Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
Volume 28 , Issue 9
Showing 1-50 articles out of 57 articles from the selected issue
Review
  • Naoki Okada
    2005 Volume 28 Issue 9 Pages 1543-1550
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    Tumor cells that generally accumulate mutations in the genome express molecules different both qualitatively and quantitatively from normal cells. An immunosurveillance system for these molecules, known as the tumor-associated antigens (TAAs), plays an important role in the elimination of cancer cells during the initial stage. Although cancer immunotherapy targeting TAAs has progressed steadily with the development of various vaccine strategies, satisfactory efficacy, such as marked tumor regression and complete response, has not been previously reported in a clinical setting. To improve the therapeutic effects of cancer immunotherapy, the application of chemokine–chemokine receptor coupling, which controls the trafficking and biodistribution of immune cells in the living body, is an attractive potential approach. This review introduces our novel “cell delivery system,” which employs an Arg-Gly-Asp (RGD) fiber-mutant adenovirus vector encoding the chemokine or chemokine receptor gene in cancer immunotherapy.
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  • Tadahide Furuno, Mamoru Nakanishi
    2005 Volume 28 Issue 9 Pages 1551-1559
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    Mast cells are widely distributed throughout the body, predominantly near blood vessels and nerves, and express effector functions in allergic reactions, inflammatory diseases, and host defense. The activation of mast cells results in secretion of the preformed chemical mediators in their granules by a regulated process of exocytosis and leads to synthesis and secretion of lipid mediators and cytokines. Their soluble factors contribute to allergic inflammation. Mast cells are associated with hypersensitivity reactions, not only in the classical immunoglobulin E (IgE)-dependent mechanism but also in an IgE-independent manner. In particular, investigations of potential anatomical and functional interactions between mast cells and the nervous system have recently attracted great interest. To understand these molecular mechanisms in mast cell activation, the ability to visualize, track, and quantify molecules and events in living mast cells is an essential and powerful tool. Recent dramatic advances in imaging technology and labeling techniques have enabled us to carry out these tasks with high spatiotemporal resolution using confocal laser scanning microscopes, green fluorescent protein and its derivatives, and image analysis systems. Here we review our investigations of the dynamic processes of intracellular signaling molecules, cellular structure, and interactions with neurons in mast cells to provide basic and valuable information for allergy and clinical immunology using these new imaging methods.
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Current Topics
  • Hiroshi Homma
    2005 Volume 28 Issue 9 Pages 1560
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
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  • Toru Nishikawa
    2005 Volume 28 Issue 9 Pages 1561-1565
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    It has now been well established that D-serine, a coagonist for the N-methyl-D-aspartate (NMDA) glutamate receptors (NR1/NR2 type), is maintained at a high concentration in mammalian brains for life and shows a brain-selective and NMDA receptor R2B subunit-related distribution, overturning the hitherto generally accepted theory that D-amino acid is not always present in mammalian tissues. D-Serine in the brain has been shown to be contained in both the glia and neurons and to have specific processes of biosynthesis, extracellular release, uptake, and degradation. Moreover, the selective elimination of D-serine reduces the NMDA receptor-mediated intracellular signaling and long-term potentiation of synaptic connections. Together with the anti-psychotic and anti-ataxic property of D-serine and the pivotal roles of the NMDA receptor in divergent higher brain functions, these observations support the view that the D-amino acid may be involved as an endogenous modulator for the NMDA receptor in various neuropsychiatric functions and their pathological conditions.
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  • Takemitsu Furuchi, Hiroshi Homma
    2005 Volume 28 Issue 9 Pages 1566-1570
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    D-Aspartate (D-Asp) is an endogenous amino acid present in nervous and endocrine tissues in mammals. A high concentration of D-Asp is observed in embryos, which disappears in nervous tissues after delivery, but increases temporarily in endocrine glands, particularly in the pituitary, pineal and adrenal glands at the specific stages. In the pineal gland, D-Asp that is apparently derived from other tissues suppresses melatonin secretion from parenchymal cells. Additionally, D-Asp levels increase in the testis just before birth and during maturation. The amino acid is presumed to be synthesized by the pituitary gland and testis. In the testis, D-Asp produced inside the seminiferous tubules acts on Leydig cells following release to enhance testosterone synthesis by activating the expression of Steroidogenic Acute Regulatory protein. Mammalian cells appear to contain all the molecular components required to regulate D-Asp homeostasis, as they can synthesize, release, take up, and degrade the amino acid. These findings collectively indicate that D-Asp is a novel type of messenger in the mammalian body.
