Kenichi Yamaguchi

Nagasaki University, Nagasaki-shi, Nagasaki-ken, Japan

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Publications (53)102.5 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: In the present study, the chemical structure and chemoprotective activity of Pyropia yezoensis protein (PYP) were investigated using sodium dodecyl sulfate (SDS)‑polyacrylamide gel electrophoresis, automated protein sequencing, matrix‑assisted laser desorption/ionization‑quadrupole ion trap‑time‑of‑flight mass spectrometry and a chemoprotective assay using a synthetic peptide. The PYP fraction was demonstrated to contain two proteins: PYP1 (10 kDa, SDS‑resistant dimer) and PYP2 (10 kDa). PYP1 is a novel protein showing sequence homology with the hypothetical function‑unknown proteins of Chondrus crispus (Rhodophyta) and Emiliania huxleyi (Haptophyceae). PYP2 is a paralog of an extrinsic protein of photosystem II found in other Rhodophyta. The synthetic peptide PYP1 (1‑20), corresponding to the N‑terminal 20 residues of PYP1 (ALEGGKSSGGGEATRDPEPT), exhibits chemoprotective activity against acetaminophen‑induced cell death in Chang liver cells, indicating that PYP1 is a chemoprotectant of the PYP fraction. A possible association between the structure of PYP and its chemoprotective activity is discussed.
    International Journal of Molecular Medicine 11/2014; · 1.96 Impact Factor
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    ABSTRACT: We evaluated the antitumor activity of crude extract and ascophyllan prepared from Ascophyllum nodosum in sarcoma-180 solid tumor-bearing mice with continuous intraperitoneal (i.p.) administration at a dose of 50 mg/kg body weight/day or oral administration at a dose of 500 mg/kg body weight/day. Ascophyllan and crude extract administered via the oral route showed greater antitumor effects than via i.p. route, and the tumor sizes in mice treated with ascopyllan and crude extract were reduced by a mean of 68.7±6.8% and 42.4±24.8% by the oral route, and 41.4±16.1% and 13.6±20.6% by i.p. route compared to control mice. Splenic natural killer cell activity in the mice treated with ascophyllan and crude extract by i.p. route was significantly enhanced, while only a slight increase of this activity was observed in orally-treated mice. Furthermore, increase in spleen weight of tumor-bearing mice was slightly suppressed by oral administration of ascophyllan, whereas i.p. administration resulted in further enlargement. Analysis of serum cytokines revealed that oral treatment with ascophyllan resulted in significant increase of tumor necrosis factor-α and interleukin-12 levels. Since ascophyllan showed no direct cytotoxic effect on sarcoma-180 cells, orally-administered ascophyllan is suggested to exhibit its antitumor activity through the activation of the host immune system.
    Anticancer research 04/2014; 34(4):1663-71. · 1.71 Impact Factor
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    Tsukasa Kuwabara, Kenichi Yamaguchi
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    ABSTRACT: http://archive.org/details/interactionism20130517
    The Joint Journal of the National Universities in Kyushu, Education and Humanitie. 10/2013; 1(1):1-11.
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    ABSTRACT: To investigate the role of sulfate groups on the macrophage-stimulating activities of ascophyllan, we prepared desulfated ascophyllan, and its effects on RAW264.7 cells were compared with native ascophyllan. The chemical structural analysis revealed that nearly 21% of sulfate groups of ascophyllan were removed by desulfation reaction, while no significant changes in the molecular mass and monosaccharide composition occurred after desulfation. NO- and cytokine- (TNF-α and G-CSF) inducing activities of the desulfated ascophyllan on RAW264.7 cells were significantly decreased as compared to native ascophyllan. Furthermore, the activity of desulfated ascophyllan to induce reactive oxygen species (ROS) generation from RAW264.7 cells decreased to almost negligible level. Our results suggest that the level of sulfate groups of ascophyllan is an important structural element responsible for the macrophage-stimulating activities. Probably, even the limited removal of sulfate residues sensitive to desulfation reaction may result in significant decrease in the bioactivities of ascophyllan.
