A mixture of extracts from natural ingredients reduces the neurotoxic polarization of microglia via modulating NF-κB/NF-E2-related factor 2 activation.

Neurotoxic microglia-provoked neuroinflammation is implicated in cognitive decline in Alzheimer's disease (AD). Supplementation with Ginkgo biloba, phosphatidylserine, Curcuma longa, and propolis is reported to improve the cognitive functions of elderly people; however, the underlying mechanisms of this combination of natural ingredients are unknown. We investigated the effects of a mixture of extracts from propolis, Coffea arabica, Gotu kola, phosphatidylserine, Ginkgo biloba, and Curcuma longa (mixture) on microglia polarization after exposure to amyloid ß1-42 (Aß1-42, 1 µM) and lipopolysaccharide from Porphyromonas gingivalis (PgLPS, 1 µg/mL), using MG6 and BV2 microglial cells. Exposure to Aß1-42 and PgLPS (AL) raised the mRNA expression of IL-1ß, TNF-α, and IL-6, nuclear translocation of p65 NF-κB in MG6 cells and BV2 cells, and mitochondrial reactive oxygen species (ROS) production in MG6 cells. The mixture dramatically suppressed the mRNA expression of IL-1ß, TNF-α, and IL-6, but significantly promoted that of IL-10, TGFß1, and BDNF in AL-exposed MG6 and BV2 cells. Furthermore, the mixture significantly suppressed the nuclear translocation of p65 NF-κB but significantly promoted that of NF-E2-related factor 2 (Nrf2) in AL-exposed MG6 and BV2 cells. Furthermore, the mixture significantly ameliorated mitochondrial ROS production but increased mitochondrial membrane potential in MG6 cells. These observations strongly suggest that the mixture demotes the neuropathic polarization of microglia by modulating NF-κB/Nrf2 activation and improving mitochondrial functions. This study supplies the potential mechanisms of the efficacy of a combination of natural ingredients that can be applied in the prevention of cognitive decline in AD and aging by targeting microglia-mediated neuroinflammation.

Royal Jelly Increases Hematopoietic Stem Cells in Peripheral Blood: A Double-Blind, Placebo-Controlled, Randomized Trial in Healthy Subjects.

Objectives: Royal jelly (RJ), produced by honeybees, influences stem cell functions, such as pluripotency maintenance of mouse embryonic stem cells and prevention of aging-related muscle stem cell functional deterioration. Thus, we hypothesized that RJ administration has various health-promoting effects based on stem cells. However, its effects are unknown in humans. In this study, we have attempted for the first time to clarify whether the administration of RJ in humans affects stem cells. Materials and Methods: This randomized, double-blind, placebo-controlled study was performed on healthy subjects (n = 90) who received protease-treated RJ at a dose of 1200 mg/day or placebo daily for four weeks. Also, the participants with a low number of hematopoietic stem cells (HSCs) in peripheral blood were preferentially selected. HSC counts, endothelial progenitor cell (EPC) counts, blood cell counts in peripheral blood, cytokines in serum, and physical conditions were evaluated. Results and Conclusion. Eligible data from 86 subjects (placebo: 42, RJ: 44) who completed the study were analyzed. There were no significant differences between the two groups regarding the changes in peripheral HSC count (p=0.103), while diastolic blood pressure showed a significant improvement in the RJ group compared to that in the placebo group (p=0.032). The subgroup analysis excluded 14 subjects who complained of cold symptoms at baseline or within five days of the four-week study. The changes in the HSC populations were significantly higher in the RJ group than those in the placebo group (p=0.042). No adverse effects were observed in any of the groups. These results suggest that RJ administration affected the peripheral HSC count and may influence stem cell functions. Further research is needed to reveal the various health-promoting benefits of RJ based on stem cells.

Difructose anhydride III enhances bioavailability of water-insoluble iron in anemic Vietnamese women.