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  • Hiroki Abe, Naoko Yoshikawa, Mohammed Golam Sarower, Shigeru Okada
    2005 Volume 28 Issue 9 Pages 1571-1577
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    Aquatic crustaceans and some bivalve mollusks contain a large amount of free D-alanine (up to 100 μmol/g wet wt.) in their tissues. Under high salinity stress, crustaceans and bivalve mollusks largely accumulate D- and L-alanine irrespective of species examined, together with L-glutamine, L-proline, and glycine of which increases are species dependent. These data indicate that D-alanine is one of the major compatible osmolytes responsible for the intracellular isosmotic regulation in the tissues of crustaceans and bivalves. Alanine racemase has been proven to catalyze the interconversion of D- and L-alanine in these invertebrates. The enzyme has been isolated to homogeneity from the muscle of black tiger prawn Penaeus monodon and its cDNA has been cloned from the muscle and hepatopancreas of kuruma prawn Penaeus japonicus for the first time in eukaryotes other than yeast. Several fish species fed on crustaceans and mollusks contain D-amino acid and D-aspartate oxidases that catalyze the decomposition of D-amino acids. A cDNA of D-amino acid oxidase has been cloned from the hepatopancreas of omnivorous common carp Cyprinus carpio. During oral administration of free D-alanine to carp, the activity and mRNA of D-amino acid oxidase increased rapidly in hepatopancreas and the increases were highest in intestine followed by hepatopancreas and kidney. These data suggest that D-amino acid oxidase is inducible in carp and an important enzyme responsible for the efficient utilization of carbon skeleton of D-alanine in their feeds.
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  • Kenji Hamase, Ryuichi Konno, Akiko Morikawa, Kiyoshi Zaitsu
    2005 Volume 28 Issue 9 Pages 1578-1584
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The determination of small amounts of D-amino acids in mammalian tissues is still a challenging theme in the separation sciences. In this review, various gas-chromatographic and high-performance liquid chromatographic methods are discussed including highly selective and sensitive column-switching procedures. Based on these methods, the distributions of D-aspartic acid, D-serine, D-alanine, D-leucine and D-proline have been clarified in the mouse brain. As the regulation mechanisms of D-amino acid amounts in mammals, we focused on the D-amino-acid oxidase, which catalyzes the degradation of D-amino acids. Using the mutant mouse strain lacking D-amino-acid oxidase activity, the effects of the enzymatic activity on the amounts and distributions of various D-amino acids have been investigated.
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  • Noriko Fujii
    2005 Volume 28 Issue 9 Pages 1585-1589
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    All living organisms on earth are composed of L-amino acids and D-sugars. Therefore the presence and function of D-amino acids in living organisms have not been studied. Recently, however, D-aspartic acid (D-Asp) which is a D-amino acid, has been detected in various proteins from human tissue samples such as eye lens, brain, skin, bone, teeth, aorta from elderly individuals. It has been explained that the presence of D-Asp is the result of racemization of Asp residues in the protein during the life span, inasmuch as the proteins in these tissues are metabolically inert. The Asp-151 and Asp-58 residues in alpha A-crystallin from elderly human lens are especially stereochemically labile and the D/L ratios of these residues were greater than 1.0. A D/L ratio greater than 1.0 is not defined as racemization, but as the inversion of configuration. This was the first observation that inversion occurred in the configuration of amino acids in vivo during the natural aging process. In this review, we summarize the D-Asp in the various tissues reported by many researchers and describe the mechanism of D-Asp formation in protein. We suggest that a chiral reaction field exists in the native higher order structure of protein which induces the inversion of L-Asp to D-Asp residue.
