We examined the effects of fucoidan, a sulfated polysaccharide, administered concurrently with the onset of a high-fat diet (HFD) for a short period in C57BL/6 N mice to evaluate its immunological effects under obesity-i...We examined the effects of fucoidan, a sulfated polysaccharide, administered concurrently with the onset of a high-fat diet (HFD) for a short period in C57BL/6 N mice to evaluate its immunological effects under obesity-inducing conditions. Numerous genes that were downregulated in the HFD group relative to the control-diet group were restored in the HFD + fucoidan group; these genes were mainly associated with inflammatory responses and cellular functions, including migration and viability, as identified by DNA microarray analysis. Upstream regulator analysis in ingenuity pathway analysis identified numerous cytokines and inflammatory mediators. Quantitative PCR confirmed that the expression of selected inflammatory cytokine genes-particularly Ifn-γ and Il-5-was decreased in the HFD group but restored in the HFD + fucoidan group. Our study suggests that fucoidan administered concurrently with an HFD attenuates HFD-induced downregulation of inflammatory and immune-related functions, including T-cell and B-cell functions, and may improve HFD-induced immune dysfunction and imbalance.
Natural products exhibit extraordinary structural diversity and play central roles in chemical ecology, biological interactions, and drug discovery. However, many natural products are available only in trace amounts, mak...Natural products exhibit extraordinary structural diversity and play central roles in chemical ecology, biological interactions, and drug discovery. However, many natural products are available only in trace amounts, making structural elucidation a major analytical challenge. This review summarizes landmark studies and recent advances in the structural analysis of natural products available in microgram- to sub-milligram quantities. It focuses primarily on NMR-based structural elucidation, including the use of high-field instrumentation, cryogenic probes, and advanced pulse sequences, while also covering approaches based on comparison with synthetic standards, DFT-assisted structure validation, the crystalline sponge method, and MicroED. Representative examples, including natural toxins, biosynthetic intermediates, and microbial signaling molecules, illustrate how integrated analytical and synthetic approaches enable reliable structure determination from limited material. These methodologies provide an important foundation not only for natural product discovery but also for understanding biosynthesis, biological activity, and chemical ecology at the molecular level.
Lactic acid bacteria enhance intestinal immunoglobulin A (IgA) secretion; however, the underlying mechanisms are often strain specific. In this study, we found a novel strain, Lentilactobacillus hilgardii H-50, that enha...Lactic acid bacteria enhance intestinal immunoglobulin A (IgA) secretion; however, the underlying mechanisms are often strain specific. In this study, we found a novel strain, Lentilactobacillus hilgardii H-50, that enhanced intestinal IgA production among L. hilgardii strains. Heat-killed H-50 strain increased IgA secretion from mouse Peyer's patch cells and elevated faecal IgA levels in vivo. Although this strain also induced the secretion of interleukin-6 (IL-6) and IL-10, IL-6 was essential for IgA production, whereas IL-10 less contributed to it. Cell surface components, including cell wall components and S-layer protein, but not nucleic acids, enhanced IL-6 induction and IgA production. Furthermore, H-50 exhibited TLR2 ligand activity, and blockade of TLR2 signalling abolished IL-6 induction and IgA production. These results indicate that H-50 enhances intestinal IgA production via a TLR2-dependent, IL-6-mediated mechanism. This strain can potentially be used as a paraprobiotics.
Sphingolipid composition was compared among selected Caryophyllales species using LC-MS/MS. Glycosylinositol phosphorylceramide molecular species profiles differed between AC and SB groups, accompanied by variation in gl...Sphingolipid composition was compared among selected Caryophyllales species using LC-MS/MS. Glycosylinositol phosphorylceramide molecular species profiles differed between AC and SB groups, accompanied by variation in glycan series distribution and the GIPC/GlcCer ratio. Differences in sphingoid base modification and ceramide structure were also observed, suggesting group-associated differences in sphingolipid metabolism and potential value for future chemotaxonomic studies.
