Tomato-fruit-derived extracellular vesicles inhibit Fusobacterium nucleatum via lipid-mediated mechanism.

Nano-sized extracellular vesicles (EV) are essential for cell communication. Studies on EV from natural sources including edible plants are gaining momentum due to the biological implications. In this study, EV from tomato fruit were isolated by ultracentrifugation and their physical and morphological features along with their biocargo profiles were analyzed. We found that tomato EV promote the growth of probiotic Lactobacillus species, while inhibiting growth of the opportunistic intestinal pathogens Clostridioides difficile and Fusobacterium nucleatum. Tomato EV reversed microbiota dysbiosis caused by F. nucleatum in a simulator of the gut microbiota fermentation model. Phospholipid analysis of tomato EV revealed that the anti-bacterial effect of tomato-EV was driven by the presence of specific lipids in the EV, as demonstrated by lipid depletion and reconstitution experiments. The findings suggest the potential of tomato-derived EV for treating gut microbiota dysbiosis and preventing intestinal bacterial infections.

Microbial Diversity of Anaerobic-Fermented Coffee and Potential for Inhibiting Ochratoxin-Produced Aspergillus niger.

Coffee flavor considerably depends on the fermentation process, with contributing factors including fermentation temperature, oxygen concentration, and microbial diversity. Efficient controlling of the fermentation can improve the quality of coffee beverages. Therefore, several studies on coffee fermentation processes have been conducted in various regions. The objective of this study was to assess the microbial diversity of coffee beans undergoing anaerobic fermentation at various temperatures (4 °C or 37 °C) and fermentation durations (12 h or 36 h) using full-length 16S rRNA sequencing. This analysis aimed to evaluate the inhibitory effects of the fermented metabolites against ochratoxin-producing Aspergillus niger. From our results, Acetobacter was identified as the dominant microbial community at higher fermentation temperatures, whereas Leuconostoc and Gluconobacter were the dominant genera at lower temperatures. However, at lower temperatures, changes in microbial communities were relatively slow. This study expands our knowledge of the microbial diversity involved in the anaerobic fermentation of coffee beans in Taiwan. The findings of this study can be used in future research to cultivate microorganisms linked to the quality and improve the quality of coffee beverages through fermentation.

Proteomic characterization of extracellular vesicles derived from lactic acid bacteria.

Lactobacillus species confer health benefits by their metabolites, secreted molecules, and population numbers. Extracellular vesicles (EVs) are nano-sized particles released from cells and mediate intercellular communications. EVs-encapsulated cargos are a crucial key to decide involved biological function. However, little is known about the composition of EVs, leaving mechanisms by which Lactobacillus-derived EVs affect recipient cells remaining unresolved. This study examined the composition of EV proteins from Lactobacillus species by using liquid chromatography coupled with tandem mass spectrometry, including L. plantarum, L. fermentum, and L. gasseri. The major proteins of EVs are associated with biological processes such as catalytic activity, gluco-neogenesis, cell wall organization, and glycolytic processes. Motif enrichment analysis revealed that EVs from L. plantarum and L. fermentum contained proteins with serine-rich motif. This is the first study to report the composition and comparison of EV proteins from Lactobacillus species, providing important information of EVs in functional food products development.

Effect of polysaccharide derived from dehulled adlay on regulating gut microbiota and inhibiting Clostridioides difficile in an in vitro colonic fermentation model.

Human body can digest only a few sugars with a low degree of polymerization. The rest of the carbohydrates become food for gastrointestinal symbiotic bacteria, affecting gut microbiota composition and human health. Adlay is a medicinal and food homologous crop. The study aims to determine whether dehulled adlay-derived polysaccharide regulates gut microbiota and barrier function to against Clostridioides difficile infection. Major molecular weight of adlay polysaccharide is 27 kDa. The growth of next-generation probiotics were promoted by adlay polysaccharides. In colonic fermentation model, the ratio of C. difficile was decreased when adding the condition medium of adlay polysaccharides-treated fecal microbiota. In addition, adlay polysaccharide promoted the expression of tight junction proteins and mucin in intestinal cells. This study shows that adlay polysaccharide can be used as prebiotics to regulate microbiota and maintain barrier function, which has the potential to be developed as novel functional food ingredients to protect intestinal health.

