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Silver nanoparticles (AgNPs) have stable reactivity and excellent optical absorption properties. They can be applied in various industries, such as environmental protection, biochemical engineering, and analyte monitoring. However, synthesizing AgNPs and determining their appropriate dosage as a coloring substance are difficult tasks. In this study, to optimize the process of AgNP synthesis and obtain a simple detection method for trace mercury in the environment, we evaluate several factors—including the reagent addition sequence, reaction temperature, reaction time, the pH of the solution, and reagent concentration—considering the color intensity and purity of AgNPs as the reaction optimization criteria. The optimal process for AgNP synthesis is as follows: Mix 10 mM of silver nitrate with trisodium citrate in a hot water bath for 10 min, then, add 10 mM of sodium borohydride to produce the AgNPs and keep stirring for 2 h, finally, adjust the pH to 12 to obtain the most stable products. For AgNP-based mercury detection, the calibration curve of mercury over the concentration range of 0.1–2 ppb exhibits good linearity (R2 &gt, 0.99). This study provides a stable and excellent AgNP synthesis technique that can improve various applications involving AgNP-mediated reactions and has the potential to be developed as an alternative to using expensive detection equipment and to be applied for the detection of mercury in food.

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.

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.

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 (Aβ) 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 Aβ-treated neural cells by 27%. Extracts of L. acidophilus (BCRC14079)-fermented mango peel treatment decreased Aβ accumulation and attenuated the increase of subG1 caused by Aβ 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.

The modulatory activity of water extracts from fresh roots of Polygonum multiflorum (WRP) on oxidative stress in two liver cell lines was investigated for the first time. WRP increased antioxidant enzyme activities in Clone 9 cells, including catalase (CAT), glutathione-S-transferase (GST), glutathione reductase (GR), and glutathione peroxidase (GPx). The intracellular glutathione (GSH) level was also increased in the Clone 9 cells owing to antioxidase activity elevating effect by WRP treatment for 24 h. On the other hand, reactive oxygen species (ROS) were generated in abundance upon WRP treatment in Hep G2 cells, resulting in the suppression of cell proliferation. Apoptosis was increased to 78% and 77% after WRP (1000 μg/mL) treatment for 24 and 48 h, respectively. Moreover, WRP stimulated the activation of caspase-8, caspase-9, and ultimately caspase-3. Hep G2 cell growth suppression and caspase induction by WRP were greater than that by emodin-like substances (WRPE), a major active compound. These results indicated that phytochemicals present in extracts of P. multiflorum root act synergistically to promote apoptosis, suggesting the possible use of these extracts for hepatoprotection.

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.

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.

PT-peptide is derived from the anti-lipopolysaccharide factor of the swimming crab Portunus trituberculatus. The peptide, consisting of 34 amino acids, contains a lipopolysaccharide binding domain. In this study, we investigated the effect of PT-peptide encapsulated in raw milk-derived extracellular vesicles (EVs), designated as EVs-PT peptide, on immune regulation. The results showed that raw milk-derived EVs efficaciously delivered the PT-peptide into monocytes and elevated immune activity, including reactive oxygen species level, superoxide anion production, and phagocytosis. PT-peptide and EVs-PT peptide also elevated the secretion of cytokines, such as interferon-&gamma, interleukin-6, and tumor necrosis factor-&alpha, in human monocytic THP-1 cells. These results suggest that the PT-peptide could be developed as an immune stimulator.

Graptopetalum paraguayense was shown to exhibit antioxidant, hepatoprotective and anti-hepatocellular carcinoma activities in a recent study. In this work, fractions of the G. paraguayense aqueous solution were extracted using n-hexane, ethyl acetate (EF) and n-butanol, and its antioxidative and anti-glycative activities were evaluated. The EF extract exhibited the highest antioxidative activity, potently inhibited the formation of advanced glycation end products (AGEs) and reduced glycation-induced protein oxidation. Additionally, the EF extract protected Hep G2 cells against AGE-induced DNA damage. Five major components were isolated and identified in the EF fraction by semi-preparative high-performance liquid chromatography, liquid chromatography-tandem mass spectroscopy and nuclear magnetic resonance: gallic acid, isoquercitrin-6-(3-hydroxy-3-methylglutarate), astragalin-6-(3-hydroxy-3-methylglutarate), isoquercitrin-2-acetyl-6-(3-hydroxy-3-methylglutarate) and astragalin-2-acetyl-6-(3-hydroxy-3-methylglutarate). G. paraguayense exhibited antioxidant and anti-glycation activities, likely because of the presence of gallic acid. The findings indicate the potential for using G. paraguayense for development of novel treatments for diabetes. Practical Applications Herbal treatments for hyperlipidemia are relatively cheap and locally available. The results indicated that the extracts of Graptopetalum paraguayense may be developed as functional foods for antidiabetic treatment mediated by attenuations of advanced glycation end product oxidation.

