Glycine max Fermented by a Novel Probiotic, Bifidobacterium animalis subsp. lactis LDTM 8102, Increases Immuno-Modulatory Function.

Many probiotic species have been used as a fermentation starter for manufacturing functional food materials. We have isolated Bifidobacterium animalis subsp. lactis LDTM 8102 from the feces of infants as a novel strain for fermentation. While Glycine max has been known to display various bioactivities including anti-oxidant, anti-skin aging, and anti-cancer effects, the immune-modulatory effect of Glycine max has not been reported. In the current study, we have discovered that the extract of Glycine max fermented with B. animalis subsp. lactis LDTM 8102 (GFB 8102), could exert immuno-modulatory properties. GFB 8102 treatment increased the production of immune-stimulatory cytokines in RAW264.7 macrophages without any noticeable cytotoxicity. Analysis of the molecular mechanism revealed that GFB 8102 could upregulate MAPK2K and MAPK signaling pathways including ERK, p38, and JNK. GFB 8102 also increased the proliferation rate of splenocytes isolated from mice. In an animal study, administration of GFB 8102 partially recovered cyclophosphamide-mediated reduction in thymus and spleen weight. Moreover, splenocytes from the GFB 8102-treated group exhibited increased TNF-α, IL-6, and IL-1ß production. Based on these findings, GFB 8102 could be a promising functional food material for enhancing immune function.

Enhancing Immunomodulatory Function of Red Ginseng Through Fermentation Using Bifidobacterium animalis Subsp. lactis LT 19-2.

Removal of sugar moieties from ginsenosides has been proposed to increase their biological effects in various disease models. In order to identify strains that can increase aglycone contents, we performed a screening using bacteria isolated from the feces of infants focusing on acid tolerance and ß-glucosidase activity. We isolated 565 bacteria and selected Bifidobacterium animalis subsp. lactis LT 19-2 (LT 19-2), which exhibited the highest ß-glucosidase activity with strong acid tolerance. As red ginseng (RG) has been known to exert immunomodulatory functions, we fermented RG using LT 19-2 (FRG) and investigated whether this could alter the aglycone profile of ginsenosides and improve its immunomodulatory effect. FRG increased macrophage activity more potently compared to RG, demonstrated by higher TNF-α and IL-6 production. More importantly, the FRG treatment stimulated the proliferation of mouse splenocytes and increased TNF-α levels in bone marrow-derived macrophages, confirming that the enhanced immunomodulatory function can be recapitulated in primary immune cells. Examination of the molecular mechanism revealed that F-RG could induce phosphorylations of ERK, p38, JNK, and NF-κB. Analysis of the ginsenoside composition showed a decrease in Rb1, Re, Rc, and Rb3, accompanied by an increase in Rd, Rh1, F2, and Rg3, the corresponding aglycone metabolites, in FRG compared to RG. Collectively, LT 19-2 maybe used as a probiotic strain to improve the bioactivity of functional foods through modifying the aglycone/glycoside profile.

Determination of Optimized Growth Medium and Cryoprotective Additives to Enhance the Growth and Survival of Lactobacillus salivarius.

The beneficial effects of lactic acid bacteria (LAB) have been intensively investigated in recent decades with special focus on modulation of the host intestinal microbiota. Numerous discoveries of effective probiotics are driven by a significantly increasing demand for dietary supplements. Consequently, technological advances in the large-scale production and lyophilization are needed by probiotic-related industries for producing probiotic LAB for commercial use. Our study had a dual objective, to determine the optimum growth medium composition and to investigate appropriate cryoprotective additives (CPAs) for Lactobacillus salivarius, and compare its responses with other Lactobacillus species. The one-factor-at-a-time method and central composite design were applied to determine the optimal medium composition for L. salivarius cultivation. The following composition of the medium was established (per liter): 21.64 g maltose, 85 g yeast extract, 1.21 ml Tween 80, 6 g sodium acetate, 0.2 g MgSO4∙7H2O, 0.02 g MnSO4∙H2O, 1 g K2HPO4, 1.5 g KH2PO4, 0.01 g FeSO4∙7H2O, and 1 g sodium citrate. A cryoprotective additive combination comprising 10% (w/v) skim milk and 10% (w/v) sucrose supplemented with 2.5% (w/v) sodium glutamate was selected for L. salivarius, and its effectiveness was confirmed using culture-independent methods in the freeze-dried cells of the Lactobacillus strains. In conclusion, the optimized medium enhanced the species-specific cultivation of L. salivarius. On the other hand, the cryoprotective effects of the selected CPA mixture may also be dependent on the bacterial strain. This study highlights the necessity for precise and advanced processing techniques for large-scale production of probiotics in the food and feed industries.

