Identification and mechanistic investigation of ellagitannins from Osbeckia octandra that attenuate liver fibrosis via the TGF-ß/SMAD signaling pathway.

Fibrosis is a major problem in chronic liver disease with limited treatment options due to its complex nature. Herbal medicines are often used as an alternative. The aim of this study was to investigate the therapeutic potential of Osbeckia octandra and to identify its active compounds and regulatory pathways. The effects of crude leaf suspension and boiled leaf extract were investigated in an animal model, and the extract was found to be the more effective treatment. Three major bioactive compounds, pedunculagin, casuarinin, and gallic acid, were isolated from the extract using the hepatic stellate cell line, LX-2-based antifibrotic effect evaluation system. The results showed that all these compounds ameliorated LX-2 in fibrotic state. This inhibitory mechanism was confirmed through the TGF-ß/SMAD signaling pathway. Collectively, the presence of these compounds in O. octandra suggests its potential as a treatment for liver fibrosis.

A new secoiridoid glucoside from Olea europaea.

A new phenolic glucoside (1), olerikaside, and other known secoiridoid glucosides [oleuropein (2), demethyl oleuropein (3), oleoside 11-methyl ester (4), oleoside 7, 11-dimethyl ester (5), 7-ß-D-glucopyranosyl 11-methyl oleoside (6), secoxyloganin (7), ilicifolioside B (8), hydroxytyrosol (9), and hydroxytyrosol glucosides (10-12)] were isolated from unprocessed olive fruits of Olea europaea cv. "Lucca". The chemical structure of olerikaside (1) was clarified based on spectroscopy and chemical analysis data.

Ferulic acid esters of glucosylglucose from Allium macrostemon Bunge.

Three new ferulic acid esters of glucosylglucose, 1-O-(E)-feruloyl-ß-d-glucopyranosyl (1-2)-[ß- d-glucopyranosyl (1-6)]-ß-d-glucopyranose (allimacronoid A, 1), 1-O-(E)-feruloyl-{ß-d-glucopyranosyl (1-4)-[ß-d-glucopyranosyl (1-2)]}-[ß- d-glucopyranosyl (1-6)]-ß-d-glucopyranose (allimacronoid B, 2), and 1-O-(E)-feruloyl-{ß-d-glucopyranosyl (1-6)-[ß-d-glucopyranosyl (1-2)]}-[ß- d-glucopyranosyl (1-6)]-ß-d-glucopyranose (allimacronoid C, 3) were isolated together with tuberonoid A (4), from the leaves of Allium macrostemon Bunge. The chemical structures were elucidated based on the analyses of the spectroscopic and chemical data.

Synthesis and antiviral activities of some 2,4,6-trisubstituted 1,3,5-triazines.

We describe the synthesis and results of biological evaluation of newly designed 2,4,6-trisubstituted symmetrical 1,3,5-triazine (TAZ) derivatives. Among the tested trisubstituted TAZ derivatives, some CS-symmetrical alkoxy-amino-substituted TAZ derivatives, including 7ggp and 6dpp, showed significant antiviral activity against herpes simplex virus type 1 (HSV-1). The compound with the highest level of antiviral activity was C3-symmetrical trialkoxy-TAZ derivative 4bbb, which showed a considerably high selectivity index (IC50/EC50=256.6). The structure-activity relationships for anti-HSV-1 activity of the tested 2,4,6-trisubstituted TAZ derivatives are also described.

RAD-Seq analysis of wild Japanese garlic (Allium macrostemon Bunge) growing in Japan revealed that this neglected crop was previously actively utilized.

Allium macrostemon Bunge, commonly referred to as "no-biru" in Japan, is a widespread wild onion species found across the country. Despite being deeply entwined in ancient Japanese culture, it remains an underutilized crop in Japan. Determining the origins of its domestic populations and understanding their genetic composition is crucial to highlighting the plant's historical significance in Japan. This study aims to bridge this knowledge gap by examining the genetic diversity of 47 A. macrostemon samples from various regions in Japan using RAD-Seq. Our analyses distinguished unique population structures, dividing the samples into three distinct groups: A, B, and C. Notably, groups A and B showed clear evidence of bulb propagation, while group C did not. Group C formed four subgroups: C1, C2, C3, and C4. Hybridization between subgroup C1 and either group A, B, or both, resulted in the emergence of subgroups C2, C3, and C4. Thus, groups A, B, and C1 are posited as the ancestral populations. Additionally, our morphological observations indicated distinct differences among these three groups. Our findings also suggest that human migration may have influenced the plant's distribution, hinting at active usage in the past that later waned, causing its current underutilized status.