    Carbohydrate research 06/2013; 380C:124-129. · 2.03 Impact Factor
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    ABSTRACT: Antioxidant activities of sulfated polysaccharide ascophyllan from Ascophyllum nodosum was investigated in vitro by various assays, and compared with those of fucoidan. A chemiluminescence (CL) analysis using a luminol analog, L-012, showed that ascophyllan scavenges superoxide, and the activity is greater than fucoidan. However, in the presence of 10 μg/ml of ascophyllan or 10 μg/ml and 100 μg/ml of fucoidan, slightly enhanced CL-responses were observed. Since EDTA-treatment resulted in disappearance of the enhancement effects, it was suggested that metal ions especially iron ions in the polysaccharides might be involved in this phenomenon. In fact, metal element analysis revealed that ascophyllan and fucoidan inherently contain iron and other metal elements. EDTA-treatment resulted in significant increase in Fe2+-chelating activities of these polysaccharides. In an electron spin resonance (ESR)-spin trapping analysis in which direct UV-radiation to hydrogen peroxide was used as a source of hydroxyl radical, ascophyllan and fucoidan showed potent hydroxyl radical scavenging activity with similar extent. Reducing power of ascophyllan was stronger than that of fucoidan. Our results indicate that ascophyllan can exhibit direct and potent antioxidant activity.
    International journal of biological macromolecules 05/2013; · 2.37 Impact Factor
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    ABSTRACT: Lactate dehydrogenase (LDH)-release assay was applied to estimate the toxic potential of harmful algal species at the cellular level. African green monkey kidney (Vero), yellowtail fin epithelia (MJF), and rainbow trout gill (RTgill-W1) cells were used as target cells. A live cell suspension of Karenia mikimotoi (SUO-1) induced the release of LDH from these cell lines, while the activity of another strain, FUK, was much lower. The cell-free culture supernatants and ruptured cell suspensions of both strains of K. mikimotoi were less effective on LDH-release assay. Exposure experiments against abalone and shrimp revealed that SUO-1 showed much stronger lethal effects on these organisms than FUK. Among six phytoplankton species, three species known to be harmful algal species induced the release of LDH to different extents depending on the cell line, whereas the other three species, known to be non-toxic, showed no effects on any cell lines. These results suggest that LDH-release assay is a useful micro-plate assay for estimation of the toxic potential of harmful phytoplankton.
    Bioscience Biotechnology and Biochemistry 02/2013; · 1.27 Impact Factor
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    ABSTRACT: In this study, we investigated the bioactivity of ascophyllan in terms of reactive oxygen species (ROS) generation. In RAW264.7 cells, we found that ascophyllan induced ROS generation in a concentration-dependent manner, but with bell-shaped profile. Immunoblot analysis demonstrated that ascophyllan promoted the translocation of cytosolic subunits (p67phox and p47phox) of NADPH oxidase to the plasma membrane. Among mitogen activated protein (MAP) kinase inhibitors tested, JNK inhibitor showed the most potent inhibitory effect on ascophyllan-induced ROS production. Consistently, significant level of phosphorylated JNK MAP kinase was detected in ascophyllan-treated RAW264.7 cells. Our findings suggest for the first time that ascophyllan can stimulate macrophages to produce ROS through the activations of NADPH oxidase and JNK MAP kinase.
    International journal of biological macromolecules 09/2012; 52:164–169. · 2.37 Impact Factor
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    ABSTRACT: Splenic natural killer (NK) cell activity against YAC-1 cells increased in mice intraperitoneally injected with ascophyllan. Ascophyllan enhanced the cytotoxicity of RAW264.7 cells toward YAC-1 cells in a concentration-dependent manner. The cytotoxicity of ascophyllan-stimulated RAW264.7 cells as to YAC-1 cells was suppressed with N(G)-nitro-L-arginine methyl ester hydrochloride (L-NAME), an inhibitor of nitric oxide (NO) synthase, suggesting the involvement of NO in the cytotoxicity of ascophyllan-stimulated RAW264.7 cells.