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  • Takahiko Shimizu, Yasuji Matsuoka, Takuji Shirasawa
    2005 Volume 28 Issue 9 Pages 1590-1596
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    Isomerization of L-aspartate and deamidation of L-asparagine in proteins or peptides dominantly give rise to L-isoaspartate by a non-enzymatic reaction via succinimide as a intermediate under physiological conditions. Isoaspartates have been identified in a variety of cellular proteins in vivo as well as pathologically deposited proteins in neurodegenerative brain tissue. We described here that the formation of isoaspartate is enhanced in amyloid-β (Aβ) peptides in Alzheimer's disease (AD). Specific antibodies recognizing isoaspartate of Aβ revealed that isomerized Aβ peptides were deposited in senile plaques as well as amyloid-bearing vessels. Moreover, it was revealed that Aβ peptides, isomerized at position 7 or 23, were differentially deposited in senile plaques and vascular amyloids in AD brains. In vitro experiments showed that the modification at position 23 greatly enhanced the aggregation of Aβ. Furthermore, systematic proline substitution analyses revealed that the β-turn structure at positions 22 and 23 of Aβ42 plays a crucial role in the aggregation and neurotoxicity of Aβ peptides. It is suggested that spontaneous isomerization at position 23 induces the conformational change to form a β-turn at position 23, which plays a pathogenic role in the deposition of Aβ peptides in sporadic AD. Protein L-isoaspartyl methyltransferase (PIMT) is a putative protein repair enzyme, which converts L-isoaspartyl residues in damaged proteins to normal L-aspartyl residues. PIMT-deficient mice manifested neurodegenerative changes concomitant with the accumulation of L-isoaspartate in the brain. We discuss here the pathological implications of the formation of isoaspartate in damaged proteins during neurodegeneration in model mice and AD.
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Biochemistry
Regular Articles
  • Kazuhito Watanabe, Tamihide Matsunaga, Toshiyuki Kimura, Tatsuya Funah ...
    2005 Volume 28 Issue 9 Pages 1743-1747
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The properties of ES46.5K, an esterase from mouse hepatic microsomes, were compared with those of carboxylesterases from rabbit and porcine liver. The inhibitory profile with a serine hydrolase inhibitor (bis-p-nitrophenylphosphate) and detergents (sodium dodecylsulfate, Emulgen 911) was different between ES46.5K and the carboxylesterases. Bis-p-nitrophenylphosphate (0.1 mM) markedly inhibited the catalytic activity of the carboxylesterases but not that of ES46.5K. Emulgen 911 (0.05—0.25%) inhibited the catalytic activity of the carboxylesterases, whereas the detergent conversely stimulated that of ES46.5K by 150%. The two carboxylesterases catalyzed the hydrolysis of acetate esters of tetrahydrocannabinol (THC) analogues with different side chain lengths (C1—C5), although ES46.5K showed marginal activity only against the acetate of Δ8-tetrahydrocannabiorcol, a methyl side chain derivative of Δ8-THC. ES46.5K hydrolyzed cannabinoid esters stereospecifically and regioselectively. The esterase hydrolyzed 8α-acetoxy-Δ9-tetrahydrocannabinol (8α-acetoxy-Δ9-THC, 5.62 nmol/min/mg protein), while the enzyme did not hydrolyze 8β-acetoxy-Δ9-THC, 7α-acetoxy-, and 7β-acetoxy-Δ8-THC at all. In contrast, the carboxylesterases from rabbit and porcine liver hydrolyzed 8β-acetoxy-Δ9-THC efficiently but not 8α-acetoxy-Δ9-THC. ES46.5K hydrolyzed side chain acetoxy derivatives of Δ8-THC at the 3′- and 4′-positions, and a methyl ester of 5′-nor-Δ8-THC-4′-oic acid. The enzyme, however, could not hydrolyze methyl esters of Δ8- and Δ9-THC-11-oic acid, while both carboxylesterases hydrolyzed side chain acetoxy derivatives of Δ8-THC and three methyl esters of THC-oic acids. These differences in stereospecificity and regioselectivity between ES46.5K and carboxylesterases suggest that the configurations of important amino acids for the catalytic activities of these enzymes are different from each other.
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  • Naomi Matsumura, Hiroshi Ishibashi, Masashi Hirano, Yukiko Nagao, Naok ...
