Common food additive carrageenan inhibits proglucagon expression and GLP-1 secretion by human enteroendocrine L-cells.

Proglucagon mRNA expression and GLP-1 secretion by cultured human L-cells (NCI-H716) were inhibited following exposure to λ-carrageenan, a commonly used additive in processed foods. Carrageenan is composed of sulfated or unsulfated galactose residues linked in alternating alpha-1,3 and beta-1,4 bonds and resembles the endogenous sulfated glycosaminoglycans. However, carrageenan has unusual alpha-1,3-galactosidic bonds, which are not innate to human cells and are implicated in immune responses. Exposure to carrageenan predictably causes inflammation, and carrageenan impairs glucose tolerance and contributes to insulin resistance. When cultured human L-cells were deprived overnight of glucose and serum and then exposed to high glucose, 10% FBS, and λ-carrageenan (1 µg/ml) for 10 minutes, 1 h, and 24 h, mRNA expression of proglucagon and secretion of GLP-1 were significantly reduced, compared to control cells not exposed to carrageenan. mRNA expression of proglucagon by mouse L-cells (STC-1) was also significantly reduced and supports the findings in the human cells. Exposure of co-cultured human intestinal epithelial cells (LS174T) to the spent media of the carrageenan-treated L-cells led to a decline in mRNA expression of GLUT-2 at 24 h. These findings suggest that ingestion of carrageenan-containing processed foods may impair the production of GLP-1, counteract the effect of GLP-1 receptor agonists and induce secondary effects on intestinal epithelial cells.

The food additive xylitol enhances the butyrate formation by the child gut microbiota developed in a dynamic colonic simulator.

The gut microbiota should be included in the scientific processes of risk assessment of food additives. Xylitol is a sweetener that shows low digestibility and intestinal absorption, implying that a high proportion of consumed xylitol could reach the colonic microbiota. The present study has evaluated the dose-dependent effects of xylitol intake on the composition and the metabolic activity of the child gut-microbiota. The study was conducted in a dynamic simulator of the colonic microbiota (BFBL Gut Simulator) inoculated with a child pooled faecal sample and supplemented three times per day, for 7 days, with increasing xylitol concentrations (1 g/L, 3 g/L and 5 g/L). Sequencing of 16S rRNA gene amplicons and group-specific quantitative PCR indicated a xylitol dose-response effect on the abundance of Lachnospiraceae, particularly the genera Blautia, Anaerostipes and Roseburia. The microbial changes observed with xylitol corresponded with a dose-dependant effect on the butyrate concentration that, in parallel, favoured an increase in epithelial integrity of Caco-2 cells. The study represents a detailed observation of the bacterial taxa that are the main contributors to the metabolism of xylitol by the child gut microbiota and the results could be relevant in the risk assessment re-evaluation of xylitol as a sweetener.

Evaluating the Effects of Chronic Oral Exposure to the Food Additive Silicon Dioxide on Oral Tolerance Induction and Food Sensitivities in Mice.

