AIMS:The aim of this study was to evaluate the antiobesity effects of heat-killed Lactiplantibacillus plantarum Shinshu N-07 (N-07) isolated from fermented Brassica rapa L. METHODS AND RESULTS:Male mice were divided into three groups (n = 10/group); normal diet, western diet (WD), or WD + N-07 (N-07) group and administered each diet for 56 days. The N-07 group showed significant suppression of body weight gain and epididymal fat, perirenal fat, and liver weights compared with the WD group. Higher levels of fecal total cholesterol, triglyceride (TG), and free fatty acid (FFA) were observed in the N-07 group than in the WD group. The mRNA expression of the cholesterol transporter ATP-binding cassette transporter G5 (ABCG5) was significantly increased in the small intestine of N-07-fed mice compared with WD-fed mice. Moreover, N-07 supplementation significantly increased the mRNA expression of ABCG5 and ABCG8 in Caco-2 cells. Furthermore, the TG- and FFA-removal ability of N-07 was confirmed to evaluate its soybean oil- and oleic acid-binding capacities in in vitro experiments. CONCLUSIONS:The antiobesity effects of N-07 might be due to its ability to promote lipid excretion by regulating cholesterol transporter expression and lipid-binding ability.

Several beneficial effects of poly-y-glutamic acid (y-PGA) have been reported. To test whether natto, a fermented soy food rich in y-PGA, can improve intestinal microbiota content and lipid metabolism in a high-fat diet, we compared the intestinal microbiota content, plasma, liver, and fecal contents, and changes in gene expression in the livers and large intestines of a group of mice fed a high-fat diet supplemented with cooked soybeans (SC group) and a group fed a high-fat diet supplemented with natto (NA group) for 42 days; high-fat diet-fed mice were used as a control (Con group). Hepatic lipid levels were significantly lower, the fecal bile acid and lipid levels were significantly greater, and the Bacteroidetes/Firmicutes ratio was significantly higher in the SC and NA groups as compared to Con group. Additionally, plasma glucose and triglyceride levels, the expression of liver fatty acid synthase, and the relative abundance of Lactobacillaceae was significantly higher in the NA group than in the Con group. Although both natto and cooked soybeans impacted the metabolic response to a high-fat diet, the addition of natto had a greater effect on glucose and lipid metabolism. y-PGA may play an important role in natto functionality.

Aims: Although physiological effects of hydrophilic- (H-) and lipophilic- (L-) antioxidant capacities (AOCs) are suggested to differ, the association of an antioxidant-rich diet and chronic kidney disease (CKD) incidence has not been examined. We therefore explored the association between the H- or L-AOC of a whole Japanese diet and CKD risk in a general population. Methods: A total of 922 individuals without CKD (69.2% women; mean age, 59.5 years old) from Ohasama Town, Japan, were examined. CKD incidence was defined as the presence of proteinuria and/or an estimated glomerular filtration rate (eGFR) of ＜60 ml/min/1.73 m2. Consumption of H-/L-AOC was determined based on the oxygen radical absorbance capacity in a specially developed Japanese food AOC database. Hazard ratios (HRs) and 95% confidence intervals (CIs) were calculated for new-onset CKD using a Cox proportional hazards model. Results: During the median follow-up of 9.7 years, 137 CKD incidents were recorded. After adjusting for potential confounding variables, the highest quartile of L-AOC was significantly associated with a 51% reduced CKD risk among only women. An increased L-AOC intake was more effective in preventing eGFR reduction than in preventing proteinuria in women. These associations were not seen for H-AOC intake in both sexes and L-AOC intake in men. Conclusions: A high intake of lipophilic antioxidants may be associated with a reduced CKD risk. The balance between dietary antioxidant intake and pro-oxidants induced by unhealthy lifestyles may be crucial for preventing future kidney deterioration.

