersed in both types of CDs to observe their fluorescence imaging effects within the zebrafish bodies. It was observed that laver-derived CDs and wakame-derived CDs exhibited similar luminescence properties but differed in terms of fish egg imaging localization. Additionally, intestinal flora sequencing revealed varying degrees of influence on the zebrafish gut microbiota by the two types of CDs, suggesting that both alga-derived CDs could enhance the abundance of intestinal flora in zebrafish.

Prediction and assessment of marine fisheries carbon sink in China based on a novel nonlinear grey Bernoulli model with multiple optimizations.

The vigorous development of marine fisheries carbon sinks (MFCS) has become a momentous pathway to mitigate global warming and effectively cope with the climate crisis. Deservedly, based on clarifying mechanism of carbon sequestration, this paper designs a research paradigm for predicting and evaluating the potential of MFCS. Specifically, a novel nonlinear grey Bernoulli model, namely MFCSNGBM(1,1), is proposed by innovatively mining the original data law through adaptive cumulative series and introducing the compound Simpson formula to optimize background values. More precisely, we utilize a heuristic Grey Wolf Optimization algorithm to find the best power index, which enhances the adaptability. To prove usefulness and robustness of MFCSNGBM(1,1) model, yields of seven common shellfishes (oyster, clam, mussel, scallop, razor clam, bloody clam, and snail) and three main algae (kelp, pinnatifid undaria, and laver) are predicted and compared with six competing models. Based on prediction results, new model has the most accurate predictions, with all prediction errors being <10 %, and thus can achieve effective prediction of shellfish and algae production from 2022 to 2025. Further, the capacity and potential of MFCS in China are scientifically evaluated using a removable carbon sink model, considering various yield levels and biological parameters of shellfish and algae. The assessment results show that during the sample period, China's marine fisheries carbon sinks steadily increased with an annual growth rate of 57,000 tons. From 2022 to 2025, with support of policy of MFCS and improvement of disaster prevention and mitigation capacity, the potential of MFCS will be further released. The growth rate of MFCS will be increased to 94,000 tons per year, and its overall scale is expected to reach 2,198,245 tons by 2025, equivalent to fixing 8.06 million tons of CO2. The carbon sink's economic value is significantly estimated to be over 400 billion yuan.

Profile change of the volatile and non-volatile compounds in dried or baked laver by photooxidation.

Effects of light or dark storage condition on the profile changes of volatile and non-volatile compounds were evaluated in dried and baked laver for 60 days. Volatile and non-volatile compounds were analyzed using gas chromatography-mass selective detection and high-performance liquid chromatography-quadrupole-time of flight-mass spectrometry, respectively. Baked laver stored in light conditions for 60 days produced the most volatile compounds, whereas dried laver stored in the dark produced the least volatile compounds. Total 11 classes of volatile compounds were detected, including alkanes, alkenes, and ketones, with aldehydes being most abundant in dried laver stored under light. Metabolite analysis of non-volatile compounds led to the selection of 12 compounds with a higher variable importance projection (VIP) value of >1.0: 6 fatty acids (VIP 1.2-2.0), 2 flavanols (VIP 1.3-1.8), hydroxybenzoic acid (VIP 1.5), hydroxycinnamic acid (VIP 2.3), a phenolic acid ester (VIP 1.9), and phloroglucinol (VIP 1.2). Generally, levels of these compounds decreased more following storage in the light than under dark, irrespective of laver preparation. The content of linolenic acid was particularly affected by storage conditions, with light conditions causing a fourfold reduction in linolenic acid level compared with dark conditions, which could result in an increased formation of aldehydes. Gallic acid and sinapinic acid were detected in dried but not baked laver, as they are destroyed by heat treatment. Therefore, laver should be baked and stored in dark conditions to prevent the development of rancidity. PRACTICAL APPLICATION: Laver is one of the representative seaweeds, and the popularity among consumers increases. Although commercially available laver is prepared in dried or baked condition, scientific studies on the changes of metabolites, including volatile and non-volatile compounds during storage, are scarce. The results of this study can be applied to improve proper storage methods to maintain the quality of laver, which can be beneficial for consumers and food industry.