    2005 Volume 28 Issue 9 Pages 1748-1751
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    We investigated the effects of nonylphenol (NP) and triclosan (TCS) on production of vitellogenin (Vg), testosterone (T), and hepatic cytochrome P450 1A and 2B activities in male South African clawed frogs (Xenopus laevis). In a 14-d waterborne exposure test, no significant differences in the level of plasma Vg synthesis in male frogs were observed among the control, 10, 50, and 100 μg/l NP and 20, 100, and 200 μg/l TCS treatment groups. Intraperitoneal injection of male frogs with 2, 20, and 200 μg/g body weight NP resulted in no significant differences in plasma Vg levels among the control and all treatment groups. However, the levels of plasma Vg in all TCS treatment groups (intraperitoneal injection of 4, 40, and 400 μg/g body weight) were lower than that in the solvent control group, and male frogs injected with high doses of NP or TCS had lower T levels than the control group. No significant differences in hepatic cytochrome P450 1A and 2B activities were observed among the all treatment groups. Male frogs injected with 20 μg/g body weight of estradiol-17β had significantly higher plasma Vg levels than the control group. These results suggest that profiles of plasma Vg and T production in male Xenopus laevis could be useful biomarkers for detecting hormonally active agents.
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Molecular and Cell Biology
Regular Articles
  • Hua Feng, Xin-Yu Li, Jia-Run Zheng, Ji-Wei Gao, Lan-Fang Xu, Mei-Yu Ta ...
    2005 Volume 28 Issue 9 Pages 1597-1602
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    We performed this study to determine the relationship between activation of nuclear factor (NF)-κB and inhibition of keratinocyte growth by anthralin, which not only might be useful for a better understanding of the role of NF-κB in the pathogenesis of psoriasis, but also indicate whether the inflammatory reaction induced by anthralin is inseparable from its antipsoriatic activity. The involvement of NF-κB was assessed using the antipsoriatic drugs leflunomide and triptolide (T0) as effectors, since they can inhibit NF-κB activation induced by anthralin. The results showed that the inhibition of keratinocyte growth by anthralin was not related to the activation of NF-κB. Using sodium salicylate, a known NF-κB inhibitor, further confirmed this conclusion. Thus it might be possible to inhibit the inflammatory response induced by anthralin via repression of NF-κB activation. We found that leflunomide or T0 could significantly inhibit the mRNA overexpression of interleukin-8 and intercellular adhesion molecule-1 in keratinocytes induced by anthralin. Taken together, our data indicate that the growth inhibition of anthralin is related to the NF-κB-independent signaling pathway, and that leflunomide or T0 could control proinflammatory cytokine expression induced by anthralin via inhibiting the activation of NF-κB.
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  • Shinji Kagaya, Naganari Ohkura, Toshihiko Tsukada, Masami Miyagawa, Yu ...
    2005 Volume 28 Issue 9 Pages 1603-1607
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    Within the nuclear receptor superfamily, Nur77, Nurr1, and NOR1 constitute the nuclear receptor subfamily 4 group A. Modulation of NOR1 function would be therapeutic potential for diseases related to dysfunction of NOR1, including extraskeletal myxoid chondrosarcoma and autoimmune diseases. By screening arachidonate metabolites for their capacity of transcriptional activation, we have identified prostaglandin (PG) A2 as a transactivator for NOR1. PGA2 acted as a potent activator of NOR1-dependent transcription through the GAL4-based reporter system. The putative ligand-binding domain (LBD) of the receptor directly bound PGA2, and LBD-deleted receptor showed little transcriptional activation by PGA2. Primary cultured spleen cells derived from transgenic mice overexpressing NOR1, showed higher sensitivity to PGA2 compared to those from wild-type mice. These observations suggest that PGA2 can serve as a transactivator of NOR1, and thus suggest a possibility of pharmacological modulation of the NOR1 pathways by PGA2-related compounds.
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Pharmacology
Regular Articles
  • Wen Chao Zhao, Jin Xia Zhu, Gui Hong Zhang, Connie Hau Yan Wong, Yiu W ...
