Safety evaluation of the excessive intake of Bacillus subtilis C-3102 in healthy Japanese adults: A randomized, placebo-controlled, double-blind, parallel-group, comparison trial.

OBJECTIVE: Continuous intake of Bacillus subtilis C-3102 (B. subtilis C-3102) has been reported to modulate the gut microbiota and increase the bone mineral density of the femur in healthy adults. This study aimed to evaluate the safety of excessive B. subtilis C-3102 intake through a randomized, placebo-controlled, double-blind, parallel-group study. METHOD: A total of 69 individuals provided an informed consent, and 44 subjects who met the inclusion criteria were allocated to either the B. subtilis C-3102 (C-3102 group, n = 22) or the placebo group (P group, n = 22). All subjects took 18 tablets containing either containing B. subtilis C-3102 or placebo per day for 4 weeks with water and without chewing. Subjects in the C-3102 group consumed 4.8 × 1010 colony forming units (cfu) per day. Physical examination, urinalysis, blood analysis, records of subjective symptoms, and a medical questionnaire administered by a clinical trial physician were performed to determine the safety of test tablets. Furthermore, bone mineral density was measured. RESULTS: The final analysis included data from 22 subjects (9 men, 13 women; age, 46.1 ±â€¯13.8 years) in the C-3102 group and 22 subjects (9 men, 13 women; age, 46.1 ±â€¯13.5 years) in the P group. The results revealed no medical-related problems in both C-3102 and P groups. CONCLUSION: This study proved the safety of 4-week continuous consumption of an excessive amount of B. subtilis C-3102 tablets.

Oral administration of Cimicifuga racemosa extract attenuates psychological and physiological stress responses.

Dried rhizomes of Cimicifuga racemosa (CR), which are known as black cohosh, have been widely used as herbal dietary supplements to treat menopausal symptoms. The present study examined the effect of CR extracts on human psychological and physiological responses to acute stress induced by mental arithmetic tests, by measuring the subjective stress intensity, the brain-wave patterns according to electroencephalography, and the concentrations of salivary chromogranin-A and cortisol. The experiments were performed double-blind and their order was counterbalanced. Treatment with CR significantly attenuated the elevated subjective perception of stress and the increased salivary chromogranin-A levels compared with placebo treatment. CR extract also rapidly recovered the decrease in alpha waveband induced by performing the mental arithmetic task. We therefore propose that CR extracts might be suitable for the prevention and treatment of stress-related disorders.

Oral administration of Cimicifuga racemosa extract affects immobilization stress-induced changes in murine cerebral monoamine metabolism.

We investigated the effects of Cimicifuga racemosa (CR) plant extracts on the changes in levels of the cerebral monoamines norepinephrine (NE), dopamine (DA), and serotonin (5-HT), the respective metabolites 3-methoxy-4-hydroxyphenylglycol (MHPG), 3,4-dihydroxyphenylacetic acid (DOPAC), and 5-hydroxyindoleacetic acid (5-HIAA), and plasma corticosterone in mice subjected to acute immobilization stress. Single oral administration of the CR extract (1,000 mg/kg) significantly attenuated plasma corticosterone levels that had been increased as a result of enforced immobilization. Acute immobilization stress caused significant changes in the corresponding amine-to-metabolite ratios in the hypothalamus, hippocampus, and cortex; however, CR-extract treatment significantly attenuated the MHPG/NE change in the hypothalamus, and the 5-HIAA/5- HT changes in each region of the brain. Our results suggest that the CR extract interacts not only with the hypothalamic-pituitary-adrenal (HPA) axis but also with the sympathetic adrenomedullary (SAM) system under stress conditions. Thus the CR extract can alleviate acute stress responses by suppressing the changes of amine-to-metabolite ratio in brain.

Oral administration of Cimicifuga racemosa extract attenuates immobilization stress-induced reactions.

Dried rhizomes of Cimicifuga racemosa (CR), known as black cohosh, have been widely used as a herbal dietary supplement in the treatment of menopausal symptoms. Here we used experimental mouse stress models to investigate the role of anti-stress food factors, and found that a CR extract had stress-relieving effects. A single oral administration of CR extract (1,000 mg/kg) significantly attenuated plasma corticosterone and aspartate aminotransferase (AST) levels that had increased as a result of enforced immobilization. Bioassay-guided fractionation of the CR extract resulted in the isolation of 10 triterpenes, among which actein, 23-epi-26-deoxyactein, and cimiracemoside F (100 mg/kg, per os) were shown to contribute to the anti-stress effects. Furthermore, the CR extract significantly prevented the development of water immersion stress-induced gastric mucosal ulcers in rats. We propose that the CR extract might be suitable for the prevention and treatment of stress-related disorders.

