Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Volume 71 , Issue 8
Showing 1-44 articles out of 44 articles from the selected issue
Award Review
  • Noriyuki YANAKA
    2007 Volume 71 Issue 8 Pages 1811-1818
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    Bacterial glycerophosphodiester phosphodiesterases (GP-PDEs), GlpQ and UgpQ, are well-characterized periplasmic and cytosolic proteins that play critical roles in the hydrolysis of deacylated glycerophospholipids to glycerol phosphate and alcohol, which are utilized as major sources of carbon and phosphate. In contrast, two novel mammalian GP-PDEs, GDE1/MIR16 and GDE3, were recently identified, and were shown to be involved in several physiological functions. GDE1/MIR16 was identified as a membrane protein interacting with RGS16, a regulator of G protein signaling, and found to hydrolyze glycerophosphoinositol preferentially. We have found that expression of GDE3 is significantly up-regulated during osteoblast differentiation and is involved in morphological changes of cells. Furthermore, five mammalian GP-PDEs were virtually identified, and very recent studies indicate that retinoic acid-induced expression of GDE2 plays essential roles in neuronal differentiation and neurite outgrowth. Thus mammalian GP-PDEs are likely to be important in controlling numerous cellular events, indicating that the GP-PDE superfamily in mammals might be a pharmacological target in the future.
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Analytical Chemistry Regular Paper
  • Ashraf AHMAD, Jahan GHASEMI
    2007 Volume 71 Issue 8 Pages 1970-1978
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    Real-time PCR is a potent technique for nucleic acid quantification for research and diagnostic purposes, the wide dynamic range being one of the advantages over other techniques like the microarray. Several additives and enhancers have been studied to expand the PCR dynamic range in order to be more efficient in quantifying low quantities of nucleic acids, increase the yield and improve reaction efficiency. Shown here is that a combination of new buffers with the regularly used Tris buffer makes it possible to expand the real-time PCR dynamic range and to improve the efficiency and correlation coefficient. Mixing HEPES, TEA or MOPS with Tris was more efficient than Tris alone. It was also found that, if the pH value of the Tris buffer was calibrated with phosphoric acid instead of hydrochloric acid, then the dynamic range was significantly improved and low quantities could be detected and quantified more efficiently. Mixing more than one compound with the Tris buffer was also effective for expanding the dynamic range and increasing the efficiency and correlation coefficient in quantitative real-time PCR.
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Organic Chemistry Regular Papers
Organic Chemistry Notes
  • Tzong-Huei LEE, Ming-Hsi YEH, Chi-I CHANG, Ching-Kuo LEE, Yi-Yuan SHAO ...
    2007 Volume 71 Issue 8 Pages 2075-2078
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    Two new lignans, including the aryltetralin-type lignan, lanceoline (1), and the diaryl butyrolactone-type lignan, 5-methoxytrachelogenin (2), together with 5-methoxywikstromol (3), were isolated from the low-polar layer of a heartwood extract of Cunninghamia lanceolata as their acetylated derivatives 1a, 2a and 3a, respectively, and were identified by spectroscopic analyses. The 13C-NMR data for 3a are reported for the first time in this paper.
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  • Mei DONG, Man-Li ZHANG, Yu-Fang WANG, Xi-Ping ZHANG, Cun-Fang LI, Qing ...
    2007 Volume 71 Issue 8 Pages 2087-2090
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    Three novel taxanes with the 10-alkoxy group were isolated from heartwood of Taxus cuspidata. The structures were identified as 2α,14β-diacetoxy-10β-ethoxytaxa-11,4(20)-dien-5α-ol (1), 2α,14β-diacetoxy-10β-methoxytaxa-11,4(20)-dien-5α-ol (2), and 2α-acetoxy-10β-ethoxytaxa-11,4(20)-diene-5α,14β-diol (3) on the basis of spectroscopic analysis. These are the first taxanes with an alkoxy moiety on the skeleton.
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Biochemistry & Molecular Biology Regular Papers
  • Ryoji MITSUI, Seiho SHINYA, Yuichi ICHIYAMA, Kenta KUDO, Takuo TSUNO, ...