New degradation mechanism of black tea pigment theaflavin involving condensation with epigallocatechin-3-O-gallate.

Theanaphthoquinone (TNQ) is the initial and main oxidation product of theaflavin, a representative black tea pigment. Nevertheless, TNQ is virtually undetected in the high-performance liquid chromatography analysis of black tea leaves using photodiode array detection. To elucidate the degradation mechanism of theaflavin in the black tea production process, this study investigated the reaction of TNQ with epigallocatechin-3-O-gallate (EGCg), which is the most abundant polyphenol in tea leaves. In citrate-phosphate buffer solution at pH 6 and room temperature, TNQ reacted nonenzymatically with EGCg to afford three products, whose structures were determined on the basis of spectroscopic data. The results indicated that the double bond of the ortho-naphthoquinone moiety in TNQ reacted with the autoxidation product of EGCg. This study demonstrates novel reactions occurring in the process of theaflavin degradation, which might be involved in the formation of thearubigins, the major black tea pigments composing oligomeric catechin oxidation products.

Genomic Epidemiology and Evolution of Scallion Mosaic Potyvirus From Asymptomatic Wild Japanese Garlic.

Scallion mosaic virus (ScaMV) belongs to the turnip mosaic virus phylogenetic group of potyvirus and is known to infect domestic scallion plants (Allium chinense) in China and wild Japanese garlic (Allium macrostemon Bunge) in Japan. Wild Japanese garlic plants showing asymptomatic leaves were collected from different sites in Japan during 2012-2015. We found that 73 wild Japanese garlic plants out of 277 collected plants were infected with ScaMV, identified by partial genomic nucleotide sequences of the amplified RT-PCR products using potyvirus-specific primer pairs. Sixty-three ScaMV isolates were then chosen, and those full genomic sequences were determined. We carried out evolutionary analyses of the complete polyprotein-coding sequences and four non-recombinogenic regions of partial genomic sequences. We found that 80% of ScaMV samples have recombination-like genome structure and identified 12 recombination-type patterns in the genomes of the Japanese ScaMV isolates. Furthermore, we found two non-recombinant-type patterns in the Japanese population. Because the wild plants and weeds may often serve as reservoirs of viruses, it is important to study providing the exploratory investigation before emergence in the domestic plants. This is possibly the first epidemiological and evolutionary study of a virus from asymptomatic wild plants.

Mesorhizobium sp. J8 can establish symbiosis with Glycyrrhiza uralensis, increasing glycyrrhizin production.

Licorice (Glycyrrhiza uralensis) is a medicinal plant that contains glycyrrhizin (GL), which has various pharmacological activities. Because licorice is a legume, it can establish a symbiotic relationship with nitrogen-fixing rhizobial bacteria. However, the effect of this symbiosis on GL production is unknown. Rhizobia were isolated from root nodules of Glycyrrhiza glabra, and a rhizobium that can form root nodules in G. uralensis was selected. Whole-genome analysis revealed a single circular chromosome of 6.7 Mbp. This rhizobium was classified as Mesorhizobium by phylogenetic analysis and was designated Mesorhizobium sp. J8. When G. uralensis plants grown from cuttings were inoculated with J8, root nodules formed. Shoot biomass and SPAD values of inoculated plants were significantly higher than those of uninoculated controls, and the GL content of the roots was 3.2 times that of controls. Because uninoculated plants from cuttings showed slight nodule formation, we grew plants from seeds in plant boxes filled with sterilized vermiculite, inoculated half of the seedlings with J8, and grew them with or without 100 µM KNO3. The SPAD values of inoculated plants were significantly higher than those of uninoculated plants. Furthermore, the expression level of the CYP88D6 gene, which is a marker of GL synthesis, was 2.5 times higher than in inoculated plants. These results indicate that rhizobial symbiosis promotes both biomass and GL production in G. uralensis.

Hypericum perforatum extract and hyperforin inhibit the growth of neurotropic parasite Toxoplasma gondii and infection-induced inflammatory responses of glial cells in vitro.

