Fumihide Takano

Scanning electron microscopy with an ionic liquid reveals the loss of mitotic protrusions of cells during the epithelial-mesenchymal transition.

Epithelial–mesenchymal transition (EMT) is a key event in cancer metastasis and is characterized by increase in cell motility, increase in expression of mesenchymal cell markers, loss of proteins from cell‐to‐cell junction complexes, and changes in cell morphology. Here, the morphological effects of a representative EMT inducer, transforming growth factor (TGF)‐β1, were investigated in human lung adenocarcinoma (A549) cells and pancreatic carcinoma (Panc‐1) cells. TGF‐β1 caused morphological changes characteristic of EMT, and immunostaining showed loss of E‐cadherin from cell‐to‐cell junction complexes in addition to the upregulation of the mesenchymal marker vimentin. During scanning electron microscopy (SEM) with an ionic liquid, we observed EMT‐specific morphological changes, including the formation of various cell protrusions. Interestingly, filopodia in mitotic cells were clearly observed by SEM, and the number of these filopodia in TFG‐β1‐treated mitotic cells was reduced significantly. We conclude that this reduction in such mitotic protrusions is a novel effect of TGF‐β1 and may contribute to EMT. Microsc. Res. Tech., 2011.

Stilbenoids from the Melinjo (Gnetum gnemon L.) Fruit Modulate Cytokine Production in Murine Peyer's Patch Cells Ex Vivo

Melinjo fruit ( Gnetum gnemon L.) has been used as a food in Southeast Asia. To investigate if this fruit has regulatory actions on ileal immune responses, we measured T-helper (Th) cytokine production, i.e., interleukin-2 (IL-2), IL-4, IL-5, and interferon-gamma (IFN- γ), in cultured Peyers patch (PP) cells from mice orally treated with a methanol extract of melinjo fruit. Oral administration of the 50 % ethanol extract at 100 mg/kg/day significantly enhanced the production of the Th1 cytokines IL-2 and IFN- γ irrespective of concanavalin-A stimulation, whereas the production of the Th2 cytokines IL-4 and IL-5 was not affected. We also isolated seven active constituents accompanied with two new stilbenoids from the ethylacetate fraction of the extracts. The structure of the new stilbene glucosides gnemonoside L (5) and gnemonoside M (7) was determined by 1D and 2D NMR and MS analyses. Five known stilbenoids were identified as resveratrol (1), isorhapontigenin (2), gnemonoside D (4), gnetins C (3) and E (6). Among these tested compounds, only new stilbenoid 7 strongly enhanced Th1 cytokine production in cultured PP cells at 10 mg/kg/day. These results indicated that this melinjo extract and its active constituent 7 potentiated T-cell-dependent immune responses in the ileal mucosa.

Anti-inflammatory constituents from Tabebuia avellanedae

Five novel compounds were isolated from the water extract of Tabebuia avellanedae, and their structures were established by analysis of NMR spectroscopy and mass spectrometry. Compounds 1-5 at 25μM showed strong inhibitory activity on the inflammatory cytokine, tumor-necrosis factor-α and interleukin-1β production in cultured human myeloma THP-1 cells co-stimulated with lipopolysaccharide without any significant cytotoxicity, and their anti-allergic and antioxidant activities were evaluated.

Oral administration of Ren-Shen-Yang-Rong-Tang 'Ninjin'yoeito' protects against hematotoxicity and induces immature erythroid progenitor cells in 5-fluorouracil-induced anemia.