Influences of dielectric constant and scan rate on hysteresis effect in perovskite solar cell with simulation and experimental analyses.

In this work, perovskite solar cells (PSCs) with different transport layers were fabricated to understand the hysteresis phenomenon under a series of scan rates. The experimental results show that the hysteresis phenomenon would be affected by the dielectric constant of transport layers and scan rate significantly. To explain this, a modified Poisson and drift-diffusion solver coupled with a fully time-dependent ion migration model is developed to analyze how the ion migration affects the performance and hysteresis of PSCs. The modeling results show that the most crucial factor in the hysteresis behavior is the built-in electric field of the perovskite. The non-linear hysteresis curves are demonstrated under different scan rates, and the mechanism of the hysteresis behavior is explained. Additionally, other factors contributing to the degree of hysteresis are determined to be the degree of degradation in the perovskite material, the quality of the perovskite crystal, and the materials of the transport layer, which corresponds to the total ion density, carrier lifetime of perovskite, and the dielectric constant of the transport layer, respectively. Finally, it was found that the dielectric constant of the transport layer is a key factor affecting hysteresis in perovskite solar cells.

Early-Life Origins of Metabolic Syndrome: Mechanisms and Preventive Aspects.

One of the leading global public-health burdens is metabolic syndrome (MetS), despite the many advances in pharmacotherapies. MetS, now known as "developmental origins of health and disease" (DOHaD), can have its origins in early life. Offspring MetS can be programmed by various adverse early-life conditions, such as nutrition imbalance, maternal conditions or diseases, maternal chemical exposure, and medication use. Conversely, early interventions have shown potential to revoke programming processes to prevent MetS of developmental origins, namely reprogramming. In this review, we summarize what is currently known about adverse environmental insults implicated in MetS of developmental origins, including the fundamental underlying mechanisms. We also describe animal models that have been developed to study the developmental programming of MetS. This review extends previous research reviews by addressing implementation of reprogramming strategies to prevent the programming of MetS. These mechanism-targeted strategies include antioxidants, melatonin, resveratrol, probiotics/prebiotics, and amino acids. Much work remains to be accomplished to determine the insults that could induce MetS, to identify the mechanisms behind MetS programming, and to develop potential reprogramming strategies for clinical translation.

Applications of Lactobacillus acidophilus-Fermented Mango Protected Clostridioides difficile Infection and Developed as an Innovative Probiotic Jam.

Clostridioides difficile infection (CDI) is a large intestine disease caused by toxins produced by the spore-forming bacterium C. difficile, which belongs to Gram-positive bacillus. Using antibiotics treatment disturbances in the gut microbiota and toxins produced by C. difficile disrupt the intestinal barrier. Some evidence indicates fecal microbiota transplantation and probiotics may decrease the risk of CDI recurrence. This study aimed to evaluate the efficacy of fermented mango by using the lactic acid bacteria Lactobacillus acidophilus and develop innovative products in the form of fermented mango jam. L. acidophilus-fermented mango products inhibited the growth of C. difficile while promoting the growth of next-generation probiotic Faecalibacterium prausnitzii. Both supernatant and precipitate of mango-fermented products prevented cell death in gut enterocyte-like Caco-2 cells against C. difficile infection. Mango-fermented products also protected gut barrier function by elevating the expression of tight junction proteins. Moreover, L. acidophilus-fermented mango jam with high hydrostatic pressure treatment had favorable textural characteristics and sensory quality.

Cardiovascular Diseases of Developmental Origins: Preventive Aspects of Gut Microbiota-Targeted Therapy.

Cardiovascular diseases (CVDs) can originate from early life. Accumulating evidence suggests that gut microbiota in early life is linked to CVDs in later life. Gut microbiota-targeted therapy has gained significant importance in recent decades for its health-promoting role in the prevention (rather than just treatment) of CVDs. Thus far, available gut microbiota-based treatment modalities used as reprogramming interventions include probiotics, prebiotics, and postbiotics. The purpose of this review is, first, to highlight current studies that link dysbiotic gut microbiota to the developmental origins of CVD. This is followed by a summary of the connections between the gut microbiota and CVD behind cardiovascular programming, such as short chain fatty acids (SCFAs) and their receptors, trimethylamine-N-oxide (TMAO), uremic toxins, and aryl hydrocarbon receptor (AhR), and the renin-angiotensin system (RAS). This review also presents an overview of how gut microbiota-targeted reprogramming interventions can prevent the developmental origins of CVD from animal studies. Overall, this review reveals that recent advances in gut microbiota-targeted therapy might provide the answers to reduce the global burden of CVDs. Still, additional studies will be needed to put research findings into practice.