BACKGROUND: The increasing prevalence of food sensitivities has been attributed to changes in gut microenvironment; however, ubiquitous environmental triggers such as inorganic nanoparticles (NPs) used as food additives have not been thoroughly investigated. OBJECTIVES: We explored the impact of the NP-structured food-grade silicon dioxide (fg-SiO2) on intestinal immune response involved in oral tolerance (OT) induction and evaluated the consequences of oral chronic exposure to this food-additive using a mouse model of OT to ovalbumin (OVA) and on gluten immunopathology in mice expressing the celiac disease risk gene, HLA-DQ8. METHODS: Viability, proliferation, and cytokine production of mesenteric lymph node (MLN) cells were evaluated after exposure to fg-SiO2. C57BL/6J mice and a mouse model of OT to OVA were orally exposed to fg-SiO2 or vehicle for 60 d. Fecal lipocalin-2 (Lcn-2), anti-OVA IgG, cytokine production, and immune cell populations were analyzed. Nonobese diabetic (NOD) mice expressing HLA-DQ8 (NOD/DQ8), exposed to fg-SiO2 or vehicle, were immunized with gluten and immunopathology was investigated. RESULTS: MLN cells exposed to fg-SiO2 presented less proliferative T cells and lower secretion of interleukin 10 (IL-10) and transforming growth factor beta (TGF-ß) by T regulatory and CD45+ CD11b+ CD103+ cells compared to control, two factors mediating OT. Mice given fg-SiO2 exhibited intestinal Lcn-2 level and interferon gamma (IFN-γ) secretion, showing inflammation and less production of IL-10 and TGF-ß. These effects were also observed in OVA-tolerized mice exposed to fg-SiO2, in addition to a breakdown of OT and a lower intestinal frequency of T cells. In NOD/DQ8 mice immunized with gluten, the villus-to-crypt ratio was decreased while the CD3+ intraepithelial lymphocyte counts and the Th1 inflammatory response were aggravated after fg-SiO2 treatment. DISCUSSION: Our results suggest that chronic oral exposure to fg-SiO2 blocked oral tolerance induction to OVA, and worsened gluten-induced immunopathology in NOD/DQ8 mice. The results should prompt investigation on the link between SiO2 exposure and food sensitivities in humans. https://doi.org/10.1289/EHP12758.

Oral exposure to food grade titanium dioxide (E171) induces intestinal and behavioural alterations in adult mice but limited effects in young mice.

BACKGROUND: Food-grade titanium dioxide (E171), a white colourant widely used in ultra-processed food products, has been banned in the European Union. However, its usage is still permitted in medicines, and in several other countries. The estimated intake of E171 in children is higher than in adults, which led us to hypothesise that E171 induces differential effects depending on age, with adult mice being the most susceptible due to age, despite the lower dose. AIM: To evaluate the effects of oral administration of E171 on intestinal permeability, ileum, and colon histology, and how these effects impact anxious and depressive behaviour in young and adult mice of both sexes. METHODS: Young and adult mice of both sexes C57BL/6 mice received 10 mg/kgbw E171/3 times per week for 3 months. E171 was administered orally in water by pipetting, while control groups only received drinking water, then intestinal permeability, histology and animal behaviour were analysed. RESULTS: E171 showed an amorphous shape, primary particles sized below 1 µm and anatase crystalline structure. Oral administration of E171 disrupted the intestinal permeability in adult male and female mice, but no effects were observed in young mice of both sexes. E171 promoted ileal adenoma formation in half of the adult female population, moreover hyperplastic crypts, and hyperplastic goblet cells at histological level in adult mice of both sexes. The colon presented hyperplastic goblet cells, hyperchromatic nuclei, increased proliferation and DNA damage in adult mice of both sexes. The anxiety and depressive behaviour were only altered in adult mice treated with E171, but no changes were detected in young animals of both sexes. CONCLUSIONS: Adult mice displayed higher susceptibility in all parameters analysed in this study compared to young mice of both sexes.

Assessing the potential of phytogenic feed additives: A comprehensive review on their effectiveness as a potent dietary enhancement for nonruminant in swine and poultry.

