Selection of initial microbial community for the alcoholic fermentation of sesame flavor-type baijiu.

The initial microbial community is critical for the production of volatile metabolites during traditional food fermentations. Selection of the initial community plays an important role in improving the quality of fermented foods. Here, we used high-throughput amplicon sequencing combined with multivariate statistical methods to explore the microbial succession in stacking and alcoholic fermentation stages in sesame flavor-type baijiu making. We proposed a selection strategy for the initial microbial community in the alcoholic fermentation stage, which determined the quality of baijiu. Results suggested that the microbial composition statistically differed between stacking and alcoholic fermentation stages (ANOSIM, Bacteria: R = 0.60, P = 0.001; Fungi: R = 0.53, P = 0.001). Microbial succession drove metabolic succession (Bacteria: r = 0.87, P < 0.05; Fungi: r = 0.56, P < 0.05) in alcoholic fermentation. The fermentation time of stacking fermentation determined the initial community for alcoholic fermentation, and it can be used as a criterion for selection of the initial microbial community for alcoholic fermentation. The succession distance of the microbial community was varied and reached the highest (Bacteria: 0.048, Fungi: 0.064) at 30 h in stacking fermentation. When we selected 30 h as stacking fermentation time, the concentration (4.58 mg/kg) and diversity (0.61) of volatile metabolites were highest at the end of alcoholic fermentation. This work developed a succession distance-guided approach to select the initial microbial community for the alcoholic fermentation of sesame flavor-type baijiu. This approach can be used to improve the quality of baijiu.

Rapid-dissolving electrospun nanofibers for intra-vaginal antibiotic or probiotic delivery.

The emergence of probiotics as an alternative and adjunct to antibiotic treatment for microbiological disturbances of the female genitourinary system requires innovative delivery platforms for vaginal applications. This study developed a new, rapid-dissolving form using electrospun polyethylene oxide (PEO) fibers for delivery of antibiotic metronidazole or probiotic Lactobacillus acidophilus, and performed evaluation in vitro and in vivo. Fibers did not generate overt pathophysiology or encourage Gardnerella growth in a mouse vaginal colonization model, inducing no alterations in vaginal mucosa at 24 hr post-administration. PEO-fibers incorporating metronidazole (100 µg MET/mg polymer) effectively prevented and treated Gardnerella infections (∼3- and 2.5-log reduction, respectively, 24 hr post treatment) when administered vaginally. Incorporation of live Lactobacillus acidophilus (107 CFU/mL) demonstrated viable probiotic delivery in vitro by PEO and polyvinyl alcohol (PVA) fibers to inhibit Gardnerella (108 CFU/mL) in bacterial co-cultures (9.9- and 7.0-log reduction, respectively, 24 hr post-inoculation), and in the presence of vaginal epithelial cells (6.9- and 8.0-log reduction, respectively, 16 hr post-inoculation). Administration of Lactobacillus acidophilus in PEO-fibers achieved vaginal colonization in mice similar to colonization observed with free Lactobacillus. acidophilus. These experiments provide proof-of-concept for rapid-dissolving electrospun fibers as a successful platform for intra-vaginal antibiotic or probiotic delivery.

Acidity drives volatile metabolites in the spontaneous fermentation of sesame flavor-type baijiu.

Environmental factors play an important role in contributing to intricate compositional dynamics and volatile metabolites in food fermentation. However, our understanding of which and how environmental factors affect volatile metabolites during sesame flavor-type baijiu fermentation is poor. Here, we examined the effects of environmental factors on the bacterial and fungal community to determine how changes in representative factors impact the microbial structure, diversity, and volatile metabolites in three fermentations. Results showed that bacterial community (ANOSIM: R = 0.79, P = 0.001), fungal community (ANOSIM: R = 0.65, P = 0.001), and volatile metabolites (ANOSIM: R = 0.84, P = 0.001) were significantly different in three fermentations. Acidity, ethanol, and moisture negatively impacted bacterial composition and diversity (P < 0.05). The fungal diversity and structure were positively and significantly affected by acidity (path coefficient, b = 0.54 for diversity, b = 0.35 for structure, P < 0.05). The fungal community rather than the bacterial community was the strongest driver of volatile metabolites. Fungal structure and diversity were equally important for the composition and content of volatile metabolites (structure: b = 0.50, diversity: b = 0.56, P < 0.05). 66 % of variations in volatile metabolites could be explained. Altogether these results indicated that acidity strongly drove volatile metabolites by modulating fungal structure and diversity. This work provides insights into managing volatile metabolites by regulating initial acidity in sesame flavor-type baijiu fermentation.

