Targeted Application of Functional Foods as Immune Fitness Boosters in the Defense against Viral Infection.

In recent times, the emergence of viral infections, including the SARS-CoV-2 virus, the monkeypox virus, and, most recently, the Langya virus, has highlighted the devastating effects of viral infection on human life. There has been significant progress in the development of efficacious vaccines for the prevention and control of viruses; however, the high rates of viral mutation and transmission necessitate the need for novel methods of control, management, and prevention. In recent years, there has been a shift in public awareness on health and wellbeing, with consumers making significant dietary changes to improve their immunity and overall health. This rising health awareness is driving a global increase in the consumption of functional foods. This review delves into the benefits of functional foods as potential natural means to modulate the host immune system to enhance defense against viral infections. We provide an overview of the functional food market in Europe and discuss the benefits of enhancing immune fitness in high-risk groups, including the elderly, those with obesity, and people with underlying chronic conditions. We also discuss the immunomodulatory mechanisms of key functional foods, including dairy proteins and hydrolysates, plant-based functional foods, fermentates, and foods enriched with vitamin D, zinc, and selenium. Our findings reveal four key immunity boosting mechanisms by functional foods, including inhibition of viral proliferation and binding to host cells, modulation of the innate immune response in macrophages and dendritic cells, enhancement of specific immune responses in T cells and B cells, and promotion of the intestinal barrier function. Overall, this review demonstrates that diet-derived nutrients and functional foods show immense potential to boost viral immunity in high-risk individuals and can be an important approach to improving overall immune health.

A secondary metabolite drives intraspecies antagonism in a gut symbiont that is inhibited by cell-wall acetylation.

The mammalian microbiome encodes numerous secondary metabolite biosynthetic gene clusters; yet, their role in microbe-microbe interactions is unclear. Here, we characterized two polyketide synthase gene clusters (fun and pks) in the gut symbiont Limosilactobacillus reuteri. The pks, but not the fun, cluster encodes antimicrobial activity. Forty-one of 51 L. reuteri strains tested are sensitive to Pks products; this finding was independent of strains' host origin. Sensitivity to Pks was also established in intraspecies competition experiments in gnotobiotic mice. Comparative genome analyses between Pks-resistant and -sensitive strains identified an acyltransferase gene (act) unique to Pks-resistant strains. Subsequent cell-wall analysis of wild-type and act mutant strains showed that Act acetylates cell-wall components, providing resistance to Pks-mediated killing. Additionally, pks mutants lost their competitive advantage, while act mutants lost their Pks resistance in in vivo competition assays. These findings provide insight into how closely related gut symbionts can compete and co-exist in the gastrointestinal tract.

Defining the Cholesterol Lowering Mechanism of Bergamot (Citrus bergamia) Extract in HepG2 and Caco-2 Cells.

Bergamot, a Mediterranean citrus fruit native to southern Italy, has been reported to have cholesterol-lowering properties; however, the mechanism of action is not well understood. Due to structural similarities with 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) inhibitors, it has been proposed that the phenolic compounds in bergamot may also inhibit HMGCR. Statins are widely used for their cholesterol-lowering properties; however, they are not universally well tolerated, suggesting there is a need to identify novel cholesterol-lowering strategies. In the present study, we investigated bergamot fruit extract (BFE) and its principal components (neoeriocitrin, naringin, neohesperidin, melitidin, and brutieridin) for their ability to regulate cholesterol levels in HepG2 and Caco-2 cells. BFE at increasing concentrations decreased the levels of total and free cholesterol in HepG2 cells. BFE and its constituents did not directly inhibit HMGCR activity. However, BFE and neohesperidin decreased HMGCR levels in HepG2 cells, suggesting that neohesperidin and BFE may downregulate HMGCR expression. An increase in AMP-kinase phosphorylation was observed in BFE and neohesperidin-treated cells. In Caco-2 cells, brutieridin exhibited a significant reduction in cholesterol uptake and decreased the level of Niemann-Pick C1 Like 1, an important cholesterol transporter. Taken together, our data suggest that the cholesterol-lowering activity of bergamot is distinct from statins. We hypothesize that BFE and its principal constituents lower cholesterol by inhibiting cholesterol synthesis and absorption.