Metabolomic characterization of a lactic-fermented pickle of nozawana (Brassica rapa L. var. hakabura) was conducted to evaluate the effects of different starter culture strains on the chemical profiles. We compared the profiles of water-soluble and volatile compounds obtained by non-targeted nuclear magnetic resonance and solid-phase microextraction gas chromatography/mass spectrometry analyses. Principal component analyses indicated that the fermented samples differed significantly in terms of the levels of various compounds, including taste- and aroma-active components, such as water-soluble residual sugars, organic acids, mannitol, ethanol, dihydroxyacetone, ornithine, γ-aminobutyric acid, choline, volatile isothiocyanates, 3,4-epithiobutyl cyanide, 2,3-butanedione, acetoin, ethyl acetate, dimethyl trisulfide, and S-methyl thioacetate. Fermentation with a Latilactobacillus curvatus culture was associated with a unique metabolite profile characterized by higher levels of isothiocyanates and hexanoic acid and lower levels of lactic acid, acetic acid, acetoin, and 2,3-butanedione. These variations in the chemical profile might be associated with different qualities in fermented nozawana pickle products.

This study aimed to evaluate the anti‐adiposity effect of heat‐killed Lactobacillus brevis KB290 originating from traditional Japanese fermented pickles in mice fed a high‐fat diet (HFD).

Natto is a traditional Japanese fermented soy product high in γ-polyglutamic acid (γ-PGA), whose beneficial effects have been reported. We prepared high-γ-PGA natto and compared the dietary influence on liver lipids and cecal microbiota in mice fed a diet containing it or a standard diet. The mice were served a 30% high-γ-PGA natto diet (PGA group) or standard diet (Con group) for 28 days. Liver lipids, fecal lipids, and fecal bile acids were quantified. Cecal microbiota were analyzed by PCR amplification of the V3 and V4 regions of 16S rRNA genes and sequenced using a MiSeq System. Additionally, the cecal short-chain fatty acid profile was assessed. The results revealed that the liver lipid and triglyceride contents were significantly lower (p<0.01) and amounts of bile acids and lipids in the feces were significantly higher in the PGA group than in the Con group. The cecal butyric acid concentration was observed to be significantly higher in the PGA group than in the Con group. Principal component analysis of the cecal microbiota revealed that the PGA and Con groups were distinct. The ratio of Firmicutes/Bacteroidetes was found to be significantly low in the PGA mice. The results revealed a significantly higher relative abundance of Lachnospiraceae (p<0.05) and significantly lower relative abundance of Coriobacteriaceae (p<0.01) in the PGA group. Analysis of the correlation between bacterial abundance and liver lipids, cecal short-chain fatty acids, fecal lipids, and fecal bile acids suggested that intestinal microbiota can be categorized into different types based on lipid metabolism. Hepatic lipid accumulation typically facilitates the onset of nonalcoholic fatty liver disease (NAFLD). Our findings suggest that high-γ-PGA natto is a beneficial dietary component for the prevention of NAFLD.

BACKGROUND:Previous Japanese studies have led to the erroneous conclusion of antioxidant capacity (AOC) intakes of the overall Japanese diet due to limitations in the number and types of food measured, especially in rice and seafood intake. The aims of the study were to construct an AOC database of foods representative of the typical Japanese diet and to clarify the high contributors to AOC intake from the overall diet of the Japanese population.METHODS:Commonly consumed foods were estimated using 3-day dietary records (DRs) over the four seasons among 55 men and 58 women in Japan. To generate an AOC database suitable for the typical Japanese diet, hydrophilic (H-)/lipophilic (L-) oxygen radical absorbance capacity (ORAC) values of foods in each food group were measured via validated methods using the food intake rankings. Subsequently, we estimated the AOC intake and the AOC characteristics of a typical Japanese diet.RESULTS:Of 989 food items consumed by the participants, 189 food items were measured, which covered 78.8% of the total food intake. The most commonly consumed types of antioxidant-containing food were tea, soybean products, coffee, and rice according to H-ORAC, and soybean products, fish and shellfish, vegetables, and algae according to L-ORAC.CONCLUSIONS:The characteristics of high AOC intake in rice and seafood more appropriately reflected the Japanese-style diet. Further studies are expected to clarify the association between food-derived AOC and its role in preventing or ameliorating lifestyle-related diseases.