When plants are infected with pathogens, they activate their immune systems, resulting in hypersensitive cell death, which to prevents the spread of pathogens. Therefore, the compounds that induce plant immunity (i.e. im...When plants are infected with pathogens, they activate their immune systems, resulting in hypersensitive cell death, which to prevents the spread of pathogens. Therefore, the compounds that induce plant immunity (i.e. immunity inducers) can protect plants from pathogens by activating the plant immune system without exhibiting antipathogenic activity, which may decrease the risk of drug resistance. However, the number of marketed plant immunity inducers is limited compared to that of marketed pesticides, and their plant protection efficiency remains insufficient. In this study, we screened for compounds that specifically potentiate pathogen-activated cell death from the Ōmura natural product library using an established high-throughput screening system with cultured suspension cells of Arabidopsis thaliana. Our result show that aurodox potentiate pathogen-activated plant cell death, but exhibits weak antibacterial activity against Pseudomonas syringae pv. tomato, suggesting the potential of aurodox as an inducer of plant immunity.
Target of rapamycin complex 1 (TORC1) integrates nutrient signals with cell growth. While its inactivation is known to trigger mitotic slippage via APC/C-Cdh1-dependent securin degradation and separase activation, the mo...Target of rapamycin complex 1 (TORC1) integrates nutrient signals with cell growth. While its inactivation is known to trigger mitotic slippage via APC/C-Cdh1-dependent securin degradation and separase activation, the molecular basis of cohesion loss during nutrient stress has remained incompletely defined. In budding yeast, we show that TORC1 inactivation elicits a noncanonical, proteasome-dependent degradation of cohesin that is independent of securin and separase. Separase was itself destabilized upon TORC1 inactivation, yet Scc1 degradation persisted even in a separase-resistant mutant. Cohesin degradation proceeds when APC/C is impaired, indicating involvement of an atypical ubiquitin ligase. These results reveal a second, aberrant route to sister chromatid dissociation upon TORC1 inactivation, operating in parallel with the previously reported APC/C-Cdh1 pathway, via the unconventional degradation of mitotic key factors.
Pteridine compounds are heterocyclic compounds that contribute to various processes in living organisms. Pterin is a representative derivative, and its analogs are widely used in biological systems. The two imine bonds w...Pteridine compounds are heterocyclic compounds that contribute to various processes in living organisms. Pterin is a representative derivative, and its analogs are widely used in biological systems. The two imine bonds within the pteridine ring confer electron donor-acceptor properties, and the oxidized form exhibits fluorescence because of its π-conjugated system. Owing to these structural and electronic features, pterins perform a variety of physiological functions, including serving as cofactors, pigments, and signaling molecules. To date, studies on the dynamics of pterins have been conducted independently from chemical and biochemical perspectives; thus, the resulting knowledge has not been sufficiently systematized. This review provides a comprehensive overview of the dynamics of naturally occurring pterins by systematically integrating existing chemical and biochemical data.
Sudachi (Citrus sudachi, a citrus fruit of the Rutaceae family) juice was found to promote neurite outgrowth in PC12 cells. Activity-guided purification of the juice led to the isolation of two furanocoumarins, byakangel...Sudachi (Citrus sudachi, a citrus fruit of the Rutaceae family) juice was found to promote neurite outgrowth in PC12 cells. Activity-guided purification of the juice led to the isolation of two furanocoumarins, byakangelicin and isopimpinellin, and a flavone, demethoxysudachitin, identifying them as neurite-outgrowth promoters for the first time. Byakangelicin and isopimpinellin showed markedly stronger neurite outgrowth-promoting activity than demethoxysudachitin, prompting a comparative evaluation of five furanocoumarins, including both isolated and related reference compounds. The results showed that the basic furanocoumarin scaffold alone is inactive, whereas substitution at either the C5 or C8 position of a furanocoumarin confers prominent activity, with dual substitution further improving activity. Among the tested compounds, byakangelicin and isopimpinellin exhibited the most potent neurite outgrowth-promoting activity. These findings identify sudachi-derived furanocoumarins as previously unrecognized neurite-outgrowth promoters and suggest their potential as neurotrophic agents for supporting cognitive function.