// Yu-Chun Lin 1, 2, * , Bao-Hong Lee 3, * , Jassey Alagie 2, 4 and Ching-Hua Su 1 1 Department of Microbiology and Immunology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan 2 Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan 3 Division of Hematology and Oncology, Department of Internal Medicine, Taipei Medical University Hospital, Taipei, Taiwan 4 International Ph.D. Program in Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan * Authors contributed equally to this work Correspondence to: Ching-Hua Su, email: curator@tmu.edu.tw Keywords: ergosterol, amphotericin B, liver cancer, necrosis, ROS Received: May 01, 2017 Accepted: July 25, 2017 Published: August 16, 2017 ABSTRACT The incidence of liver cancer, the second leading cause of cancer-related deaths has increased over the past few decades. Although recent treatments such as sorafenib are promising in patients with advanced hepatocellular carcinoma (HCC), the response rates remain poor thereby warranting the identification of novel therapeutic agents against liver cancer. Herein, we investigated the anti-cancer effect of ergosterol (a secondary metabolite in medicinal fungus) pretreatment followed by amphotericin B (AmB) treatment on liver cancer cell lines. We demonstrated that pretreatment with a nontoxic dose of ergosterol synergistically enhanced the cytotoxicity of AmB in both Hep3B and HepJ5 cells. The combination treatment-mediated suppression of cancer cell viability occurred through necrosis characterized by disrupted cell membrane and significant amounts of debris accumulation. In addition, we also observed a concomitant increase in reactive oxygen species (ROS) and LC3-II levels in HepJ5 cells treated with ergosterol and AmB. Our results suggest that ergosterol-AmB combination treatment effectively induced necrotic cell death in cancer cells, and deserves further evaluation for development as an anti-cancer agent.

BACKGROUND Ice plant (Mesembryanthemum crystallinum) has been used as an anti-diabetic agent in Japan because it contains d-pinitol. The efficacy of ice plant in the regulation of blood glucose is unclear at present. Recently, memory impairment and development of Alzheimer's disease found in diabetic patients are thought to be caused by high blood glucose. The mechanism by which ice plant protects against the impairment of memory and learning abilities caused by high blood glucose remains unclear. The aim of this study was to evaluate the protection of ice plant water extracts (IPE) and d-pinitol against memory impairments in a Wistar rat model of streptozotocin (STZ)-induced diabetes. We hypothesised that IPE and d-pinitol could suppress blood glucose and elevate insulin sensitivity in these rats. RESULTS For memory evaluation, IPE and d-pinitol also improved the passive avoidance task and the working memory task. In addition, inhibition of acetylcholinesterase activity in hippocampus and cortex was found in this rat model administered IPE or d-pinitol. IPE and d-pinitol also markedly elevated superoxide dismutase activity against oxidative stress and reduced malondialdehyde production in hippocampus and cortex of the rats. CONCLUSION These findings indicated that IPE and d-pinitol possess beneficial effects for neural protection and memory ability in a rat model of diabetes. © 2013 Society of Chemical Industry

Fagopyrum tataricum is used for the treatment of type 2 diabetes mellitus in Taiwan. The aim of this study was to evaluate the inhibitory effects of 75 % ethanol extract of buckwheat (EEB) and rutin on carbohydrate-metabolized enzymes, including α-amylase and α-glucosidase, which are related to hyperglycemia. The rutin dosage (40 μg/mL) was equivalent to that of EEB (200 μg/mL). In addition, the antioxidant and antiglycation activities of EEB and rutin were investigated. Results showed that both EEB and rutin exerted free radical (DPPH and ABTS) scavenging activity. They also attenuated protein glycation to lower the generation of advanced glycation end-products (AGEs) through the suppression of fructosamine and α-dicarbonyl compounds. Moreover, EEB and rutin also inhibited α-amylase and α-glucosidase activity. Taken together, these findings suggest that EEB and rutin may reduce oxidative stress, AGEs formation, and carbohydrate-metabolized enzymes hence EEB may use as protection agent in diabetic patients.

Abnormal cellular accumulation of the dicarbonyl metabolite methylglyoxal (MG) results in cell damage, inflammation, and oxidative stress. It is also associated with increased protein linkage to form advanced glycation end products (AGEs) or induce DNA strand breaks. The association between peroxisome proliferator-activated receptor-γ (PPARγ) and nuclear factor-erythroid 2-related factor 2 (Nrf2) is unclear. This study investigated Nrf2 activator protection against PPARγ phosphorylation and degradation to maintain pancreatic function. MG was used at a noncytotoxic concentration (200 μM) to induce protein kinase C (PKC) and PPARγ phosphorylation in pancreatic RINm5F cells. For in vivo studies, MG (60 mg/kg bw) was intraperitoneally (IP) injected into Balb/C mice for 28 d to induce pancreas damage, at which point we investigated the effect of monascin protection (PPARγ and Nrf2 activator), rosiglitazone (PPARγ activator), allyl isothiocyanate (AITC; Nrf2 activator), or N-acetylcysteine (NAC) on pancreatic function. The in vitro and in vivo results indicated that MG leads to marked PPARγ phosphorylation (serine 82); this effect led to reduction in pancreatic and duodenal homeobox-1 (PDX-1), glucokinase (GCK), and insulin expression. However, monascin and rosiglitazone may protect PPARγ degradation by elevating PDX-1, GCK, and as a result, insulin expression. Monascin and AITC can attenuate PKC activation to suppress PPARγ phosphorylation caused by oxidative stress through the Nrf2 pathway. Similarly, the N-acetylcysteine (NAC) antioxidant also improved oxidative stress and pancreatic function. This study examined whether MG caused impairment of PDX-1, GCK, and insulin through PPARγ phosphorylation and degradation. MG and AGE accumulation improved on Nrf2 activation, thereby protecting against pancreas damage. Taken together, PPARγ activation maintained pancreatic PDX-1, GCK, and insulin expression levels to regulate blood glucose levels.