Combined Luteolin and Indole-3-Carbinol Synergistically Constrains ERα-Positive Breast Cancer by Dual Inhibiting Estrogen Receptor Alpha and Cyclin-Dependent Kinase 4/6 Pathway in Cultured Cells and Xenograft Mice.

The high concentrations of individual phytochemicals in vitro studies cannot be physiologically achieved in humans. Our solution for this concentration gap between in vitro and human studies is to combine two or more phytochemicals. We screened 12 phytochemicals by pairwise combining two compounds at a low level to select combinations exerting the synergistic inhibitory effect of breast cancer cell proliferation. A novel combination of luteolin at 30 µM (LUT30) and indole-3-carbinol 40 µM (I3C40) identified that this combination (L30I40) synergistically constrains ERα+ breast cancer cell (MCF7 and T47D) proliferation only, but not triple-negative breast cancer cells. At the same time, the individual LUT30 and I3C40 do not have this anti-proliferative effect in ERα+ breast cancer cells. Moreover, this combination L30I40 does not have toxicity on endothelial cells compared to the current commercial drugs. Similarly, the combination of LUT and I3C (LUT10 mg + I3C10 mg/kg/day) (IP injection) synergistically suppresses tumor growth in MCF7 cells-derived xenograft mice, but the individual LUT (10 mg/kg/day) and I3C (20 mg/kg/day) do not show an inhibitory effect. This combination synergistically downregulates two major therapeutic targets ERα and cyclin dependent kinase (CDK) 4/6/retinoblastoma (Rb) pathway, both in cultured cells and xenograft tumors. These results provide a solid foundation that a combination of LUT and I3C may be a practical approach to treat ERα+ breast cancer cells after clinical trials.

Identification of key odorants responsible for cooked corn-like aroma of green teas made by tea cultivar 'Zhonghuang 1'.

Fangping green tea (FPGT) produced by Zhonghuang 1 (C. sinensis var. sinensis cv. Zhonghuang 1), a new tea variety, has a classical cooked corn-like aroma, which is completely different from the green tea aroma. In order to illustrate the aroma characteristics of the green tea, the volatiles of FPGT was analyzed with gas chromatography-mass spectrometry (GC-MS) and gas chromatography-olfactometry (GC-O). The results showed that odor activity value (OAV) of dimethyl sulfide (DMS) was 1195.21 and the odor intensity about DMS was 6.2 in FPGT. Aroma recombination experiment also confirmed the important contribution of DMS to cooked corn-like aroma. Aroma character impact (ACI) values of DMS in tea processed by Zhonghuang 1 and Fudingdabai were 72.01% and 37.86%, respectively. This indicated that the high proportion of DMS was the dominant character of green tea with cooked corn-like aroma. In addition, the S-methylmethionine (SMM) content in fresh leaves of Zhonghuang 1 (0.21 mg/g) was significantly higher than that of Fudingdabai (0.16 mg/g), which was an important reason for high DMS content.

How Does Ginsenoside Rh2 Mitigate Adipogenesis in Cultured Cells and Obese Mice?