The purpose of this study was to investigate the efficacy of four different Japanese and Chinese herbal prescriptions, Ren-Shen-Yang-Rong-Tang (Ninjin’yoeito, NYT), Chai-Hu-Gui-Zhi-Gan-Jiang-Tang (Saikokeishikankyoto, SKKT), Si-Jun-Zi-Tang (Shikunshito, SKT) and Si-Wu-Tang (Shimotsuto, SMT), which are traditionally used for anemia and fatigue, against hematotoxicity in mice treated with 5-fluorouracil (5-FU). NYT 1–100 mg kg–1 day–1 injected orally for 7 consecutive days before and after 5-FU injection significantly suppressed reductions in red blood cell, white blood cell and platelet counts in peripheral blood, and accelerated their recovery. Administration of SKKT also produced a slight but significant improvement in 5-FU-induced erythrocytopenia, whereas SMT and SKT could not prevent anemia. Oral injection of NYT also inhibited 5-FU-induced decreases in peripheral reticulocyte and bone marrow cell counts on day 10, and markedly hastened their recovery on day 20, in a dose-dependent manner. Erythroid progenitor colonies, such as colony forming units-erythroid and burst forming units-erythroid, formed by marrow cells from mice treated with 5-FU were significantly increased by oral administration of NYT. These findings suggest that NYT has the potential to protect against hematotoxicity, and also has hematopoietic activity, through stimulation of immature erythroid progenitor cell differentiation.

Constituents of Rhodiola rosea showing inhibitory effect on lipase activity in mouse plasma and alimentary canal.

As a methanol extract of the rhizome of Rhodiola rosea inhibits the activity of lipase in isolated mouse plasma in vitro and in the mouse gastrointestinal tube in vivo, the active components in this plant were investigated. After fractionation and separation processes, rhodionin and rhodiosin were isolated as active ingredients. Their IC50 values were 0.093 mM and 0.133 mM in vitro, respectively. Both compounds significantly suppressed the elevation of the postprandial blood triglyceride level, e.g., by 45.6 % (150 mg/kg, 60 min after oral administration) and 57.6 % (200 mg/kg, 180 min after oral administration), respectively. Consequently, we anticipate the application of this plant and its constituents to the treatment of lifestyle-related diseases such as hyperlipidemia and exogeneous obesity and to health foods.

Bioactive Phenylpropanoid Glycosides from Tabebuia avellanedae

Three novel phenylpropanoid glycosides 2, 5, 6 were isolated from water extract of Tabebuia avellanedae, together with three known phenylpropanoid glycosides 1, 3, 4. All compounds were identified on the basis of spectroscopic analysis and chemical methods and, for known compounds, by comparison with published data. All isolated compounds showed strong antioxidant activity in the DPPH assay, and compound 5 give the highest antioxidant activity among all compounds, with an IC50 of 0.12 µM. All compounds exhibited moderate inhibitory effect on cytochrome CYP3A4 enzyme.

Phenolic lipid ingredients from cashew nuts

Five new phenolic lipids, 2-(8″Z-eicosenoyl)-6-(8′Z-pentadecenyl) salicylic acid (3), 2-(9″Z-hexadecenoyl)-6-(8′Z, 11′Z-pentadecadienyl) methyl salicylate (5), 2-(10″Z, 13″Z-nonadecadienoyl)-6-(8′Z, 11′Z-pentadecadienyl) salicylic acid (6), 2-(16″Z-pentacosenoyl)-6-(8′Z-pentadecenyl) salicylic acid (7) and 2-(9″Z-octadecenoyl)-6-(8′Z, 11′Z-pentadecadienyl) methyl salicylate (8), and three known compounds, cardols (1), anacardic acid (2) and cardanols (4), were isolated from the nuts of the cashew, Anacardium occidentale L. The structures were established on the basis of detailed MS and NMR spectroscopic analyses. Compound 1 highly enhanced both Th-1 (IL-2, IFN-γ) and Th-2 (IL-4, IL-5) cytokine production, and compounds 7 and 8 highly increased cytokine IL-2 and IFN-γ production in response to concanavalin A in cultured murine Peyer’s patch cells ex vivo. The isolated compounds showed moderate inhibitory activities on cytochrome CYP3A4 enzyme.

Carotenoids modulate cytokine production in Peyer's patch cells ex vivo.

