Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Volume 68 , Issue 4
Showing 1-39 articles out of 39 articles from the selected issue
Analytical Chemistry Note
  • Masakazu TSUKAMOTO, Yoshiyuki MURATA, Mikiro TADA, Yasuaki SHIMOISHI
    2004 Volume 68 Issue 4 Pages 920-923
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    A simple and sensitive specrophotometric method combined with solid-phase extraction (SPE) for the simultaneous determination of sodium linear-dodecylbenzenesulfonate (DBS) and sodium dodecyl sulfate (SDS) is described. The C2 (ethyl group bonded silicagel) cartridge could be repeatedly used more than 500 times for SPE, and it enabled the anionic surfactants to be concentrated by 50-fold. The calibration graph for DBS was linear in the range from 1.6×10−8 M to 5.0×10−7 M and for SDS from 2.0×10−9 M to 3.0×10−7 M. The relative standard deviation (n=5) for 5.0×10−7 M DBS was 3.1% and for 2.5×10−7 M SDS was 1.7%. The proposed method was applied to the simultaneous determination of DBS and SDS in river-water samples.
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Organic Chemistry Regular Papers
  • Satomi KOYA-MIYATA, Iwao OKAMOTO, Shimpei USHIO, Kanso IWAKI, Masao IK ...
    2004 Volume 68 Issue 4 Pages 767-773
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    We have previously shown that royal jelly (RJ) promoted collagen production by skin fibroblasts in the presence of ascorbic acid-2-O-alpha-glucoside (AA-2G). In this study, we purified the honeybee RJ-derived collagen production-promoting factor (HBRJ-CPF) from an alkali-solubilized fraction of RJ by C18 reverse-phase column chromatography. The elution profile by the C18 column chromatography and the molecular mass of the purified HBRJ-CPF material coincided with those of 10-hydroxy-2-decenoic acid (10H2DA). We then examined the collagen production-promoting activities of several commercially available fatty acids contained in RJ. We found that 10H2DA and 10-hydroxydecanoic acid increased the collagen production in a dose-dependent manner. Furthermore, 10H2DA induced the fibroblast cell line, NHDF, to produce transforming growth factor-β1 (TGF-β1) which is an important factor for collagen production. As expected, the collagen production-promoting activity of 10H2DA was neutralized by the anti-TGF-β1 antibody. These result suggest that HBRJ-CPF identified as 10H2DA promoted the collagen production of AA-2G-treated fibroblasts by inducing TGF-β1 production.
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  • Yoshiro SAITO, Akira SHIGA, Yasukazu YOSHIDA, Takuya FURUHASHI, Yoji F ...
    2004 Volume 68 Issue 4 Pages 781-786
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    Rosemary is commonly used as a spice and a flavoring agent in food processing. Although the antioxidative properties of its extracts have been investigated, there have been few reports on the volatile components of rosemary. We designed a novel antioxidative system which can generate the volatile constituents in the gaseous phase from a rosemary extract and evaluated the gaseous antioxidative activities against both lipid peroxidation and cell death induced by nitrogen dioxide and ultraviolet radiation. The antioxidative effects of the major volatile components on the oxidation of linoleic acid induced by azo compounds were also investigated in a solution.
    The volatile components in the novel antioxidative system suppressed the Jurkat cell death induced by nitrogen dioxide and the intracellular formation of reactive oxygen species in fibroblast cells induced by ultraviolet radiation. 1,8-Cineole among the volatile components exerted an antioxidative effect against the oxidation of linoleic acid in a solution induced by azo compounds and ultraviolet radiation. These data suggest that the volatile constituents of a rosemary extract had antioxidative properties and that gaseous exposure antioxidant is a promising method for promoting health.