Ginsenoside Rh2, an intermediate metabolite of ginseng, but not naturally occurring, has recently drawn attention because of its anticancer effect. However, it is not clear if and how Rh2 inhibits preadipocytes differentiation. In the present study, we hypothesized that ginsenoside Rh2 attenuates adipogenesis through regulating the peroxisome proliferator-activated receptor gamma (PPAR-γ) pathway both in cells and obese mice. Different concentrations of Rh2 were applied both in 3T3-L1 cells and human primary preadipocytes to determine if Rh2 inhibits cell differentiation. Dietary Rh2 was administered to obese mice to determine if Rh2 prevents obesity in vivo. The mRNA and protein expression of PPAR-γ pathway molecules in cells and tissues were measured by real-time polymerase chain reaction (RT-PCR) and Western blot, respectively. Our results show that Rh2 dose-dependently (30-60 µM) inhibited cell differentiation in 3T3-L1 cells (44.5% ± 7.8% of control at 60 µM). This inhibitory effect is accompanied by the attenuation of the protein and/or mRNA expression of adipogenic markers including PPAR-γ and CCAAT/enhancer binding protein alpha, fatty acid synthase, fatty acid binding protein 4, and perilipin significantly (p < 0.05). Moreover, Rh2 significantly (p < 0.05) inhibited differentiation in human primary preadipocytes at much lower concentrations (5-15 µM). Furthermore, dietary intake of Rh2 (0.1 g Rh2/kg diet, w/w for eight weeks) significantly (p < 0.05) reduced protein PPAR-γ expression in liver and hepatic glutathione reductase and lowered fasting blood glucose. These results suggest that ginsenoside Rh2 dose-dependently inhibits adipogenesis through down-regulating the PPAR-γ pathway, and Rh2 may be a potential agent in preventing obesity in vivo.

Rapid-Release Griffithsin Fibers for Dual Prevention of HSV-2 and HIV-1 Infections.

The biologic griffithsin (GRFT) has recently emerged as a candidate to safely prevent sexually transmitted infections (STIs), including human immunodeficiency virus type 1 (HIV-1) and herpes simplex virus 2 (HSV-2). However, to date, there are few delivery platforms that are available to effectively deliver biologics to the female reproductive tract (FRT). The goal of this work was to evaluate rapid-release polyethylene oxide (PEO), polyvinyl alcohol (PVA), and polyvinylpyrrolidone (PVP) fibers that incorporate GRFT in in vitro (HIV-1 and HSV-2) and in vivo (HSV-2) infection models. GRFT loading was determined via enzyme-linked immunosorbent assay (ELISA), and the bioactivity of GRFT fibers was assessed using in vitro HIV-1 pseudovirus and HSV-2 plaque assays. Afterwards, the efficacy of GRFT fibers was assessed in a murine model of lethal HSV-2 infection. Finally, murine reproductive tracts and vaginal lavage samples were evaluated for histology and cytokine expression, 24 and 72 h after fiber administration, to determine safety. All rapid-release formulations achieved high levels of GRFT incorporation and were completely efficacious against in vitro HIV-1 and HSV-2 infections. Importantly, all rapid-release GRFT fibers provided potent protection in a murine model of HSV-2 infection. Moreover, histology and cytokine levels, evaluated from collected murine reproductive tissues and vaginal lavage samples treated with blank fibers, showed no increased cytokine production or histological aberrations, demonstrating the preliminary safety of rapid-release GRFT fibers in vaginal tissue.

Sustained-release Griffithsin nanoparticle-fiber composites against HIV-1 and HSV-2 infections.

Human immunodeficiency virus (HIV-1) and herpes simplex virus 2 (HSV-2) affect hundreds of millions of people worldwide. The antiviral lectin, Griffithsin (GRFT), has been shown to be both safe and efficacious against HSV-2 and HIV-1 infections in vivo. The goal of this work was to develop a multilayered nanoparticle (NP)-electrospun fiber (EF) composite to provide sustained-release of GRFT, and to examine its safety and efficacy in a murine model of lethal HSV-2 infection. Composites were fabricated from polycaprolactone (PCL) fibers surrounding polyethylene oxide (PEO) fibers that incorporated methoxy poly(ethylene glycol)-b-poly(lactide-co-glycolide) (mPEG-PLGA) GRFT NPs. GRFT loading and release were determined via ELISA, showing that NP-EF composites achieved high GRFT loading, and provided sustained-release of GRFT for up to 90 d. The in vitro efficacy of GRFT NP-EFs was assessed using HIV-1 pseudovirus assays, demonstrating complete in vitro protection against HIV-1 infection. Additionally, sustained-release NP-EFs, administered 24 h prior to infection, prevented against a lethal dose of HSV-2 infection in a murine model. In parallel, histology and cytokine expression from murine reproductive tracts and vaginal lavages collected 24 and 72 h post-administration were similar to untreated mice, suggesting that NP-EF composites may be a promising and safe sustained-delivery platform to prevent HSV-2 infection. Future work will evaluate the ability to provide prolonged protection against multiple virus challenges, and different administration times with respect to infection.