Immunomodulation and mechanisms of fucoidan from Cladosiphon okamuranus ameliorates atopic dermatitis symptoms.

Atopic dermatitis (AD) is a long-term allergic skin disorder that occurs most frequently in children. Currently, the common treatment of AD is corticosteroids; however, the drugs cause serious side effects. Therefore, there are many patients who seek complementary and alternative treatments such as healthy food. We report that fucoidan from Cladosiphon okamuranus (COP) exhibit exceptional immuno-modulatory effects significantly improving atopic dermatitis (AD) at both in vitro and in vivo levels: First, we performed the P815 cell degranulation assay, of which the results revealed that COP possesses anti-degranulation activity suggesting COP is very conducive to relieving allergic reactions of AD. Next, we performed the animal model examination, of which AD was significantly improved, suggesting COP can focally and globally modulate the immune systems of animals. The systemic improvements were manifested clearly by decreased epidermal hyperplasia, reduced infiltration of eosinophils, and decreased expression of AD-associated cytokines. Notably, COP reduced epidermal hyperplasia by downregulating the expression of IL-22. COP displayed therapeutic effects, which is comparable to corticosteroids but lack corticosteroid side effects, such as weight loss in our animal study. COP is multitudinous immunomodulatory abilities to serve as a healthy food supplement at the current stage, not least beneficial to atopic dermatitis.

Polysaccharides Obtained from Cordyceps militaris Alleviate Hyperglycemia by Regulating Gut Microbiota in Mice Fed a High-Fat/Sucrose Diet.

Polysaccharides isolated from fungus Cordyceps militaris display multi-biofunctions, such as immunostimulation, down-regulation of hyperlipidemia, and anti-cancer function. The occurrence of obesity and metabolic syndrome is related to the imbalance of gut microbiota. In this study, the effects of C. militaris and its fractions on modifying metabolic syndrome in mice were evaluated. Mice were fed a high-fat/high-sucrose diet (HFSD) for 14 weeks to induce body weight increase and hyperlipidemia symptoms in mice, and then the mice were simultaneously given a HFSD and C. militaris samples for a further 8 weeks. The results indicated that the fruit body, polysaccharides, and cordycepin obtained from C. militaris had different efficacies on regulating metabolic syndrome and gut microbiota in HFSD-treated mice. Polysaccharides derived from C. militaris decreased the levels of blood sugar and serum lipids in mice fed HFSD. In addition, C. militaris-polysaccharide treatment obviously improved intestinal dysbiosis through promoting the population of next generation probiotic Akkermansia muciniphila in the gut of mice fed HFSD. In conclusion, polysaccharides derived from C. militaris have the potential to act as dietary supplements and health food products for modifying the gut microbiota to improve the metabolic syndrome.

The Protection of Lactic Acid Bacteria Fermented-Mango Peel against Neuronal Damage Induced by Amyloid-Beta.

Mango peels are usually discarded as waste; however, they contain phytochemicals and could provide functional properties to food and promote human health. This study aimed to determine the optimal lactic acid bacteria for fermentation of mango peel and evaluate the effect of mango peel on neuronal protection in Neuron-2A cells against amyloid beta (Aß) treatment (50 µM). Mango peel can be fermented by different lactic acid bacteria species. Lactobacillus acidophilus (BCRC14079)-fermented mango peel produced the highest concentration of lactic acid bacteria (exceeding 108 CFU/mL). Mango peel and fermented mango peel extracts upregulated brain-derived neurotrophic factor (BDNF) expression for 1.74-fold in Neuron-2A cells. Furthermore, mango peel fermented products attenuated oxidative stress in Aß-treated neural cells by 27%. Extracts of L. acidophilus (BCRC14079)-fermented mango peel treatment decreased Aß accumulation and attenuated the increase of subG1 caused by Aß induction in Neuron-2A cells. In conclusion, L. acidophilus (BCRC14079)-fermented mango peel acts as a novel neuronal protective product by inhibiting oxidative stress and increasing BDNF expression in neural cells.