Pharmacokinetic Analysis of Carnosic Acid and Carnosol in Standardized Rosemary Extract and the Effect on the Disease Activity Index of DSS-Induced Colitis.

Rosemary extract (RE) is an approved food preservative in the European Union and contains dietary phytochemicals that are beneficial for gastrointestinal health. This study investigated the effects of RE on dextran sodium sulfate (DSS)-induced colitis and also determined the pharmacokinetics of dietary phytochemicals administered to mice via oral gavage. Individual components of rosemary extract were separated and identified by LC-MS/MS. The pharmacokinetics of two major diterpenes from RE, carnosic acid (CA) and carnosol (CL), administered to mice via oral gavage were determined. Then, the effect of RE pre-treatment on the disease activity index (DAI) of DSS-induced colitis in mice was investigated. The study determined that 100 mg/kg RE significantly improved DAI in DSS-induced colitis compared to negative control. Sestrin 2 protein expression, which increased with DSS exposure, was reduced with RE treatment. Intestinal barrier integrity was also shown to improve via fluorescein isothiocyanate (FITC)-dextran administration and Western blot of zonula occludens-1 (ZO-1), a tight junction protein. Rosemary extract was able to improve the DAI of DSS-induced colitis in mice at a daily dose of 100 mg/kg and showed improvement in the intestinal barrier integrity. This study suggests that RE can be an effective preventative agent against IBD.

Covalent Modification of Proteins by Plant-Derived Natural Products: Proteomic Approaches and Biological Impacts.

Plant-derived natural products (NPs) with electrophilic functional groups engage various subsets of the proteome via covalent modification of nucleophilic cysteine residues. This electrophile-nucleophile interaction can change protein conformation, alter protein function, and modulate their biological action. The biological significance of these covalent protein modifications in health and disease is increasingly recognized. One way to understand covalent NP-protein interactions is to utilize traditional proteomics and modern mass spectrometry (MS)-based proteomic strategies. These strategies have proven effective in uncovering specific NP protein targets and are critical first steps that allow for a much deeper understanding of the ability of NPs to modulate cellular processes. Here, plant-derived NPs that covalently modify proteins are reviewed, the biological significance of these covalent modifications, and the different proteomic strategies that have been employed to study these NP-protein interactions.

Prenylated xanthones from mangosteen (Garcinia mangostana) activate the AhR and Nrf2 pathways and protect intestinal barrier integrity in HT-29 cells.

Xanthones from the tropical fruit mangosteen (Garcinia mangostana) display anti-inflammatory and anti-oxidative activities. Here, we isolate and identify potential inducers of the aryl hydrocarbon receptor (AhR) and nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathways from mangosteen using a bioassay-guided strategy. Mangosteen fruit pericarp extracts were subjected to sequential solvent extractions, followed by chromatography coupled with NMR spectroscopy and mass spectrometric analyses for identification and isolation of pure compounds. Isolation of active fractions led to seven prenylated xanthones that were identified and subsequently evaluated for bioactivity. In vitro luciferase reporter cellular assays using H1L6.1c3 (AhR induction) and HepG2-ARE (Nrf2 induction) were used to identify AhR and Nrf2 activators. All seven prenylated xanthones displayed AhR inducing activity, whereas only five xanthones activated Nrf2. Garcinone D (GarD) significantly upregulated AhR/Cyp1a1 and Nrf2/HO-1 protein expression and enhanced zonula occludens-1 and occludin protein levels in HT-29 cells. In addition, GarD inhibited oxidative stress-induced intestinal epithelial barrier dysfunction by enhancing tight junction (TJ) proteins and inhibition of reactive oxygen species production. Inhibition of AhR by pretreating cells with an AhR antagonist revealed that the AhR pathway is required for the improved epithelial barrier functions of GarD. These results highlight a dual mechanism by GarD that confers protection against intestinal epithelial barrier dysfunction.

Inhibition of CDK2/CyclinE1 by xanthones from the mangosteen (Garcinia mangostana): a structure-activity relationship study.