Combination of curcumin and luteolin synergistically inhibits TNF-α-induced vascular inflammation in human vascular cells and mice.

Emerging evidence shows that phytochemicals, the secondary plant metabolites present in a large variety of foods, have the potential ability in reducing the risk of cardiovascular diseases. However, the dosages of phytochemicals in the cellular and animal studies are too high to reach in humans by relevant foods or dietary supplement intake. The aims of this study were to investigate whether and how combined curcumin and luteolin synergistically inhibit tumor necrosis factor-alpha (TNF-α)-induced monocytes adhesion endothelium, a crucial step of the development of endothelial dysfunction, both in human vascular cells and mouse aortic endothelium. Our results show that combined curcumin (1 µM) and luteolin (0.5 µM) synergistically (combination index is 0.60) inhibited TNF-α-induced monocytes adhesion to human EA.hy926 endothelial cells while the individual chemicals did not have such effect at the selected concentrations. We also found that TNF-α-enhanced protein expressions of vascular cell adhesion molecule 1 (VCAM-1), monocyte chemotactic protein-1 (MCP-1) and nuclear factor (NF)-κB translocation were synergistically reduced by the combined curcumin and luteolin in EA.hy 926 cells while the individual chemical did not have this inhibitory effect. Consistently, 2 weeks dietary intake of combined curcumin (500 mg/kg) and luteolin (500 mg/kg) in C57BL/6 mice synergistically prevented TNF-α-stimulated adhesion of mouse monocytes to aortic endothelium ex vivo as well as the TNF-α-increased aortic protein expression of MCP-1 and VCAM-1. Therefore, combined curcumin and luteolin at physiological concentrations synergistically inhibits TNF-α-induced monocytes adhesion to endothelial cells and expressions of MCP-1 and VCAM-1 via suppressing NF-κB translocation into the nucleus.

Dietary epicatechin improves survival and delays skeletal muscle degeneration in aged mice.

We recently reported that epicatechin, a bioactive compound that occurs naturally in various common foods, promoted general health and survival of obese diabetic mice. It remains to be determined whether epicatechin extends health span and delays the process of aging. In the present study, epicatechin or its analogue epigallocatechin gallate (EGCG) (0.25% w/v in drinking water) was administered to 20-mo-old male C57BL mice fed a standard chow. The goal was to determine the antiaging effect. The results showed that supplementation with epicatechin for 37 wk strikingly increased the survival rate from 39 to 69%, whereas EGCG had no significant effect. Consistently, epicatechin improved physical activity, delayed degeneration of skeletal muscle (quadriceps), and shifted the profiles of the serum metabolites and skeletal muscle general mRNA expressions in aging mice toward the profiles observed in young mice. In particular, we found that dietary epicatechin significantly reversed age-altered mRNA and protein expressions of extracellular matrix and peroxisome proliferator-activated receptor pathways in skeletal muscle, and reversed the age-induced declines of the nicotinate and nicotinamide pathway both in serum and skeletal muscle. The present study provides evidence that epicatechin supplementation can exert an antiaging effect, including an increase in survival, an attenuation of the aging-related deterioration of skeletal muscles, and a protection against the aging-related decline in nicotinate and nicotinamide metabolism.-Si, H., Wang, X., Zhang, L., Parnell, L. D., Admed, B., LeRoith, T., Ansah, T.-A., Zhang, L., Li, J., Ordovás, J. M., Si, H., Liu, D., Lai, C.-Q. Dietary epicatechin improves survival and delays skeletal muscle degeneration in aged mice.