Curcuma zedoaria may serve as a promising material for preventing and/or improving lifestyle-related diseases, considering its known in vitro adiponectin-elevating effects. Here, we aimed to determine the effects of a C....Curcuma zedoaria may serve as a promising material for preventing and/or improving lifestyle-related diseases, considering its known in vitro adiponectin-elevating effects. Here, we aimed to determine the effects of a C. zedoaria ethanol extract (CZE) on high-fat diet (HFD)-induced obesity in C57BL/6 N mice, to elucidate its effects on lifestyle-related diseases. CZE treatment with an HFD (HFD-CZE) reduced liver weight, subcutaneous fat, mesenteric fat, and hepatic lipid accumulation, along with alleviating the levels of plasma triglyceride, total-cholesterol, low density lipoprotein-cholesterol, aspartate aminotransferase, and alanine aminotransferase relative to those in the HFD group. Decreased mRNA expressions of Tnfa and Il6 and increased mRNA expression of adiponectin were observed in epididymal white adipose tissue of the HFD-CZE group. CZE also reduced hepatic Tnfa and Fabp4 mRNA expressions. Thus, C. zedoaria may ameliorate obesity-induced whole-body inflammation, potentially aiding in preventing and improving obesity and subsequent lifestyle-related diseases.
Alternative polyadenylation (APA) is a key post-transcriptional regulatory mechanism in eukaryotes, but its role in adipogenesis remains poorly understood. Here, we investigated how insulin-induced gene 2 (Insig2) APA re...Alternative polyadenylation (APA) is a key post-transcriptional regulatory mechanism in eukaryotes, but its role in adipogenesis remains poorly understood. Here, we investigated how insulin-induced gene 2 (Insig2) APA regulates adipogenesis. Functional assays showed that Insig2 knockdown promoted 3T3-L1 preadipocyte differentiation, whereas Insig2 overexpression inhibited it. During adipogenesis, Insig2 generated two APA isoforms with distinct 3'UTR lengths, and the short 3'UTR isoform was more abundant than the long isoform. Mechanistically, miR-96-5p specifically targeted the unique extension region of the long Insig2 3'UTR isoform to suppress its expression, thereby alleviating its inhibitory effect on adipogenesis. In contrast, the short Insig2 isoform lacked the miR-96-5p binding site and escaped miR-96-5p-mediated repression, thus exerting a stronger inhibitory effect on adipogenesis. Collectively, these findings reveal an isoform-specific Insig2 APA/miR-96-5p regulatory axis that dynamically controls adipogenesis at the post-transcriptional level. Our results revealed a new regulatory mechanism of Insig2 APA in adipogenesis.
The filamentous fungus Fusarium sp. D2 strain produces hydroxy fatty acids via the hydration of unsaturated fatty acids under anaerobic or microaerobic conditions. In this study, the oleate hydratase gene (D2Ohy) derived...The filamentous fungus Fusarium sp. D2 strain produces hydroxy fatty acids via the hydration of unsaturated fatty acids under anaerobic or microaerobic conditions. In this study, the oleate hydratase gene (D2Ohy) derived from Fusarium sp. D2 was cloned for the first time from a fungal source. The amino acid sequence of D2Ohy was up to 69.7% homologous to bacterial oleate hydratases (Ohys) and contained the flavin adenine dinucleotide (FAD)-binding motif, FAD-lid, and active loop, all of which are crucial for hydration activity. D2Ohy was stable over a temperature range of 40-50°C and a pH range of 6.0-8.0, demonstrating properties distinct from previously reported bacterial-derived Ohys. Compared to bacterial Ohys, D2Ohy efficiently utilized oleic, palmitoleic, linoleic, γ-linolenic, and ricinoleic acids as substrates. This study reports novel functional properties of D2Ohy, identified for the first time in fungi.