Phytogenic feed additives (PFAs) often referred to as phytobiotics or botanical feed additives, are natural compounds derived from various plants, herbs, spices and other botanical sources. These feed additives are intended to serve a variety of purposes, including an immune system regulator, an antimicrobial, an antimutagenic, an antioxidant and a growth promoter. They are composed of bioactive compounds extracted from plants, including essential oils, polyphenols, terpenoids and flavonoids. They are mostly utilized as substitute antibiotic growth promoters in nonruminant (swine and poultry) livestock production, owing to the prohibition of antibiotic usage in the feed industry. It has been thoroughly examined to ascertain their impact on intestinal health and activity, correlation with animals' effective health and well-being, productivity, food security and environmental impact. The potential uses of these feed additives depend on the properties of herbs, the comprehension of their principal and secondary components, knowledge of their mechanisms of action, the safety of animals and the products they produce. They are gaining recognition as effective and sustainable tools for promoting animal health and performance while reducing the reliance on antibiotics in nonruminant nutrition. Their natural origins, multifaceted benefits and alignment with consumer preferences make them a valuable addition to modern animal farming process. However, because of their inconsistent effects and inadequate knowledge of the mechanisms of action, their usage as a feed additive has been limited. This review offers a comprehensive assessment of the applications of PFAs as an effective feed supplement in swine and poultry nutrition. In summary, this comprehensive review provides current knowledge, identifies gaps in research and emphasizes the potential of phytogenic additives to foster sustainable and healthier livestock production systems while addressing the global concerns associated with antibiotic use in livestock farming.

Trilobatin attenuates cerebral ischaemia/reperfusion-induced blood-brain barrier dysfunction by targeting matrix metalloproteinase 9: The legend of a food additive.

BACKGROUND AND PURPOSE: Blood-brain barrier (BBB) breakdown is one of the crucial pathological changes of cerebral ischaemia-reperfusion (I/R) injury. Trilobatin (TLB), a naturally occurring food additive, exerts neuroprotective effects against cerebral I/R injury as demonstrated in our previous study. This study was designed to investigate the effect of TLB on BBB disruption after cerebral I/R injury. EXPERIMENTAL APPROACH: Rats with focal cerebral ischaemia caused by transient middle cerebral artery occlusion were studied along with brain microvascular endothelial cells and human astrocytes to mimic BBB injury caused by oxygen and glucose deprivation/reoxygenation (OGD/R). KEY RESULTS: The results showed that TLB effectively maintained BBB integrity and inhibited neuronal loss following cerebral I/R challenge. Furthermore, TLB increased tight junction proteins including ZO-1, Occludin and Claudin 5, and decreased the levels of apolipoprotein E (APOE) 4, cyclophilin A (CypA) and phosphorylated nuclear factor kappa B (NF-κB), thereby reducing proinflammatory cytokines. TLB also decreased the Bax/Bcl-2 ratio and cleaved-caspase 3 levels along with a reduced number of apoptotic neurons. Molecular docking and transcriptomics predicted MMP9 as a prominent gene evoked by TLB treatment. The protective effects of TLB on cerebral I/R-induced BBB breakdown was largely abolished by overexpression of MMP9, and the beneficial effects of TLB on OGD/R-induced loss of BBB integrity in human brain microvascular endothelial cells and astrocyte co-cultures was markedly reinforced by knockdown of MMP9. CONCLUSIONS AND IMPLICATIONS: Our findings reveal a novel property of TLB: preventing BBB disruption following cerebral I/R via targeting MMP9 and inhibiting APOE4/CypA/NF-κB axis.

The impact of antimicrobial food additives and sweeteners on the growth and metabolite production of gut bacteria.

Metabolic disorders caused by the imbalance of gut microbiota have been associated with the consumption of processed foods. Thus, this study aimed to evaluate the effects of antimicrobial food additives (benzoate, sorbate, nitrite, and bisulfite) and sweeteners (saccharin, stevia, sucralose, aspartame, and cyclamate) on the growth and metabolism of some gut and potentially probiotic bacterial species. The effects on the growth of Bifidobacterium longum, Enterococcus faecium, Lactobacillus acidophilus, and Lactococcus lactis subsp. lactis cultures were investigated using a turbidimetric test and by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). To evaluate the metabolic activity, the cultures were exposed to compounds with the highest antimicrobial activity, subjected to cultivation with inulin (1.5%), and analyzed by liquid chromatography for the production of short-chain fatty acids (acetate, propionate, and butyrate). The results showed that potassium sorbate (25 mg/mL), sodium bisulfite (0.7 mg/mL), sodium benzoate, and saccharin (5 mg/mL) presented greater antimicrobial activity against the studied species. L. lactis and L. acidophilus bacteria had reduced short-chain fatty acid production after exposure to saccharin and sorbate, and B. longum after exposure to sorbate, in comparison to controls (acetic acid reduction 1387 µg/mL and propionic 23 µg/mL p < 0.05).