The effects of dimerumic acid (DMA) on apoptosis prevention and insulin production in the glucose metabolite methylglyoxal (MG)-treated pancreatic RINm5F cells were evaluated. MG was found to induce apoptotic signal molecules mediated by oxidative stress in RINm5F cells, including caspase-3 and caspas-9, resulting in cell death and decreasing insulin production. In addition, DMA elevated insulin synthesis via up-regulating mammalian homologue of avian MafA/L-Maf (Maf-A) and pancreatic-duodenal homeobox-1 (PDX-1) and inhibited MG-mediated CCAAT/enhancer binding proteins-β (C/EBPβ) expression, which is a negative regulator for insulin production. Nuclear factor-erythroid2-related factor 2 (Nrf2) is essential for antioxidant responsive element (ARE)-mediated induction. Results indicated that nuclear translocation of Nrf2 was promoted by DMA in MG-treated RINm5F cells to increase heme oxygenase-1 (HO-1) and glutamate-cysteine ligase (GCL) expressions. We confirmed that DMA protected PDX-1, Maf-A, and insulin production against MG-induced oxidative stress mediating by Nrf2 in Nrf2-knockdown RINm5F cells. These results suggest that DMA exerts antioxidative ability to attenuate MG-induced pancreatic cell damage and elevates insulin level to improve diabetes.

The anti-obesity activity of Lactobacillus paracasei subsp. paracasei NTU 101 and Lactobacillus plantarum NTU 102 and their soy milk fermented products (SM101 and SM102) were investigated. Results indicated that the inhibition of 3T3-L1 differentiation and the accumulation of free fatty acids markedly increased in rats treated with SM101 and SM102. Moreover, the up-regulation and down-regulation of lipolysis and heparin-releasable lipoprotein lipase, respectively, were observed in the 3T3-L1 adipocytes of the SM101 and SM101 groups, and these effects of SM101 and SM102 were greater than unfermented soy milk (USM). We also found that SM101 and SM102 both improved obesity in Wistar rats fed with a high-fat diet (HFD) and that this improvement was stronger than that observed for USM. The level of serum leptin in HFD-induced rats was significantly elevated by the 5-week administration of SM101 and SM102 (106–1010 CFU/mL per rat per day); however, this activity was not promoted by USM. The anti-obesity activity of SM101 and SM102 may result from the increased daidzein and genistein levels that were observed during the fermentation by L. paracasei subsp. paracasei NTU 101 and L. plantarum NTU 102.

Hyperglycemia is associated with advanced glycation end products (AGEs). This study was designed to evaluate the inhibitory effects of monascin on receptor for advanced glycation end product (RAGE) signal and THP-1 monocyte inflammation after treatment with S100b, a specific ligand of RAGE. Monascin inhibited cytokine production by S100b-treated THP-1 monocytes via up-regulation of nuclear factor-erythroid 2-related factor-2 (Nrf2) and alleviated p47phox translocation to the membrane. Methylglyoxal (MG, 600 mg/kg bw) was used to induce diabetes in Wistar rats. Inhibitions of RAGE and p47phox by monascin were confirmed by peripheral blood mononuclear cells (PBMCs) of MG-induced rats. Silymarin (SM) was used as a positive control group. It was found that monascin promoted heme oxygenase-1 (HO-1) expression mediated by Nrf2. Suppressions of AGEs, tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-β) in serum of MG-induced rats were attenuated in the monascin administration group treated with retinoic acid (RA). RA treatment resulted in Nrf2 inactivation by increasing RA receptor-α (RARα) activity, suggesting that RA acts as an inhibitor of Nrf2. The results showed that monascin exerted anti-inflammatory and antioxidative effects mediated by Nrf2 to prevent the development of diseases such as type 2 diabetes caused by inflammation.

Monascus has traditionally been used for the preparation of fermented products and as a food additive in Asia and is also a traditional Chinese medicine. The present review summarizes the underlying biochemical and molecular mechanisms that can explain anti-inflammation, antioxidation, hepatoprotection, inhibition of cancer cell invasion, and detoxification of dimerumic acid (DMA) isolated from Monascus-fermented products. DMA is an antioxidant that effectively inhibits liver damage and inflammation by attenuating oxidative stress in vitro and in vivo, which is stronger than silymarin, resveratrol, or curcumin. In addition, several lines of evidence have indicated that reactive oxygen species (ROS) promotes cancer cell invasion and suppress tumor development. DMA has been reported to attenuate ROS-induced invasion of SW620 (colorectal adenocarcinoma cells) greater than silymarin, resveratrol, and curcumin. These findings suggest that DMA has potential for development as a novel antioxidant for functional foods to act against several diseases-associated oxidative stress.