    Bioscience Biotechnology and Biochemistry 08/2012; 76(8):1573-6. · 1.27 Impact Factor
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    ABSTRACT: A time-course analysis of reactive oxygen species (ROS) generation in fertilized eggs of the devil stinger (Inimicus japonicus) from 0 h post-fertilization (hpf) to the early larval stage indicated that the ROS level was highest in the 22 hpf embryo, and declined thereafter. Phorbol myristate acetate (PMA) had no effect on ROS generation by the 22 hpf embryo, whereas PMA significantly increased larval ROS generation, suggesting that the ROS generation mechanisms of the 22 hpf embryo and larva are different at least in terms of PMA-responsiveness. Our results suggest the presence of a specific ROS generation system in devil stinger embryo which can be transitionally activated during embryogenesis.
    Bioscience Biotechnology and Biochemistry 08/2012; 76(8):1561-4. · 1.27 Impact Factor
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    ABSTRACT: The antioxidant and macrophage-stimulating activities of polyguluronic acid (PG) and polymannuronic acid (PM) prepared from alginate were examined. A chemiluminescence (CL) method using a luminol analog, L-012, showed that both PM and PG scavenge superoxide produced by hypoxanthine-xanthine oxidase system in a concentration-dependent manner. At 100 μg/ml, PG showed slightly stronger superoxide scavenging activity than PM. In an electron spin resonance (ESR)-spin trapping method in which the Fenton reaction was used as hydroxyl radical generation system, we found that both PM and PG showed potent hydroxyl radical scavenging activity to a similar extent. Because PM and PG showed no chelating activity on Fe(2+), it was confirmed that PM and PG can directly scavenge hydroxyl radical. No significant scavenging activity of PM and PG toward hydrogen peroxide was observed. Interestingly, the macrophage-stimulation activity of PG as measured by nitric oxide (NO)-production from mouse macrophage cell line RAW264.7 cells was evidently stronger than that of PM. Our results suggest that RAW264.7 cells might be able to distinguish the conformational differences between PM and PG, and respond differently to them, whereas the effects of such structural differences between PM and PG on the radical scavenging activities may not be so significant.
    Carbohydrate research 02/2012; 352:88-93. · 2.03 Impact Factor
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    Zedong Jiang, Yoichiro Hama, Kenichi Yamaguchi, Tatsuya Oda
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    ABSTRACT: Porphyran, extracted from an edible red alga (Porphyra yezoensis), is a sulphated polysaccharide with a wide variety of biological activities including anti-tumour, antioxidant and immuno-modulating activities. In this study, we examined the effect of porphyran on nitric oxide (NO) production in mouse macrophage cell line RAW264.7 cells. Although no significant activity of porphyran to induce NO or tumour necrosis factor-α (TNF-α) production in RAW264.7 cells was observed at the concentration range tested (10-500 µg/ml), it was found for the first time that porphyran inhibited NO production and expression of inducible nitric oxide synthase (iNOS) in RAW264.7 cells stimulated with lipopolysaccharide (LPS). In the presence of 500 µg/ml porphyran, NO production and expression of iNOS in LPS-treated RAW264.7 cells were completely suppressed. On the other hand, porphyran showed only a marginal effect on the secretion of TNF-α from LPS-stimulated RAW264.7 cells. Electrophoretic mobility shift assay (EMSA) using infrared dye labelled oligonucleotide with nuclear factor-κB (NF-κB) consensus sequence suggested that porphyran inhibited the LPS-induced NF-κB activation. The LPS-inducible nuclear translocation of p65, and the phosphorylation and degradation of IκB-α were also inhibited by the pre-treatment with porphyran. Our results obtained in in vitro analysis suggest that porphyran suppresses NO production in LPS-stimulated macrophages by the blocking of NF-κB activation.