Difructose anhydride III (DFAIII) is an indigestible disaccharide and has been shown to enhance iron absorption in animal studies; however, the effect has not been investigated in anemic subjects. We investigated the efficacy of co-administration of DFAIII with water-insoluble iron in the treatment of iron deficiency anemia in Vietnamese women. One hundred sixty-eight moderately anemic women (80 g/L

Low activities of intestinal lactase suppress the early phase absorption of soy isoflavones in Japanese adults.

BACKGROUND & AIMS: The contribution of lactase to isoflavone bioavailability has not been clarified. We evaluated the association between lactase activity and the bioavailability of isoflavone glucosides in Japanese adults. METHODS: Twenty-six Japanese adult participants completed a study that included tests of breath hydrogen after a lactose load, orocecal transit time, and isoflavone glucoside absorption. Lactose malabsorbers were defined as those with an increase in breath hydrogen level (DeltaH(2)) of more than 20 ppm after a load of lactose (20 g). Participants ingested 200 ml soymilk, and serum isoflavones were analyzed until 480 min. RESULTS: Serum daidzein and genistein levels increased rapidly until 60 min, then slowly increased. The increases of serum isoflavones in the early phase, but not the later phase, were suppressed in lactose malabsorbers. DeltaH(2 max) after a load of lactose inversely correlated with serum daidzein levels at 30 min. The percentage of equol producers tended to be greater among lactose malabsorbers (P=0.067). CONCLUSIONS: Lactase may be involved in absorption of isoflavone glucosides in the small intestine, but the bacterial deglycosylation in the large intestine compensates the reduction of isoflavone absorption. The beneficial effect of isoflavones may not be impaired by low lactase activity.

Dietary melibiose regulates th cell response and enhances the induction of oral tolerance.

We examined how dietary melibiose affected the T-helper (Th) cell responses induced by an orally fed antigen in ovalbumin (OVA)-specific T cell receptor transgenic mice (OVA 23-3). Dietary melibiose markedly decreased the Th2 type responses as shown by a significant decrease in the interleukin (IL)-4 production and T cell proliferative response induced by sensitization from the 7-d oral administration of OVA. It was additionally observed that the Th1 type responses tended to decrease. We therefore examined the effect of melibiose feeding on the induction of immunological tolerance induced by the oral administration of an antigen (oral tolerance). The Th cell responses induced in BALB/c mice by subcutaneous immunization with OVA were suppressed by the prior oral administration of OVA. Such responses in the OVA-fed and immunized mice were further diminished by dietary melibiose. These results suggest that dietary melibiose strongly affected the Th cell responses to an ingested antigen, and further demonstrate the potential of melibiose to enhance the induction of oral tolerance.

Ingestion of difructose anhydride III enhances absorption and retention of calcium in healthy men.

We examined the effects of a nondigestible disaccharide difructose anhydride III (DFAIII) on calcium absorption and retention by means of a human balance study of single-blind crossover design. Twelve healthy male subjects ingested 250 mg of shell powder as calcium carbonate (corresponding to 100 mg of calcium) with or without 1.0 g DFAIII three times a day for 13 d. In the last 4 d as a balance period, all urine and feces were collected and evaluated for calcium excretion. The apparent calcium absorption (mg/d) and rate of absorption (%) were higher, and those of retention were much higher, in the DFAIII group than in the control group. Furthermore, serum osteocalcin increased after the experimental period in the DFAIII group but not in the control group. These results indicate that DFAIII ingestion enhances intestinal calcium absorption, which might be beneficial for bone metabolism.

Different effects of difructose anhydride III and inulin-type fructans on caecal microbiota in rats.