Ankaflavin (AK) is a yellow pigment isolated from Monascus-fermented product. The potential mechanism of AK-regulated dyslipidemia in high-fat diet hamsters was investigated. The results showed that AK treatment reduced plasma total cholesterol (TC) level in hamsters, elevated faecal TC and bile acid concentration, and protected high density apolipoprotein-cholesterol (HDL-C) from diminishing. We further used hepatocytes to demonstrate that AK stabilized HDL levels via nuclear factor-erythroid 2 related factor 2 (Nrf2) activation and downstream molecule haem oxygenase-1 (HO-1) expression, as well as increasing liver x receptor-α (LXRα) mRNA level to promote ATP-binding cassette transporter (ABCA1) and apolipoprotein-A1 (apo-A1) expression. AK increased low density apolipoprotein-cholesterol (LDL-C) expression to promote cholesterol absorption in hepatocytes, and resulted in cholesterol metabolism by cytochrome P450 7A1 (CYP7A1), leading to the conversion of cholesterol into bile acid. Taken together, AK may serve as a cholesterol lowering agent to improve dyslipidemia.

The effects of ankaflavin (AK) on adipokines level and insulin sensitivity were investigated. Mice were fed with high-fat diet (HFD) and AK to evaluate hyperglycemia and hyperlipidemia. Additionally, mature differentiated 3T3-L1 adipocytes were treated with AK or monascin (MS) to determine insulin signaling. Results showed that AK down-regulated serum and hepatic triacylglyceride (TG) and total cholesterol (TC) levels in mice, ameliorating hyperglycemia and hyperinsulinemia symptoms elicited by the HFD. AK elevated serum adiponectin levels in HFD-induced mice and mature 3T3-L1 adipocytes. The effect of AK on insulin sensitivity and adipokines secretion were abolished by peroxisome proliferator activated receptor-γ PPAR-γ antagonist GW9662. Moreover, we made the novel discovery that AK markedly promoted insulin receptor and Akt phosphorylation, and increased glucose uptake in mature 3T3-L1 adipocytes to a greater extent than did MS. Taken together, AK attenuated insulin resistance in HFD-induced mice and promoted insulin sensitivity in 3T3-L1 adipocytes through PPAR-γ activation.

Miracle fruit (Synsepalum dulcificum) belongs to the Sapotaceae family. It can change flavors on taste buds, transforming acidic tastes to sweet. We evaluated various miracle fruit extracts, including water, butanol, ethyl acetate (EA), and hexane fractions, to determine its antioxidant effects. These extracts isolated from miracle fruit exerted potential for reduction of uric acid and inhibited xanthine oxidase activity in vitro and in monosodiumurate (MSU)-treated RAW264.7 macrophages. Moreover, we also found that the butanol extracts of miracle fruit attenuated oxonic acid potassium salt-induced hyperuricaemia in ICR mice by lowering serum uric acid levels and activating hepatic xanthine oxidase. These effects were equal to those of allopurinol, suggesting that the butanol extract of miracle fruit could be developed as a novel anti-hyperuricaemia agent or health food.

Polygonum multiflorum is known as a medicinal plant. It has been used as a folk medicine which showed antioxidative property.Protective effects of the water extracts (w/v:1/10) from fresh P. multiflorum (WEP) on carbon tetrachloride (CCl(4))-induced liver damage in rats were investigated.CCl(4) was used for inducing liver damage of SD rats, and WEP and emodin were fed for eight consecutive weeks.We found that emodin levels in fresh WEP was higher than that in ripening WEP. Rats were administered WEP and emodin, the main active compound, for 56 consecutive days. WEP significantly lowered the serum levels of hepatic enzyme markers, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) and reduced the generation of malonaldehyde. Treatment with WEP recovered glutathione S-transferase and catalase activity in rats as compared to treatment with CCl(4) alone. In addition, serum tumor necrosis factor-α, an inflammatory marker, was found to decrease in rats treated with WEP. In histopathological evaluation, fatty degeneration and necrosis were found to be significantly decreased in the CCl(4) plus WEP treatment group.WEP may be effective in attenuating liver damage by reducing lipid peroxidation as well as by positively modulating inflammation.

Gamma-aminobutyric acid (GABA) and angiotensin-converting enzyme inhibitor (ACEI) are compounds which can influence hypertension. The goal of this study is to optimize the culture condition for GABA and ACEI production by Lactobacillus plantarum NTU 102 fermented skim milk. In this study, we used 3-factor-3-level Box–Behnken design combining with response surface methodology, where the 3 factors represent the concentration of skim milk, the concentration of monosodium glutamate, and culture temperature. Best conditions for GABA and ACEI production differed. The results indicated that L. plantarum NTU 102 produced the highest combined levels of GABA and ACEI at 37 °C, in milk having 8% to 12% nonfat solids supplemented with 0.6% to 1% MSG. Agitation of the medium during fermentation had no effect on GABA or ACEI production but extended incubation (up to 6 d) increases levels of the bioactive compounds. L. plantarum NTU 102 fermented products may be a potential functional food source for regulating hypertension.