    2007 Volume 71 Issue 8 Pages 1858-1864
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    Bacillus cereus 809A and Burkholderia sp. 711C were isolated from soil. These strains demonstrate hydrolysis activity towards prochiral 2-phenyl-1,3-propanediol diacetate and accumulated the corresponding chiral monoacetates into the reaction mixture. When 2-phenyl 1,3-propanediol diacetate was used as a substrate, the produced monoacetates with Burkholderia sp. 711C were obtained in a racemic form but that produced by Bacillus cereus 809A showed an excess of the (S)-form. The resting cell reaction revealed that for Bacillus cereus 809A, there was an enrichment of one of the enantiomers of the monoacetate such that the enantiomeric excess (e.e.) of the (S)-form was over 95%. The purified enzyme from Bacillus cereus 809A hydrolyzed diacetate to monoacetate, and the e.e. value of the (S)-form increased by prolonged reaction in a way similar to the resting cell reaction. From N-terminal amino acids, this esterase is conserved in some strains of Bacillus for which the genomic sequences have been reported.
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  • Tohru SAEKI, Yuki MUNETAKA, Kazumitsu UEDA, Kimikazu IWAMI, Ryuhei KAN ...
    2007 Volume 71 Issue 8 Pages 1865-1872
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    Although ileal and hepatic Na+-dependent bile acid transporters (SLC10A2 and SLC10A1 respectively) share structural similarities, the mutation of conserved amino acids often has distinct effects on them. We have identified two Cys residues in mouse Slc10a2 (Cys51 and Cys106) the replacement of which by Ala remarkably reduces taurocholic acid (TCA) transport. Although Cys51 is conserved in Slc10a1 as Cys44, Ala substitution gave no apparent difference in TCA uptake. Here, we further analyzed the kinetics of TCA uptake and cell surface localization of these mutants. The C51A and C106A mutants of Slc10a2 showed significantly reduced TCA uptake, while no apparent difference in TCA uptake was observed for the Slc10a1-C44A mutant. The Km values for TCA uptake by these mutants were comparable, suggesting that these residues are not involved in the interaction with TCA.
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  • Shizuko TACHIBANA, Maki HIRANO, Takashi HIRATA, Michinori MATSUO, Ikuo ...
    2007 Volume 71 Issue 8 Pages 1886-1895
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    In this study we analyzed functions of ATP-binding cassette (ABC) transporters involved in sterol transport from Caco-2 cells. Treatment with a synthetic liver x receptor ligand elevated both mRNA and protein levels of ABCG5, G8, and ABCA1. The ligand stimulated cholesterol efflux, suggesting that ABC transporters are involved in it. To identify the acceptors of cholesterol, potential molecules such as apolipoprotein A-I, glycocholic acid, phosphatidylcholine, and bile acid micelles were added to the medium. Apo A-I, a known acceptor of cholesterol transported by ABCA1, elevated cholesterol efflux on the basal side, whereas the others raised cholesterol efflux on the apical side. Moreover, bile acid micelles preferentially augmented plant sterol efflux rather than cholesterol. Finally, in HEK293 cells stably expressing ABCG5/G8, bile acid micelle-mediated sterol efflux was significantly accelerated. These results indicate that ABCG5/G8, unlike ABCA1, together with bile acids should participate in sterol efflux on the apical surface of Caco-2 cells.
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  • Akari INADA, Hidenori ARAI, Kojiro NAGAI, Jun-ichi MIYAZAKI, Yuichiro ...
    2007 Volume 71 Issue 8 Pages 1920-1926
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    Few studies have been done to examine gender differences in diabetic mouse models. Here we examined a gender difference in Inducible cAMP Early Repressor (ICER) transgenic (Tg) mice, a diabetic mouse model. Longitudinal changes in diabetes and nephropathy were investigated in male and female Tg mice. Both male and female Tg mice developed severe diabetes early in life due to severely impaired insulin synthesis and decreased β-cell numbers, but only female Tg mice became less hyperglycemic later in life, and most female Tg mice did not develop diabetic nephropathy. Even some female Tg mice that remained hyperglycemic showed less renal expansion than age-matched male Tg mice. Thus the gender difference in the severity of diabetes and diabetic nephropathy was evident with age in this model. This study indicates that sex hormones may play a role in glucose metabolism in diabetic conditions.
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  • Atsushi TERADA, Takeshi YOSHIDA, Makoto KIMURA
    2007 Volume 71 Issue 8 Pages 1940-1945
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    Ribonuclease P (RNase P) is involved in the processing of the 5′ leader sequence of precursor tRNA (pre-tRNA). We have found that RNase P RNA (PhopRNA) and five proteins (PhoPop5, PhoRpp21, PhoRpp29, PhoRpp30, and PhoRpp38) reconstitute RNase P activity with enzymatic properties similar to those of the authentic ribozyme from the hyperthermophilic archaeon Pyrococcus horikoshii OT3. We report here that nucleotides A40, A41, and U44 at helix P4, and G269 and G270 located at L15/16 in PhopRNA, are, like the corresponding residues in Esherichia coli RNase P RNA (M1RNA), involved in hydrolysis by coordinating catalytic Mg2+ ions, and in the recognition of the acceptor end (CCA) of pre-tRNA by base-pairing, respectively. The information reported here strongly suggests that PhopRNA catalyzes the hydrolysis of pre-tRNA in approximately the same manner as eubacterial RNase P RNAs, even though it has no enzymatic activity in the absence of the proteins.