Lactobacillus plantarum Shinshu N-07 (N07) and Lactobacillus curvatus #4G2 (#4G2) were isolated from fermented Brassica rapa L. and selected as promising probiotics with anti-adiposity activities based on in vitro assays. The anti-adiposity effects of these two strains were investigated using a diet-induced obesity animal model. Epididymal adipose tissue weight and adipocyte area were significantly lower and serum triglycerides and glucose tended to be lower in mice fed the high-fat diet supplemented with N07 compared with those fed the unsupplemented high-fat diet. Strain N07 suppressed hepatic steatosis, with accompanying downregulation of lipogenic genes in the liver. Expression of inflammatory cytokines and macrophage infiltration markers tended to be suppressed by N07 supplementation. Upregulation of uncoupling protein-1 in epididymal adipose tissue by N07 suggested that the transformation of white adipose tissue to brown might have been induced. Intestinal microbiota analysis revealed that a decrease in abundance of family S24-7 (phylum Bacteroidetes) following ingestion of the high-fat diet was partly recovered by supplementation with N07. Changes in those parameters were not observed in mice fed the high-fat diet supplemented with strain #4G2, suggesting strain specificities. Thus, N07 is a potential probiotic strain that could be used to develop functional foods that attenuate visceral fat accumulation after an appropriate human intervention trial.

To provide reliable data for high quality epidemiological studies examining the relationship between health and antioxidant intake from daily foods, 107 plant-derived food items (12 rice, bread and noodles, 5 potatoes and starches, 9 pulses, 6 nuts/seeds, 29 vegetables, 22 fruits, 5 mushrooms, 7 algae, and 12 beverages) were selected as commonly consumed foods in Japan based on dietary records, and their antioxidant capacities were evaluated by validated hydrophilic- and lipophilic-oxygen radical absorbance capacity (H-ORAC and L-ORAC) methods. The food items covered more than 60% of total food intake for each category on a weight basis. The H-ORAC and L-ORAC values were widely distributed at 0-210 and 0-30 μmol-Trolox equivalent/g, respectively. The foods possessing potent antioxidant capacities were found in vegetables and fruits as well as other plant-derived foods. In most foods measured, the H-ORAC values were much larger than the L-ORAC values, except for certain kinds of pulses, nuts/seeds, mushrooms, and algae. The ORAC data shown here is sufficient to accurately estimate the antioxidant intake from plant-derived foods in Japan, and should be useful in future epidemiological studies aiming to clarify the biological significance of ORAC values.

The functionality and content of functional components in foods need to be measured precisely. We have improved methods to evaluate the antioxidant capacity of foods based on oxygen radical absorbance capacity, and the content of quercetin in onions and isoflavones in soybeans. The precision of these methods was validated by inter-laboratory tests in accordance with internationally standardized protocols. These methods enable precise evaluation of antioxidant capacity and content, and the values are comparable. In this review article, the validated methods are introduced and the possible applications of the methods are discussed.

Lactic acid bacteria (LAB) exert beneficial health effects by regulating immune responses. Brassica rapa L., known as Nozawana, is commonly consumed as a lactic acid-fermented food called nozawana-zuke. Few studies have investigated changes in the bacterial community and cytokine production activities during the fermentation of B. rapa L. In order to obtain more detail information, we herein conducted a study on fresh B. rapa L. fermented for 28 d. An amplicon analysis of the 16S rRNA gene revealed that Lactobacillales predominated during fermentation, and the microbiota became less diverse on day 7 or later. Fermented B. rapa L. promoted the production of interferon (IFN)-γ and interleukin (IL)-10 by mouse spleen cells more than non-fermented vegetables. Lactobacillus curvatus was the predominant species during fermentation, followed by L. plantarum and L. brevis. L. sakei was occasionally detected. A correlation analysis showed that IFN-γ concentrations positively correlated with the numbers of L. curvatus and L. plantarum, while those of IL-10 correlated with the numbers of L. sakei in addition to these 2 species. Significantly higher levels of IFN-γ and IL-10 were induced by fermented B. rapa L. when isolated Lactobacillus strains were added as starter cultures. These results suggest that the Lactobacillus species present in fermented B. rapa L. are beneficial for manufacturing vegetables with immunomodulatory effects.