Resveratrol Butyrate Esters Inhibit Obesity Caused by Perinatal Exposure to Bisphenol A in Female Offspring Rats.

Resveratrol butyrate esters (RBE) are derivatives of resveratrol (RSV) and butyric acid and exhibit biological activity similar to that of RSV but with higher bioavailability. The aim of this study was designed as an animal experiment to explore the effects of RBE on the serum biochemistry, and fat deposits in the offspring rats exposed to bisphenol A (BPA), along with the growth and decline of gut microbiota. We constructed an animal model of perinatal Bisphenol A (BPA) exposure to observe the effects of RBE supplementation on obesity, blood lipids, and intestinal microbiota in female offspring rats. Perinatal exposure to BPA led to weight gain, lipid accumulation, high levels of blood lipids, and deterioration of intestinal microbiota in female offspring rats. RBE supplementation reduced the weight gain and lipid accumulation caused by BPA, optimised the levels of blood lipids, significantly reduced the Firmicutes/Bacteroidetes (F/B) ratio, and increased and decreased the abundance of S24-7 and Lactobacillus, respectively. The analysis of faecal short-chain fatty acid (SCFA) levels revealed that BPA exposure increased the faecal concentration of acetate, which could be reduced via RBE supplementation. However, the faecal concentrations of propionate and butyrate were not only significantly lower than that of acetate, but also did not significantly change in response to BPA exposure or RBE supplementation. Hence, RBE can suppress BPA-induced obesity in female offspring rats, and it demonstrates excellent modulatory activity on intestinal microbiota, with potential applications in perinatological research.

The applications of Lactobacillus plantarum-derived extracellular vesicles as a novel natural antibacterial agent for improving quality and safety in tuna fish.

Bacteria release membrane vesicles into the extracellular environment but which activity is unclear. We investigated the applications of extracellular vesicles (EVs) isolated from probiotic Lactobacillus plantarum to protect tuna fish against spoilage and quality loss in this study. A significant difference was found in EVs size obtained from L. plantarum after 8, 24, and 48 hr incubation. The L. plantarum-derived EVs were collected and used to confirm the anti-bacterial activity versus Shewanella putrefaciens. Finally, the tuna fish was stored at 4 °C for 5 days after coating with EVs or sodium erythorbate, and the quality indexes were assayed. Results indicated that EVs markedly inhibited oxidation reaction, total volatile base nitrogen (TVBN), peroxide value (PV), malondialdehyde (MDA), and bacteria levels. These results finding out that EVs from L. plantarum may have potential for application in food storage technology. Overall, we indicated this new material may be developed as an anti-bacterial agent for prolonging the shelf life of tuna fish.

Polysaccharides obtained from mycelia of Cordyceps militaris attenuated doxorubicin-induced cytotoxic effects in chemotherapy.

OBJECTIVES: Fungus Cordyceps militaris has been used as a herbal tonic in traditional Chinese medicine, which could be surface liquid-cultured for mycelia production. To evaluate the potential of polysaccharides obtained from mycelia of Cordyceps militaris (PS-MCM) for attenuation of side-effects of chemotherapy. METHODS: Doxorubicin was used to induce cytotoxicity in THP-1 monocytes and EL-4 T cells, and the effects of PS-MCM on cell viability and cytokine production were detected on doxorubicin-treated THP-1 and EL-4 cells. RESULTS: PS-MCM reduced doxorubicin-induced cell death and promoted cell proliferation in THP-1 and EL-4 cells. Moreover, PS-MCM elevated the level of cytokines associated with immune-modulation of THP-1 and EL-4 cells. CONCLUSION: These findings indicate that PS-MCM has potential for development as a functional food to counteract side effects of chemotherapy.

Effects of long-term exercise on arrhythmogenesis in aged hypertensive rats.