Anti-adipogenic Effects and Mechanisms of Ginsenoside Rg3 in Pre-adipocytes and Obese Mice.

Red or black ginseng has been reported more powerful than white/fresh ginseng in dealing with various diseases/conditions including obesity. The major reason is that heating/steaming, the process of making red or black ginseng, produces large amount of bioactive compounds including ginsenoside Rg3 (Rg3), which are trace in fresh or white ginseng. In the present study, Rg3 was applied both in pre-adipocytes and obese mice to investigate the anti-adipogenic effects and relevant mechanisms. Our results show that Rg3 dose-dependently inhibited cell differentiation both in 3T3-L1 cells (30, 50, and 100 µM) and human primary pre-adipocytes (10, 20, and 30 µM). This inhibitory effect is accompanied by the attenuation of the expressions of adipogenic markers including peroxisome proliferator-activated receptor gamma (PPAR-γ), CCAAT/enhancer binding protein alpha (C/EBP-α), fatty acid synthase (FAS), fatty acid binding protein 4 (FABP4) and perilipin. Although dietary intake of Rg3 (0.1 mg Rg3/kg diet, 8 weeks) did not significantly affect body weight gain, fat pads and food intake as well as of PPAR-γ expression in fat tissues, we found that hepatic PPAR-γ and C/EBP-α protein expressions and hepatic glutathione reductase and glutathione S-transferase, two major antioxidants molecules were significantly reduced by Rg3. These results suggest that ginsenoside Rg3 may be a potential agent in reducing/preventing obesity.

Ginseng and obesity: observations and understanding in cultured cells, animals and humans.

Ginseng, a traditional medical herb, has been reported having beneficial effects in fatigue, heart diseases, diabetes, immune function and erectile dysfunction. In recent years, increasing investigations have been conducted on ginseng in preventing and treating of obesity, one of the major worldwide escalating public health concerns. However, the effect and the relevant mechanisms behind how ginseng works as an antiobesity treatment are still controversial. In this review, we briefly discussed the chemical structures, metabolism and pharmacokinetics of ginseng and its major bioactive components ginsenosides. The major focus is on the antiobesity effects and the physiological, cellular and molecular mechanisms of ginseng and its ginsenosides in cultured cells, animal models and humans. We particularly compared the ginsenosides profiles, the antiobesity effects and the mechanisms between Asian ginseng (Panax ginseng) and American ginseng (Panax quinquefolius), the two major ginseng species having opposite medical effects in traditional Chinese medicine. Our unpublished data on the ginseng antiobesity in cultured cells and mice were also included. We further addressed the current problems and future directions of the ginseng antiobesity research.

Regulation of metabolic flux in Lactobacillus casei for lactic acid production by overexpressed ldhL gene with two-stage oxygen supply strategy.

This study describes a novel strategy to regulate the metabolic flux for lactic acid production in Lactobacillus casei. The ldhL gene encoding L-lactate dehydrogenase (L-LDH) was overexpressed in L. casei, and a two-stage oxygen supply strategy (TOS) that maintained a medium oxygen supply level during the early fermentation phase, and a low oxygen supply level in the later phase was carried out. As a consequence, a maximum L-LDH activity of 95.6 U/ml was obtained in the recombinant strain, which was over 4-fold higher than that of the initial strain. Under the TOS for L. casei (pMG-ldhL), the maximum lactic acid concentration of 159.6 g/l was obtained in 36 h, corresponding to a 62.8% increase. The results presented here provide a novel way to regulate the metabolic flux of L. casei for lactic acid production in different fermentation stages, which is available to enhance organic acid production in other strains.

High corn oil dietary intake improves health and longevity of aging mice.