The effects of different kinds of inulin-type fructans on caecal microbiota were evaluated in rats. Four groups of male Wistar rats were fed either a control diet, or diets containing 5% inulin, 5% fructooligosaccharides (FOS), or 5% difructose anhydride III (DFAIII) for two weeks. In the DFAIII group, caecal propionate, butyrate, counts of bifidobacteria, and total anaerobes were lower than in the inulin group, while caecal propionate, succinate, counts of bifidobacteria, and total anaerobes were lower than in the FOS group. Compared to controls, in the DFAIII group the counts of clostridia in caecum were increased by 3 log units. However, this change was statistically not significant. There were no differences between inulin and FOS groups for the pool of short chain fatty acids in caecum and bacterial counts. Results indicate that DFAIII has different effects on caecal microbiota compared to inulin and FOS and that these differences are most likely due to the alpha(3-->2) bonds in DFAIII.

Supplementation of difructose anhydride III enhanced elevation of plasma equol concentrations and lowered plasma total cholesterol in isoflavone-fed rats.

Equol, a derivative of daidzein produced by enterobacteria, has greater activity as a phyto-oestrogen compared with daidzein. Difructose anhydride III (DFAIII) is a newly manufactured non-digestible disaccharide with unique fermentation properties. The present study evaluated the prebiotic effects of DFAIII on equol production and on plasma cholesterol concentrations related to the changes in equol production. We compared plasma equol concentrations at 10.00 and 18.00 hours and faecal isoflavone excretion in three groups of seven rats (male Wistar-ST strain, 6 weeks old) fed a basal diet or a DFAIII or fructooligosaccharide (15 g/kg diet) diet containing 1 g soya isoflavones/kg diet for 20 d. Equol concentrations in the DFAIII group were higher than in the control and fructooligosaccharides groups, especially in the later phase of the light period (18.00 hours) throughout the experiment. Daizein and genistein concentrations did not change between the diet groups. The faecal ratios of equol:daidzein were very high in all groups, but the ratios were higher in the DFAIII group than the control and fructooligosaccharide groups on day 3, and this tendency continued throughout the experiment. On day 20, the plasma total cholesterol concentration was lowest in the DFAIII group. Additionally, the cholesterol concentrations were inversely correlated to plasma equol concentration in all the rats. In conclusion, DFAIII efficiently enhanced plasma equol concentrations, which may be associated with an increase in equol production and a decrease in equol degradation by enterobacteria. Higher plasma equol concentrations may contribute to the hypocholesterolaemic effect of DFAIII feeding.

Difructose anhydride III does not contribute to body energy accumulation in rats.

We evaluated the body energy accumulation as fat and protein from ingestion of difructose anhydride III (DFAIII). Male Wistar rats were fed 0, 0.25, 0.5, 1.0, or 1.5 g per d of sucrose or DFAIII added to a 7 g of basal diet for 20 d. Supplements of DFAIII did not increase whole body or peripheral fat or total body energy, whereas sucrose increased them in a dose-dependent manner. Dose-dependent increases in body water were observed in both groups. The body protein was influenced by the dose of sugars. The estimated available energy value of DFAIII was 0.263 kcal per gram; this value is one-fifteenth that of sucrose. Ingestion of DFAIII dose-dependently increased the cecal SCFA pool. DFAIII was not detected in feces, showing complete degradation of DFAIII in the intestine. These results indicate that DFAIII is a fermentable saccharide with quite low available energy for fat accumulation.

Two-week feeding of difructose anhydride III enhances calcium absorptive activity with epithelial cell proliferation in isolated rat cecal mucosa.

OBJECTIVE: Ingestion of difructose anhydride III (DFAIII) enhances calcium (Ca) absorption in rats. The present study investigated the mechanism involved in increased Ca transport by DFAIII ingestion. The short-term and long-term effects of DFAIII feeding on Ca transport were determined by using isolated epithelium from the small and large intestine in rats. METHODS: Male Sprague-Dawley rats were fed an 8% cellulose or 5% cellulose plus 3% DFAIII diet for 14 d. Net epithelial Ca transport in the small intestine, cecum, and colon was compared between the two diet groups by using an Ussing chamber. The contents and epithelial tissues in the cecum were analyzed. RESULTS: There were no differences in basal and luminal DFAIII-induced Ca transport in the isolated small intestinal and colonic mucosa between the two diet groups. Basal and lumen DFAIII-induced Ca transport in the cecum in the DFAIII-fed group was higher than that in the control group. A decrease in pH and an increase in Ca pools, short-chain fatty acids, or organic acids in the cecal contents and in the depth and number of cells in crypts in cecal tissue were observed. CONCLUSIONS: The increase in Ca transport involved two mechanisms: the presence of DFAIII in the small intestine directly affected the epithelial tissue and caused increased Ca absorption as a short-term effect, and the degradation of DFAIII by microbial fermentation produced short-chain fatty acids and subsequently enhanced Ca absorption in the large intestine as a long-term effect.