Chronic hypertension is a multifactorial disease that is highly associated with cardiovascular disorders. Physical activity, such as long-term exercise, is advocated as a treatment for hypertension, but the responses of different age groups to long-term exercise are unknown. We used aged spontaneous hypertensive rats (SHRs, 80 weeks old) to test the hypothesis that long-term exercise compensated for deficient autonomic control and reduced susceptibility to ventricular tachycardia (VT) and ventricular fibrillation (VF) in this animal model. The aged SHRs were divided into control and voluntary exercise groups. Ambulatory electrocardiography was recorded for the heart rate variability (HRV) analysis. Programmed stimulation was applied to exposed hearts to induce ventricular arrhythmia in situ. Then, the hearts were isolated for an optical mapping study. The results showed that increased HRV indices were broadly related to vagal dominance in the high-intensity exercise group. Exercise altered the electrical propagation dynamic properties, such as the action potential duration restitution (APDR). Furthermore, the VF inducibility decreased with increased exercise intensity. Taken together, our results suggest that long-term exercise reduces the risk of arrhythmogenesis in aged SHRs through enhanced vagal control and stabilized electrical dynamics.

Optimal conditions for cordycepin production in surface liquid-cultured Cordyceps militaris treated with porcine liver extracts for suppression of oral cancer.

Cordycepin is one of the most crucial bioactive compounds produced by Cordyceps militaris and has exhibited antitumor activity in various cancers. However, industrial production of large amounts of cordycepin is difficult. The porcine liver is abundant in proteins, vitamins, and adenosine, and these ingredients may increase cordycepin production and bioconversion during C. militaris fermentation. We observed that porcine liver extracts increased cordycepin production. In addition, air supply (2 h/d) significantly increased the cordycepin level in surface liquid-cultured C. militaris after 14 days. Moreover, blue light light-emitting diode irradiation (16 h/d) increased cordycepin production. These findings indicated that these conditions are suitable for increasing cordycepin production. We used these conditions to obtain water extract from the mycelia of surface liquid-cultured C. militaris (WECM) and evaluated the anti-oral cancer activity of this extract in vitro and in vivo. The results revealed that WECM inhibited the cell viability of SCC-4 oral cancer cells and arrested the cell cycle in the G2/M phase. Oxidative stress and mitochondrial dysfunction (mitochondrial fission) were observed in SCC-4 cells treated with WECM for 12 hours. Furthermore, WECM reduced tumor formation in 7,12-dimethylbenz[a]anthracene-induced hamster buccal pouch carcinogenesis through the downregulation of proliferating cell nuclear antigen, vascular endothelial growth factor, and c-fos expression. The results indicated that porcine liver extracts irradiated with blue light light-emitting diode and supplied with air can be used as a suitable medium for the growth of mycelia and production of cordycepin, which can be used in the treatment of oral cancer.

A Method for Estimating the Filling Rate of the Mold Cavity in Nanoimprint Lithography.

When the dimensions of a microelectronic structure decrease, high manufacturing costs are inevitable. A low cost and high throughput manufacturing technique for nanostructures is desired. Nanoimprint lithography involves patterning the resist through physical deformation by using a mold at nanoscale and has the potential to meet these expectations. Therefore, nanoimprint lithography has been extensively studied in recent years. Many real time measurements have been proposed for enhancing the yield of nanoimprint lithography. Among these measurements, the application of surface plasmon resonance has the advantage of quick, highly accurate analysis. In surface plasmon resonance application, the mold contains a gold film for exciting surface plasmon resonance and an adhesion layer is applied to both sides of the gold film to increase the lifespan of the mold. However, the effect of the geometric characteristics of the adhesive layer on the surface plasmon resonance spectrum and the mechanical strength of the mold has not been extensively studied. To improve the detection accuracy and reliability of the measurement, this study investigated the aforementioned effect. Analytical and experimental investigations confirmed that the shape of the spectrum is influenced by the surface roughness and thickness of the titanium adhesion layer. To maintain the sharpness of the resonance dip, we suggest reducing the thickness of the titanium adhesion layer to below 6 nm and maintaining the surface roughness below 3 nm. Moreover, the proposed mold structure conforms to these requirements and is applied to estimate the filling rate. The measurement results demonstrate that the surface plasmon resonance spectrum is clearly affected by the mold filling. Specifically, the change in the surface plasmon resonance spectrum curve and resonance angle can indicate the quality of the imprinted pattern. This study demonstrates the effectiveness and high sensitivity of the proposed technique for estimating the filling rate of the mold cavity in nanoimprint lithography.