Development of putative probiotics as feed additives: validation in a porcine-specific gastrointestinal tract model.

Enforced restrictions on the use of antibiotics as growth promoters (AGPs) in animal production have prompted investigations into alternative feed additives in recent decades. Probiotics are currently the main feed additive used in livestock. However, the selection of probiotic candidates relies on human-based methods and little is known about the verification criteria for host-specific selection. We investigated the probiotic potential of Lactobacillus salivarius strains isolated from fed pig feces for their use as porcine feed additives. Two methods were developed that simulated the pig gastrointestinal (GI) tract and the intestinal epithelium, and these were compared with human-based in vitro methods and used for selecting porcine probiotics. Lactobacillus salivarius strain LS6 was identified as a promising probiotic strain for potential use as a porcine feed additive. This strain prevented disruption of the epithelial integrity of pig small intestine (PSI) cells by inhibiting the adherence of enterotoxigenic Escherichia coli K88. It also showed high survival rates in the in vitro pig GI tract model and good adhesion to PSI cells. We propose that host target-specific screening and validation methods are important tools in the development of effective probiotic feed additives, and this approach may support future-oriented agriculture.

Enhanced Absorption Study of Ginsenoside Compound K (20-O-ß-(D-Glucopyranosyl)-20(S)-protopanaxadiol) after Oral Administration of Fermented Red Ginseng Extract (HYFRG™) in Healthy Korean Volunteers and Rats.

To evaluate the pharmacokinetics of compound K after oral administration of HYFRG and RG in humans, an open-label, randomized, single-dose, fasting, and one-period pharmacokinetic study was conducted. After oral administration of a single 3 g dose of HYFRG and RG to 24 healthy Korean males, the mean (±SD) of AUC0-t and C max of compound K from HYFRG were 1466.83 ± 295.89 ng·h/mL and 254.45 ± 51.20 ng/mL, being 115.2- and 80-fold higher than those for RG (12.73 ± 7.83 ng·h/mL and 3.18 ± 1.70 ng/mL), respectively; in case of Sprague Dawley rats the mean (±SD) of AUC0-t and C max of compound K from HYFRG was 58.03 ± 32.53 ng·h/mL and 15.19 ± 10.69 ng/mL, being 6.3- and 6.0-fold higher than those from RG (9.21 ± 7.52 ng·h/mL and 2.55 ± 0.99 ng/mL), respectively. T max of compound K in humans and rats was 2.54 ± 0.92 and 3.33 ± 0.50 h for HYFRG and 9.11 ± 1.45 and 6.75 ± 3.97 hours for RG, respectively. In conclusion, the administration of HYFRG resulted in a higher and faster absorption of compound K in both humans and rats compared to RG.

Triglyceride-Lowering Effects of Two Probiotics, Lactobacillus plantarum KY1032 and Lactobacillus curvatus HY7601, in a Rat Model of High-Fat Diet-Induced Hypertriglyceridemia.

The triglyceride-lowering effect of probiotics Lactobacillus plantarum KY1032 and Lactobacillus curvatus HY7601 were investigated. Male SD Wistar rats were randomly divided into three groups and fed high-fat diet (HFD), HFD and probiotics (5 X 10(9) CFU/day of L. plantarum KY1032 and 5 X 10(9) CFU/day of L. curvatus HY7601), or normal diet for 6 weeks. Probiotic treatment significantly lowered the elevated plasma triglyceride and increased plasma free fatty acid, glycerol, and plasma apolipoprotein A-V (ApoA-V) levels. The probiotic-treated group showed elevated hepatic mRNA expression of PPARα, bile acid receptor (FXR), and ApoA-V. These results demonstrate that L. plantarum KY1032 and L. curvatus HY7601 lower triglycerides in hypertriglyceridemic rats by upregulating ApoA-V, PPARα, and FXR.