Monascus-fermented products offer valuable therapeutic benefits and have been extensively used for centuries in East Asia. Dioscorea has been proved to have anti-cancer effect. The aim of this study is to investigate the anti-tumor ability of the ethanol extract of red mold dioscorea (RMDE) on 7,12-dimethyl-1,2-benz[a]anthracene (DMBA)-induced hamster buccal pouch carcinogenesis. We induced oral squamous cell carcinoma (OSCC) in the buccal pouch of male Syrian golden hamsters by painting with 0.5% DMBA three times a week for 14 weeks. From 9 to 14 weeks, a dose of 50, 100, and 200 mg RMDE per kg body weight were painting with the hamsters for 6 weeks on days alternate to the DMBA application. The results demonstrated that RMDE decreased nitric oxide (NO), reactive oxygen species (ROS), and prostaglandin E2 (PGE2) overexpression in hamster buccal pouches in the DMBA treatment group and increased p53, serum tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) to significantly stimulate caspase-8 and -3 activities, indicating that RMDE reduced oxidative damage causing by DMBA and induced apoptosis in oral cancer cells. Therefore, RMDE may have therapeutic potentials against OSCC.

Products fermented with lactic acid bacteria may show antihypertensive effects via substances such as angiotensin I-converting enzyme inhibitor (ACEI) and γ-aminobutyric acid (GABA). It was previously found that milk fermented with Lactobacillus paracasei subsp. paracasei NTU 101 (101FM) or Lactobacillus plantarum NTU 102 (102FM) has ACEI and GABA activities. This study aimed to investigate the antihypertensive effects of 101FM and 102FM orally administered to spontaneously hypertensive rats (SHRs). Eight hours after a single oral administration or after 8 weeks of weekly (chronic) administration, 101FM and 102FM significantly decreased systolic and diastolic blood pressures in the SHRs. Microscopic examination of aortic tissue demonstrated that 101FM and 102FM reduced the disorganization of the media layer. These findings suggest that orally administered 101FM and 102FM have antihypertensive effects, possibly via ACEI and GABA activity, in SHRs. Therefore, 101FM and 102FM may be useful ingredients in physiologically functional foods to prevent hypertension.

BACKGROUND:Monascus-fermented products are among the most commonly used traditional food supplements. Dioscorea is known to exhibit anticancer properties. In this study the effects of the ethanol extract of red mold dioscorea (RMDE) on cell proliferation, cell cycle and apoptosis in human oral cancer cells were investigated. RESULTS: RMDE exercised growth inhibition on squamous cell carcinoma-25 (SCC-25) cells. RMDE-mediated G2/M phase arrest was associated with the down-regulation of NF-κB, resulting in the inhibition of cyclin B1 and CDK1 expression; this may be the mechanism by which RMDE inhibits cancer cells. Furthermore, the proapoptotic activity of RMDE was revealed by the Annexin V-FITC/PI double-staining assay. In addition, the proapoptotic effect of RMDE was evident by the inhibition of Bax expression in the mitochondria, resulting in the activation of caspase-9 and caspase-3 and subsequent triggering of the mitochondrial apoptotic pathway. RMDE also enhanced caspase-8 activity, indicating the involvement of the death receptor pathway in RMDE-mediated SCC-25 cell apoptosis. CONCLUSION: RMDE treatment inhibited the growth of SCC-25 cells by arresting cell cycle at the G2/M phase and induced apoptosis in a time- and dose-dependent manner. Therefore RMDE may be a good candidate for development as a dietary supplement against oral cancer. Copyright © 2010 Society of Chemical Industry

Methylglyoxal (MG) is a reactive dicarbonyl compound generated as an intermediate of glycolysis during the physical glycation in the diabetic condition. MG itself has been commonly implicated in the development of diabetic neuropathy. Several active compounds in Actinidia callosa have been found to inhibit glycation and MG-protein reaction.This study investigated the protective effects of A. callosa (kiwi fruits) peel ethanol extracts (ACE) on MG-induced Neuro-2A cell apoptosis.The Neuro-2A cells pre-treated by ACE (50-200 μg/mL) or allyl-isothiocyanate (AITC) (50 μM) for 6 h, in turn, the cells were treated with MG (250 μM) for 24 h.ACE or AITC treatment markedly inhibited the generation of reactive oxygen species (ROS) and the elevation of caspase-3 and capase-9 levels induced by MG in Neuro-2A cells. ACE and AITC elevated Bcl2 and inhibited Bax expressions in MG-induced Neuro-2A cells. ACE elevated Nrf2 transcriptional activity and nuclear translocation in MG-induced Neuro-2A cells. Nrf2 down-stream molecules including HO-1 and GCL were elevated by ACE or AITC treatment in MG-induced Neuro-2A cells. The protective effects of ACE on MG-induced Neuro-2A apoptosis were attenuated while Nrf2 knockdown.We established the first evidence that ACE might contribute to the prevention of the development of diabetic neuropathy by blocking the MG-mediated intracellular glycation system.

