Identification of components in coriander (Coriandrum sativum L.) inhibiting degranulation of RBL-2H3 cells.

We found that a water-soluble extract of coriander (Coriandrum sativum L.) (leaves, petioles and stems) inhibits antigen-induced degranulation of RBL-2H3 cells, a rat basophil leukemia cell line. The aim of this study was to elucidate the anti-degranulation active components in the extract. The methanol-eluate fraction obtained by fractionation of the water-soluble extract using MCI gel column chromatography had strong activity, and eight components were isolated and identified. Two of them were identified as new compounds, (3S)-3-methyl-6-hydroxyisocoumarin 8-O-ß-D-glucopyranoside (compound 1) and (7S,8R)-7,8-dihydro-8-ß-D-glucopyranosyloxy-4-methoxy-7-methyl-5H-fro[2,3-g][2]benzopyran-5-one (compound 2). As a result of evaluation of anti-degranulation activity of eight components, seven of them, such as tryptophan, phenylalanine, dihydroxycoumarin glucoside, quercetin glycoside, rutin, compound 1, and compound 2, had the activity. These results indicated that the water-soluble extract of coriander contains several anti-degranulation substances.

Hydroxy-ß-sanshool isolated from Zanthoxylum piperitum (Japanese pepper) shortens the period of the circadian clock.

We showed that an ethanol extract from Zanthoxylum piperitum can shorten the circadian rhythm at the cellular level and that this activity was due to hydroxy-ß-sanshool, a secondary metabolite in this plant. An ethanol extract of Z. piperitum was repeatedly fractionated using solid phase extraction and reverse-phase HPLC, then the circadian rhythms of cells to which the fractions were loaded were monitored using real-time reporter gene assays. We purified one HPLC peak and identified it as hydroxy-ß-sanshool using liquid chromatography (LC)-precision-mass spectrometry (MS). This compound shortened the period of Bmal1 and Per2 at the cellular level. Incubation cells for 24 h with hydroxy-ß-sanshool resulted in upregulated Per2 promoter activity. Hydroxy-ß-sanshool also dose-dependently upregulated expression of the clock genes Bmal1, Per1, Per2 and Cry1 and the clock-controlled oxidative stress responsive genes Gpx1and Sod2.

Umbelliferose Isolated from Cuminum cyminum L. Seeds Inhibits Antigen-Induced Degranulation in Rat Basophilic Leukemia RBL-2H3 Cells.

Cuminum cyminum L. (cumin) is an annual plant of the Umbelliferae family native to Egypt. We previously showed that the aqueous extract of cumin seeds suppresses degranulation by downregulating the activation of antigen-induced intracellular signaling molecules in rat basophilic leukemia RBL-2H3 cells. However, the active substances in the extract have not yet been identified. Accordingly, herein, we aimed to ascertain the water-soluble substances present in cumin seeds that inhibit degranulation, which led to the identification of umbelliferose, a characteristic trisaccharide present in plants of the Umbelliferae family. Our study is the first to reveal the degranulation-suppressing activity of umbelliferose, and quantification studies suggest that cumin seed powder contains 1.6% umbelliferose. Raffinose, an isomer of umbelliferose, was also found to significantly suppress antigen-induced degranulation, but less so than umbelliferose. Both umbelliferose and raffinose contain sucrose subunits in their structures, with galactose moieties bound at different sites. These differences in structure suggest that the binding of galactose to the sucrose subunit at the α1-2 bond contributes to its strong degranulation-inhibiting properties.

Effects of Continuous Intake of Rosemary Extracts on Mental Health in Working Generation Healthy Japanese Men: Post-Hoc Testing of a Randomized Controlled Trial.

We previously performed a 4 week interventional trial that suggested that continuous intake of rosemary extract improves the mood states, fatigue, and cognitive function of working generation healthy adult Japanese men. However, the severity of depression in participants in our previous study was relatively mild. Therefore, in the present study, a post-hoc analysis of our previous study was conducted, limited to participants whose total mood disturbance (TMD) scores, which indicate greater mood disturbance, were above the median at baseline, to evaluate whether rosemary extract was effective for individuals with poor mental health. Following the intervention, the scores of TMD and "Confusion-Bewilderment" were significantly decreased (both p < 0.05), and scores of "Vigor-Activity" were significantly increased in the rosemary group (n = 8) compared with those in the control group (n = 13; p < 0.01). When comparing the scores from pre- and post-intervention, significant improvements in "Tension-Anxiety", "Vigor-Activity", "Fatigue on awakening", "Daytime sleepiness", and "Psychomotor speed" were observed in the rosemary group only (all p < 0.05). Based on these results, it was expected that rosemary extracts were effective for improving the mental energy and sleep quality of work-age men with poor mental health.