    2005 Volume 28 Issue 9 Pages 1608-1611
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The present study investigated the effect of ferulic acid, a compound purified from traditional Chinese herbal medicine Chuanxiong and Awei, on anion secretion by human colonic cells (T84) using the short circuit current (ISC) and microspectrofluorimetric technique. Basolateral administration of sodium ferulate (SF) produced a concentration-dependent increase of ISC in T84 cells with an EC50 of 1.2 mM. The SF-induced increase in ISC contained a transient peak followed by a sustained plateau. Removal of extracellular Cl, basolateral addition of bumetanide, an inhibitor of the Na+–K+–Cl cotransporter (NKCC) and apical pretreatment with DPC, a Cl channels blocker, decreased the SF-induced increase in ISC by 94% (p<0.001), 84% (p<0.001) and 85% (p<0.001) respectively. Pretreatment with thapsigargin, a specific microsomal Ca2+-ATPase inhibitor, in combination with EGTA, a Ca2+ chelator, decreased the SF-induced peak by 52% (p<0.01) and inhibited the SF-induced plateau by 60% (p<0.05). Pretreatment with MDL12330A, an adenylate cyclase inhibitor, blocked the SF-induced ISC plateau by 87% (p<0.01) but did not affect the SF-induced ISC peak. Microspectrofluorimetric measurements show that SF induced a sustained increase in [Ca2+]i. The results suggested that SF could induce Cl secretion in T84 cells via Ca2+ and cAMP-dependent pathways.
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  • Jing Ming Jia, Chun Fu Wu, Wen Liu, Hai Yu, Yue Hao, Jian Hua Zheng, Y ...
    2005 Volume 28 Issue 9 Pages 1612-1614
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The tissue culture of Saussurea involucrata (TCSauI) KAR. et KIR. was studied to determine its antiinflammatory and analgesic activities in experimental animals. Similar to wild S. involucrata, TCSauI at doses of 75—300 mg/kg i.g. for 7 d markedly inhibited hindpaw edema induced by carrageenin in rats, ear edema induced by dimethylbenzene, and increased capillary permeability in the mouse abdominal cavity induced by acetic acid. Moreover, TCSauI had inhibitory activities against the writhing reaction induced by acetic acid and the hot plate reaction in mice. The present study provided evidence first that TCSauI has antiinflammatory and analgesic activities, suggesting the potential of the tissue culture technique to substitute for wild S. involucrata in the pharmaceutical industry.
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  • Joo-Hyon Kim, Keun-Ho Ryu, Ki-Won Jung, Chang-Kyun Han, Wie-Jong Kwak, ...
    2005 Volume 28 Issue 9 Pages 1615-1620
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    SKI306X was previously reported to have good anti-inflammatory and analgesic efficacy in various studies. To determine its mode of action, an investigation was carried out for some representative mediators. Arachidonic acid metabolism and its products are particularly important in inflammation and pain. The pro-inflammatory cytokines, especially interleukin-1 (IL-1) and tumor necrosis factor (TNF-α), and induced nitric oxide (NO) appear to be most involved in the inflammatory process such as osteoarthritis (OA). Thus SKI306X was tested to determine the effects on enzymes related to arachidonic acid metabolism and the release or synthesis of inflammatory mediators. SKI306X did inhibit the expression of cyclooxygenase-2 (COX-2) enzyme without affecting COX-1 and COX-2 activity. Leukotriene B4 (LTB4) production also was inhibited by SKI306X (IC50=98.7±4.26 μg/ml). SKI306X inhibited significantly TNF-α release (IC50=97.6±17.8 μg/ml) and NO production (IC50=280±17.8 μg/ml). But IL-1α release was not attenuated by SKI306X. This study suggests that inhibition of these mediators by SKI306X may be one of the mechanisms responsible for its anti-inflammatory effects.
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  • Hwa-Young Cha, Jeong-Hill Park, Jin-Tae Hong, Hwan-Soo Yoo, Sukgil Son ...
    2005 Volume 28 Issue 9 Pages 1621-1625
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    In a previous study, we reported that ginseng extract has anxiolytic-like effects in the elevated plus-maze model and that the ginseng saponin fraction plays an important role. This experiment was performed to investigate the anxiolytic-like effects of ginsenosides Rb1, Rg1, Rg3-R, and Rg3-S, and the Rg5 and Rk mixture isolated from the ginseng saponin fraction in the elevated plus-maze. Furthermore, the anxiolytic-effects of Rb1, Rg1, Rg3-R, Rg3-S, and the Rg5 and Rk mixture were compared with those of a well-known active anxiolytic drug (diazepam). The oral administration of ginsenoside Rb1 significantly increased the number of open arm entries and the time spent on the open arm compared with those in the vehicle-treated group. Ginsenoside Rg1 and the Rg5 and Rk mixture also significantly increased the number of open arm entries and the time spent on the open arm. However, ginsenosides Rg3-R and Rg3-S did not increase the number of open arm entries or the time spent on the open arm. On the other hand, ginsenoside Rb1 and the Rg5 and Rk mixture decreased locomotor activity in a manner similar to diazepam. These data indicate that ginsenosides Rb1, Rg1, and the Rg5 and Rk mixture have anxiolytic-like effects, but ginsenosides Rg3-R and Rg3-S do not in this model. We provide evidence that some ginsenosides may be useful for the treatment of anxiety.