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  • J. Joe HULL, Ryosuke KAJIGAYA, Kiyohiro IMAI, Shogo MATSUMOTO
    2007 Volume 71 Issue 8 Pages 1993-2001
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    In most female moths, pheromone biosynthesis activating neuropeptide (PBAN) regulates sex pheromone production by stimulating an influx of extracellular Ca2+. Little is known about the plasma membrane channel or how the PBAN stimulus is communicated to the channel. Fluorescent Ca2+ imaging techniques confirmed PBAN-induced Ca2+ influx in the silkworm, Bombyx mori, and showed that the PBAN response is reduced with repeated stimulation. Compounds known to impact Ca2+ signaling were examined for their effects on sex pheromone production. These experiments demonstrated that the PBAN signal is likely mediated by a store-operated channel (SOC). SOC blockers, SKF-96365 and 2-aminoethoxydiphenyl borate, abolished sex pheromone production, as did flufenamic acid, a blocker of transient receptor potential (TRP) channels. Thapsigargin mimicked the pheromonotropic effects of PBAN. Similar results were seen when PBAN-induced lipase activity was assayed. Conversely, 1-oleoyl-2-acetyl-sn-glycerol and arachidonic acid, activators of diacylglycerol-dependent Ca2+ channels, had no effect on bombykol production.
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  • Siew Ping LIM, Niran ROONGSAWANG, Kenji WASHIO, Masaaki MORIKAWA
    2007 Volume 71 Issue 8 Pages 2002-2009
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    Fluorescent Pseudomonas sp. MIS38 produces a cyclic lipopeptide, arthrofactin. Arthrofactin is synthesized by a unique nonribosomal peptide synthetase (NRPS) with dual C/E-domains. In this study, another class of cyclic peptide, pyoverdine, was isolated from MIS38, viz., Pvd38. The main fraction of Pvd38 had an mz value of 1,064.57 and contained Ala, Glu, Gly, (OHOrn), Ser, and Thr at a ratio of 2:1:1:(1):1:1 in the peptide part, suggesting a new structure compound. A gene encoding NRPS for the chromophore part of Pvd38 was identified, and we found that it contained a conventional E-domain. Gene disruption completely impaired the production of Pvd38, demonstrating that the synthetase is functional. This observation allows us to conclude that different NRPS systems with dual C/E-domains (in arthrofactin synthetase) and a conventional E-domain (in pyoverdine synthetase) are both functional in MIS38.
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  • Verawat CHAMPREDA, Pattanop KANOKRATANA, Rutchadaporn SRIPRANG, Sutipa ...
    2007 Volume 71 Issue 8 Pages 2010-2020
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    An endophytic fungus, Fusicoccum sp. BCC4124, showed strong amylolytic activity when cultivated on multi-enzyme induction enriched medium and agro-industry substrates. α-Amylase and α-glucosidase activities were highly induced in the presence of maltose and starch. The purified target α-amylase, Amy-FC1, showed strong hydrolytic activity on soluble starch (kcat/Km=6.47×103 min−1(ml/mg)) and selective activity on γ- and β-cyclodextrins, but not on α-cyclodextrin. The enzyme worked optimally at 70 °C in a neutral pH range with t1⁄2 of 240 min in the presence of Ca2+ and starch. Maltose, matotriose, and maltotetraose were the major products from starch hydrolysis but prolonged reaction led to the production of glucose, maltose, and maltotriose from starch, cyclodextrins, and maltooligosaccharides (G3–G7). The amylase showed remarkable glucose tolerance up to 1 M, but was more sensitive to inhibition by maltose. The deduced protein primary structure from the putative gene revealed that the enzyme shared moderate homology between α-amylases from Aspergilli and Lipomyces sp. This thermotolerant, glucose tolerant maltooligosaccharide-forming α-amylase is potent for biotechnological application.
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  • Yohtaro SAITO, Hiroki ASHIDA, Chojiro KOJIMA, Haruka TAMURA, Hiroyoshi ...