Inhibitory effect of aqueous extract of Cuminum cyminum L. seed on degranulation of RBL-2H3 cells and passive cutaneous anaphylaxis reaction in mice.

Cuminum cyminum L. (cumin) seed is used as a spice in various countries. Although several functions of the components in cumin seed have been reported, the anti-allergic effect of the water-soluble component in cumin seed has not been reported yet. In this study, we focused on the suppressive effect of cumin seed aqueous extract on degranulation in order to reveal the anti-allergic effect of cumin. Cumin seed aqueous extract significantly suppressed the antigen-induced degranulation of rat basophilic leukemia cell line RBL-2H3 cells in a dose-dependent manner without cytotoxicity. The extract also inhibited the elevation of the intracellular calcium ion concentration induced by antigen. Immunoblot analysis revealed that the extract suppresses phosphorylation of phosphatidylinositol 3-kinase, Bruton's tyrosine kinase, phospholipase C-γ1/2, and Akt in the signaling pathways activated by antigen induction via FcεRI. Furthermore, the extract suppressed microtubule formation induced by antigen. In addition, oral administration of cumin seed aqueous extract significantly suppressed the passive cutaneous anaphylaxis reaction in BALB/c mice. Our findings suggest that cumin seed contains water-soluble components with the anti-allergic effect. Therefore, cumin seed has potential as anti-allergic functional food.

Donor-Donor'-Acceptor Triads Based on [3.3]Paracyclophane with a 1,4-Dithiafulvene Donor and a Cyanomethylene Acceptor: Synthesis, Structure, and Electrochemical and Photophysical Properties.

Donor-donor'-acceptor triads (1, 2), based on [3.3]paracyclophane ([3.3]PCP) as a bridge, with electron-donating properties (D') using 1,4-dithiafulvene (DTF; TTF half unit) as a donor and dicyanomethylene (DCM; TCNE half unit) or an ethoxycarbonyl-cyanomethylene (ECM) as an acceptor were designed and synthesized. The pulse radiolysis study of 1 a in 1,2-dichloroethane allowed the clear assignment of the absorption bands of the DTF radical cation (1 a.+ ), whereas the absorption bands due to the DCM radical anion could not be observed by γ-ray radiolysis in 2-methyltetrahydrofuran rigid glass at 77 K. Electrochemical oxidation of 1 a first generates the DTF radical cation (1 a.+ ), the absorption bands of which are in agreement with those observed by a pulse radiolysis study, followed by dication (1 a2+ ). The ESR spectrum of 1 a.+ showed a symmetrical signal with fine structure and an ESR simulation predicted that the spin of 1 a.+ is delocalized over S and C atoms of the DTF moiety and the central C atom of the trimethylene bridge bearing the DTF moiety. Pulse radiolysis, ESR, and electrochemical studies indicate that the DTF radical cation of 1 a.+ is more stable than that of 6.+ , and the latter shows a strong tendency to dimerize. This result indicates that the [3.3]PCP moiety as a bridge can stabilize the DTF radical cation more than the 1,3-diphenylpropane moiety because of kinetic stability due to its rigid structure and the weak electronic interaction of DTF and DCM moieties through [3.3]PCP.

Immunostimulatory effect of aqueous extract of Coriandrum sativum L. seed on macrophages.

BACKGROUND: Coriandrum sativum L. seed is generally used as a spice and crude drug. Although many functions of the various components in C. sativum L. seed have been reported, the immunostimulatory effect of water-soluble components in C. sativum L. seed has not been studied. In the present study, we focused on the immunostimulatory effect of C. sativum L. seed aqueous extract (CAE) on macrophages as a novel health function of C. sativum L. seed components. RESULTS: CAE significantly enhanced the production of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in both RAW264.7 cells and peritoneal macrophages by enhancing the expression levels of these cytokine genes. CAE also stimulated nitric oxide (NO) production and the phagocytosis activity in RAW264.7 cells. We suggest that the activity of CAE is a result of the upregulation of mitogen-activated protein kinase and nuclear factor-κB cascades via TLR4. In addition, IL-6 production by peritoneal macrophages collected from CAE-administered mice was significantly enhanced, suggesting that CAE could stimulate macrophage activity in vivo. CONCLUSION: The findings of the present study suggest that CAE contains a novel water-soluble component with an immunostimulatory effect on macrophages. CAE would contribute to activating host defense against pathogens by stimulating the innate immunity. © 2017 Society of Chemical Industry.

Effect of royal jelly on bisphenol A-induced proliferation of human breast cancer cells.

Royal jelly is known as a functional food containing many useful minerals. In this study, we found an anti-environmental estrogen activity of royal jelly. Bisphenol A (BPA) is an environmental estrogen that stimulates proliferation of human breast cancer MCF-7 cells. Royal jelly inhibited the growth-promoting effect of BPA on MCF-7 cells, even though it did not affect the proliferation of cells in the absence of BPA. In addition, this inhibiting effect of royal jelly was heat-stable.