Effects of intravenous betaine on methionine-loading-induced plasma homocysteine elevation in rats.

An intravenous methionine-loading model was characterized, and the suppressive effect of betaine on plasma homocysteine elevation induced by methionine loading was examined in rats. The plasma homocysteine concentrations significantly increased 5-120 minutes after 0.34 mmol/kg of methionine loading and then returned to the baseline within 240 minutes. Betaine was then intravenously administered at the same time as the methionine loading. The total increment of plasma homocysteine was assessed using the positive incremental area under the plasma homocysteine concentration curve over the 240-minute post-methionine-loading period (DeltaAUC(0-240)). Betaine reduced DeltaAUC(0-240) dose-dependently: 81% of the control by 1.7 mmol/kg of betaine and 33% by 3.4 mmol/kg. The effects of glycine and methylglycine, analogues of betaine, were also investigated. As observed for betaine, methylglycine decreased DeltaAUC(0-240) to 44% of the control, whereas glycine showed no significant effect on DeltaAUC(0-240), indicating that methyl groups of betaine and dimethylglycine were necessary to suppress plasma homocysteine elevation. These results suggest that betaine contributes to the suppression of plasma homocysteine elevation by promoting homocysteine metabolism, and seems to work as a methyl donor.

Comparative effect of repeated ingestion of difructose anhydride III and palatinose on the induction of gastrointestinal symptoms in humans.

We evaluated the safety and change in fermentability from repeated ingestion of difructose anhydride III (DFAIII) in humans. A randomized controlled single-blind crossover study with thirteen subjects was conducted. Each subject ingested 5 g of DFAIII or palatinose daily for 12 days, before and after which the subject was loaded with 10 g of DFAIII and had breath hydrogen measured from 0 to 9 h (DL test) to evaluate the fermentability of DFAIII. The defecation frequency and abdominal symptom score were the same between each ingestion period. Moreover, DFAIII ingestion had no influence on blood test results. Only the breath hydrogen excretion in post-DFAIII ingestion was slightly higher at h 8 than the pre-ingestion. Consequently, repeated ingestion of DFAIII for 12 days was as safe as palatinose ingestion, especially with respect to abdominal symptoms and blood test results, and its high resistance to enterobacterial fermentation in humans was not impaired.

Effect of difructose anhydride III on calcium absorption in humans.

The effects of difructose anhydride III (DFAIII) on stimulating calcium absorption was investigated in humans. We studied changes in the time-course of characteristics urinary calcium excretion in 12 healthy men given 0.3, 1.0 or 3.0 g of DFAIII and 300 mg of calcium as calcium carbonate. In addition, urinary excretion and urine concentrations of creatinine and deoxypyridinoline were determined. Urine calcium excretion every 2 hours after the intake were higher over than that of the control subjects. The total amount of urinary calcium excretion for 10 hours was significantly greates in the subjects given 1.0 g or 3.0 g of DFAIII than that of the control subjects. However, there were no differences in the urine concentrations of creatinine and deoxypyridinoline between the subjects given DFAIII and the control subjects. These findings suggests that low dose of DFAIII had a stimulating effect on calcium absorption in humans.

Indigestible disaccharides open tight junctions and enhance net calcium, magnesium, and zinc absorption in isolated rat small and large intestinal epithelium.