This study investigated the effects of carotenoid and capsaicin constituents of Capsicum on intestinal immune responses in mice. Peyers patch (PP) cells were isolated from mice orally administered with capsaicin, or one of three carotenoids (beta-carotene, beta-cryptoxanthin, or lycopene), at 5 mg/kg/day for 7 consecutive days. Collagenase-separated PP cells were then cultured in the presence or absence of concanavalin A (Con A). PP cells from mice treated with capsaicin, beta-carotene, or beta-cryptoxanthin all showed significantly enhanced interleukin (IL)-2 and interferon (IFN)-gamma production when costimulated with 5 microg/mL Con A, with capsaicin having the greatest effect (approximately two times greater than in normal mice). No increase in the production of IL-2 or IL-4 was observed when PP cells from mice were cultured without Con A. We further tested the combined efficacy of carotenoids and capsaicin on intestinal T-cell cytokine production. Oral administration of capsaicin with beta-carotene, both at 5 mg/kg/day for 7 days, increased IFN-gamma and IL-2 production in cultured PP cells costimulated with Con A. In contrast, oral administration of beta-cryptoxanthin counteracted the stimulatory effect of capsaicin treatment on T-helper cytokine production. Flow cytometric analysis revealed that the population of IFN-gamma(+) and IL-4(+) cells in PPs from mice administered capsaicin and/or carotenoids did not change, which suggested that the effects of carotenoids and capsaicin on cytokine production were not due to changes in the lymphoid population in PPs. These results indicate that carotenoids and capsaicin, which are common components of foods such as Capsicum, mutually modulate T-cell immune responses to exogenous or endogenous inducers such as antigens in PPs, without changing the lymphoid population. Carotenoids modulate the potentiality of cytokine production in T cells or indirectly activate T cells but have no triggering effect such as Con A.

Efficacy of porcine placental extract on climacteric symptoms in peri- and postmenopausal women

ABSTRACT Objectives Injections of human placental extract have long been used to treat menopausal symptoms. Recently, porcine placental extract (PPE), an oral supplement, has been developed for this purpose. The aim of this study was to assess whether PPE has an impact on climacteric symptoms in perimenopausal and postmenopausal women. Methods Seventy-six women with climacteric symptoms were enrolled into this open-label, randomized, controlled study. The control group (n = 38) underwent 24 weeks of open treatment with Toki-shakuyaku-san (TJ23), an oral herbal remedy used to alleviate climacteric symptoms. The PPE group (n = 38) received three capsules of PPE/day orally for the initial 12 weeks and six capsules/day for the next 12 weeks. Climacteric symptoms were evaluated in both groups using the Simplified Menopausal Index (SMI) score, Zungs Self-Rating Depression Scale (ZSDS) and the Spielberger State-Trait Anxiety Inventory (STAI) before commencing treatment, after 12 weeks of treatment and on completion of treatment. Results Treatment with PPE was significantly (p < 0.01) more effective in reducing the SMI, ZSDS and STAI measures at 12 and 24 weeks than TJ23 treatment alone. Treatment with PPE was also significantly effective (p < 0.01) in reducing the subscale scores of the SMI for items such as hot flushes, insomnia, irritability, depression, fatigue and joint pain. PPE treatment had no significant adverse effects. Conclusion Oral PPE treatment is another possible option for treating perimenopausal and postmenopausal women with climacteric symptoms.

Two new acetylated flavonoid glycosides from Centaurium spicatum L

Two new acetylated flavonol glycosides, quercetin 3-O-[(2,4-diacetyl-α-l-rhamnopyranosyl)-(1→6)]-2,4-diacetyl-β-d-galactopyranoside (1) and quercetin 3-O-[(2,4-diacetyl-α-l-rhamnopyranosyl)-(1→6)]-3,4-diacetyl-β-d-galactopyranoside (2), in addition to two known acetylated quercetin glycosides quercetin 3-O-[(2,3,4-triacetyl-α-l-rhamnopyranosyl)-(1→6)-β-d-galactopyranoside (3) and quercetin 3-O-[(2,3,4-triacetyl-α-l-rhamnopyranosyl)-(1→6)-3-acetyl-β-d-galactopyranoside (4), were isolated from the aerial part of Centaurium spicatum (L.) Fritsch (Gentianaceae). Structure elucidation, especially the localization of the acetyl groups, and complete 1H and 13C NMR assignments of these biologically active compounds were carried out using one- and two-dimensional NMR measurements, including 1H- and 13C-NMR, DEPT-135, H–H COSY, HMQC and HMBC, in addition to HR-FAB/MS experiments.