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  • Tomoyuki FUJITA, Hideo HAYASHI
    2004 Volume 68 Issue 4 Pages 820-826
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    Three new brasiliamide congeners, brasiliamides C, D and E, were isolated from okara fermented with Penicillium brasilianum Batista JV-379. Their structures were elucidated on the basis of spectral data and chemical evidence. NMR spectra of these brasiliamides exhibited a mixture of four or two conformers due to the restricted rotation of an amide bond in a solution. The 1H- and 13C-NMR spectral data were analyzed for a major rotamer at an appropriate temperature, since the signals were broadened at room temperature. Both brasiliamides C and D showed convulsive activity against silkworms with an ED50 value of 400 μg/g of diet, whereas brasiliamide E showed less activity than the others.
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  • Ken-ichi ONODERA, Takuya FUKATSU, Nozomi KAWAI, Yukio YOSHIOKA, Tetsuj ...
    2004 Volume 68 Issue 4 Pages 848-852
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    A novel fatty acid derivative named zooxanthellactone (ZL) was isolated from several strains of symbiotic microalgae, dinoflagellates of the genus Symbiodinium. The metabolite is structurally related to docosahexaenoic acid (DHA) and seems to be biosynthesized by oxidation and subsequent lactonization. The absolute stereochemistry was determined from the specific rotation of the perhydro derivative. The distribution of ZL within several Symbiodinium isolates was quantitatively analyzed by HPLC techniques and suggested a relationship between the productivity of this metabolite and the Symbiodinium phylogeny. The cytotoxicity of ZL was evaluated by using human squamous cell carcinoma cell lines in comparison with that of DHA and other common fatty acids, suggesting that the long unsaturated chain was important rather than the γ-lactone moiety.
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  • Toshikazu KOMODA, Madoka KISHI, Naoki ABE, Yasumasa SUGIYAMA, Akira HI ...
    2004 Volume 68 Issue 4 Pages 903-908
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    Three novel lipoxygenase inhibitors, tetrapetalone B (2, C28H35NO9), C (3, C26H34NO8), and D (4, C28H36NO10), were isolated from a culture broth of Streptomyces sp. USF-4727 that produced a lipoxygenase inhibitor tetrapetalone A (1) simultaneously. Each chemical structure was revealed by spectroscopic evidence, this suggests that these three compounds are structurally related to 1. They had a tetracyclic skeleton and a β-D-rhodinosyl moiety. Tetrapetalone B, C, and D inhibited soybean lipoxygenase with IC50: 320, 360, and 340 μM respectively.
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Organic Chemistry Note
Biochemistry & Molecular Biology Regular Papers
  • Akira KURIOKA, Fujie KURIOKA, Masayoshi YAMAZAKI
    2004 Volume 68 Issue 4 Pages 774-780
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    Acid-degraded sericin powder (AC-SP) was prepared from aqueous solution containing citric acid-degraded sericin polypeptides of Bombyx mori. The morphological and biochemical properties of AC-SP were compared with those of alkali-degraded sericin powder (AL-SP) and hot-water degraded sericin powder (HW-SP). Based on an SEM analysis, AC-SP showed a thin film structure of 10–100 μm with good dispersity while AL-SP and HW-SP had a much larger thin film structure (<500 μm). The extract of AC-SP showed stronger trypsin inhibitor activity due to cocoon shell trypsin inhibitor (CSTI-IV) than that of HW-SP. The extract of AL-SP showed no CSTI-IV activity. It was found that AC-SP was a trypsin inhibitor complex powder and that the release of CSTI-IV from AC-SP depended on pH and ion strength. Similar powder materials were obtained when such organic acids as tartaric acid and succinic acid were used. These results suggest that the acid-degraded sericin polypeptides work as a protein matrix to which CSTI-IV may bind ionically.