Effects of barium sulfate on rheological properties and IDDSI flow consistency of liquid stimuli prepared using commercial thickening powders.

During videofluoroscopic swallowing study (VFSS), barium sulfate (BaSO4 ) is commonly added into food samples as a radiopaque contrast media for bolus visualization and examination. Accordingly, the consistency and flow behavior of barium stimuli can differ significantly from their non-barium counterparts. Such differences may have a subsequent impact on the validity of VFSS. Therefore, in this study, effects of barium sulfate on the shear and extensional rheological properties and IDDSI (International Dysphagia Diet Standardization Initiative) flow consistency of liquids prepared using various commercial thickening powders were investigated. Results showed that all barium stimuli exhibited shear thinning behavior but with significantly higher shear viscosity compared to the non-barium counterparts. A shift factor of viscosity at shear rate 50 s-1 with values in range of 1.21-1.73 could be used to describe the increase in the viscosity for samples thickened with gum-based thickeners. However, the change in the viscosity was not invariant for the stimuli prepared starch-based thickener. The addition of BaSO4 had a negative impact on extensional properties of samples by demonstrating a faster filament rupture. The extent of impact on the decrease in filament breakup time was more pronounced in xanthan > guar gum ≈ tara gum-based thickeners. Based on the IDDSI flow test, no significant effect of BaSO4 was found on the gum-based thickeners, whereas there was a marked effect in the starch-based sample. These results can be used beneficially to assist clinicians in the dysphagia diagnosis for matching rheological properties of the barium stimuli to enhance effectiveness dysphagia interventions.

A 13-week subchronic toxicity study of heme iron in SD rats.

Heme iron (HI) has been widely used as a food additive and supplement to support iron fortification. However, no sufficient toxicological data to evaluate the safety of HI have been reported. In the current study, we performed a 13-week subchronic toxicity study of HI in male and female Crl:CD(SD) rats. Rats were orally administered HI in the diet at concentrations of 0%, 0.8%, 2%, and 5%. Observations of general condition, body weight (bw) and food consumption, urinalysis, hematology, serum biochemistry, and macroscopic and histopathological examination were performed. The results showed that HI had no adverse effects on any of the examined parameters. Therefore, we concluded that the no-observed-adverse-effect level (NOAEL) for HI was estimated to be 5% for both sexes (2,890 mg/kg bw/day for males and 3,840 mg/kg bw/day for females). Since the iron content of HI used in this study was in a range of 2.0-2.6%, iron content at NOAEL for HI was calculated to be 57.8-75.1 mg/kg bw/day for males and 76.8-99.8 mg/kg bw/day for females.

Protection of Mice from Controlled Cortical Impact Injury by Food Additive Glyceryl Tribenzoate.

Despite intense investigations, no effective therapy is available to halt the pathogenesis of traumatic brain injury (TBI), a major health concern, which sometimes leads to long-term neurological disability, especially in war veterans and young adults. This study highlights the use of glyceryl tribenzoate (GTB), a flavoring ingredient, in ameliorating the disease process of controlled cortical impact (CCI)-induced TBI in mice. Oral administration of GTB decreased the activation of microglia and astrocytes to inhibit the expression of inducible nitric oxide synthase (iNOS) in hippocampus and cortex of TBI mice. Accordingly, GTB treatment protected and/or restored synaptic maturation in the hippocampus of TBI mice as revealed by the status of PSD-95, NR-2A and GluR1. Furthermore, oral GTB also reduced the size of lesion cavity in the brain of TBI mice. Finally, GTB treatment improved locomotor functions and protected spatial learning and memory in TBI mice. These results outline a novel neuroprotective property of GTB which may be beneficial in treatment of TBI.