Uncontrolled regulation of cyclin dependent kinases (CDKs) has negative implications in many cancers and malignancies and has recently led to the approval of select CDK inhibitors. Herein we present data reporting that xanthones, a class of compounds isolated from the purple mangosteen (Garcinia mangostana) fruit, can inhibit CDK2/CyclinE1. We evaluated nine different xanthones, including α-mangostin, ß-mangostin, γ-mangostin, gartanin, 8-desoxygartanin, garcinone C, garcinone D, 9-hydroxycalabaxanthone, and 3-isomangostin for toxicity in 22Rν1 (prostate cancer cells) and MDA-MB-231 (breast cancer cells). All compounds dose-dependently inhibited the viability of both cell lines. A cell free biochemical assay was performed to determine if selected phytochemicals inhibited CDK2/CyclinE1. γ-Mangostin and α-mangostin were the strongest inhibitors, respectively. The results suggest that the position of key functional groups including hydroxyl and isoprenyl groups contribute to the CDK2 inhibitory effect. Taken together, the evidence suggests that xanthones can directly target CDK2 providing a possible explanation for their therapeutic potential.

Valorization of pomelo (Citrus grandis Osbeck) peel: A review of current utilization, phytochemistry, bioactivities, and mechanisms of action.

Citrus grandis Osbeck, commonly known as "pomelo" or "shaddock," is the largest citrus fruit, the peel of which is a well-known agricultural residual waste. Pomelo peel offers a wide range of components such as essential oils, polysaccharides, and phytochemicals with potential food applications. Utilization of pomelo peel to recover these components is an important step toward agricultural sustainability. This review covers pomelo peel utilization opportunities beyond conventional composting and animal feed production, and critically examines value-added uses via the recovery of potentially bioactive components. The peel of pomelo accounts for approximately 30% of the total fruit weight and contains phytochemicals, including aroma-active volatiles, pectin, flavonoids, phenolic acids, carotenoids, coumarins, and polysaccharides. Recovery of these phytochemicals offers an opportunity for value-added utilization such as the development of enriched or functional foods and nutraceuticals. The health-promoting and therapeutic potential of pomelo peel extracts and isolated pure compounds have been evaluated through numerous in vitro and in vivo studies that revealed a wide range of bioactivities, including hypolipidemic, hypoglycemic, antioxidant, antimicrobial, anti-inflammatory, and anticancer effects. Preclinical evidence highlights multifaceted molecular and signaling events that possibly underlie the said bioactive potential. Overall, the pomelo processing industry offers a great opportunity to recover or produce valuable products from the large amounts of residual wastes it generates. It is envisaged that a thorough understanding of the bioactive components of pomelo peel, their functional and nutraceutical applications, and mode of actions will benefit the food industry.

S-1-propenylmercaptocysteine protects murine hepatocytes against oxidative stress via persulfidation of Keap1 and activation of Nrf2.

The onion-derived metabolite, S-1-propenylmercaptocysteine (CySSPe), protects against oxidative stress and exhibits anti-inflammatory effects by modulating cellular redox homeostasis. We sought to establish whether CySSPe activates nuclear factor erythroid 2-related factor 2 (Nrf2) and whether activation of Nrf2 by CySSPe involves modification of the Kelch-like ECH-associated protein-1 (Keap1) to manifest these effects. We found that CySSPe stabilized Nrf2 protein and facilitated nuclear translocation to induce expression of antioxidant enzymes, including NQO1, HO-1, and GCL. Moreover, CySSPe attenuated tert-butyl hydroperoxide-induced cytotoxicity and dose-dependently inhibited reactive oxygen species production. Silencing experiments using Nrf2-siRNA confirmed that CySSPe conferred protection against oxidative stress by activating Nrf2. CySSPe enhanced cellular pool of reduced glutathione (GSH) and improved GSH:GSSG ratio. Pretreatment of cells with l-buthionine-S,R-sulfoximine (BSO) confirmed that CySSPe increases de novo synthesis of GSH by upregulating expression of the GSH-synthesizing enzyme GCL. Treatment of cells with CySSPe elevated hydrogen sulfide (H2S) production. Inhibition of H2S-synthesizing enzymes, cystathionine-gamma-lyase (CSE) and cystathionine-beta-synthase (CBS), by pretreating cells with propargylglycine (PAG) and oxyaminoacetic acid (AOAA) revealed that H2S production was partially dependent on a CSE/CBS-catalyzed ß-elimination reaction with CySSPe that likely produced 1-propenyl persulfide (RSSH). Depleting cells of their GSH pool by exposure to BSO and diethylmaleate attenuated H2S production, suggesting a GSH-dependent formation of H2S, likely via the reduction of RSSH by GSH. Finally, treatment of cells with CySSPe persulfidated Keap1, which may be the mechanism involved for the stabilization of Nrf2 by CySSPe. Taken together, our results showed that attenuation of oxidative stress by CySSPe is associated with its ability to produce H2S or RSSH, which persulfidates Keap1 and activates Nrf2 signaling. This study provides insights on the potential of CySSPe as an onion-derived dietary agent that modulates redox homeostasis and combats oxidative stress.