Is happier music groovier? The influence of emotional characteristics of musical chord progressions on groove.

Specific rhythmic patterns in music have been reported to induce an urge to move with feelings of pleasure or enjoyment, called "groove." However, it is unclear how the emotional characteristics of music (e.g., happiness or sadness) affect groove. To address this issue I investigated the effects of the emotional characteristics of music on groove by altering the chord progressions accompanying drum breaks composed by a professional composer while manipulating independent tempo and rhythmic patterns. An online listening experiment was conducted using pieces composed by a professional composer but comprising different types of chord progressions that lead to happiness or sadness. Participants evaluated the nine items on a 7-point scale, including urge to move (i.e., groove), felt emotions, nori, and liking. The experiment found that: (1) chord progressions that evoke happiness were more likely to induce groove, (2) emotional characteristics did not interact with tempi and syncopation in terms of groove ratings, and (3) the accompaniment of drum breaks enhanced groove in both happy and sad chord progressions. Musical pieces with chord progressions that induce happiness were more likely to evoke groove, namely the urge to move. This implies that considering the emotional characteristics of musical pieces and rhythms is crucial when creating music for movement during rehabilitation, therapy, or dance.

Characterization and comparative analysis of the complete organelle genomes of three red macroalgae species (Neoporphyra dentata, Neoporphyra seriata, and Neopyropia yezoensis) and development of molecular makers for their identification.

BACKGROUND: Many species of red algae belonging to the phylum Rhodophyta are consumed by humans as raw materials for nutrition and medicine. As the seaweed market grows, the importance of the laver species has increased. The classification of red algal species has changed significantly, and the accuracy of this classification has improved significantly in recent years. Here, we report the complete circular genomes of the chloroplasts (cp) and mitochondria (mt) of three laver species (Neoporphyra dentata, Neoporphyra seriata, and Neopyropia yezoensis). OBJECTIVE: This study aims to assemble, annotate, and characterize the organization of the organelle genomes of three laver species, conduct comparative genomic studies, and develop molecular markers based on SNPs. METHODS: We analyzed organelle genome structures, repeat sequences, sequence divergence, gene rearrangements, and phylogenetic relationships of three laver species. RESULTS: The chloroplast genomes of the three species contained an average of 212 protein-coding genes (PCGs), while the mitochondrial genomes contained an average of 25 PCGs. We reconstructed the phylogenetic trees based on both chloroplast and mitochondrial genomes using 201 and 23 PCGs (in cp and mt genomes, respectively) shared in the class Bangiophyceae (and five species of Florideophyceae class used as an outgroup). In addition, 12 species-specific molecular markers were developed for qRT-PCR analysis. CONCLUSIONS: This is the first report of Neoporphyra seriata complete organellar genomes. With the results, this study provides useful genetic information regarding taxonomic discrepancies, the reconstruction of phylogenetic trees, and the evolution of red algae. Moreover, the species-specific markers can be used as fast and easy methods to identify a target species.

Ultrasound-assisted fermentation of Porphyra yezoensis sauce at different growth stages using Lactiplantibacillus plantarum: Metabolic response and biological activity.

This study first employed ultrasonic-assisted fermentation of seaweed foot material with Lactiplantibacillus plantarum to produce Porphyra yezoensis sauce. The aim was to examine L. plantarum's growth and metabolism of nutritional components at different growth stages under low- (133.99 W/L) and high-ultrasonic power densities (169.17 W/L). After 24-h fermentation, L. plantarum exhibited a 21.32 % increase in the sonicated P. yezoensis sauce at 133.99 W/L and the logarithmic growth phase compared to that at 169.17 W/L. In addition, compared to the non-sonicated sauce, total phenolic and flavonoid contents increased by around 58 % and 27 % in sonicated sauce at 133.99 W/L, reaching 92.38 mg GEA/g DW and 111.08 mg RE/g DW, respectively. Principal Component Analysis (PCA) of the evaluation criteria for different fermentation stages under 133.99 W/L power ultrasonication revealed that the P. yezoensis sauce generated more phenolic compounds and exhibited stronger antioxidant capabilities in the sonicated sample at the logarithmic phase of L. plantarum. Compared to the traditional treated P. yezoensis sauce, the content of free amino acids was significantly increased in sonicated sauce, especially for logarithmic phase. Finally, GC-IMS analysis demonstrated that the ultrasonication at logarithmic phase released more volatile compounds compared to the non-sonicated sauce. This led to a reduction in the fishy odour of the Porphyra yezoensis sauce and an improved release of favourable flavour compounds.