Yellow pigment monascin (MS) is a secondary metabolite isolated from Monascus -fermented products and has numerous physiological activities. However, the potential use of MS for immunomodulation remains unclear. We showed that MS and the synthetic peroxisome proliferator-activated receptor (PPAR)-γ ligand rosiglitazone (RG) significantly inhibited the production of Th2 cytokines, including IL-4, IL-5, and IL-13, in PMA/ionomycin-activated mouse EL-4 T cells. Moreover, we showed that this was due to cellular PPAR-γ translocation. These results indicate that MS and RG promote PPAR-γ-DNA interactions and suggest that the regulatory effects of MS and RG on Th2 cytokine production could be abolished with PPAR-γ antagonist treatment. MS and RG also suppressed Th2 transcription factor translocation (e.g., GATA-3 and nuclear factor of activated T cells) by preventing the phosphorylation of protein kinase C and signal transducer and activator of transcription 6.

Monascin (MS) is a yellow compound isolated from Monascus-fermented products that has pancreatic protective, anti-inflammatory, anti-oxidative, and hypolipidemic activity. We recently found that MS also acts as a peroxisome proliferator-activated receptor-gamma (PPARγ) agonist, thereby promoting insulin sensitivity in C2C12 cells. However, the attenuation of hyperglycemia by MS treatment in vivo remains uncertain. In the present study, both MS and pioglitazone significantly down-regulated blood glucose and hyperinsulinemia in fructose-rich diet (FRD)-induced C57BL/6 mice (8 weeks). In addition, inhibitions of inflammatory factor production, serum dyslipidemia, and hepatic fatty acid accumulation by MS and pioglitazone were attenuated by GW9662 (PPARγ antagonist). These results were mediated by MS-suppressing FRD-elevated lipogenic transcription factors, including sterol regulatory element-binding protein-1c (SREBP-1c), carbohydrate response element-binding protein (ChREBP), PPARγ coactivator-1α (PGC-1α), and PPARγ coactivator-1β (PGC-1β). Taken together, de novo lipogenesis results in hyperlipidemia and hyperglycemia by fructose induction thereby leading to diabetes development; we found that MS may inhibit lipogenesis in FRD-induced mice. These findings suggest that MS acts as an antidiabetic agent and thus may have therapeutic potential for prevention of diabetes.

Hyperglycemia is associated with advanced glycation end products (AGEs). Recently, AGEs were found to cause pancreatic damage, oxidative stress, and hyperglycemia through the AGE receptor. Carboxymethyllysine (CML) is an AGE but whether it induces pancreatic dysfunction remains unclear. Graptopetalum paraguayense, a vegetable consumed in Taiwan, has been used in folk medicine and is an antioxidant that protects against liver damage. We investigated the protective properties of G. paraguayense 95% ethanol extracts (GPEs) against CML-induced pancreatic damage. The results indicated that resveratrol, GPE, and gallic acid (the active compound of GPE) increased insulin synthesis via upregulation of pancreatic peroxisome proliferator activated-receptor-γ (PPARγ) and pancreatic-duodenal homeobox-1 (PDX-1) but inhibited the expression of CML-mediated CCAAT/enhancer binding protein-β (C/EBPβ), a negative regulator of insulin production. Moreover, resveratrol and GPE also strongly activated nuclear factor-erythroid 2-related factor 2 (Nrf2) to attenuate oxidative stress and improve insulin sensitivity in the liver and muscle of CML-injected C57BL/6 mice and resulted in reduced blood glucose levels. Taken together, these findings suggested that GPE and gallic acid could potentially be used as a food supplement to protect against pancreatic damage and the development of diabetes.

Methylglyoxal (MG) is a toxic-glucose metabolite and a major precursor of advanced glycation endproducts (AGEs). MG has been reported to result in inflammation by activating receptor for AGEs (RAGE). We recently found that Monascus-fermented metabolite monascin acts as a novel natural peroxisome proliferator-activated receptor-γ (PPARγ) agonist that improves insulin sensitivity. We investigated the metabolic, biochemical, and molecular abnormalities characteristic of type 2 diabetes in MG-treated Wistar rats treated with oral administration of monascin or rosiglitazone. Monascin (a novel PPARγ agonist) activated nuclear factor-erythroid 2-related factor 2 (Nrf2) and down-regulated hyperinsulinmia in oral glucose tolerance test (OGTT). Monascin was able to elevate glyoxalase-1 expression via activation of hepatic Nrf2, hence, resulting in MG metabolism to d-lactic acid and protected from AGEs production in MG-treated rats. Rosiglitazone did not activate Nrf2 nor glyoxalase expression to lower serum and hepatic AGEs levels. Monascin acts as a novel natural Nrf2 activator with PPARγ-agonist activity were confirmed by Nrf2 and PPARγ reporter assays in Hep G2 cells. These findings suggest that monascin acts as an anti-diabetic and anti-oxidative stress agent to a greater degree than rosiglitazone and thus may have therapeutic potential for the prevention of diabetes.