Oligomeric procyanidins in apple polyphenol are main active components for inhibition of pancreatic lipase and triglyceride absorption.

Inhibitory effects of apple polyphenol extract (AP) and procyanidin contained in AP on in vitro pancreatic lipase activity and in vivo triglyceride absorption in mice and humans were examined. AP and procyanidin considerably inhibited in vitro pancreatic lipase activity. However, polyphenols, except for procyanidin, in AP (i.e., catechins, chalcones, and phenol carboxylic acids) showed weak inhibitory activities on pancreatic lipase. Procyanidins separated by normal-phase chromatography according to the degree of polymerization were also examined. Inhibitory effects of procyanidins increased according to the degree of polymerization from dimer to pentamer. On the other hand, pentamer or greater procyanidins showed maximal inhibitory effects on pancreatic lipase. These results suggested that with respect to in vitro pancreatic lipase inhibition, the degree of polymerization was an important factor and oligomeric procyanidin mainly contributed. Next, we performed a triglyceride tolerance test in mice and humans. Simultaneous ingestion of AP and triglyceride significantly inhibited an increase of plasma triglyceride levels in both models. These results suggested that the oligomeric procyanidins contained in AP inhibited triglyceride absorption by inhibiting pancreatic lipase activity in mice and humans.

Safety evaluation of polyphenols extracted from hop bracts.

Hop bract polyphenols contain polyphenols as promising functional ingredients. To assess the safety of topical hop bract polyphenols, Hopsphenon, we examined acute, 14-day, 28-day and 90-day toxicity tests in rats, and mutagenicity tests using Ames test and micronucleus test in mice. The acute, 14-day, 28-day and 90-day toxicity tests revealed that Hopsphenon produced no symptoms of significant injury. The lethal dose of hop bract polyphenols is greater than 2000 mg/kg. The Ames test in the absence of S9 mix for TA98 and in the presence of S9 mix for TA1537 revealed that Hopsphenon had slight mutagenicity at a high dose of 5000 microg/plate; however, in the micronucleus test, Hopsphenon was negative. These tests demonstrated that hop bract polyphenols are safe and do not cause any detrimental effects in vivo under the conditions investigated in this study.

Neocimicigenosides A and B, cycloartane glycosides from the rhizomes of Cimicifuga racemosa and their effects on CRF-stimulated ACTH secretion from AtT-20 cells.

Two new cycloartane glycosides, named neocimicigenosides A (1) and B (2), were isolated from the rhizomes of Cimicifuga racemosa. The structures of 1 and 2 were determined on the basis of extensive spectroscopic analysis and enzymatic hydrolysis followed by chromatographic and spectroscopic analyses to be (16S,23R,24S)-24-acetoxy-16,23:16,25-diepoxy-15alpha-hydroxycycloartan-3beta-yl alpha-L-arabinopyranoside (1) and (16S,23R,24S)-24-acetoxy-16,23:16,25-diepoxy-15alpha-hydroxycycloartan-3beta-yl beta-D-xylopyranoside (2), respectively. Neocimicigenosides A and B enhanced CRF-stimulated ACTH secretion from AtT-20 cells.

Comparison of ingestive effects of brewer's yeast, casein, and soy protein on bioavailability of dietary iron.

The effects of brewer's yeast, casein, and soy protein intakes on the absorption and retention as well as the incorporation into hemoglobin and systemic iron stores of dietary iron were examined in an animal experiment with growing rats. Relative biological values (RBV) of iron in the rats fed casein (C), soy protein (SP), and yeast (Y) diets were 1.00, 0.31, and 1.77, respectively. The apparent absorption of iron in Y-diet-fed rats was significantly higher than that in C- or SP-diet-fed rats. The hemoglobin regeneration efficiency (HRE) of iron in Y group was significantly higher than those in C and SP groups. As a result of search for iron-absorptive enhancers (IAE) in yeast, RBV and HRE of the yeast-cell-wall-including diet turned out to be significantly higher than those of its lacking diet. These results suggest that IAE occurring in the yeast cell wall may be effective for iron absorption.

Iron absorption in rats increased by yeast glucan.

The effects of brewer's yeast cell walls and two of its components, glucan and mannan, on the absorption of 59Fe by anemic rats were investigated. After administration of the label, the percentage of 59Fe taken up into the blood of group given glucan was generally similar to that of a group given yeast cell walls, both values were higher than in controls. The incorporation of 59Fe into the small intestines was higher in the group given glucan than in the controls or a group given a glucan-mannan mixture. Glucan is the main substance in yeast cell walls that increases iron absorption.