The effects of three indigestible disaccharides on net calcium (Ca), magnesium (Mg), and zinc (Zn) transport in isolated rat jejunal, ileal, cecal, and colonic epithelium were determined. Permeability of fluorescein isothiocynate-dextran-4 (FD4) and transepithelial electrical resistance (TEER), which vary according to tight junction (TJ) activity in the intestinal mucosa, were also determined. The addition of 1-100 mM melibiose, difructose anhydride (DFA) III, or DFA IV to the mucosal medium increased the net absorption of the three minerals and FD4 permeability, while decreasing TEER dose dependently in the four intestinal portions. Positive linear relations were found between the net transport of the three minerals and FD4 passage in all portions of the intestine, whereas negative linear relations were observed between net absorption of the three minerals and TEER. We concluded that the three indigestible saccharides directly affect the epithelial tissue and open TJs, thereby promoting Ca, Mg, and Zn absorption in the small and large intestine in vitro.

The effects of di-D-fructofuranose-1,2':2,3'-dianhydride (DFA III) administration on human intestinal microbiota.

Di-D-fructofuranose-1,2':2,3'-dianhydride (DFA III) was shown to enhance Ca absorption in rat and human intestine. The effects of DFA III administration (9 g per day for 4 weeks that corresponded to 3-fold the optimal dosage of DFA III) on human intestinal microbiota were studied using denaturing gradient gel electrophoresis (DGGE). The major groups of human intestinal microbiota reported previously: the Bacteroides, the Clostridium coccoides group (Clostridium cluster XIVa), the Clostridium leptum group (Clostridium cluster IV), and the Bifidobacterium group were detected. The similarity of 30 DGGE profiles based on the V3 region (before and after administration to the 15 subjects) of the 16S rDNA were calculated using Pearson's correlation based on numbers, positions and intensity of bands, and then a dendrogram of DGGE profiles was constructed by the unweighted pair group method using arithmetic average (UPGMA) clustering method. By these analyses, no difference in DGGE profiles after DFA III administration was observed in healthy subjects, while two subjects with chronic constipation showed different profiles, namely on numbers, positions and the intensity of some bands. Their stools were softer and stool frequencies increased and they obtained relief from constipation.

(-)-hydroxycitrate ingestion increases fat oxidation during moderate intensity exercise in untrained men.

We examined the effects of (-)-Hydroxycitrate (HCA) ingestion on fat oxidation during moderate intensity exercise in untrained men. Six subjects ingested 500 mg of HCA or a placebo for 5 days and did endurance exercise. Blood FFA concentrations were significantly increased and respiratory exchange ratio (RER) decreased by HCA ingestion. These results suggested short-term HCA ingestion increases fat oxidation in untrained men.

(-)-Hydroxycitric acid ingestion increases fat utilization during exercise in untrained women.

(-)-Hydroxycitric acid (HCA) is a competitive inhibitor of the enzyme ATPcitrate-lyase, which inhibits lipogenesis in the body. Moreover, HCA increases endurance exercise performance in trained mice and athletes. However, had not been investigated in untrained animals and humans. Therefore, we investigated the effects of short-term HCA ingestion on endurance exercise performance and fat metabolism in untrained women. In two experiments designed as a double-blind crossover test, six subjects ingested 250 mg of HCA or placebo (same amount of dextrin) via capsule for 5 d and then participated in cycle ergometer exercise. They cycled at 40% VO2max for 1 h and then the exercise intensity was increased to 60% VO2max until exhaustion on day 5 of each experiment. HCA tended to decrease the respiratory exchange ratio (RER) and carbohydrate oxidation during 1 h of exercise. In addition, exercise time to exhaustion was significantly enhanced (p<0.05). These results suggest that HCA increases fat metabolism, which may be associated with a decrease in glycogen utilization during the same intensity exercise and enhanced exercise performance.

Absorptive activity of calcium in the isolated cecal epithelium adaptively increased by 2 week's feeding of difructose anhydride III in rats.