Immunostimulation effects of proteose-peptone component 3 fragment on human hybridomas and peripheral blood lymphocytes.

Fat-free bovine milk fermented by 12 kinds of lactic acid bacteria and yeast enhanced monoclonal antibody production of human hybridoma HB4C5 cells 2.8-fold in serum-free medium. Immunoglobulin production of human peripheral blood lymphocytes (PBL) was also stimulated in vitro. IgM and IgG production of human PBL was accelerated up to 2.8-fold and 5.4-fold, respectively. Interferon-gamma production of human PBL was also accelerated 6.0-fold by 50 microg/ml of the fermented milk. However, interleukin-4 production of PBL was not affected, and tumor necrosis factor-alpha production was suppressed. The activity was enhanced 2.5-fold by the thermal treatment for 30 min at 65 degrees C and was completely lost by trypsin digestion. The findings suggested that the active substance in the fermented milk was heat stable protein. Gel-filtration and the SDS-PAGE analysis revealed that the molecular weight of the active substance was estimated as 19.0 kDa, which was not detected in fat-free bovine milk before fermentation. N-terminal amino acid sequence of the 19.0 kDa protein was highly homologous to proteose-peptone component 3 (PP3). Since molecular weight of PP3 is 28 kDa, it is suggested that the 19.0 kDa protein is derived from degradation of PP3 during fermentation of fat-free milk. Moreover, PP3 purified from fat-free milk also enhanced IgM production of HB4C5 cells.

Identification of Shiga toxins in Shiga toxin-producing Escherichia coli using immunoprecipitation and high-performance liquid chromatography-electrospray ionization mass spectrometry.

We developed a rapid and reliable identification method for Shiga toxins in Shiga toxin-producing Escherichia coli (STEC) using immunoprecipitation and high-performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS). Polyclonal antisera specific for Shiga toxin 1 (Stx1) and Shiga toxin 2 (Stx2) were raised in rabbits so as to be used for the immunoprecipitation. The immunoprecipitaion was carried out by mixing sample solutions with 50 microl each of the antisera to Stx1 and Stx2 followed by allowing the mixed solutions to stand for 30 min. The quantity required to obtain the immunoprecipitate was more than 0.5 microg of Shiga toxins. HPLC-ESI-MS analysis of the resulting immunoprecipitates provided accurate molecular weight information on Shiga toxins, leading to direct evidence for the presence of these toxins. It requires at most two days to perform our procedure from toxin extraction to measurement of HPLC-ESI-MS whereas the previous method using isolation procedures required about two weeks to complete. The usefulness of the present method has been demonstrated by identifying Stx1, Stx2 and a variant of Stx2 (Stx2e) in the immunoprecipitates prepared from STEC strains.

Helicobacter pylori seropositivity and IL-1B C-31T polymorphism among Japanese Brazilians.

We reported previously that anti-Helicobacter pylori antibody seropositivity (HP+) had an association with interleukin 1B (IL-1B) C-31T genotype, especially among smokers. This study examined the association for Japanese Brazilians. In this cross-sectional study, voluntary participation was announced through Japanese Brazilian communities in Sao Paulo, Curitiba, Mogi das Cruzes, and Mirandopolis; 963 Japanese Brazilians (399 males and 564 females) aged 33-69 years participated. Lifestyle data and peripheral blood were collected. An anti-HP IgG antibody test and genotyping for IL-1B C-31T and IL-1RN 86 bp VNTR were independently conducted. The genotype frequency of the IL-1B polymorphism among 947 individuals was 23.9% for C/C genotype, 45.6% for C/T genotype, and 30.5% for T/T genotype. Sex-age-adjusted odds ratio (aOR) of HP+ was 1.30 (95% confidence interval, 0.94-1.81) for C/T genotype and 1.45 (1.02-2.07) for T/T genotype relative to C/C genotype. The aOR for 127 current smokers was 2.45 (0.91-6.55) for C/T and 3.49 (1.17-10.46) for T/T, while that for 667 never smokers was 1.21 (0.82-1.78) and 1.36 (0.90-2.05), respectively. The corresponding figures were 2.42 (1.16-5.02) and 3.00 (1.33-6.78) for 226 current drinkers, and 1.21 (0.82-1.78) and 1.36 (0.90-2.05) for 667 non-drinkers. The difference in the OR was observed for milk consumption, salty pickled vegetable eating, and physical exercise practice. 4/4 Genotype of IL-1RN 86 bp VNTR was 84.8%, and had no association with the HP seropositivity. The observed association between HP+ and IL-1B -31TT indicated that the genetic trait also influences the susceptibility to HP for Japanese in Brazil.