    2007 Volume 71 Issue 8 Pages 2021-2028
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    The product of the mtnA gene of Bacillus subtilis catalyzes the isomerization of 5-methylthioribose 1-phosphate (MTR-1-P) to 5-methylthioribulose 1-phosphate (MTRu-1-P). The catalysis of MtnA is a novel isomerization of an aldose phosphate harboring a phosphate group on the hemiacetal group. This enzyme is distributed widely among bacteria through higher eukaryotes. The isomerase reaction analyzed using the recombinant B. subtilis enzyme showed a Michaelis constant for MTR-1-P of 138 μM, and showed that the maximum velocity of the reaction was 20.4 μmol min−1 (mg protein)−1. The optimum reaction temperature and reaction pH were 35 °C and 8.1. The activation energy of the reaction was calculated to be 68.7 kJ mol−1. The enzyme, with a molecular mass of 76 kDa, was composed of two subunits. The equilibrium constant in the reversible isomerase reaction [MTRu-1-P]/[MTR-1-P] was 6. We discuss the possible reaction mechanism.
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  • Kazushi KADOMURA, Sayaka NARUSE, Shin SUGIHARA, Kenichi YAMAGUCHI, Tat ...
    2007 Volume 71 Issue 8 Pages 2029-2033
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    Previous studies have indicated that the devil stinger produces reactive oxygen species (ROS) during early development from fertilized egg to larva. To determine whether ROS generation is a common feature in marine fish species, we conducted chemiluminescence analysis using ROS specific probe (L012) on larvae of six marine fish species. Marbled rockfish, black rockfish, and devil stinger showed higher levels of chemiluminescence response (CR), whereas the levels of CR of sevenband grouper, tiger puffer, and red seabream were fairly lower. These CRs were inhibited by the addition of superoxide dismutase. Hypersensitive photon-counting microscopic observation of black rockfish suggested that ROS production was concentrated in the head area. Our results suggest that the larvae of these six marine fishes produce ROS to considerably different extents depending on species, and that rockfish species, belonging to ovoviviparous fish, tend to produce much higher levels of ROS especially at the later larval stage.
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Biochemistry & Molecular Biology Notes
Biochemistry & Molecular Biology Communications
  • Tsuyoshi NAKAGAWA, Takamasa SUZUKI, Satoko MURATA, Shinya NAKAMURA, Ta ...
    2007 Volume 71 Issue 8 Pages 2095-2100
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    We made a series of improved Gateway binary vectors (pGWBs) for plant transformation. Fifteen different reporters and tags, sGFP, GUS, LUC, EYFP, ECFP, G3GFP, mRFP, 6×His, FLAG, 3×HA, 4×Myc, 10×Myc, GST, T7, and TAP, were employed. Some vectors carry the 2×35S-Ω promoter for higher-level expression. The kanamycin- and hygromycin-resistant markers are independently available for each of the 43 types of vectors, thus an additional transformation of once-transformed plants can be carried out easily. Their small size and high-copy number in Escherichia coli make possible easier handling at plasmid preparation and sequencing. Improved pGWBs should be a powerful tool for transgenic research in plants.
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  • Mamoru NISHIMOTO, Motomitsu KITAOKA
    2007 Volume 71 Issue 8 Pages 2101-2104
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    A one-pot enzymatic reaction to produce lacto-N-biose I (LNB), which is supposed to represent the bifidus factor in human milk oligosaccharides, was demonstrated. Approximately 500 mM of LNB was generated in 10-liter of reaction mixture initially containing 660 mM of sucrose and 600 mM of GlcNAc by the concurrent actions of four enzymes, sucrose phosphorylase, UDP-glucose—hexose-1-phospate uridylyltransferase, UDP-glucose 4-epimerase, and lacto-N-biose phosphorylase, in the presence of UDP-Glc and phosphate, indicating a reaction yield of 83%. LNB was isolated from the mixture by crystallization after yeast treatment. Finally, 1.4 kg of LNB of 99.6% purity was recovered after recrystallization.
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Food & Nutrition Science Regular Papers
  • Takamitsu CHAKI, Noriyoshi KAJIMOTO, Hiroshi OGAWA, Takashi BABA, Nozo ...