In vitro-simulated intestinal flora fermentation of Porphyra haitanensis polysaccharides obtained by different assisted extractions and their fermented products against HT-29 human colon cancer cells.

Herein, we studied the in vitro-simulated intestinal flora fermentation of Porphyra haitanensis polysaccharides (PHPs) with microwave, ultrasonic, ultra-high pressure-assisted extraction and the protective effect of their fermented products against HT-29 human colon cancer cells. The results showed that PHPs were largely degraded at the 18 h stage of ascending colon fermentation, further greatly increasing the contents of reducing sugars and short-chain fatty acids (p < 0.05). Particularly, the PHPs subjected to ultra-high pressure-assisted extraction (UHP-PHP) showed the highest reducing sugar content of 1.68 ± 0.01 mg mL-1 and butyric acid content of 410.77 ± 7.99 mmol mL-1. Moreover, UHP-PHP showed a better effect in increasing the ratio of Bacteroidetes/Firmicutes and decreasing the abundance of Proteobacteria and Escherichia coli. PHPs could protect against HT-29 cells by increasing the ROS levels in a concentration-dependent manner, especially UHP-PHP fermented in a descending colon for 24 h. This was related to the up-regulated apoptosis-related genes (Bax and Bak), down-regulated protein expression of Bcl-2 and activation of the p-AKT protein, thereby promoting the apoptosis of HT-29 cells. Our results can facilitate the modification of PHPs and their practical application in the development of intestinal health improving products.

Enhancing Immunity and Modulating Vaginal Microflora Against Candidal Vaginitis Through Nanoemulsion Supplemented with Porphyra Oligosaccharide as an Intravaginal Vaccine Adjuvant.

Background: Intravaginal vaccination is an encouraging approach to prevent infectious vaginitis, with nanoemulsions showing effectiveness as mucosal adjuvants. Purpose: This study aimed to formulate a nanoemulsion incorporating Porphyra oligosaccharide (PO@NE) and assess its effectiveness as a mucosal adjuvant in intravaginal vaccines against candidal vaginitis. Materials and Methods: PO@NE was prepared, and the stability, immunomodulatory activity and cytotoxicity were screened in vitro. Further, the preventive effect of PO@NE as adjuvants for heat-killed Candida albicans (HK-CA) vaccines was explored in a murine model of candidal vaginitis, in comparison with those supplemented with polysaccharide (PP@NE). The mice were intravaginally vaccinated with 106 HK-CA cells, suspended in 1% NE without or with either PO or PP at a final concentration of 6.5 µg/mL, in a total volume of 20 µL. This vaccination was intravaginally administered once a week for 3 weeks. One week following the final vaccination, the mice underwent an intravaginal challenge with 107 C. albicans cells. One week after the challenge, the mice were euthanized to isolate serum, spleen, vaginal washes, and vaginal tissues for analysis. Results: PP@NE and PO@NE, with diameters approximately around 100 nm, exhibited exceptional stability at 4°C and low cytotoxicity when used at a concentration of 1% (v/v). Intravaginal vaccination with HK-CA adjuvanted with PO@NE effectively protected against candidal vaginitis evidenced by less Candida hyphae colonization, milder mucosal damage and cell infiltration. Moreover, enhanced mucosal antibody production, induction of T helper (Th)1 and Th17-related immune responses, enlarged the population of CD8+ cells, and elevated vaginal microflora diversity were observed in vaccinated mice. Interestingly, the potency was rather attenuated when PO@NE was replaced with PP@NE. Conclusion: These findings indicate PO@NE as a HK-CA vaccine adjuvant for candidal vaginitis prevention via enhancement of both cellular and humoral immunity and modulation of vaginal microflora, emphasizing further intravaginal vaccination development.