We compared net Ca absorption and Lucifer Yellow (LY), a paracellular passage dye, permeability in the epithelium isolated from the rat small intestine, cecum, and colon after feeding with control and difructose anhydride (DFA) III diets for 14 days using the Ussing chamber system. Feeding of DFA III increased net Ca transport and LY passage in the cecal but not in small intestinal or colonic epithelium. Ability of paracellular Ca passage via Tight-junction (TJ) in the cecum was changed adaptively by feeding of DFA III. Changes in microbial fermentation may affect the functional changes of Ca transport in cecal epithelium itself.

Effects of garcinia cambogia (Hydroxycitric Acid) on visceral fat accumulation: a double-blind, randomized, placebo-controlled trial.

BACKGROUND: (-)-Hydroxycitric acid (HCA) is an active ingredient extracted from the rind of the Indian fruit Garcinia cambogia. It inhibits adenosine triphosphate citrate lyase and has been used in the treatment of obesity. OBJECTIVE: The primary end point of this study was the effects of 12 weeks of G cambogia extract administration on visceral fat accumulation. The secondary end points were body indices (including height, body weight, body mass index [BMI], waist and hip circumference, and waist-hip ratio) and laboratory values (including total cholesterol, triacylglycerol, and free fatty acid). METHODS: This study was performed according to a double-blind, randomized, placebo-controlled, parallel-group design. Subjects aged 20 to 65 years with a visceral fat area >90 cm(2) were enrolled. Subjects were randomly assigned to receive treatment for 12 weeks with G cambogia (containing 1000 mg of HCA per day) or placebo. At the end of the treatment period, both groups were administered placebo for 4 weeks to assess any rebound effect. Each subject underwent a computed tomography scan at the umbilical level at -2, 0, 12, and 16 weeks. RESULTS: Forty-four subjects were randomized at baseline, and 39 completed the study (G cambogia group, n = 18; placebo group, n = 21). At 16 weeks, the G cambogia group had significantly reduced visceral, subcutaneous, and total fat areas compared with the placebo group (all indices P<0.001). No severe adverse effect was observed at any time in the test period. There were no significant differences in BMI or body weight at week 12, but there were slight numeric decreases in body weight and BMI in men. There were no signs of a rebound effect from week 12 to week 16. CONCLUSION: G cambogia reduced abdominal fat accumulation in subjects, regardless of sex, who had the visceral fat accumulation type of obesity. No rebound effect was observed. It is therefore expected that G cambogia may be useful for the prevention and reduction of accumulation of visceral fat.

Evidence suggesting that difructose anhydride III is an indigestible and low fermentable sugar during the early stages after ingestion in humans.

We investigated the influences of difructose anhydride III (DFAIII), a novel commercially available disaccharide, on sugar metabolism, breath hydrogen and serum acetate in the early stages after ingestion to determine whether DFAIII is an indigestible sugar and to what degree it is fermentable in humans. This study was designed as a randomized controlled single-blind crossover test with 9 healthy subjects, who drink a 200 mL water solution containing 10 g of DFAIII, lactulose or sucrose following overnight fasting. Blood samples (for analysis of glucose, fructose, insulin, triacylglycerol, free fatty acids, and acetate) were collected at 0, 0.5, 1, 2, 4, 8 h after the ingestion and breath samples (for analysis of hydrogen and methane gases) were collected at 1 h intervals until 8 h after the ingestion. We also interviewed each subject hourly about the incidence and severity of specific abdominal complaints and other symptoms. The results revealed that ingestion of 10 g of DFAIII did not change the serum levels of glucose, fructose, and insulin, similarly to the case with lactulose, and no increase in breath hydrogen excretion was comparable to the case with sucrose. The incidence of specific abdominal symptoms tended to be lower after DFAIII ingestion than after lactulose ingestion. It thus turned out that DFAIII was indigestible and low fermentable in the early stages after ingestion.