Corn oil has been recommended as a replacement for saturated fats because of its high levels of poly- and mono-unsaturated fatty acids. In the present study, we tested whether very high levels of corn oil (58.6% fat-derived calories, FDC) intake improve health and longevity of aging mice. Twelve month old male C57BL/6 mice were fed a normal diet (10% FDC of corn oil, N) or a high fat diet (58.6% FDC of corn oil, HF) for 13-15 months. Our results show that a HF diet significantly increased the longevity of the aged mice (at 25 months of age, 53.8% of mice died in the N group, whereas the mortality rate was only 23.2% in the HF group). High corn oil also reversed aging-increased blood lipids including triglyceride, total cholesterol and LDL. Similarly, high corn oil intake overturned aging-raised pro-inflammatory markers including IL-1ß, IL-6, and monocyte chemotactic protein-1 (MCP-1) in the blood. In addition, corn oil intake reversed aging-damaged rotarod performance and liver function. Interestingly, the HF group was significantly heavier than the N group (53.6g/mouse vs. 41.3g/mouse); however, both HF and N groups had the same calorie intake (12.48 kcal/d/mouse vs. 12.24 kcal/d/mouse). Although, the HF group's food consumption was lower than that of the N group (2.4 g/d/mouse vs. 3.4 g/d/mouse). These results suggest that if total calorie consumption stays in the normal range, very high levels of corn oil intake improve health and longevity of aging mice.

[Role of histone deacetylase in inhibiting invasion of human gastric carcinoma cell line SGC-7901 by PPARgamma-mediated pathway].

BACKGROUND AND OBJECTIVE: Histone deacetylase (HDAC) can attenuate the function of peroxisome proliferator-activated receptor gamma (PPARgamma) to drive adipocyte differentiation. PPARgamma activation is confirmed to inhibit the development and metastasis of a variety of malignant cells. This study was to investigate the role of HDAC in inhibiting the invasion of human gastric carcinoma SGC-7901 cells through PPARgamma-mediated pathway, and explore potential mechanism. METHODS: SGC-7901 cells were treated with different concentrations of Trichostatin A (TSA) and Rosiglitazone (ROZ) respectively to select the best combination through assessing cell proliferation by MTT assay. Then cells were randomly divided into control group, TSA group, ROZ group, and combination group. Cell proliferation was detected by MTT assay after 48 h; cell invasion was detected by Boyden chamber invasion test. The mRNA levels of PPARgamma and matrix metalloproteinase-2 (MMP-2) were assessed by reverse transcription-polymerase chain reaction (RT-PCR), and the protein level of MMP-2 was evaluated by Western blot. RESULTS: Both TSA and ROZ inhibited the proliferation of SGC-7901 cells in a dose-dependent manner. A combination of 20 nmol/L TSA and 5 mumol/L ROZ synergistically inhibited the invasion of SGC-7901 cells (q=1.41). ROZ down-regulated the mRNA and protein expression of MMP-2. TSA and ROZ in combination reduced MMP-2 expression more obviously than ROZ alone. TSA up-regulated the expression of PPARgamma mRNA. CONCLUSIONS: HDAC suppresses the activation of PPARgamma through a series of molecular mechanisms. The activity of ROZ in inhibiting invasion of human gastric carcinoma cells can be enhanced after the activity of HDAC is inhibited by TSA.

[Performance optimization of property-improved biodiesel manufacturing process coupled with butanol extractive fermentation].

The products concentrations in traditional acetone-butanol (AB) fermentation are too low that large amount of energy has to be consumed in the distillation and product recovery process. Aiming at direct utilization of the fermentation products, in this study, optimization of property-improved biodiesel manufacturing process coupled with AB extractive fermentation was conducted, under the condition of using the biodiesel originated from waste cooking oil as the extractant and high concentrated corn flour medium. The effect of biodiesel/broth volume ratio, waste supernatant recycle ratio, and electronic carrier addition on the major process performance index was carefully investigated. Under the optimized condition, the biodiesel quality was improved with the cetane value increased from 51.4 to 54.4; "actual butanol yield" reached to a level of 18%, and waste supernatant recycle ratio exceeded 50%. In this way, elimination of energy-consuming product recovery process and realization of "energy-saving & waste minimization" industrial production target advocated by the state government, could be potentially expected.