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  • Katsuhiko TAGUCHI, Masaki MOTOYAMA, Toshiaki KUDO
    2004 Volume 68 Issue 4 Pages 787-795
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    Rhodococcus rhodochrous K37, a Gram-positive bacterium grown under alkaline conditions, was isolated for its ability to metabolize PCBs. Analysis revealed that it has eight genes encoding extradiol dioxygenase, which has 2,3-dihydroxybiphenyl 1,2-dioxygenase activity, and these genes were designated bphC1 to bphC8. According to the classification of extradiol dioxygenases [Eltis, L. D., and Bolin, J. T., J. Bacteriol., 178, 5930–5937 (1996)], BphC3 and BphC6 belong to the type II enzyme group. The other six BphCs were classified as members of the type I extradiol dioxygenase group. BphC4 and BphC8 were classified into a new subfamily of type I, family 3. Two linear plasmids, 200 kb and 270 kb in size, were found in K37, and the bphC6 and bphC8 genes were located in the 200 kb linear plasmid. Northern hybridization analysis revealed that the bphC1, bphC2, and bphC7 genes were induced in the presence of testosterone, the bphC6 gene was induced by fluorene, and the bphC8 gene was induced by biphenyl. All eight BphC products exhibited much higher substrate activity for 2,3-dihydroxybiphenyl than for catechol, 3-methylcatechol, or 4-methylcatechol.
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  • Sachiko ONO, Tsuneyuki TANAKA, Yuriko WATAKABE, Kazuyuki HIRATSUKA
    2004 Volume 68 Issue 4 Pages 803-807
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    In order to develop a rapid and versatile assay system suitable for the analysis of regulated expression of tobacco pathogenesis-related protein 1a (PR-1a) gene, we investigated the use of the transient gene expression system in tobacco BY-2 cells by microprojectile bombardment. Using dual luciferase assay as a reporter gene expression detection system, we observed significant induction of PR-1a promoter activity by salicylic acid (SA) treatment. On the other hand, treatment with 4-hydroxybenzoic acid (4-HBA) resulted in no detectable increase in luciferase activity. Co-expression of a trans-acting factor, the NPR1/NIM1 protein of Arabidopsis, resulted in the induction of higher expression levels of the PR-1a promoter. These results suggest that the assay system is applicable for the analysis of factors involved in the regulated expression of SA-inducible defense-related genes.
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  • Kenji KASHIWAGI, Kaoru FUKAMI-KOBAYASHI, Kiyotaka SHIBA, Ken NISHIKAWA
    2004 Volume 68 Issue 4 Pages 808-813
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    The respective type-1 and type-2 periplasmic binding proteins (PBPs) MglB and ArgT are believed to have evolved from a common ancestor into siblings showing topological differences in their main chain connectivity. At first glance, they show similar structure. But, more detailed examination reveals that the chain connectivity of ArgT is more convoluted than that of MglB. Reflecting that complexity, the folding of ArgT is complicated and involves intermediate folds. On the other hand, the folding of MglB is a simple two-state transition. In the present study, we constructed and characterized several chimeras made up of various subdomains of MglB and ArgT with the aim of gaining insight into the evolution of protein folding and protein structure. Although these chimeras did not fold as compactly as their parental proteins, some did exhibit cooperative folding, which suggests that novel proteins with new connectivity and new folding pathways could have emerged at a fairly high rate throughout the evolution of proteins.
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  • Le Huyen Ai THUY, Krisana PHUCHAROEN, Akira IDENO, Tadashi MARUYAMA, T ...
    2004 Volume 68 Issue 4 Pages 814-819
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    A catalase gene, ohktA, from an alkali- and halo-tolerant bacterium, Halomonas sp. SK1, on the pKK223-3, was expressed in the catalase-lacking Escherichia coli strain UM2. Highly purified catalase showing a single band on SDS-PAGE was obtained by two liquid chromatography steps on DEAE-Toyopear1 and Chelating-Sepharose Fast Flow. The enzyme, oHktA, shows high catalase activity with a pH optimum at 10, and the activity was stable in 4 M KC1. This enzyme is thermo-sensitive, showing a significant loss of activity within 5 minutes at 37 °C. To modify the stability of the catalase, the addition of domain II of the heat stable Mn catalase from Thermus thermophilus to the C-terminus was made. When coexpressed with a chaperone (PhFKBP29) gene product, peptidyl–prolyl cistrans isomerase, from a thermophilic bacterium, a chimeric catalase was produced in the soluble fraction. The stability of this catalase in the range of 37°–45 °C was improved and it was stable for more than 1 h at 37 °C.