    Journal of Biochemistry 01/2012; 151(1):65-74. · 3.07 Impact Factor
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    ABSTRACT: Alginate is a natural acidic linear polysaccharide that is produced by brown seaweeds. It is currently used in a broad range of commercial enterprises, such as the food and medical products industries. Recent evidence has demonstrated that alginate oligosaccharides may function as growth promoting agents for certain plant cells, including those of some green algae. Chlamydomonas reinhardtii is a green alga that is used as a model organism in fundamental molecular biology studies; it is also a producer of biohydrogen. In the present study, we examined effects of two types of alginate oligosaccharide mixtures (AOMs), which were prepared by either enzymatic degradation (ED) or acid hydrolysis (AH), on the growth of C. reinhardtii. Growth was significantly promoted by AOM (ED) in a concentration-dependent manner. The maximum effect was observed on day 4 of treatment. The fatty acid composition of C. reinhardtii was also influenced by AOM (ED); the levels of C16:0, C18:2 cis and C18:3 n-3 increased in treated cells. AOM (AH) and the other saccharides that we tested did not affect the growth of C. reinhardtii. The effects that we identified could promote efficient biomass production by reducing culture times and by changing cellular fatty acid levels.
    Journal of Bioscience and Bioengineering 01/2012; 113(1):112-6. · 1.74 Impact Factor
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    Zedong Jiang, Takasi Okimura, Kenichi Yamaguchi, Tatsuya Oda
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    ABSTRACT: Ascophyllan isolated from the brown alga Ascophyllum nodosum is a fucose-containing sulfated polysaccharide, which has similar but distinct characteristic monosaccharide composition and entire chemical structure to fucoidan. In this study, we examined the effects of ascophyllan, fucoidan isolated from A. nodosum (A-fucoidan), and fucoidan from Sigma (S-fucoidan) as a representative fucoidan derived from other source (Fucus vesiculosus) on mouse macrophage cell line RAW264.7 cells. No significant cytotoxic effects of ascophyllan and A-fucoidan on RAW264.7 cells were observed up to 1000μg/ml, while S-fucoidan showed cytotoxic effect in a concentration-dependent manner. Ascophyllan induced extremely higher level of nitric oxide (NO) production from RAW264.7 cells than those induced by fucoidans over the concentration range tested (0-200μg/ml). Reverse transcription polymerase chain reaction (RT-PCR) and western blot analysis revealed that expression level of inducible NO synthase (iNOS) in ascophyllan-treated RAW264.7 cells was much higher than the levels detected in the cells treated with fucoidans. Furthermore, the activities of ascophyllan to induce the secretion of tumor necrosis factor-α (TNF-α) and granulocyte colony-stimulating factor (G-CSF) from RAW264.7 cells were also greater than those induced by fucoidans especially at lower concentration range (3.1-50μg/ml). The activities of ascophyllan to induce NO and cytokine production in mouse peritoneal macrophages were also stronger than those of fucoidans. Electrophoretic mobility shift assay (EMSA) using infrared dye labeled nuclear factor-kappa B (NF-κB) and AP-1 consensus sequences suggested that ascophyllan can strongly activate these transcription factors. Marked increase in the nuclear translocation of p65, and the phosphorylation and degradation of IκB-α were also observed in ascophyllan-treated RAW264.7 cells. Analysis using mitogen-activated protein (MAP) kinase inhibitors and western blot analysis suggested that c-Jun N-terminal kinase (JNK) and p38 MAP kinase are mainly involved in ascophyllan-induced NO production.
    Nitric Oxide 11/2011; 25(4):407-15. · 3.27 Impact Factor
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    ABSTRACT: The effects of cytotoxic lectins, modeccin and phytohemagglutinin (PHA) on mouse macrophage cell line RAW264.7 was studied by detecting the induction of inflammatory mediators. Results showed that modeccin induced the release of tumor necrosis factor-α (TNF-α) from RAW264.7 cells with a bell-shape concentration-dependent profile. PHA that showed no significant cytotoxicity on RAW264.7 cells up to 100,000 ng/ml induced much higher level of TNF-α than modeccin. PHA simultaneously induced the secretion of granulocyte colony stimulation factor (G-CSF) from RAW264.7 cells with even much higher level than that of TNF-α, whereas modeccin did not. Furthermore, PHA induced the secretion of nitric oxide (NO) in RAW264.7 cells, while no significant level of NO was detected in the modeccin-treated cells. NH₄Cl (a lysomotoropic agent) and cycloheximide (a ribosome inhibitor) strongly inhibited modeccin-induced TNF-α secretion, but no significant inhibitory effects of these reagents on the PHA-induced TNF-α secretion were observed. Contrary to modeccin-induced TNF-α secretion, even slightly increased TNF-α secretion was observed in PHA-treated cells in the presence of 10 mM NH₄Cl. In addition, the inhibition profiles of modeccin-induced TNF-α secretion by various kinase inhibitors were different from those of PHA. These results suggested that the action mode of modeccin to stimulate RAW264.7 cells leading to the secretion of inflammatory molecules, including TNF-α, is distinct from that of PHA. On the other hand, significantly increased translocation of activator protein-1 (AP-1), a crucial transcription factor involved in expression of inflammatory molecules, into nucleus was observed in RAW264.7 cells treated with PHA and modeccin.