The bacterial community structure in 29 naturally fermented samples of sunki, an unsalted lactic-fermented pickle in Japan, was determined by 16S rRNA gene-targeted metagenomic analysis. The data revealed that genus Lactobacillus was dominant in all samples and various bacterial species, related to Lactobacillus delbrueckii, Lactobacillus plantarum, Lactobacillus buchneri, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus crispatus, Lactobacillus intestinalis, and Lactobacillus gasseri, showed a range of dominance depending on the samples. Comparative analysis of the bacterial composition by principal coordinate analysis and hierarchical clustering classified the varied bacterial composition of the 29 samples into three types of bacterial community structure. These types comprised lactobacilli belonging to different phylogenetic groups: type A had a certain ratio of Lactobacillus fermentum (70.3-22.1%, average 41.2%) in combination with several species belonging to Lactobacillus delbrueckii-phylogenetic group, type B comprised remarkably high levels of species Lactobacillus delbrueckii (average 89.5%), and type C had combinations of species belonging to Lactobacillus plantarum- and Lactobacillus buchneri-phylogenetic groups. Interestingly, these types differed in the compositional profiles of water-soluble and volatile compounds, and statistically significant differences were observed in the levels of acetic acid, succinic acid, ethanol, ethyl acetate, glutamic acid, gamma-aminobutyric acid, and isovaleraldehyde. Furthermore, metabolomic analysis revealed a correlation of Lactobacillus fermentum dominance with pH value and lactic and acetic acid levels, with high R values of 0.643, -0.642, and 0.528, respectively. The data reported in this study showed the characteristics of the bacterial composition in the unsalted sunki pickle and its potential relationship with the compositional profile. (C) 2019, The Society for Biotechnology, Japan. All rights reserved.

Changes in quercetin content and antioxidant capacity during cooking were compared between the quercetin-rich onion cultivar 'Quer-Gold' and the commonly cultivated 'Kitamomiji 2000'. Irrespective of the cultivar, the quercetin content and antioxidant capacity were not significantly changed by sauteing, and were higher for 'Quer-Gold'. Two kinds of onion-rich dishes were prepared, and the quercetin content was compared between the cultivars. The quercetin content of the dishes was higher when 'Quer-Gold' was used.

This study aimed to evaluate both the lipophilic and hydrophilic oxygen radical absorbance capacities (ORAC) in pressed oils made from sesame (Sesamum indicum L.) cultivar "Maruhime" and rapeseed (Brassica napus L.) cultivar "Nanaharuka." For this reason, we slightly modified the extraction method for antioxidant compounds using an accelerated solvent extractor system, and the multi-laboratory validated lipophilic ORAC method previously reported. Acceptable recovery was observed when four matrices of oils and fats fortified with a-tocopherol and gallic acid were subjected to the modified extraction method. Moreover, it was confirmed that the lipophilic ORAC value of a-tocopherol, as determined by the modified method, was within the range of the mean reproducibility standard deviation in the multi-laboratory comparison study. In the pressed oil from sesame cultivar "Maruhime," L-ORAC value was 12.6 to 33.8 times higher than H-ORAC, and the mean L-ORAC and H-ORAC values were 2 926 and 130 mu mol-Trolox equivalent/100 g, respectively. Also, L-ORAC value in the pressed oil from rapeseed cultivar "Nanaharuka" was 2.2 to 13.1 times higher than H-ORAC, and the mean L-ORAC and H-ORAC values were 552 and 87 mu mol-Trolox equivalent/100 g, respectively. The contribution of tocopherols to L-ORAC value of the pressed oil from rapeseed cultivar "Nanaharuka" ranged from 24 to 55 %; however, the contribution was smaller, ranging from 7 to 22 %, in the pressed oil from sesame cultivar "Maruhime."

We investigated how different processing of soy foods impacts their hypolipidemic properties. Male rats were fed a diet containing 300 g/kg of roasted soybean, powdered soy milk, dried fresh tofu, or kori tofu for 21 days. Roasted soybean, a fiber-rich food, significantly increased the fecal total lipid content. The fecal excretion of bile acids was significantly stimulated by fresh tofu and kori tofu, both of which contain coagulated protein. Soy milk and tofu products, which are relatively rich in protein, partly suppressed hepatic lipogenesis compared to roasted soybean. Unfortunately, the impact of these differences was not apparent in reducing lipid levels in serum and liver. Given these observations, constant consumption of each soy food may modulate intestinal lipid absorption and hepatic lipogenesis differently.