    2007 Volume 71 Issue 8 Pages 1819-1825
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    Sodium alginate oligosaccharides (NaAOs) consisting of a mixture of eight oligosaccharides have previously been reported to lower blood pressure. We investigated in this study the excretion of NaAOs into the urine or feces, and attempted to elucidate the mechanism for lowering blood pressure by using isolated mesenteric arteries from the rabbit. The recovery rate of P8, which is the main component of NaAOs, was 5.2% and 58.9% over 48 hours in the urine and feces, respectively. The mechanism for lowering blood pressure appeared to be NaAOs having calcium antagonist activity, especially voltage-operated calcium channels. Our results suggest that NaAOs are substantially excreted into the feces, although some of them may be absorbed internally, exerting antagonist activity towards the calcium channels, especially voltage-operated calcium channels.
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  • Moussa NDONG, Mariko UEHARA, Shinichi KATSUMATA, Shigeru SATO, Kazuhar ...
    2007 Volume 71 Issue 8 Pages 1826-1833
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    The effects of Moringa oleifera (MO), Moringaceae on hyperlipidemia and hepatocyte ultrastructural changes caused by iron deficiency were investigated. Four-week-old male Wistar-strain rats were fed a control diet based on AIN-93G (C), an iron deficient diet (FeD), a FeD + 0.5% MO (FeD-m) diet, or a FeD + MO 1% (FeD-M) diet for 4 weeks. It was found that MO reduced iron-deficient diet-induced increases in serum and hepatic lipids with dose-dependent increases of serum quercetin and kaempherol, but did not prevent anemia. By electron microscopy, in iron deficient hepatocytes, slightly swollen mitochondria and few glycogen granules were observed, but glycogen granules increased and mitochondria were normalized by treatment with MO. Furthermore, lipoproteins were observed in the Golgi complex under treatment with MO. These results suggest a possible beneficial effect of MO in the prevention of hyperlipidemia and ultrastructural changes in hepatocytes due to iron-deficiency.
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  • Guei-Jane WANG, Shyr-Yi LIN, Wen-Chun WU, Wen-Chi HOU
    2007 Volume 71 Issue 8 Pages 1873-1878
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    Wild, liquid state culture and solid state culture of Taiwanofungus camphoratus (Chang-chih) were sequentially extracted with cold water, methanol, and hot water to get cold water soluble, methanol soluble, and hot water soluble extracts respectively. The extracts from three Chang-chih were used to determine 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, semicarbazide sensitive amine oxidase inhibitory, and cytotoxic activities against B16-F10 and HT-1080 cell lines. It was found that extracted fractions from three Chang-chih exhibited the different levels of biological activities.
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  • Yumi AOKI, Tomoko TANIGAWA, Hiroko ABE, Yoko FUJIWARA
    2007 Volume 71 Issue 8 Pages 1879-1885
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    To investigate the new physiological functions of oolong tea, the effects on melanogenesis were studied. An oolong tea extract inhibited melanogenesis without affecting cell growth in B16 mouse melanoma cells. However, the oolong tea extract hardly showed any inhibitory effect on mushroom tyrosinase in a cell-free system. The effects of an oolong tea extract on the intracellular tyrosinase level in B16 cells were therefore studied. All the levels of activity, protein and mRNA were decreased in the oolong tea extract-treated cells. We also investigated the inhibitory effects of oolong tea on the pigmentation induced by ultraviolet B (UVB) by using brownish guinea pigs in vivo. The number of 3,4-dihydroxyphenylalanine (DOPA)-positive melanocytes increased by UVB was repressed by an oral administration of oolong tea. These results imply that oolong tea might be effective in whitening and that its inhibitory effect on melanogenesis was involved in the decrease of intracellular tyrosinase at the mRNA level.
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  • Tsuyoshi TSUZUKI, Akira SHIBATA, Yuki KAWAKAMI, Kiyotaka NAKAGAYA, Ter ...
    2007 Volume 71 Issue 8 Pages 1902-1910
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    The anti-angiogenic effects of conjugated docosahexaenoic acid (CDHA), which was prepared by an alkaline treatment of docosahexaenoic acid and contained conjugated double bonds, were investigated in vitro and in vivo. CDHA inhibited tube formation by the bovine aortic endothelial cell (BAEC), and also inhibited the proliferation of BAEC at a concentration of CDHA that suppressed tube formation, but did not influence cell migration. The inhibition of BAEC growth caused by CDHA was accompanied by a marked change in cellular morphology. Nuclear condensation and brightness were observed in Hoechst 33342-stained cells treated with CDHA, indicating that CDHA induced apoptosis in BAEC. We also evaluated the angiogenesis inhibition of CDHA in vivo. The vessel formation which was triggered by tumor cells was clearly suppressed in mice orally given CDHA. Our findings suggest that CDHA has potential use as a therapeutic dietary supplement for minimizing tumor angiogenesis.