ETHNOPHARMACOLOGICAL RELEVANCE: Hypericum perforatum L. has been widely used as a natural antidepressant. However, it is unknown whether it is effective in treating infection-induced neuropsychiatric disorders. AIM OF THE STUDY: In order to evaluate the effectiveness of H. perforatum against infection with neurotropic parasite Toxoplasma gondii, which has been linked to neuropsychiatric disorders, this study investigated the anti-Toxoplasma activity using in vitro models. MATERIALS AND METHODS: Dried alcoholic extracts were prepared from three Hypericum species: H. perforatum, H. erectum, and H. ascyron. H. perforatum extract was further separated by solvent-partitioning. Hyperforin and hypericin levels in the extracts and fractions were analyzed by high resolution LC-MS. Anti-Toxoplasma activities were tested in vitro, using cell lines (Vero and Raw264), murine primary mixed glia, and primary neuron-glia. Toxoplasma proliferation and stage conversion were analyzed by qPCR. Infection-induced damages to the host cells were analyzed by Sulforhodamine B cytotoxicity assay (Vero) and immunofluorescent microscopy (neurons). Infection-induced inflammatory responses in glial cells were analysed by qPCR and immunofluorescent microscopy. RESULTS: Hyperforin was identified only in H. perforatum among the three tested species, whereas hypericin was present in H. perforatum and H. erectum. H. perforatum extract and hyperforin-enriched fraction, as well as hyperforin, exhibited significant anti-Toxoplasma property as well as inhibitory activity against infection-induced inflammatory responses in glial cells. In addition, H. perforatum-derived hyperforin-enriched fraction restored neuro-supportive environment in mixed neuron-glia culture. CONCLUSIONS: H. perforatum and its major constituent hyperforin are promising anti-Toxoplasma agents that could potentially protect neurons and glial cells against infection-induced damages. Further study is warranted to establish in vivo efficacy.

Teadenol A in microbial fermented tea acts as a novel ligand on GPR120 to increase GLP-1 secretion.

Post-fermented teas, produced by microbial fermentation, are attracting attention due to their health benefits that reduce the risk of hyperlipidemia and atherosclerosis. Although several novel polyphenols have been identified from post-fermented teas, their biological activities have not yet been fully elucidated. In this study, we found that teadenol A, a polyphenol recently isolated from Japanese post-fermented tea, acts as a novel ligand on a long-chain fatty acid receptor, GPR120. Teadenol A activated GPR120 was over-expressed in 293T cells, and this activation was inhibited by the GPR120 antagonist AH7614. Additionally, teadenol A induced Erk1/2 phosphorylation and increased the intracellular Ca2+ concentration in 293T cells, and these effects were completely dependent on GPR120 expression. Our results suggest that teadenol A binds and activates GPR120 directly. Furthermore, teadenol A enhanced the secretion of GLP-1 from intestinal endocrine STC-1 cells. GLP-1 suppresses appetite and increases insulin secretion, exhibiting anti-diabetic effects. GPR120/GLP-1 signaling is attracting attention as a potential target for pharmaceuticals against type 2 diabetes. Our results suggest that teadenol A is a key molecule in post-fermented tea responsible for beneficial effects on metabolic syndrome.

Hypericum erectum alcoholic extract inhibits Toxoplasma growth and Entamoeba encystation: an exploratory study on the anti-protozoan potential.

Hypericum erectum is an important ethnobotanical medicine in East Asian tradition. To explore the anti-parasitic potential of H. erectum, inhibitory effects on the growth of intracellular parasite Toxoplasma and on the encystation of intestinal parasite Entamoeba were examined. The constituents in H. erectum alcoholic extracts and fractions separated by solvent-partitioning were analysed by high resolution LC-MS. Toxoplasma gondii growth inhibition assay was performed using GFP-labelled T. gondii strain PTG-GFP by measuring the fluorescence intensity. Anti-Toxoplasma drug pyrimethamine was used as a positive control. T. gondii-induced immune reaction was assessed by quantitative PCR and fluorescence microscopy, using co-culture of PTG-GFP and monocyte-macrophage cell line Raw264. The inhibitory effect on the encystation of Entamoeba invadens was measured by flow-cytometry, where paromomycin was used as a positive control. H. erectum methanol (MeOH) extract (50 µg/mL) and ethyl acetate (EtOAc) fraction (50 µg/mL) inhibited the growth of T. gondii, while 50%MeOH extract and hydrophilic fractions were ineffective. Co-culture with T. gondii reduced the viability of macrophages, however macrophages were protected in the presence of H. erectum MeOH extract or EtOAc fraction (above 10 µg/mL). The MeOH extract and EtOAc fraction also effectively suppressed the encystation of E. invadens at 1 mg/mL. Hypericine, a major constituent in MeOH extract and EtOAc fraction, inhibited T. gondii growth and E. invadens encystation. Our results demonstrated that H. erectum effectively inhibited Toxoplasma growth and Entamoeba encystation. These activities are partly mediated by hypericin. In addition, it was suggested the extract and fraction may protect innate immune cells from Toxoplasma-induced damages, thereby enhancing parasite clearance. Further investigation is warranted to address the in vivo effectiveness of H. erectum as an anti-protozoal medicine.