Gut Symbionts Lactobacillus reuteri R2lc and 2010 Encode a Polyketide Synthase Cluster That Activates the Mammalian Aryl Hydrocarbon Receptor.

A mechanistic understanding of microbe-host interactions is critical to developing therapeutic strategies for targeted modulation of the host immune system. Different members of the gut symbiont species Lactobacillus reuteri modulate host health by, for example, reduction of intestinal inflammation. Previously, it was shown that L. reuteri activates the aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor that plays an important role in the mucosal immune system, by the production of tryptophan catabolites. Here, we identified a novel pathway by which L. reuteri activates AhR, which is independent of tryptophan metabolism. We screened a library of 36 L. reuteri strains and determined that R2lc and 2010, strains with a pigmented phenotype, are potent AhR activators. By whole-genome sequencing and comparative genomics, we identified genes unique to R2lc and 2010. Our analyses demonstrated that R2lc harbors two genetically distinct polyketide synthase (PKS) clusters, functionally unknown (fun) and pks, each carried by a multicopy plasmid. Inactivation of pks, but not fun, abolished the ability of R2lc to activate AhR. L. reuteri 2010 has a gene cluster homologous to the pks cluster in R2lc with an identical gene organization, which is also responsible for AhR activation. In conclusion, we identified a novel PKS pathway in L. reuteri R2lc and 2010 that is responsible for AhR activation.IMPORTANCE Temporary changes in the composition of the microbiota, for example, by oral administration of probiotics, can modulate the host immune system. However, the underlying mechanisms by which probiotics interact with the host are often unknown. Here, we show that Lactobacillus reuteri R2lc and 2010 harbor an orthologous PKS gene cluster that activates the aryl hydrocarbon receptor (AhR). AhR is a ligand-activated transcription factor that plays a key role in a variety of diseases, including amelioration of intestinal inflammation. Understanding the mechanism by which a bacterium modulates the immune system is critical for applying rational selection strategies for probiotic supplementation. Finally, heterologous and/or optimized expression of PKS is a logical next step toward the development of next-generation probiotics to prevent and treat disease.

Data on chromatographic isolation of cysteine mixed-disulfide conjugates of Allium thiosulfinates and their role in cellular thiol redox modulation.

This data article contains experimental data on the preparation and semi-preparative isolation of S-Alk(en)ylmercaptocysteine (CySSRs, R = allyl, "A", 1-propenyl, "Pe" or methyl, "Me") generated through conjugation reactions between allyl and 1-propenyl enriched thiosulfinates (TS) and cysteine. The data presented are related to the research article "S-Alk(en)ylmercaptocysteine suppresses LPS-induced pro-inflammatory responses in murine macrophages through inhibition of NF-κB pathway and modulation of thiol redox status" (Tocmo and Parkin, in press). In this data article, we included a detailed procedure for CySSR preparation, their purification through semi-preparative chromatography and their toxicity profiles in RAW 264.7 macrophages. Data included also highlight, the ability of CySSRs to modulate intracellular thiol redox status.

S-Alk(en)ylmercaptocysteine suppresses LPS-induced pro-inflammatory responses in murine macrophages through inhibition of NF-κB pathway and modulation of thiol redox status.