The Impact of Food Additives on the Abundance and Composition of Gut Microbiota.

The gut microbiota has been confirmed as an important part in human health, and is even take as an 'organ'. The interaction between the gut microbiota and host intestinal environment plays a key role in digestion, metabolism, immunity, inflammation, and diseases. The dietary component is a major factor that affects the composition and function of gut microbiota. Food additives have been widely used to improve the color, taste, aroma, texture, and nutritional quality of processed food. The increasing variety and quantity of processed food in diets lead to increased frequency and dose of food additives exposure, especially artificial food additives, which has become a concern of consumers. There are studies focusing on the impact of food additives on the gut microbiota, as long-term exposure to food additives could induce changes in the microbes, and the gut microbiota is related to human health and disease. Therefore, the aim of this review is to summarize the interaction between the gut microbiota and food additives.

Natural compound glycyrrhetinic acid protects against doxorubicin-induced cardiotoxicity by activating the Nrf2/HO-1 signaling pathway.

BACKGROUND: As one of the most classic antineoplastic agents, doxorubicin (Dox) is extensively used to treat a wide range of cancers. Nevertheless, the clinical outcomes of Dox-based therapies are severely hampered due to the significant cardiotoxicity. Glycyrrhetinic acid (GA) is the major biologically active compound of licorice, one of the most well-known food additives and medicinal plants in the world. We previously demonstrated that GA has the potential capability to protect mice from Dox-induced cardiac injuries. However, the underlying cardioprotective mechanism remains unexplored. PURPOSE: To investigate the cardioprotective benefits of GA against Dox-induced cardiotoxicity and to elucidate its mechanisms of action. STUDY DESIGN/METHODS: H9c2 cardiomyoblasts and AC16 cardiomyocytes were used as the cell models in vitro. A transgenic zebrafish model and a 4T1 mouse breast cancer model were applied to explore the cardioprotective effects of GA in vivo. RESULTS: In vitro, GA inhibited Dox-induced cell death and LDH release in H9c2 and AC16 cells without affecting the anti-cancer effects of Dox. GA significantly alleviated Dox-induced ROS generation, mitochondrial dysfunction, and apoptosis in H9c2 cells. Moreover, GA abolished the expression of pro-apoptotic proteins and restored Nrf2/HO-1 signaling pathway in Dox-treated H9c2 cells. On the contrary, Nrf2 knockdown strongly abrogated the cardioprotective effects of GA on Dox-treated H9c2 cells. In vivo, GA attenuated Dox-induced cardiac dysfunction by restoring stroke volume, cardiac output, and fractional shortening in the transgenic zebrafish embryos. In a 4T1 mouse breast cancer model, GA dramatically prevented body weight loss, attenuated cardiac dysfunction, and prolonged survival rate in Dox-treated mice, without compromising Dox's anti-tumor efficacy. Consistently, GA attenuated oxidative injury, reduced cardiomyocytes apoptosis, and restored the expressions of Nrf2 and HO-1 in Dox-treated mouse hearts. CONCLUSION: GA protects against Dox-induced cardiotoxicity by suppressing oxidative stress, mitochondrial dysfunction, and apoptosis via upregulating Nrf2/HO-1 signaling pathway. These findings could provide solid evidence to support the further development of GA as a feasible and safe adjuvant to Dox chemotherapy for overcoming Dox-induced cardiotoxicity.

Calcium Chloride Toxicology for Food Safety Assessment Using Zebrafish (Danio rerio) Embryos.