Barley and rye are major cereals worldwide and contain proteinaceous α-amylase inhibitors for biological defence against insects. Inhibiting mammalian α-amylase could potentially be a promising functional food characteri...Barley and rye are major cereals worldwide and contain proteinaceous α-amylase inhibitors for biological defence against insects. Inhibiting mammalian α-amylase could potentially be a promising functional food characteristic for suppressing postprandial blood glucose levels. This study examined in vitro mammalian α-amylase inhibitory effects and the physicochemical and functional properties of barley and rye albumins, including thermal stability, solubility, emulsifying and foaming properties, to evaluate their potential applications in various foods. Both barley and rye albumins inhibited mammalian α-amylase and maintained their activity even after heating. High solubility was observed at pH 3.0-6.0, which was maintained after heating at 80 °C for 20 min. Furthermore, barley and rye albumins exhibited high emulsifying and foaming properties at pH 3.0-6.0. These findings suggest that barley and rye albumins are promising materials for suppressing postprandial blood glucose elevation and can be used in various functional foods.
The gut microbiome is a potential source of non-invasive cancer biomarkers. We evaluated six fecal microbial markers and developed targeted qPCR-based logistic models for colorectal cancer (CRC) and pancreatic cancer (PC...The gut microbiome is a potential source of non-invasive cancer biomarkers. We evaluated six fecal microbial markers and developed targeted qPCR-based logistic models for colorectal cancer (CRC) and pancreatic cancer (PC). Using LASSO with the 1-standard-error rule, four markers were selected for CRC (afb, nan, fsr, and 5ar) and three for PC (but, fsr, and saa). In post-selection leave-one-out cross-validation of fixed model structures, the CRC and PC models yielded AUCs of 0.824 and 0.780, respectively. Fixed-model application yielded AUCs of 0.716 for colorectal adenoma and 0.540 for the pancreatic high-risk group. In an exploratory Early PC versus high-risk comparison, the fecal qPCR score showed a higher AUC point estimate than CA19-9, while the difference was not statistically significant. Overall, the disease-specific model performance and fixed-model behavior across clinically related groups support further evaluation of model-derived cancer probability scores as exploratory cancer-assessment tools.
This study investigated the mechanisms of glucose-induced β-glucosidase (Bgl) activation in Bacillus subtilis strains 168 and Miyagino. Distinct Bgl-related genes were upregulated by glucose in the two strains. These inc...This study investigated the mechanisms of glucose-induced β-glucosidase (Bgl) activation in Bacillus subtilis strains 168 and Miyagino. Distinct Bgl-related genes were upregulated by glucose in the two strains. These increases were suppressed in the ΔpfkA and ΔfbaA mutants, but not Δpyk, mutant, suggesting the involvement of upper glycolytic flux changes. Additionally, we found that soybean broth medium may contain non-sugar Bgl-activating factors.
Food allergy is increasing worldwide, and sensitization through the skin is an important route of disease onset. However, the principal molecules driving epicutaneous sensitization to cashew nut remain unclear. Here, we...Food allergy is increasing worldwide, and sensitization through the skin is an important route of disease onset. However, the principal molecules driving epicutaneous sensitization to cashew nut remain unclear. Here, we established a BALB/c mouse model of epicutaneous sensitization and investigated major reactive antigens. Cashew nut-specific IgE was significantly higher in the cashew nut group at weeks 4 and 5, and cashew nut-specific IgG1 was significantly higher at weeks 3, 4, and 5. Immunoblotting showed antibody binding across multiple molecular weight regions. Immunoreactivity-guided fractionation followed by LC-MS/MS identified Ana o 3 (2S albumin; ∼10 kDa) and Ana o 2 (11S globulin; ∼50 kDa) as major IgG1-reactive antigens. Cross-reactivity was strongest with pistachio. Heating (160°C, 30 min) reduced immunoreactivity of the recoverable extract in coated-antigen assays under the present extraction conditions. These findings identify Ana o 2 and Ana o 3 as major reactive antigens in this mouse model.
The defatted residue of Perilla frutescens seeds, a by-product of edible oil production, remains largely underutilized in food applications. This study investigated the effects of a hot-water extract derived from this re...The defatted residue of Perilla frutescens seeds, a by-product of edible oil production, remains largely underutilized in food applications. This study investigated the effects of a hot-water extract derived from this residue on osteoclast differentiation and bone-related changes. In bone marrow-derived macrophages, the extract inhibited receptor activator of nuclear factor-κB ligand-induced osteoclast differentiation, as evidenced by the reduced formation of tartrate-resistant acid phosphatase-positive multinucleated cells and decreased tartrate-resistant acid phosphatase activity, without significantly affecting cell viability. In ovariectomized mice, the extract increased trabecular region length and cortical bone thickness. In a separate experiment, it enhanced femoral mechanical strength in a three-point bending test. High-performance liquid chromatography analysis identified rosmarinic acid and salviaflaside in the extract. These findings suggest that defatted Perilla frutescens seed residue may be a functional resource for improving bone health.