The present study investigated the antiallergic and anti-inflammatory effects of 10-hydroxy-cis-12-octadecenoic acid (HYA), a novel gut microbial metabolite of linoleic acid, in NC/Nga mice, a model of atopic dermatitis (AD). Feeding HYA decreased the plasma immunoglobulin E level and skin infiltration of mast cells with a concomitant decrease in dermatitis score. HYA feeding decreased TNF- and increased claudin-1, a tight junction protein, levels in the mouse skin. Cytokine expression levels in the skin and intestinal Peyer's patches cells suggested that HYA improved the Th1/Th2 balance in mice. Immunoglobulin A concentration in the feces of the HYA-fed mice was approximately four times higher than that in the control mice. Finally, denaturing gradient gel electrophoresis of the PCR-amplified 16S rRNA gene of fecal microbes indicated the modification of microbiota by HYA. Taken together, the alterations in the intestinal microbiota might be, at least in part, associated with the antiallergic effect of HYA.

Singlet oxygen absorption capacity (SOAC) is an assay method used to evaluate the singlet oxygen quenching capacity. We evaluated the precision of the SOAC assay method in an inter-laboratory validation study using 3 antioxidant solutions and 3 food extracts. This study involved 14 participating laboratories, with data sets from 8 successfully participating laboratories subjected to statistical analysis. In accordance with the harmonized protocol, this study showed that the intermediate precision relative standard deviation (RSDint) and the reproducibility relative standard deviation (RSDRR) ranged from 3.4 to 9.9%, and from 6.2 to 14.0%, respectively, and the HorRat values ranged from 0.87 to 1.97. Therefore, the SOAC assay method was validated by this inter-laboratory study in accordance with the internationally harmonized protocol.

A lipophilic oxygen radical absorbance capacity (L-ORAC) assay is an evaluation of the antioxidant capacity of solutions of lipophilic compounds. The concentrations of fluorescein k]radical generator k]and Trolox standard solutions were optimized to improve the precision of the assay. An interlaboratory study using two antioxidant solutions and three food extracts as test samples conducted in accordance with harmonized protocol demonstrated satisfactory L-ORAC measurements; the intermediate precision relative standard deviations (RSRDint) ranged from 7.0 to 16.7% k]the reproducibility relative standard deviations (RSDR) ranged from 14.8 to 19.4% k]and the HorRat values ranged from 1.35 to 1.78.

Two cultivars of Japanese parsley were harvested in different seasons; their antioxidant capacities were evaluated by oxygen radical absorbance capacity (ORAC) methods, and the contents of hydrophilic and lipophilic antioxidants were compared. Japanese parsley possessed potent antioxidant capacities both in hydrophilic and lipophilic extracts when evaluated by ORAC methods. LC/MS/MS analyses revealed that chlorogenic acid and four kinds of quercetin glycosides were major antioxidants in the hydrophilic extract. Lutein was the main contributor to the antioxidant capacity of the lipophilic extract. Antioxidant capacities of the hydrophilic extracts of both cultivars tended to be higher in winter because of the increase in the contents of chlorogenic acid and quercetin glycosides. An obvious trend in the lipophilic antioxidant capacities or lutein contents was not observed irrespective of the cultivar.

Oxygen radical absorbance capacity (ORAC) is a method for evaluating antioxidant capacities of both hydrophilic (H-ORAC) and lipophilic compounds (L-ORAC). In this study, we evaluated the antioxidant capacities of eight herbs (Italian parsley, coriander, celery white, watercress, rocket, mustard green, basil, and corn salad) by ORAC. Basil showed the highest antioxidant capacity among the herbs assessed in this study. The harvest season affected both the H- and L-ORAC values of the herbs, except for the H-ORAC value of Italian parsley. The ratios of H- to L-ORAC values of the herbs ranged from 36 - 86%. Total polyphenol contents of herbs were positively correlated with H-ORAC values, and were also affected by harvest season. We found that isoquercitrin and rutin but not ferulic acid contributed to the seasonal change in the H-ORAC value of basil.