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  • Ningjia HE, Yoichi ASO, Hiroshi FUJII, Yutaka BANNO, Kohji YAMAMOTO
    2004 Volume 68 Issue 4 Pages 835-840
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    Various chymotrypsin inhibitors occur in the hemolymph of silkworm larvae. Interaction of chymotrypsin inhibitor b1 (CI-b1) with Escherichia coli was examined from the viewpoint of action against invading bacteria. Injection of dead E. coli cells into larva reduced the CI-b1 content of the hemolymph, suggesting in vivo binding of CI-b1 to the outer membrane of the cell. Results from incubation of E. coli in cell-free hemolymph in the presence or absence of lipopolysaccharide indicated that CI-b1 is the only CI bound to E. coli and that it interacts with lipopolysaccharide. CI-b1 formed a complex with lipopolysaccharide in vitro; the value of the dissociation constant was relatively large. Inhibitory activity of CI-b1 changed insignificantly in mixture with lipopolysaccharide. CI-b1 affected the growth of E. coli but never worked lethally. CI-b1 is speculated to be a mediator that scavenges intruding bacteria rather than a direct anti-bacterial factor. This is the first report confirming that CI-b1 is a lipopolysaccharide binding protein.
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  • Koh AMANO, Masahiro FUJIHASHI, Akikazu ANDO, Kunio MIKI, Yoshiho NAGAT ...
    2004 Volume 68 Issue 4 Pages 841-847
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    Since the involvement of Tyr residues in the fucose-binding of Aleuria aurantia lectin (AAL) was proved by chemical modification using the Tyr-specific reagent tetranitromethane, site-directed mutagenesis was attempted. Since the tertiary structure of AAL was determined recently to be a six-bladed β-propeller fold, and five fucose-binding sites per subunit were found, based on positions of Tyr residues in the tertiary structure, three classes of mutants were constructed: 1) Tyr on the 2nd β-strand of each blade (β-2 mutants), 2) Tyr or Trp on the 3rd β-strand (β-3 mutants), and 3) Tyr outside of binding sites (other-Y mutants). The mutagenized cDNA was expressed in Escherichia coli as His-tag-AAL, and the hemagglutinating activity was assayed. Among 14 mutants, three β-2 mutants (Y26A, Y79A, and Y181A), and three β-3 mutants (Y92A, W149A, and Y241A) showed decreased activity. These mutated residues resided at Sites 1, 2, and 4, at the same locations relatively in the binding sites. Mutagenesis of Tyr or Trp at the corresponding locations in Sites 3 and 5 did not lead to a reduction in activity. Results indicate that the properties of Sites 1, 2, and 4 are different from those of Sites 3 and 5, and that the contribution of these two sites to the hemagglutination reaction was minor.
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  • Koichi MORIMOTO, Saori KUNII, Kaori HAMANO, Ben’ichiro TONOMURA
    2004 Volume 68 Issue 4 Pages 861-867
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    Pepsin-hydrolyzed collagen (atelocollagen) is a trimer, consisting of α1 and α2 monomers, and shows molecular species corresponding to a monomer, dimer (β chain), and trimer (γ chain) by SDS-polyacrylamide gel electrophoresis. Atelocollagen was purified from yellowfin tuna (Thunnus albacares) by salt precipitation and cation-exchange chromatography. Enzymatic hydrolysis of the atelocollagen by actinidain, a cysteine protease purified from kiwifruit, was analyzed by SDS-polyacrylamide gel electrophoresis. The triple helical structure unique to collagen was retained in the atelocollagen as judged by circular dichroism spectra. The actinidain-processed atelocollagen showed only monomeric α1 and α2 chains, with no β and γ chains, by SDS-polyacrylamide gel electrophoresis; nevertheless, it retained the typical triple helical structure. It is suggested that actinidain cleaved the atelocollagen molecule at specific sites on the inside of the inter-strand cross-linking peptides.
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  • Tomoyuki NAKAMURA, Seiichi MATSUGO, Yasuyuki UZUKA, Sumitaka MATSUO, H ...