The intestinal microbiota is essential for maintaining intestinal homeostasis by regulating mucosal immune responses. Identifying bacterial traits that support stable host interactions is therefore crucial for developing...The intestinal microbiota is essential for maintaining intestinal homeostasis by regulating mucosal immune responses. Identifying bacterial traits that support stable host interactions is therefore crucial for developing next-generation mucosal probiotics. RNase T2 is a conserved ribonuclease with species-specific functions. In Escherichia coli, RNase T2 (RNase I) suppresses biofilm formation, a key process for intestinal colonization. We previously showed that an RNase I-deficient E. coli (Δrna) strain stably colonizes the mouse intestine. Here, we analyzed the immunogenic properties of the Δrna strain. Heat-killed Δrna induced an early IL-1β and TNF-α-skewed cytokine response in RAW264.7 macrophages, with reduced IL-6 expression compared to wild type. In mice, Δrna colonization caused no intestinal tissue injury but enhanced IL-6 and IL-22 expression in the cecum and colon. Selected TLR-specific reporter assays identified TLR5-dependent signaling driven by increased FliC expression. These findings show that RNase I deficiency modulates immune activation while maintaining non-pathogenic mucosal interactions.
Vitamin D (VD) and Wnt signaling both play important roles in bone metabolism and are targeted in osteoporosis therapy. Combination treatment with VD derivatives and anti-sclerostin antibodies that activate Wnt signaling...Vitamin D (VD) and Wnt signaling both play important roles in bone metabolism and are targeted in osteoporosis therapy. Combination treatment with VD derivatives and anti-sclerostin antibodies that activate Wnt signaling improves clinical outcomes, although the molecular basis of this cooperative effect remains unclear. Because β-catenin functions as a transcriptional co-regulator in canonical Wnt signaling, we examined whether β-catenin modulates vitamin D receptor (VDR)-mediated transcription. Reporter assays were performed in Saos-2 osteoblastic cells using vitamin D response element (VDRE) and TCF/LEF-responsive reporters. Activation of Wnt signaling by CHIR99021 or expression of constitutively active β-catenin did not enhance VDR transcriptional activity but modestly suppressed VD-induced transcription. Similar results were obtained in HCT116 and COS-1 cells. These findings indicate that VDR and Wnt/β-catenin signalings are not cross-talked at the transcriptional level.
Tuliposides (Pos), major secondary metabolites in tulip, are classified into monoacyl- and diacyl-type compounds. Herein, we report the first total synthesis of naturally occurring diacyl-type PosD and PosF and their iso...Tuliposides (Pos), major secondary metabolites in tulip, are classified into monoacyl- and diacyl-type compounds. Herein, we report the first total synthesis of naturally occurring diacyl-type PosD and PosF and their isomers diOH-PosD and isoPosF, respectively, from D-glucose in five steps. Under neutral conditions, the non-natural Pos isomers were significantly less stable than their natural counterparts.
In recent years, there has been increasing evidence that plant-derived vesicles, i.e. exosome-like nanovesicles (ENs), have biological activities. We investigated the ingredients in green tea "Sencha" infusion-derived ex...In recent years, there has been increasing evidence that plant-derived vesicles, i.e. exosome-like nanovesicles (ENs), have biological activities. We investigated the ingredients in green tea "Sencha" infusion-derived exosome-like nanovesicle (TIEN) fractions and found that gallic acid-type polyphenolics were detected disproportionately in these fractions. Furthermore, it is thought that plant-derived ENs arise mainly from vacuoles, endoplasmic reticulum, and Golgi apparatus, etc. However, our findings suggest that TIENs may originate from chloroplasts.