Apple Pomace Extract Improves Endurance in Exercise Performance by Increasing Strength and Weight of Skeletal Muscle.

Ursolic acid is a lipophilic pentacyclic triterpenoid found in many fruits and herbs and is used in several herbal folk medicines for diabetes. In this study, we evaluated the effects of apple pomace extract (APE; ursolic acid content, 183 mg/g) on skeletal muscle atrophy. To examine APE therapeutic potential in muscle atrophy, we investigated APE effects on the expression of biomarkers associated with muscle atrophy and hypertrophy. We found that APE inhibited atrophy, while inducing hypertrophy in C2C12 myotubes by decreasing the expression of atrophy-related genes and increasing the expression of hypertrophy-associated genes. The in vivo experiments using mice fed a diet with or without APE showed that APE intake increased skeletal muscle mass, as well as grip strength and exercise capacity. In addition, APE significantly improved endurance in the mice, as evidenced by increased exhaustive running time and muscle weight, and reduced the expression of the genes involved in the development of muscle atrophy. APE also decreased the concentration of serum lactate and lactate dehydrogenase, inorganic phosphate, and creatinine, the indicators of accumulated fatigue and exercise-induced stress. These results suggest that APE may be useful as an ergogenic functional food or dietary supplement.

The inhibitory effect of Lactobacillus plantarum KY1032 cell extract on the adipogenesis of 3T3-L1 Cells.

Some probiotics and their cell components are known to modulate lipid metabolism in vitro and/or in vivo. This study was carried out to investigate possible anti-adipogenic action of a probiotic cell extract, Lactobacillus plantarum KY1032 cell extract (KY1032-CE), in vitro using 3T3-L1 cells. Lipid regulation in the cell culture system was assessed by AdipoRed assay and Oil red O staining of intracellular lipids and real-time polymerase chain reaction and western blot analysis of adipogenesis-related factors. AdipoRed assay revealed that KY1032-CE treatment significantly decreased lipid accumulation in maturing 3T3-L1 preadipocytes in a dose-dependent manner. Oil red O staining demonstrated that KY1032-CE reduced the number of lipid-containing rounded cells. KY1032-CE down-regulated the mRNA and protein expression of four adipocyte-specific genes: peroxisome proliferator-activated receptor-γ2, CCAAT/enhancer binding protein-α, fatty acid synthase, and adipocyte-fatty acid binding protein. Accordingly, these results indicate that KY1032-CE can reduce fat mass by modulating adipogenesis in maturing preadipocytes. Further studies are needed to elucidate its mode of actions in efficacy tests of KY1032-CE in vivo.

Protective and therapeutic effects of an extract mixture of alder tree, labiate herb, milk thistle green bean-rice bran fermentation, and turnip against ethanol-induced toxicity in the rat.

An herbal extract mixture and yogurt added to the herbal extract mixture were tested for their protective and therapeutic effects on ethanol-induced liver injury. The herbal extract mixture, yogurt and commercial drugs were used for treatment for two weeks prior to administering a single oral dose of ethanol (3 g/kg body weight). The herbal extract mixture and yogurt added to the herbal extract mixture were found to provide protection against ethanol-induced toxicity comparable to the commercial drug treatment, according to the serum and histopathological analysis. It was also shown that co-treatment with herbal extract mixture and yogurt against a triple oral dose of ethanol (2 g/kg body weight, over one week) provided protection against ethanol toxicity. After the initial set of experiments, the herbal extract mixture and yogurt treatments were extended for three more weeks. When compared to the positive control, further treatment with both the herbal extract and yogurt significantly reduced liver injury and resulted in a lower grade of lipid deposition.