Chemical and molecular bases of dome formation in human colorectal cancer cells mediated by sulphur compounds from Cucumis melo var. conomon.

Colorectal cancer was the third most commonly diagnosed malignant tumor and the fourth leading cause of cancer deaths worldwide in 2012. A human colorectal cancer cell line, RCM-1, was established from a colon cancer tissue diagnosed as a well-differentiated rectum adenocarcinoma. RCM-1 cells spontaneously form 'domes' (formerly designated 'ducts') resembling villiform structures. Two sulphur-containing compounds from Cucumis melo var. conomon (Katsura-uri, or Japanese pickling melon), referred to as 3-methylthiopropionic acid ethyl ester (MTPE) and methylthioacetic acid ethyl ester (MTAE), can induce the differentiation of the unorganized cell mass of an RCM-1 human colorectal cancer cell culture into a dome. However, the underlying molecular mechanisms of such dome formation have not been previously reported. Here, we performed a structure-activity relationship analysis, which indicated that methylthioacetic acid (MTA) was the lowest molecular weight compound with the most potent dome-inducing activity among 37 MTPE and MTAE analogues, and the methylthio group was essential for this activity. According to our microarray analysis, MTA resulted in down-regulation of 537 genes and up-regulation of 117 genes. Furthermore, MTA caused down-regulation of many genes involved in cell-cycle control, with the cyclin E2 (CCNE2) and cell division cycle 25A (CDC25A) genes being the most significantly reduced. Pharmacological analysis showed that the administration of two cell-cycle inhibitors for inactivating CDC25A phosphatase (NSC95397) and the cyclin E2/cyclin-dependent kinase 2 complex (purvalanol A) increased the dome number independently of MTA. Altogether, our results indicate that MTA is the minimum unit required to induce dome formation, with the down-regulation of CDC25A and possibly CCNE2 being important steps in this process.

First Genome Sequence of Shallot Latent Carlavirus from Allium macrostemon Bunge.

A wild Japanese garlic plant (Allium macrostemon Bunge, wild onion) with leaves showing chlorotic stripes was collected in Saitama Prefecture, Japan. Genome sequencing showed that it was infected with shallot latent carlavirus. The genomic sequence of this virus is reported for the first time from wild onion.

Ferulic-Acid Esters of Oligo-glucose from A-ilium macrostemon.

Two new ferulic acid esters of oligo-glucose, 1-Ο-(E)-feruloyl-ß--D-gentiobioside (1) and 1-Ο-(E)feruloyl-{ß-D-glucopyranosy (1->6)-[ß-D-glucopyranosyl (I--2)]}-ß-D-glucopyranoside.(allimacronoid D, 2) were isolated together with 1-Ο-(E)-feruloyl-ß-D-glucopyranoside (3) and trans-ferulic acid (4) from the leaves of Allium macrostemon Bunge. The chemical structures were elucidated based on the analyses of the spectroscopic and chemical data.

A New Flavonoid from Camellia sinensis Fermented Tea.

A new flavonoid, quercetin 3-0-α-L-rhamnopyranosyl-(l-6)-(2-E-p-coumaroyl)-D-D-glucopyranoside-4'-O-α-L-rhamnopyranoside, was isolated from the fermented tea of Camellia sinensis L. (Pu-erh tea) from China. The chemical structure was elucidated based on analyses of the spectroscopic data.

Polyacetylene glycosides from Pratia nummularia cultures.