The salt calcium chloride (CaCl2) is widely used in industry as a food additive; levels for human consumption are regulated by international or governmental agencies. Generally, the food industry relies on toxicity studies conducted in mammals such as mice, rats, and rabbits for determining food safety. However, testing in mammals is time-consuming and expensive. Zebrafish have been used in a range of toxicological analyses and offer advantages with regard to sensitivity, time, and cost. However, information in not available with regard to whether the sensitivity of zebrafish to CaCl2 is comparable to the concentrations of CaCl2 used as food additives. The aim of this study was to compare the CaCl2 tolerance of zebrafish embryos and larvae with concentrations currently approved as food additives. Acute toxicity, embryotoxicity, cardiotoxicity, and neurotoxicity assays were used to determine the threshold toxic concentration of CaCl2 in zebrafish embryos and larvae. The data showed that doses above 0.4% had toxic effects on development and on the activity of the cardiac and neuronal systems. Furthermore, all embryos exposed to 0.8 and 1.6% of CaCl2 died after 24 hpf. These findings are consistent with the limits of CaCl2 concentrations approved by Codex Alimentarius. Therefore, zebrafish embryos could be suitable for screening food additives.

THE FEATURES OF THE NORMAL ULTRASTRUCTURE OF THE RAT DUODENUM AND UNDER THE COMBINED EFFECT OF THE FOOD ADDITIVES COMPLEX.

OBJECTIVE: The aim: Investigation of ultrastructural changes in the elements of rats' duodenal mucosa in norm and exposed to a complex of food additives (monosodium glutamate, sodium nitrite and Ponceau 4R). PATIENTS AND METHODS: Materials and methods: 70 rats of the experimental groups was administered 0.6 mg/kg of sodium nitrite, monosodium glutamate at a dose of 20 mg/kg, Ponceau 4R at a dose of 5 mg/kg in 0.5 ml of distilled water once daily per os. The doses of food additives were twice lower the allowable normal rate in food products. Animals were removed from the experiment at 1, 4, 8, 12 and 16 weeks. RESULTS: Results: The effect of the complex of food additives on the mucous membrane of the duodenum was manifested by the development of edema and increased local immune response. In the later stages of observation, dystrophic changes in epithelial cells were determined. Vacuoles were found in the cytoplasm. CONCLUSION: Conclusions: The use of a complex of food additives led to general ultramicroscopic changes in the mucous membrane of rats' duodenum, triggering the morphological mechanisms of nonspecific inflammation in the form of dystrophic changes and the development of apoptosis.

MORPHOLOGICAL FEATURES OF THE LIVER PARENCHYMA IN THE EXPERIMENTAL SUPPLEMENTATION OF RATION WITH THE FOOD ADDITIVES.

OBJECTIVE: The aim: The aim of the paper was the experimental study of the morphological features of albino rat hepatocytes after the consumption of the complex of food additives (monosodium glutamate, sodium nitrite, Ponceau 4R) supplemented into the ration and consumed for four weeks. PATIENTS AND METHODS: Materials and methods: The study was performed on 30 outbred albino rats of both genders, weighing 204±0.67 g. The ration of the experimental animals, supplemented with a combination of food additives, namely, monosodium glutamate, Ponceau 4R, sodium nitrate, was consumed for 1 and 4 weeks. The study of the structure of hepathocytes was carried out on traditional histological preparations and preparations stained with Best's carmine. RESULTS: Results: Supplementation of ration with the complex of food additives for one week showed the phenomena of fatty degeneration that dominated in hepatocytes, and in a longer consumption of food additives in the ration (for four weeks), the number of liver cells with the phenomena of hydropic degeneration significantly increased, while individual hepatocytes had signs of irreversible destructive changes. CONCLUSION: Conclusions: Consumption of the complex of food additives supplemented into the standard ration of laboratory animals for 4 weeks leads to a significant change in the dimensions of the liver cells, a decrease in their glycogen content, and a progressive increase in the number of hepatocytes with alterations.

Beneficial effects of dietary capsaicin in gastrointestinal health and disease.