The neuronal protection of a zinc-binding protein isolated from oyster.

Mitochondrial function is applied as oxidative stress and neuronal damage index. In this study, d-galactose was used to induce free radicals production and neuronal damage in HN-h cells, and the effect of novel 43 kDa protein isolated from oyster on anti-mitochondrial dysfunction and zinc-binding ability were evaluated. Crystal violet stain results indicated zinc-binding protein of oyster (ZPO) attenuated neuronal cell death induced by 100 mM of d-galactose on HN-h cells in a dose-dependent manner. ZPO alleviated mitochondrial inactivation, mitochondrial membrane potential decreasing, oxidative stress, and fusion/fission state in non-cytotoxic concentration of d-galactose (50 mM)-treated HN-h cells. ZPO treatment recovered metallathionein-3 (MT-3) decrease and inhibited ß- and γ-secretase as well as amyloid beta (Aß) accumulation in HN-h cells caused by d-galactose induction. These results suggest ZPO could avoid oxidative stress and is a functional protein for zinc concentration maintainability, which has potential for development of functional foods for neuronal protection.

Dual degradation signals destruct GLI1: AMPK inhibits GLI1 through ß-TrCP-mediated proteasome degradation.

Overexpression of the GLI1 gene has frequently been found in various cancer types, particularly in brain tumors, in which aberrant GLI1 induction promotes cancer cell growth. Therefore, identifying the molecular players controlling GLI1 expression is of clinical importance. Previously, we reported that AMPK directly phosphorylated and destabilized GLI1, resulting in the suppression of the Hedgehog signaling pathway. The current study not only demonstrates that AMPK inhibits GLI1 nuclear localization, but further reveals that ß-TrCP plays an essential role in AMPK-induced GLI1 degradation. We found that activation of AMPK promotes the interaction between ß-TrCP and GLI1, and induces ß-TrCP-mediated GLI1-ubiquitination and degradation. Inhibiting AMPK activity results in the dissociation of the ß-TrCP and GLI1 interaction, and diminishes ß-TrCP-mediated-GLI1 ubiquitination and degradation. On GLI1, substitution of AMPK phosphorylation sites to aspartic acid (GLI13E) results in stronger binding affinity of GLI1 with ß-TrCP, accompanied by enhanced GLI1 ubiquitination and later degradation. In contrast, the GLI1 alanine mutant (GLI13A) shows weaker binding with ß-TrCP, which is accompanied by reduced ß-TrCP-mediated ubiquitination and degradation. Together, these results demonstrate that AMPK regulates GLI1 interaction with ß-TrCP by phosphorylating GLI1 and thus both post-translational modifications by AMPK and ß-TrCP ultimately impact GLI1 degradation.

Antimetastatic effects of cordycepin mediated by the inhibition of mitochondrial activity and estrogen-related receptor α in human ovarian carcinoma cells.

Cordycepin (3'-deoxyadenosine) is a compound for antitumor, which has been found to exert antiangiogenic, antimetastatic, and antiproliferative effects, as well as inducing apoptosis. However, the association between cancer metastasis and mitochondrial activity in cordycepin-treated ovarian carcinoma cells remains unclear. The 50 and 100 µM of cordycepin inhibits mitochondrial fusion and induces mitochondrial fission, respectively. These suggested that cordycepin showed the down-regulation of mitochondrial function and limitation of energy production. Because of activation of mitochondria and generation of energy are needed in cancer cell migration/invasion. After 24 h treatment, cordycepin suppresses epithelial-mesenchymal transition and migration in ovarian carcinoma cells through inhibiting estrogen-related receptor (ERR)-α. The ERRα is a co-transcription factor for gene expressions associated with mitochondrial fusion. Our results indicate that cordycepin suppresses metastasis and migration of ovarian carcinoma cells via inhibiting mitochondrial activity in non-toxic concentrations, and cordycepin has potential benefits in ovarian cancer therapy.