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  • Yoshiaki MIYAKE, Mika MOCHIZUKI, Miki OKADA, Masanori HIRAMITSU, Yasuj ...
    2007 Volume 71 Issue 8 Pages 1911-1919
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    Phenolic glucosides having radical scavenging activity were examined from the fraction eluted with 20% methanol on Amberlite XAD-2 resin applied to lemon (Citrus limon) juice by using reversed phase chromatography. Four phenolic glucosides were identified as 1-feruloyl-β-D-glucopyranoside, 1-sinapoyl-β-D-glucopyranoside, 6,8-di-C-glucosylapigenin and 6,8-di-C-glucosyldiosmetin by 1H-NMR, 13C-NMR, and MS analyses. They exhibited radical scavenging activity for 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide, although the activity was low in comparison with eriocitrin, a potent antioxidant in lemon fruit, and the eriodictyol of its aglycone. The phenolic compounds in lemon juice were examined for their suppressive effect on the expression of blood adhesion molecules by measuring the expression of intercellular adhesion molecule-1 (ICAM-1) in human umbilical vein endothelial cells (HUVECs) induced by necrosis factor-α (TNF-α). 6,8-Di-C-glucosylapigenin, apigenin, and diosmentin of the flavones were found to significantly suppress the expression of ICAM-1 at 10 μM (P<0.05). The phenolic glucosides isolated in this study were contained in comparative abundance in daidai (Citrus aurantium) and niihime (Citrus unshiu × Citrus tachibana) among the sour citrus juices.
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  • Shuichi SEGAWA, Yoshihiro TAKATA, Yoshihisa WAKITA, Takafumi KANEKO, H ...
    2007 Volume 71 Issue 8 Pages 1955-1962
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    The clinical effects of an oral administration of a hop water extract (HWE) on the improvement of Japanese cedar pollinosis (JCPsis) symptoms were investigated. In a double-blind, placebo-controlled trial, 39 subjects took a drink containing either 100 mg of HWE or a placebo for 12 weeks during the pollen season. Nasal symptoms (sneezing attacks, nasal discharge, and nasal obstruction) were assessed from the subjects’ diaries. A clinical examination and blood sampling were carried out before and 4, 8 and 12 weeks after the initiation of treatment. As a result, a significant difference was observed in the symptom score and in the symptom-medication score 10 weeks after the intervention in comparison with the placebo group. Improvements were observed in nasal swelling, nasal color, amount of nasal discharge, and characteristics of nasal discharge in the intervention group 12 weeks after the treatment. No significant eosinophil infiltration into the nasal discharge was apparent in the intervention group throughout the study period, although it was observed in the placebo group. These findings indicate that an oral administration of HWE may be effective in alleviating the allergic symptoms related to JCPsis.
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  • Yeon Suk JUNG, Shin-ei MATSUMOTO, Makiko YAMASHITA, Kosuke TOMIMATSU, ...
    2007 Volume 71 Issue 8 Pages 1963-1969
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    We have established an in vitro immunization protocol whereby human peripheral blood mononuclear cells (PBMCs) are initially treated with L-leucyl-L-leucine methyl ester (LLME) and subsequently sensitized with antigen in the presence of interleukin (IL)-2, IL-4, and adjuvant. This protocol resulted in the production of antigen-specific antibodies. PBMCs are potentiated to react with exogenous antigens upon treatment with LLME. We are using this system to investigate the immunomodulatory activity of additives. In the present study, we aimed to evaluate the immunomodulatory activity of Propionibacterium acnes (P. acnes), which is known to exhibit various immunomodulatory effects in murine models, using this in vitro immunization protocol. P. acnes was found to augment the production of antigen-specific antibodies by PBMC, possibly through increased production of inflammatory cytokines and/or increased T-B cell interaction. P. acnes hence appears to act as an adjuvant in the antibody response in in vitro immunization.