Chili pepper and its major active compound capsaicin have long been used not only a daily food additive but also medication worldwide. Like in other human organs and systems, capsaicin has multiple actions in gastrointestinal (GI) physiology and pathology. Numerous studies have revealed that capsaicin acts on GI tract in TRPV1-dependent and -independent manners, mostly depending on its consumption concentrations. In this review, we will focus on the beneficial role of capsaicin in GI tract, a less highlighted aspect, in particular how dietary capsaicin affects GI health, the mechanisms of actions and its preventive/therapeutic potentials to several GI diseases. Dietary capsaicin affects GI tract not only via TRPV1-derpendent and independent manners, but also via acute and chronic effects. Although high dose intake of dietary capsaicin is harmful to human health sometimes, current literatures suggest that appropriate dose intake is likely beneficial to GI health and is preventive/therapeutic to GI disease in most cases as well. With extensive and intensive studies on its GI actions, capsaicin, as a daily consumed food additive, has potential to become a safe drug for the treatment of several GI diseases.

Comprehensive Assessment of Functional Effects of Commonly Used Sugar Substitute Sweeteners on Ex Vivo Human Gut Microbiome.

The composition and function of the human gut microbiome are often associated with health and disease status. Sugar substitute sweeteners are widely used food additives, although many studies using animal models have linked sweetener consumption to gut microbial changes and health issues. Whether sugar substitute sweeteners directly change the human gut microbiome functionality remains largely unknown. In this study, we systematically investigated the responses of five human gut microbiomes to 21 common sugar substitute sweeteners, using an approach combining high-throughput in vitro microbiome culturing and metaproteomic analyses to quantify functional changes in different taxa. Hierarchical clustering based on metaproteomic responses of individual microbiomes resulted in two clusters. The noncaloric artificial sweetener (NAS) cluster was composed of NASs and two sugar alcohols with shorter carbon backbones (4 or 5 carbon atoms), and the carbohydrate (CHO) cluster was composed of the remaining sugar alcohols. The metaproteomic functional responses of the CHO cluster were clustered with those of the prebiotics fructooligosaccharides and kestose. The sugar substitute sweeteners in the CHO cluster showed the ability to modulate the metabolism of Clostridia. This study provides a comprehensive evaluation of the direct effects of commonly used sugar substitute sweeteners on the functions of the human gut microbiome using a functional metaproteomic approach, improving our understanding of the roles of sugar substitute sweeteners on microbiome-associated human health and disease issues. IMPORTANCE The human gut microbiome is closely related to human health. Sugar substitute sweeteners as commonly used food additives are increasingly consumed and have potential impacts on microbiome functionality. Although many studies have evaluated the effects of a few sweeteners on gut microbiomes using animal models, the direct effect of sugar substitute sweeteners on the human gut microbiome remains largely unknown. Our results revealed that the sweetener-induced metaproteomic responses of individual microbiomes had two major patterns, which were associated with the chemical properties of the sweeteners. This study provided a comprehensive evaluation of the effects of commonly used sugar substitute sweeteners on the human gut microbiome.

Phenolipids as new food additives: from synthesis to cell-based biological activities.

Increasing interest has been shown in phenolic compounds for enhancing food quality, but their hydrophilicity restricts application in lipophilic systems. Therefore, in this study, twelve hydroxycinnamates derivatives (alkyl and steryl esters of sinapic acid (SA), caffeic acid (CA), and ferulic acid [FA]) were synthesised and evaluated for antioxidant and cytotoxic characteristics. CA esters had the highest radical scavenging activity (RSA) analysed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays. Values of inhibitory concentration (IC50) of synthesised compounds were related to their structure and lipophilicity. The effect of these hydroxycinnamic acid esters on the antioxidant potential of real samples (rapeseed oil, margarine and mayonnaise) was estimated. None of the investigated derivatives significantly affected the viability of the model intestinal cells Caco2, while the octyl esters demonstrated a toxic effect at low concentrations. The synthesised esters exerted cytotoxic and anti-proliferative effects against transformed cell lines (HeLa and A549). Octyl esters were potent anticancer compounds on two human cancer cell lines. The synthesised phenolipids, as valuable and safe antioxidant additives, can find broader applications in the production of fat-based products to prevent oxidation processes, extend their shelf life and improve quality.