The Allium vegetable-derived metabolite, S-alk(en)ylmercaptocysteine (CySSR), has been reported to modulate oxidative stress and inflammatory responses. However, the underlying mechanisms of action and structure-activity relationships are not completely understood. We investigated the mechanistic basis of the protective effects of CySSR on pro-inflammatory responses involving redox/oxidative stress induced by E. coli lipopolysaccharide (LPS) using RAW 264.7 cells. CySSR (R = allyl, "A" or 1-propenyl, "Pe") pre-treatments conferred concentration-dependent reductions in cytokines (TNF-α, IL-1ß and IL-6), NO production and iNOS (inducible nitric synthase) overexpression, and attenuated oxidant production in LPS-stimulated RAW 264.7 cells where viability remained > 90%. These protective effects were manifested through inhibited activation of the nuclear factor-kappa B (NF-κB) signaling pathway via suppression of the IκB kinases (IKK) phosphorylation possibly by transforming growth factor ß-activated kinase 1 or a kinase further upstream the canonical NF-κB signaling pathway. The attenuation of LPS-induced inflammation by CySSRs was associated with enhanced levels of cellular cysteine (CySH) and glutathione (GSH) mediated by cellular import/reduction of CySSR and the induction of glutamate cysteine ligase (GCL), one of > 200 nuclear factor erythroid 2-related factor 2 (Nrf2) regulated proteins. The reduction of anti-inflammatory effect of CySSR following pretreatment of cells with L-buthionine-S,R-sulfoximine (BSO) implicates GSH having a major role in reducing inflammation, likely in the context of other Nrf2-regulated antioxidant enzymes that scavenge H2O2 and peroxides using GSH as co-substrate. The anti-inflammatory effect of CySSPe was significantly greater than CySSA for almost all indicators measured, and cell metabolites of CySSRs may have a role in attenuating NF-κB signaling.

Boiling enriches the linear polysulfides and the hydrogen sulfide-releasing activity of garlic.

Garlic is rich in polysulfides, and some of them can be H2S donors. This study was conducted to explore the effect of cooking on garlic's organopolysulfides and H2S-releasing activity. Garlic bulbs were crushed and boiled for a period ranging from 3 to 30min and the solvent extracts were analyzed by GC-MS/FID and HPLC. A cell-based assay was used to measure the H2S-releasing activity of the extracts. Results showed that the amounts of allyl polysulfides increased in crushed garlic boiled for 6-10min; however, prolonging the thermal treatment to 20 or 30min decreased their concentrations. Data of the H2S-releasing activity, expressed as diallyl trisulfide equivalents (DATS-E), parallel this trend, being significantly higher at 6 and 10min boiling. Our results showed enhancement of H2S-releasing activity upon moderate boiling, suggesting that shorter cooking time may maximize its health benefits as a dietary source of natural H2S donors.

Data on the effect of boiling on the organosulfides and the hydrogen sulfide-releasing activity of garlic.

This article contains experimental data on the identification and quantification of the organosulfides on boiled garlic extracts. Data included are related to the research article "Boiling enriches the linear polysulfides and the hydrogen sulfide-releasing activity of garlic" (R. Tocmo, Y. Wu, D. Liang, V. Fogliano, D. Huang, 2016) [1]. Characterization was carried out by GC-MS and HPLC. Dose-response curves obtained from the cell-based H2S-releasing capacity assay of allicin transformation products, namely vinyl dithiins and ajoene are also included. DATS-E values were calculated from these dose-response curves to quantify the contribution of the individual polysulfides to the H2S-releasing capacity of boiled extracts.

Organosulfide profile and hydrogen sulfide-releasing capacity of stinky bean (Parkia speciosa) oil: Effects of pH and extraction methods.

Stinky beans (Parkia speciosa) were hydrodistilled and solvent-extracted and the oil obtained was analyzed by GC-MS/FID. Nine cyclic and one acyclic organosulfides were identified comprising 36% of total volatiles. Solvent extracts contained significantly (p < 0.05) higher total organosulfides (680 ppm) as compared to distilled oil (444 ppm). The concentrations of organosulfides are highly dependent on the pH values of the matrix, with control sample (pH 5.40) giving the highest total organosulfides (424 ppm) followed by that of pH 7.0 (234 ppm), pH 9.0 (195 ppm), and pH 3.0 (152 ppm). The H2S-releasing capacity, expressed as diallyl trisulfide equivalents (DATS-E in mmol DATS/g), corresponded well with the differences in organosulfide concentrations as affected by pH with control having the highest value (24.35) followed pH 7.0 (7.27), pH 9.0 (3.27), and pH 3.0 (1.80). We conclude that stinky bean oil is a potent H2S-releasing agent that could have health-beneficial properties.