    Acta Biochimica et Biophysica Sinica 01/2011; 43(1):52-60. · 1.81 Impact Factor
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    ABSTRACT: We used bi-algal culture experiments to investigate and verify the roles of growth interaction between Heterocapsa circularisquama and Prorocentrum dentatum in monospecific bloom formation. Growth of H. circularisquama was slightly inhibited when inoculated at 102 cells mL–1 along with P. dentatum at 104 cells mL–1. In other combinations of inoculation densities, P. dentatum density rapidly decreased to extremely low levels in the presence of H. circularisquama. We used a mathematical model to simulate growth and interactions of H. circularisquama and P. dentatum in bi-algal cultures. The model indicates that one species will always inhibit the growth of the other and that the relative initial cell densities of the species are critical in determining the outcome. When cultured together under conditions without cell contact, growth of H. circularisquama and P. dentatum was not inhibited. As with P. dentatum, the growth of Heterosigma akashiwo and Skeletonema costatum was inhibited in intact cell suspensions with H. circularisquama, but a nontoxic species, Heterocapsa triquetra, did not affect the growth of P. dentatum or the other species. Similarly, cell suspensions of H. circularisquama showed hemolytic activity toward rabbit erythrocytes, but those of H. triquetra did not. In addition, the cell-free supernatant of H. circularisquama cultures showed no significant hemolytic activity. These results suggest that H. circularisquama causes lethality in P. dentatum by direct cell contact in which live-cell-mediated hemolytic activity might be a contributing factor.
    Journal of Sea Research 01/2011; · 1.86 Impact Factor
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    ABSTRACT: Heterocapsa circularisquama showed much higher toxic effects on short-necked clams than Chattonella marina. Clams exposed to H. circularisquama exhibited morphological changes concomitant with an accumulation of mucus-like substances in the gills, a profound reduction in filtration activity, and lysosomal destabilization in hemocytes. Chattonella marina was less effective than H. circularisquama, and Heterocapsa triquetra was almost harmless in all these criteria. These results suggest that H. circularisquama exerted its lethal effect on short-necked clams through gill tissue damage and subsequent induction of physiological stress.
    Bioscience Biotechnology and Biochemistry 01/2011; 75(10):2052-5. · 1.27 Impact Factor
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    ABSTRACT: The biological toxic potentials of aqueous extracts from the dinophycean flagellates Gymnodinium impudicum and Alexandrium affine and the raphidophycean flagellate Chattonella ovata were examined in both in vitro and in vivo systems. Interestingly, the extract from A. affine was the only one that showed potent cytotoxicities towards HeLa, Vero, and Neuro-2a cells in a concentration-dependent manner. Mice given intraperitoneal injections of the extracts revealed that none of the extracts exhibited serious toxicities in mice. However, temporal body weight loss was observed in the mice injected with the extract from A. affine during the early stage, and the dramatic enlargement of spleens was also observed in the mice on the 7th day after injection. Since A. affine extract showed potent hemolytic activity in vitro towards mouse erythrocytes, hemolytic anemia may be a possible mechanism responsible for the splenomegaly in the mice injected with A. affine extract. Similar marginal effects were observed in the mice injected with the extract from C. ovata; however, no significant toxic or detrimental effects were detected in the mice injected with the extract from G. impudicum. These results suggest that the extract from G. impudicum may not be contaminated with detectable levels of biologically hazardous compounds and may be relatively safe compared with the other two extracts.