This study was designed to evaluate the effects of dimerumic acid (DMA) on receptor for advanced glycation endproducts (RAGE) signal activation and THP-1 monocyte inflammation treated with S100b, a specific ligand of RAGE. We found that DMA inhibited inflammatory cytokine production via upregulation of nuclear factor-erythroid 2-related factor 2 (Nrf2) and alleviated oxidative stress through attenuation of p47phox translocation to the membrane of S100b-treated THP-1 monocytes. We found that DMA activated Nrf2 mediated by the p38 kinase pathway in THP-1 monocytes. However, anti-inflammatory activity of DMA was attenuated by Nrf2 siRNA treatment. In an animal model, methylglyoxal (MG; 200mg/kg bw) was chosen to induce diabetes in Balb/C mice (6 weeks) in this work. The in vivo verification of anti-inflammation in peripheral blood mononuclear cells by DMA treatment was confirmed by tumor necrosis factor-α and interleukin-1β measurements. Oral glucose tolerance test, insulin tolerance test, hyperinsulinemia, and hyperglycemia were improved in MG-treated mice by DMA treatment and these effects were greater than those of silymarin and N-acetylcysteine. Furthermore, DMA increased hepatic glyoxalase mRNA and glutathione mediated by Nrf2 activation to metabolize MG into d-lactic acid, thereby reducing serum and hepatic AGE levels and suppressing inflammatory factor generation in MG-treated mice. However, DMA did not exert the antiglycation activity in MG-bovine serum albumin incubation. Taken together, the results indicate that DMA is a novel antioxidant and Nrf2 activator that lowers AGE levels and may prove to be an effective treatment for diabetes.

Ankaflavin (AK) is an active compound having anti-inflammatory, anti-cancer, antiatherosclerotic, and hypolipidemic effects. We have previously reported that AK acts as an antioxidant and antidiabetic drug; however, the mechanism by which AK prevents diabetes remains unknown. Hyperglycemia is associated with protein glycation, which produces advanced glycation end-products (AGEs). Methylglyoxal (MG)—a metabolite of carbohydrates—is believed to cause insulin resistance by inducing inflammation and pancreas damage. In this work, diabetes was induced in Wistar rats (4 weeks of age) by treating them with MG (600 mg/kg bw) for 4 weeks. We observed that AK (10 mg/kg bw) exerted peroxisome proliferator-activated receptor-γ (PPARγ) agonist activity, thereby enhancing insulin sensitivity (as indicated by hepatic GLUT2 translocation, PTP1B suppression, and glucose uptake) by downregulating blood glucose and upregulating pancreatic and duodenal homeobox-1 and Maf-A expression and increasing insulin production in MG-induced rats. However, these effects were abolished by the administration of GW9662 (PPARγ antagonist), but the expression of hepatic heme oxygenase-1 (HO-1) and glutamate–cysteine ligase (GCL) was not suppressed in MG-induced rats. Therefore, the nuclear factor erythroid-related factor-2 (Nrf2) activation was investigated. AK did not affect hepatic Nrf2 mRNA or protein expression but significantly increased Nrf2 phosphorylation (serine 40), which was accompanied by increased transcriptional activation of hepatic HO-1 and GCL. These data indicated that AK protected rats from oxidative stress resulting from MG-induced insulin resistance. In contrast, these effects were not detected when the rats were treated with the antidiabetic drug rosiglitazone (10 mg/kg bw). Moreover, we found that AK did not inhibit the generation of AGEs in vitro; however, the glutathione (GSH) levels in liver and pancreas of MG-induced rats were elevated in rats administered AK. Therefore, we believe that GSH may lower the MG level, which attenuates the formation of AGEs in the serum, kidney, liver, and pancreas of MG-induced rats. We also found that AK treatment reduced the production of inflammatory factors, such as tumor necrosis factor-α and interleukin-1β. Taken together, the results of our mechanistic study of MG-induced rats suggest that the protective effects of AK against diabetes are mediated by the upregulation of the signaling pathway of Nrf2, which enhances antioxidant activity and serves as a PPARγ agonist to enhance insulin sensitivity.

Fermentation products of the fungus Monascus offer valuable therapeutic benefits and have been used extensively for centuries in Asia. The aim of this study is to investigate the inhibitory effect of the Monascus -fermented metabolite monascin (MS) on the molecular mechanism of ovalbumin (OVA)-induced inflammation in the human THP-1 monocyte cell line. We found that 1, 5, and 25 μM of MS significantly attenuated several proinflammatory mediators, including inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression as well as nitric oxide (NO) and prostaglandin E 2 (PGE 2 ) formation caused by OVA stimulation. Further, 5 and 25 μM of MS significantly reduced the generation of tumor necrosis factor-α (TNF-α) and interleukin 6 (IL-6) at both the protein and mRNA levels. MS (5 and 25 μM) decreased OVA-induced phosphorylation of mitogen-activated protein kinase (MAPK) c-Jun NH 2 -terminal kinase (JNK), but not that of extracellular signal-regulated kinase (ERK) or p38 kinase. We used the peroxisome proliferator activated receptor-γ (PPAR-γ) antagonist GW9662 to show that MS inhibit JNK phosphorylation through increased expression of PPAR-γ. Thus, the metabolites from Monascus fermentation may serve as a dietary source of anti-inflammatory agents.