    2004 Volume 68 Issue 4 Pages 868-872
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    All the fractions of Phellinus linteus mycelia showed anti-tumor activity toward solid tumors planted in mice. The highest anti-tumor activity of 81.2% was observed in the protein–glucan complex obtained by precipitating the 24% NaOH extract at pH 6.0. This protein–glucan complex consisted of 39.3% polysaccharide and 49.4% protein. Its 13C- and 1H-NMR data showed that the main glucan part of the complex was simple α-1,3-glucan chains.
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  • Kazutoshi SAWADA, Morifumi HASEGAWA, Lena TOKUDA, Junko KAMEYAMA, Osam ...
    2004 Volume 68 Issue 4 Pages 873-880
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    The rice Oryza sativa selenium-binding protein homologue (OsSBP) gene encodes a homologue of mammalian selenium-binding proteins, and it has been isolated as one of the genes induced by treating a plant with a cerebroside elicitor from rice blast fungus. The possible role of OsSBP in plant defense was evaluated by using a transgenic approach. Plants overexpressing OsSBP showed enhanced resistance to a virulent strain of rice blast fungus as well as to rice bacterial blight. The expression of defense-related genes and the accumulation of phytoalexin after infection by rice blast fungus were accelerated in the OsSBP overexpressors. A higher level of H2O2 accumulation and reduced activity of such scavenging enzymes as ascorbate peroxidase and catalase were seen when the OsSBP-overexpressing plants were treated with the protein phosphatase 1 inhibitor, calyculin A. These results suggest that the upregulation of OsSBP expression conferred enhanced tolerance to different pathogens, possibly by increasing plant sensitivity to endogenous defense responses. Additionally, the OsSBP protein might have a role in modulating the defense mechanism to biotic stress in rice.
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  • Fengzhi LIU, Hiroto SHINOMIYA, Teruo KIRIKAE, Hajime HIRATA, Yoshihiro ...
    2004 Volume 68 Issue 4 Pages 894-902
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    Such phagocytic leukocytes as macrophages and neutrophils are the key cellular components of innate immunity. The actin cytoskeleton is essential for their recruitment and activation in infected tissues. We have previously identified p65/L-plastin with Ca2+-, calmodulin-, and β-actin-binding domains in macrophages. In order to further investigate the p65/L-plastin-involved cellular functions, we cloned the cDNA for murine grancalcin, a possible binding partner of p65/L-plastin. According to the sequence, grancalcin is a member of the penta-EF-hand protein family. We prepared recombinant (r) grancalcin for functional studies and found that it exhibited Ca2+-dependent precipitation. High-titer antibodies against the protein enabled us to detect intracellular grancalcin. A flow cytometric analysis revealed grancalcin to be highly expressed in macrophages and neutrophils. The protein was particularly abundant in those cells recovered from bacteria-infected sites. Immunohistochemical studies clarified that grancalcin was translocated to the actin cytoskeleton in macrophages upon exposure to bacterial lipopolysaccharide. These findings suggest that grancalcin plays a key role in leukocyte-specific functions that are responsible for host defense.
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  • Hirofumi KOYAMA, Taka-aki NAGAO, Tomomi INAI, Kohji MIYAHARA, Yasufumi ...
    2004 Volume 68 Issue 4 Pages 909-919
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    RSC is a nucleosome-remodeling complex of Saccharomyces cerevisiae essential for growth that can alter histone–DNA interaction by using the energy of ATP hydrolysis. Nps1p/Sth1p is an ATPase subunit of RSC. A mutation in the conserved ATPase domain of Nps1p causes a sporulation defect with decreased expression of early meiotic genes, especially IME2. This defect is partially suppressed by the overexpression of either IME1 or IME2. A homozygous diploid of a novel temperature-sensitive nps1 mutation, nps1-13, harboring amino acid substitutions within the bromodomain, was unable to sporulate. Overexpression of IME, IME2, or both of these genes allowed the completion of meiosis I and meiosis II in nps1-13 but not the formation of mature asci. In nps1-13 carrying YEpIME1, the expression of a group of sporulation-specific genes, which express at the middle stages of sporulation and are required for spore-wall formation, notably diminished, and several late sporulation genes expressed at the early stages of sporulation. These results suggest that Nps1p/RSC plays important roles during the spore development process by controlling gene expression for initiating both meiosis and spore morphogenesis, and ensures proper expression timing of late meiotic genes.