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  • Tsuyoshi TSUZUKI, Ikuo IKEDA
    2007 Volume 71 Issue 8 Pages 2034-2040
    Published: August 23, 2007
    Released: August 23, 2007
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    We have previously shown that the 9c,11t-conjugated linoleic acid (CLA) concentration was always significantly higher than the 10t,12c-CLA concentration following the administration of these compounds to mice and rats, and considered that structural differences between the conjugated double bonds in these isomers affected absorption in the small intestine. This study investigates the absorption of CLA in the rat intestine by a lipid absorption assay of lymph from the thoracic duct. In Study 1, we used safflower oil and a triacylglycerol form of CLA (CLA-TG), while in Study 2, we used 9c,11t-CLA and 10t,12c-CLA. The cumulative recovery of CLA was lower than that of linoleic acid until two hours after sample administration. There was no difference in the extent of lymphatic recovery of 9c,11t-CLA and 10t,12c-CLA after the administration of CLA-TG, 9c,11t-CLA, and 10t,12c-CLA to the rats, suggesting that geometrical and positional isomerism of the conjugated double bonds did not influence the absorption.
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Food & Nutrition Science Notes
Food & Nutrition Science Communication
  • Sayaka IKEUCI, Yuka MIYAMOTO, Tsuyoshi KATOH, Kimio NISHIMURA
    2007 Volume 71 Issue 8 Pages 2091-2094
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    The influence of L-ascorbic acid at 40 °C incubation on the subfragment-1 and rod regions, prepared by chymotryptic digestion of myosin, and myosin was investigated by SDS–polyacrylamide gel electrophoresis and transmission electron microscopy respectively. It was observed that L-ascorbic acid acted more readily on the subfragment-1 region of myosin. Further, circular dichroism measurement indicated that L-ascorbic acid did not affect the structure of myosin. These results suggest that L-ascorbic acid acts more readily on the myosin subfragment-1 region and promotes the gelation of myosin without producing a conformational change in this protein.
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Microbiology & Fermentation Technology Regular Papers
  • Harri MÄKIVUOKKO, Hannele KETTUNEN, Markku SAARINEN, Tatsuya KAMI ...
    2007 Volume 71 Issue 8 Pages 1834-1843
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
    JOURNALS FREE ACCESS
    Effects of cocoa mass and supplemented dietary fiber (polydextrose) on microbial fermentation were studied by combining digestion simulations of stomach and small intestine with multi-staged colon simulations. During the four phases of digestion, concentrations of available soluble proteins and reducing sugars reflected in vivo absorption of nutrients in small intestine. In colon simulation vessels, addition of polydextrose to digested cocoa mass significantly increased concentrations of total short-chain fatty acids and butyric acid, from 103 to 468 mM (P<0.01) and from 12 to 22 mM (P<0.01), respectively. Long-chain fatty acid concentrations (decreasing from 1,222 to 240 mM) were mainly affected by the presence of digested cocoa mass. Cocoa mass with or without polydextrose addition significantly decreased production of cadaverine (P<0.02) and branched-chain fatty acids compared to control during colon simulations. Results indicate beneficial effects on metabolism of colonic microbiota after digestion of cocoa mass, and even more so with polydextrose addition.
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  • Riko HATAKEYAMA, Tomoyuki NAKAHAMA, Yujiro HIGUCHI, Katsuhiko KITAMOTO
    2007 Volume 71 Issue 8 Pages 1844-1849
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    In the filamentous fungus Aspergillus oryzae, there has been no report on photoreaction. Here we investigated the effect of light in A. oryzae and found that conidiation was repressed by white light. This reaction is contrary to that of other Aspergilli, which show abundant conidiation under light. Moreover, red light also caused reduced conidiation. Genome sequencing of A. oryzae indicated the existence of homologs of some light-related genes in other filamentous fungi. To approach the molecular mechanism of this photoresponse, the effect of red light on the expression level of several genes putatively responsible for conidiation or photoperception, i.e., brlA, a gene known to be required for conidiation, AofphA, the putative homolog of the A. nidulans phytochrome gene fphA, and AoveA, the putative homolog of the negative regulator gene in conidiation in A. nidulans, was examined. These three genes showed no significant response to red light at the transcriptional level. The results indicate that A. oryzae perceives and responds to red light in a manner independent of the transcriptional regulation of these genes.
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  • Yukio TAMAI, Keiko KANAI, Yoshinobu KANEKO
    2007 Volume 71 Issue 8 Pages 1850-1857
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    A novel HO gene (Uv-HO) was cloned from the Saccharomyces bayanus var. uvarum (abbreviated as S. uvarum in this study) type strain. The coding region of Uv-HO showed relatively high homology (95%) to that of the Sb-HO gene (S. bayanus var. bayanus HO), but not to the HO genes of other Saccharomyces sensu stricto species. However, the 5′ and 3′ non-coding region of Uv-HO showed less similarity (79% and 76% respectively) even to those of the most homologous gene Sb-HO. Motifs of the mating-type control and the cell-cycle control were conserved in the 5′ non-coding region of Uv-HO, but numbers and positions of motifs were different from those of Sb-HO. CHEF-Southern analysis showed that all tested strains of S. bayanus species, including S. uvarum, carried the HO gene on the 1,100-kb chromosome. By HO-typing PCR using mixed primers, which provided a rapid and convenient tool for yeast identification, either the Uv-HO gene or the Sb-HO gene was detected in strains of S. bayanus species, but two strains were found to have both types of HO gene in each genome. These results suggest that S. uvarum has a unique sequence, but might share the same chromosome constitution within S. bayanus species, and that S. bayanus is a heterogeneous species, of which some strains might be natural hybrid.