BACKGROUND: Immunomodulation by probiotic microorganisms has become a topic of increasing interest in food microbiology. Polysaccharides are broadly used in the food industry as gelling, thickening, stabilizing, or emulsifying agents. Some probiotics such as lactic acid bacteria also produce exopolysaccharides that stimulate macrophage production of cytokines. The aim of this study was to characterize the effects of exopolysaccharides of Lactobacillus paracasei subsp. paracasei NTU 101 (101EP) and Lactobacillus plantarum NTU 102 (102EP) exopolysaccharides on antioxidant activity and immunomodulation in vitro. RESULTS: The sugar composition (including arabinose, galactose, glucose, fructose, mannose, and maltose) of 101EP and 102EP was quantified by high-performance anion-exchange chromatography. Cytokine production (including IL-6, TNF-α, and IL-1β) was induced by 101EP and 102EP in Raw 264.7 in a dose-dependent manner (5-500 µg mL−1). 101EP and 102EP also demonstrated potential antioxidant properties (1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, chelation of ferrous ions, inhibition of linoleic acid peroxidation, and reducing power) in vitro. CONCLUSION: 101EP and 102EP stimulate cell proliferation and may be useful as a mild immune modulator of macrophages. Copyright © 2011 Society of Chemical Industry

Monascus -fermented products offer valuable therapeutic benefits and have been extensively used in East Asia. This study investigated the prevention of oral tumor formation and antioxidative ability of the ethanol extracts from red mold dioscorea (RMDE) on 7,12-dimethyl-1,2-benz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis. The HBP was painted with DMBA three times per week for 14 weeks, and animals were painted with celecoxib, RMDE (50, 100, and 200 mg/kg of bw), and ethanol extracts from dioscorea (200 mg/kg of bw) on days alternate to the DMBA application. The results demonstrated that RMDE attenuated tumor formation by elevating the antioxidase activity and suppressing the overproduction of reactive oxygen species, nitric oxide, prostaglandin E(2), and pro-inflammatory cytokines in the HBP caused by DMBA induction. These results indicated that RMDE exerted anti-inflammatory and antioxidative activity to prevent oral cancer. Therefore, the metabolite from Monascus fermentation may serve as a possible functional edible substance for the prevention of oral cancer.

Monascus-fermented products offer valuable therapeutic benefits and have been extensively used in East Asia. However, the polysaccharide obtained from Monascus-fermented products has never been investigated. This study evaluated the effects of dioscorea polysaccharide (DPS) and red mold dioscorea polysaccharide (RMDPS) on differentiation of leukemic THP-1 cells. DPS and RMDPS inhibited THP-1 cells proliferation in dose- and time-dependent manners. The differentiation induction (macrophage-like cells) was observed when THP-1 cells were treated with DPS and RMDPS for 5 days. Superoxide anion production, phagocytic capacity, and cytokine secretion confirmed activity for differentiating THP-1 cells. Results indicated that RMDPS elevated reactive oxygen species production and immune activity, including phagocytosis, interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and interferon-γ (IFN-γ) productions in THP-1 cells, which was greater than that seen with DPS. These results may be attributed to Monascus-fermentation altering the carbohydrate components and polysaccharide structure. RMDPS may serve as a novel material and functional ingredient to exert anticancer capacity.

Because of the high incidence of cardiovascular diseases in Asian countries, traditional fermented foods from Asia have been increasingly investigated for antiatherosclerotic effects. This study investigated the production of nattokinase, a serine fibrinolytic enzyme, in pigeon pea by Bacillus subtilis fermentation. B. subtilis 14714, B. subtilis 14715, B. subtilis 14716, and B. subtilis 14718 were employed to produce nattokinase. The highest nattokinase activity in pigeon pea was obtained using B. subtilis 14715 fermentation for 32 hours. In addition, the levels of antioxidants (phenolics and flavonoids) and angiotensin converting enzyme inhibitory activity were increased in B. subtilis 14715-fermented pigeon pea, compared with those in nonfermented pigeon pea. In an animal model, we found that both water extracts of pigeon pea (100 mg/kg body weight) and water extracts of B. subtilis-fermented pigeon pea (100 mg/kg body weight) significantly improved systolic blood pressure (21 mmHg) and diastolic blood pressure (30 mmHg) in spontaneously hypertensive rats. These results suggest that Bacillus-fermented pigeon pea has benefits for cardiovascular health and can be developed as a new dietary supplement or functional food that prevents hypertension.