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Biochemistry & Molecular Biology Notes
Biochemistry & Molecular Biology Communications
Food & Nutrition Science Regular Papers
  • Yasuko MANABE, Hitoshi UTSUNOMIYA, Kuniyasu GOTOH, Takeyuki KUROSU, To ...
    2004 Volume 68 Issue 4 Pages 796-802
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    Selection of foods by animals largely depends on their physiological condition. In this paper, we reported how physiological changes in rats after ingestion of sake (Japanese rice wine) affected their preference for different kinds of sake. Rats could discriminate among various kinds of sakes in a two-bottle choice test, even after adjustment of the glucose concentration or alcohol concentration of the sake, suggesting that the choice of a sake by animals was based on more than one ingredient. To identify effect of a rat’s physiological condition on the selection of sake, we monitored the levels of blood glucose, nonesterified fatty acids (NEFA), and ketone bodies in serum in mice after forced intragastric ingestion of sake that had previously been offered to the rats in a two-bottle choice test. Blood glucose levels in mice were not different between the rats fed the palatable and unpalatable sakes, and the NEFA level and ketone level were high in rats fed the unpalatable sake. To further clarify the relationship between physiological condition and preference for sake, rats were offered eight kinds of Junmai-syu, which is made with only rice and no sub-ingredient. Rats could still discriminate among Junmai-syu. Furthermore, with two exceptions, the preference for a sake was significantly correlated with the ketone level. The order in the ratio of blood insulin level to blood glucagon level, which is an indicator of metabolism, was correlated with the order in preference to sake. These results suggest that some physiological factors besides oral stimulation also are important factors in the selection of a sake.
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  • Wataru MIZUNOYA, Shinichi OYAIZU, Akimori HIRAYAMA, Tohru FUSHIKI
    2004 Volume 68 Issue 4 Pages 827-834
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    We investigated the effects of physical fatigue produced by swimming exercise on learning the Morris water maze in BALB/c mice. We measured the escape latency in the maze immediately after the swimming exercise. The control group was soaked in the water but not fatigued. For easier tasks, like one with an obvious cue flag, the escape latency was not changed by exercise fatigue. However, escape latency was increased after exercise fatigue for more difficult tasks of spatial learning. These results appear to suggest that physical fatigue impaired learning performance. The effects of swimming exercise fatigue on learning efficiency were then investigated. Mice were continuously fatigued during the spatial learning period. This increased escape latency between the first and third sessions. The results suggest that learning efficiency was impaired by exercise fatigue. This system may be useful for screening new foods used to enhance brain function during exercise.
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  • Hiroyuki WAKABAYASHI, Hidefumi KUWATA, Koji YAMAUCHI, Susumu TERAGUCHI ...
    2004 Volume 68 Issue 4 Pages 853-860
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    We investigated the transfer of dietary bovine lactoferrin (LF) and its functional lactoferricin (LFcin) B-containing fragments to the portal blood of healthy adult rats by using several techniques. After a single administration of 125I-labeled LF, radioactive bands were detected in autoradioluminograms of the portal blood, but similar bands were also observed after the administration of [125I]NaI. Although ovalbumin was detected by ELISA at 3–18 ng/ml in the portal blood plasma after an overnight administration, no LF was detected (≤1.5 ng/ml). The antibody-captured ovalbumin fragments, but not the LF fragments, were detected in the plasma by surface-enhanced laser desorption/ionization affinity mass spectrometry (SELDI affinity MS). We finally attempted to detect the LFcin B-containing fragments by SELDI affinity MS with on-chip LFcin B-conversion, but could not detect them (≤1 ng/ml) in the portal blood after the LF ingestion. The level of LF or its functional fragments transferred to the portal blood was therefore extremely low, if any.