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  • Katsuhiko BABASAKI, Hitoshi NEDA, Hitoshi MURATA
    2007 Volume 71 Issue 8 Pages 1927-1939
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
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    Molecular studies on the evolution and systematics of fungi have been established primarily based on the neutral theory by analyzing neutral mutations in some defined segments of housekeeping genes as genetic markers. Such an approach is, however, hardly applicable for analyzing ancient evolutionary radiations. In the present study, we looked for DNA sequences characterizing higher taxa, and discovered a unique macroevolutionary genomic marker, megB1, that specifies the phylum Basidiomycota. megB1 is an approximately 500-bp DNA element, which is defined by terminal sequences and five internal segments conserved throughout the phylum. megB1 resides on the rDNA intergenic spacer 1 (IGS1) from 27 species of 10 Basidiomycota genera examined. While megB1 was not found in IGS1 from the other 92 species of the 27 Basidiomycota genera, several genera representing them carry megB1 in some other genomic regions. No known taxonomic criteria fit into the classification on the basis of whether megB1 resides on rDNA. Neighbor-joining analysis of the megB1 sequence, however, properly assigned species to their respective genera. Thus far, megB1 has not been found in any genomic or genetic databases currently available for other phyla. These results suggest that megB1 may have emerged upon the occurrence of Basidiomycota, and that this phylum evolved thereafter leaving this element conserved throughout their further differentiation. megB1 may be a novel genomic marker useful in the analysis of ancient through the latest evolutionary radiation in Basidiomycota.
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  • Shun IWATANI, Takeshi ZENDO, Fuminori YONEYAMA, Jiro NAKAYAMA, Kenji S ...
    2007 Volume 71 Issue 8 Pages 1984-1992
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
    JOURNALS FREE ACCESS
    A novel bacteriocin, lacticin Z, produced by Lactococcus lactis QU 14 isolated from a horse’s intestinal tract was identified. Lacticin Z was purified through a three step procedure comprised of hydrophobic-interaction, cation-exchange chromatography, and reverse-phase HPLC. ESI-TOF MS determined the molecular mass of lacticin Z to be 5,968.9 Da. The primary structure of lacticin Z was found to consist of 53 amino acid residues without any leader sequence or signal peptide. Lacticin Z showed homology to lacticin Q from L. lactis QU 5, aureocin A53 from Staphylococcus aureus A53, and mutacin BHT-B from Streptococcus rattus strain BHT. It exhibited a nanomolar range of MICs against various Gram-positive bacteria, and the activity was completely stable up to 100 °C. Unlike many of other LAB bacteriocins, the stability of lacticin Z was emphasized under alkaline conditions rather than acidic conditions. All the results indicated that lacticin Z belongs to a novel type of bacteriocin.
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  • Hideyuki TAMEGAI, Eriko IKEDA, Chiaki KATO, Koki HORIKOSHI
    2007 Volume 71 Issue 8 Pages 2041-2045
    Published: August 23, 2007
    Released: August 23, 2007
    [Advance publication] Released: August 07, 2007
    JOURNALS FREE ACCESS
    The nap gene cluster encoding periplasmic nitrate reductase was identified from Pseudomonas sp. strain MT-1, a deep-sea denitrifier isolated from the Mariana Trench. The ORFs identified were highly homologous with those of Pseudomonas stutzeri, but the cluster included only four ORFs (napDABC), less than those in other organisms. For other bacteria, some additional small ORFs (such as napE, napF, napG, napH, and napK) are found in the nap gene cluster, although their physiological function is still unclear. The soluble fraction of MT-1 grown under denitrifying condition showed significant nitrate reductase activity. This observation suggests that the periplasmic nitrate reductase encoded by the gene cluster identified in this study is functional. The activity was highest when the organism was grown under denitrifying conditions, suggesting that the enzyme participates in dissimilatory nitrite reduction.
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