Trilobatin rescues cognitive impairment of Alzheimer's disease by targeting HMGB1 through mediating SIRT3/SOD2 signaling pathway.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder with cognitive impairment that currently is uncurable. Previous study shows that trilobatin (TLB), a naturally occurring food additive, exerts neuroprotective effect in experimental models of AD. In the present study we investigated the molecular mechanisms underlying the beneficial effect of TLB on experimental models of AD in vivo and in vitro. APP/PS1 transgenic mice were administered TLB (4, 8 mg· kg-1 ·d-1, i.g.) for 3 months; rats were subjected to ICV injection of Aß25-35, followed by administration of TLB (2.5, 5, 10 mg· kg-1 ·d-1, i.g.) for 14 days. We showed that TLB administration significantly and dose-dependently ameliorated the cognitive deficits in the two AD animal models, assessed in open field test, novel object recognition test, Y-maze test and Morris water maze test. Furthermore, TLB administration dose-dependently inhibited microglia and astrocyte activation in the hippocampus of APP/PS1 transgenic mice accompanied by decreased expression of high-mobility group box 1 (HMGB1), TLR4 and NF-κB. In Aß25-25-treated BV2 cells, TLB (12.5-50 µM) concentration-dependently increased the cell viability through inhibiting HMGB1/TLR4/NF-κB signaling pathway. HMGB1 overexpression abrogated the beneficial effects of TLB on BV2 cells after Aß25-35 insults. Molecular docking and surface plasmon resonance assay revealed that TLB directly bound to HMGB1 with a KD value of 8.541×10-4 M. Furthermore, we demonstrated that TLB inhibited Aß25-35-induced acetylation of HMGB1 through activating SIRT3/SOD2 signaling pathway, thereby restoring redox homeostasis and suppressing neuroinflammation. These results, for the first time, unravel a new property of TLB: rescuing cognitive impairment of AD via targeting HMGB1 and activating SIRT3/SOD2 signaling pathway.

Natural food flavour (E)-2-hexenal, a potential antifungal agent, induces mitochondria-mediated apoptosis in Aspergillus flavus conidia via a ROS-dependent pathway.

Natural food flavour (E)-2-hexenal, a green leaf volatile, exhibits potent antifungal activity on Aspergillus flavus, but its antifungal mechanism has not been fully elucidated. In this study, we evaluated (E)-2-hexenal-induced apoptosis in A. flavus conidia and explored the underlying mechanisms of action. Evidence of apoptosis in A. flavus conidia were investigated by methods including fluorescent staining, flow cytometry, confocal laser scanning microscope, and spectral analysis. Results indicated that 4.0 µL/mL (minimum fungicidal concentration, MFC) of (E)-2-hexenal application induced early markers of apoptotic cell death in A. flavus conidia with a rate of 38.4% after 6 h exposure. Meanwhile, typical hallmarks of apoptosis, such as decreased mitochondrial membrane potential (MMP), activated metacaspase activity, fragmented DNA, mitochondrial permeability transition pore (MPTP) opening and cytochrome c (Cyt C) release from mitochondria to the cytosol were also confirmed. Furthermore, intracellular ATP levels were reduced by 63.3 ± 3.6% and reactive oxygen species (ROS) positive cells increased by 31.1 ± 3.1% during A. flavus apoptosis induced by (E)-2-hexenal. l-Cysteine (Cys), an antioxidant, could strongly block the excess ROS generation caused by (E)-2-hexenal, which correspondingly resulted in a significant inhibition of MPTP opening and decrease of apoptosis in A. flavus, indicating that ROS palys a pivotal role in (E)-2-hexenal-induced apoptosis. These results suggest that (E)-2-hexenal exerts its antifungal effect on A. flavus conidia via a ROS-dependent mitochondrial apoptotic pathway.