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  • Michihiro KOHGUCHI, Toshio KUNIKATA, Hikaru WATANABE, Naoki KUDO, Taka ...
    2004 Volume 68 Issue 4 Pages 881-887
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    The immuno-potentiating effects of the antler-shaped fruiting body of Ganoderma lucidum (Rokkaku-Reishi, RR), which has been used as a traditional supplement for human health, were investigated in mice. BALB/c mice were administered orally with RR for 3 days at a dose of 50 mg/kg or 500 mg/kg, and interferon-gamma (IFN-γ) production by splenocytes in response to lipopolysaccharide (LPS) was examined on day 4. The oral administration of 500 mg/kg of RR resulted in a significant increase (p<0.05) in IFN-γ production. Stimulation of splenic adherent cells from these mice with LPS also resulted in a significant increase (p<0.05) in interleukin-12 (IL-12) production compared with that from the control mice, suggesting that splenic macrophages were activated by RR administration. Furthermore, 500 mg/kg of RR administered for 14 days resulted in a significant increase (p<0.05) in IFN-γ production by splenocytes in response to both LPS and concanavalin A (Con A). These results suggest that not only splenic macrophages but also T cells were activated by the long-term treatment with RR in vivo. On the other hand, the production of interleukin-4 (IL-4), which is known as an allergic disease-related cytokine, was not affected by the long-term treatment with RR. Our results suggest that the oral administration of RR resulted in Th1-associated immuno-potentiating activities in vivo.
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  • Ryuhei KANAMOTO, Kousuke FUJITA, Megumi KUMASAKI, Saeko IMAI, Makoto K ...
    2004 Volume 68 Issue 4 Pages 888-893
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    Rats of different ages (3 to 15-wk-old) were fed on a 25% casein diet for one week, and the nitrogen balance and liver serine dehydratase (SDH, EC 4.2.1.13) activity were then determined. The value for nitrogen balance decreased with the age of the rats, while the liver SDH activity increased. A statistical analysis showed clear inverse correlation between the two factors (R2 = 0.7372, p < 0.01). This result suggests that SDH was induced by response to the amount of surplus amino acids from dietary protein taken beyond the body’s requirement. The increase in SDH activity was accompanied by an increase in the level of SDH mRNA. Since the half-life of this mRNA did not change significantly, the induction was mainly controlled at the level of transcription. In addition, the induction seems not to be related to gluconeogenesis, since the mRNA levels of tyrosine aminotransferase (TAT) and phosphoenolpyruvate carboxykinase (PEPCK), other gluconeogenic enzymes, were not changed under these experimental conditions.
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Food & Nutrition Science Notes
Microbiology & Fermentation Technology Note
Microbiology & Fermentation Technology Communication
  • Shunsuke KUBOTA, Ikuko TAKEO, Kazunori KUME, Muneyoshi KANAI, Atsunori ...
    2004 Volume 68 Issue 4 Pages 968-972
    Published: 2004
    Released: April 23, 2004
    JOURNALS FREE ACCESS
    The budding yeast Saccharomyces cerevisiae has been used in the fermentation of various kinds of alcoholic beverages. But the effect of ethanol on the cell growth of this yeast is poorly understood. This study shows that the addition of ethanol causes a cell-cycle delay associated with a transient dispersion of F-actin cytoskeleton, resulting in an increase in cell size. We found that the tyrosine kinase Swe1, the negative regulator of Cdc28-Clb kinase, is related to the regulation of cell growth in the presence of ethanol. Indeed, the increase in cell size due to ethanol was partially abolished in the SWE1-deleted cells, and the amount of Swe1 protein increased transiently in the presence of ethanol. These results indicated that Swe1 is involved in cell size control in the presence of ethanol, and that a signal produced by ethanol causes a transient up-regulation of Swe1. Further we investigated comprehensively the ethanol-sensitive strains in the complete set of 4847 non-essential gene deletions and identified at least 256 genes that are important for cell growth in the presence of ethanol.
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