Two polyacetylene glycosides, lobetyol 9-O-glc(6)-(1)rha (pratialin-A) and lobetyol 9-O-glc(6)-(1)glc(6)-(1)glc (pratialin-B), were isolated from Pratia nummularia (Campanulaceae) callus and hairy root cultures and their chemical structures were determined by analysis of spectroscopic data. From the methanol extract of the hairy root cultures, together with the known polyacetylene constituents lobetyol, lobetyolin, and lobetyolinin, tryptophan was also isolated. This report is the first example of the isolation and structure elucidation of rutinoside (pratialin-A) and triglucoside (pratialin-B) derivatives of polyacetylene constituents.

Antioxidative capacity of extracts and constituents in Cornus capitata adventitious roots.

Radical scavenging activities of extracts and constituents in Cornus capitata adventitious root cultures were evaluated by using 1,1-diphenyl-2-pycrylhydrazyl (DPPH) and superoxide anion radicals. Inhibitory activity against peroxidation of linoleic acid was assayed by using the thiobarbituric acid (TBA) method. Ethyl acetate and aqueous fractions were prepared from adventitious roots cultured in Murashige-Skoog liquid medium with 0.1 microM Cu(2+) (0.1CuMS) or 10 microM Cu(2+) (10CuMS). The highest scavenging activities on DPPH and superoxide anion radicals were observed in the ethyl acetate fraction from 0.1CuMS. In the inhibitory activity against linoleic acid oxidation, the ethyl acetate fraction from 10CuMS was highest among the fractions tested. The ethyl acetate fraction of adventitious roots cultured in 0.1CuMS contained mainly galloylglucoses (1,2,3,6-tetragalloylglucose and 1,2,3,4,6-pentagalloylglucose). The ethyl acetate fraction of adventitious roots cultured in 10CuMS contained mainly ellagic acid derivatives [3,3'-di-O-methylellagic acid 4-(5"-acetyl)-alpha-L-arabinofuranoside and stenophyllin H1]. Aqueous fractions prepared from both media contained iridoid glycosides (dihydrocornin and cornin). Tetra- and pentagalloylglucoses showed strong inhibitory activities (61.9 and 85.2%, respectively) against linoleic acid oxidation relative to those of butylated hydroxytoluene (BHT) (91.1%) or alpha-tocopherol (49.5%) at 50 microM concentration. Although both ellagic acid derivatives had weak activities (<50%) on DPPH and superoxide anion radical scavenging, 3,3'-di-O-methylellagic acid 4-(5"-acetyl)-alpha-L-arabinofuranoside was stronger (74.7%) than alpha-tocopherol (49.5%) in inhibiting linoleic acid oxidation at 50 microM concentration. Iridoid glycosides exhibited little activity against DPPH and superoxide anion radicals or against oxidation of linoleic acid.

Tropane alkaloids in adventitious root cultures of Physochlaina physaloides.

Adventitious root cultures of Physochlaina physaloides were established and their growth and tropane alkaloid production were investigated. Although the roots cultured in Murashige-skoog liquid medium containing 1.0-3.0 mg/l IBA grew well, the addition of lower concentrations of auxins was more suitable for alkaloid production. The best alkaloid yield was obtained with 0.01 mg/l NAA in 1/2 MS liquid medium. In addition, the alkaloid differences between the cultured roots and the regenerated plant cultivated in the soil was studied. Cultured roots in this condition showed higher level of alkaloids when compared to leaf, stem and roots from the regenerated plant (3 months in field conditions).

Tropane alkaloids in auxin-independent root cultures of Physochlaina physaloides.

Adventitious and hairy root cultures of Physochlaina physaloides were established. These roots grew well and produced high amounts of tropane alkaloids (particularly hyoscyamine and 6 beta-hydroxyhyoscyamine) in auxin-free culture medium. The effects of basal media and temperature on the growth and alkaloid production of these roots were investigated. Both root cultures produced highest amount of tropane alkaloids in B5 medium though the optimum temperature for hairy roots were lower than that for adventitious roots.

New phenolic compounds from Camellia sinensis L. fermented leaves.

Two new phenolic compounds (1 and 2, named as teasperol and teasperin, respectively) were isolated from fermented tea (Camellia sinensis L.) products, together with known phenolic compounds. Teasperol (1) was isolated from Chinese traditional post-fermented tea leaves (Liu-pao tea) and teasperin (2) was from a Japanese tea product which was selectively fermented with Aspergillus sp. The chemical structures of 1 and 2 were elucidated based on the analyses of their spectroscopic data.