    The Journal of Toxicological Sciences 08/2010; 35(4):591-9. · 1.38 Impact Factor
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    ABSTRACT: The effects of fucose-containing sulfated polysaccharides, ascophyllan and fucoidan, isolated from the brown alga Ascophyllum nodosum, on the growth of various cell lines (MDCK, Vero, PtK(1), CHO, HeLa, and XC) were investigated. In a colony formation assay, ascophyllan and fucoidan showed potent cytotoxic effects on Vero and XC cells, while other cell lines were relatively resistant to these polysaccharides. Almost no significant effects of these polysaccharides were observed in the cell lines tested using the Alamar blue cytotoxicity assay over 48 h with varying initial cell densities (2500-20,000 cells/well) in growth medium. Interestingly, a significant growth promoting effect of ascophyllan on MDCK cells was observed, whereas treatment with fucoidan showed growth suppressive effects on this cell line under the same experimental conditions. These results suggest that ascophyllan is distinguishable from fucoidan in terms of their bioactivities. This is the first report of the growth promoting effects of a sulfated fucan on a mammalian cell line under normal growth conditions.
    Journal of Bioscience and Bioengineering 07/2010; 110(1):113-7. · 1.74 Impact Factor
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    ABSTRACT: We investigated the effects of two strains (SUO-1 and FUK) of the dinoflagellate Karenia mikimotoi on the rotifer Brachionus plicatilis. The SUO-1 strain was highly toxic to rotifers, whereas the FUK strain was less toxic. After 10-h incubations, the survivorship of rotifers exposed to SUO-1 and FUK was 20% and 95%, respectively. Both the cell-free culture supernatant and the ruptured cell suspension prepared from these strains were not toxic to rotifers. Furthermore, when direct contact between K. mikimotoi and rotifers was interrupted with a cell-impermeable membrane (3-μm pores), the toxicity of both the SUO-1 and FUK strains of K. mikimotoi to rotifers were completely inhibited even after a 24-h exposure. Cell suspensions of SUO-1 showed hemolytic activity toward horse erythrocytes, but the FUK strain did not. The cell-free supernatant and the ruptured cell suspension of SUO-1 showed no significant hemolytic activity. These results suggest that this highly toxic strain of K. mikimotoi causes lethality in rotifers by direct contact in which live cell-mediated hemolytic activity might be a contributing factor.
    Harmful Algae 05/2010; · 2.90 Impact Factor
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    ABSTRACT: An N-acetylgalactosamine (GalNAc)-specific Ca(2+)-dependent lectin (C-type lectin), isolated from the marine invertebrate Holothuroidea (Cucumaria echinata), CEL-I, showed potent mitogenic activity toward normal mouse spleen cells. The mitogenic activity of CEL-I, which reached a maximum at 100 microg/ml, was inhibited by GalNAc in a concentration-dependent manner. The mitogenic effect of CEL-I at 10 microg/ml on T cell- enriched splenocytes was at a similar level due to a well-known T cell mitogen, concanavalin A (Con A), at 10 microg/ml. Furthermore, CEL-I evoked a mitogenic response from nude mouse spleen cells, while no significant effects of Con A on this cell population were observed over a wide range of concentrations. These results suggest that CEL-I is a potent mitogenic lectin with the ability to stimulate both T and B cells.
    Bioscience Biotechnology and Biochemistry 01/2010; 74(8):1613-6. · 1.27 Impact Factor

Publication Stats

375 Citations
102.50 Total Impact Points

Institutions

  • 2003–2013
    • Nagasaki University
      • • Department of Biochemistry
      • • Department of Science and Technology
      Nagasaki-shi, Nagasaki-ken, Japan
  • 2012
    • National Fisheries University
      Simonoseki, Yamaguchi, Japan
  • 2009–2012
    • Korea Basic Science Institute KBSI
      • Jeju Center
      Seoul, Seoul, South Korea
  • 2008
    • Cheju Halla University
      Tse-tsiu, Jeju, South Korea
  • 2007
    • National Research Institute of Fisheries and Environment of Inland Sea
      Hirosima, Hiroshima, Japan
  • 2004
    • Kyushu University
      Hukuoka, Fukuoka, Japan