Chemical and biochemical mechanisms underlying the cardioprotective roles of dietary organopolysulfides.

Foods that are rich in organosulfides are highly regarded for their broad range of functions in disease prevention and health promotion since ancient time yet modern scientific study, particularly clinical studies could not agree with traditional wisdom. One of the complexities is due to the labile nature of organosulfides, which are often transformed to different structures depending on the processing conditions. The recent evidence on polysulfides as H2S donors may open up a new avenue for establishing structure and health promotion activity relationship. To put this development into perspective, we carried out a review on the recent progress on the chemistry and biochemistry of organopolysulfides with emphasis on their cardioprotective property. First, we briefly surveyed the foods that are rich in polysulfides and their structural diversity. This is followed by in-depth discussion on the chemical transformations of polysulfides under various processing conditions. We further reviewed the potential action mechanisms of polysulfides in cardioprotection through: (a) hydrogen sulfide releasing activity; (b) radical scavenging activity; and (c) activity in enzyme inhibition and intervention of gene regulation pathways. Based on the literature trend, we can conclude that the emerging concept of organopolysulfides as naturally occurring H2S donors is intriguing and warrants further research to establish the structure and activity relationship of the organopolysulfides as H2S donors.

Effect of processing conditions on the organosulfides of shallot (Allium cepa L. Aggregatum group).

There is a growing account of the health benefits of H2S as an endogenous cell-signaling molecule. H2S from organic polysulfides, in particular, is increasingly gaining attention for their beneficial effects to cardiovascular health. Here, we studied shallot as a potential dietary source of organic polysulfides and examined the effects of processing conditions on its polysulfide profiles. Boiling, autoclaving, and freeze-drying were tested on whole and crushed shallot bulbs, analyzing their effect on the yield of organosulfides. Seventeen organosulfides, including disulfides, trisulfides, and cyclic polysulfides, were identified. Significant differences in the quantitative and qualitative profiles of organosulfides in the hydrodistilled and solvent extracted oils were observed. Freeze-drying retained the majority of the organosulfides, but the whole-autoclaved and whole-boiled shallots lost more than 95% of their organic polysulfides. Crushed-boiled and crushed-autoclaved shallot lost 76-80% of their organosulfides, likely due to the thermal sensitivity of these compounds. The organosulfide profiles are sensitive to the pH values of the processing media. In general, disulfides increased at basic pH (pH 9.0) while trisulfides and cyclic organosulfides are much higher at the acidic to neutral pH values (pH 3.0-5.0). Our results provide important information on the effects of processing conditions that are relevant for optimizing extraction of organosulfides from shallot for further studies evaluating their H2S-releasing activity.

Listeria monocytogenes in Vacuum-Packed Smoked Fish Products: Occurrence, Routes of Contamination, and Potential Intervention Measures.

The occurrence of Listeria monocytogenes in ready-to-eat (RTE) fish products is well documented and represents an important food safety concern. Contamination of this pathogen in vacuum-packed (VP) smoked fish products at levels greater than the RTE food limit (100 CFU/g) has been traced to factors such as poor sanitary practices, contaminated processing environments, and temperature abuse during prolonged storage in retail outlets. Intervention technologies including physical, biological, and chemical techniques have been studied to control transmission of L. monocytogenes to these products. High-pressure processing, irradiation, and pulsed UV-light treatment have shown promising results. Potential antilisterial effects of some sanitizers and combined chemical preservatives have also been demonstrated. Moreover, the concept of biopreservation, use of bioactive packaging, and a combination of different intervention technologies, as in the hurdle concept, are also under consideration. In this review, the prevalence, routes of contamination, and potential intervention technologies to control transmission of L. monocytogenes in VP smoked fish products are discussed.