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  • Kota Ishida, Mikio Murata, Masatoshi Kato, Iku Utsunomiya, Keiko Hoshi ...
    2005 Volume 28 Issue 9 Pages 1626-1629
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    Using an in vivo intra-striatal microdialysis technique, we examined the effects of systemically administered β-phenylethylamine (β-PEA), a psychomotor stimulating trace amine, on striatal acetylcholine release in freely moving rats. Infusion of N-methyl-D-aspartic acid (NMDA; 10−5 M) significantly increased acetylcholine release. In addition, locally applied amino-3-hydroxy-5-methylisozasole-4-propionic acid (AMPA; 10−5 M) significantly increased acetylcholine release in the striatum. Intra-striatal application of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 10−5 M), an AMPA-type glutamatergic receptor antagonist, had little effect on acetylcholine release, while application of MK-801 (10−5 M, 10−6 M), an NMDA-type glutamatergic receptor antagonist, significantly reduced acetylcholine release. Acetylcholine within striatal perfusate was significantly increased by intraperitoneal administration of β-PEA in a dose-dependent manner. This increase in acetylcholine release was completely blocked by application of CNQX (10−5 M) through the microdialysis probe into the striatum. However, increased acetylcholine response to systemic β-PEA was unaltered by addition of MK-801 to the perfusion medium. These results suggest a regulatory function of β-PEA, mediated by AMPA-type glutamatergic receptors, on the release of acetylcholine in the rat striatum.
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  • Rong Lin, Juntian Liu, Ning Peng, Guangde Yang, Weijie Gan, Weirong Wa ...
    2005 Volume 28 Issue 9 Pages 1630-1634
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The role of C-reactive protein (CRP) in atherogenesis has been supported by more recent data. Some studies have demonstrated marked up-regulation inflammatory responses in endothelial cells subjected to CRP. The nuclear factor-κB (NF-κB) signal transduction is known to play a key role in the expression of these proatherogenic entities. Statins have anti-inflammatory properties independent of their cholesterol-lowering effects. Therefore, we studied the effects of CRP and lovastatin on NF-κB activation in human umbilical vein endothelial cells (HUVECs). By using an electrophoretic mobility shift assays (EMSA), we found that CRP (50 μg/ml) increased activation of NF-κB and degradation of inhibitory kappa B (IκB) in HUVECs, reaching a maximal effect after the incubation with CRP for 1 h. Lovastatin (10−5 mol/l) diminished NF-κB activation induced by CRP. Furthermore, lovastatin may block NF-κB activation by causing a stabilization of the IκB-α in cellular cytoplasm with western blotting analysis. Preincubation of HUVECs with pyrrolidinethiocarbamate (PDTC, NF-κB inhibitor) diminished CD40 expression induced by CRP with flow cytometry. Our results suggest that CRP increases activation of NF-κB and induces CD40 expression in HUVECs partly via activation of NF-κB. Lovastatin, through the inhibition of NF-κB activation, reduces the inflammation involved in the pathogenesis of atherosclerosis.
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  • Yuka Shimeda, Yoshihiko Hirotani, Youko Akimoto, Kyoko Shindou, Yoshio ...
    2005 Volume 28 Issue 9 Pages 1635-1638
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    Cisplatin-induced nephrotoxicity is related to an increase in lipid peroxidation and oxygen free radicals in a kidney. In the present study, we investigated the effect of the dietary antioxidants, capsaicin (Cap), against cisplatin-induced lipid peroxidation and nephrotoxicity in rats. Nephrotoxicity induced by treatment with a single dose of cisplatin (5 mg/kg body weight i.p.). The animals were divided into 4 groups. Cap (10 mg/kg/d) was given by gavage from the same day of cisplatin injection. Cisplatin administration resulted in significant increases in the kidney weight as a percentage of the total body weight, urine volume, serum creatinine, and blood urea nitrogen by about 132, 315, 797, and 556% in comparison with the control rats, respectively (p<0.05). Also, the renal tissue from the cisplatin-treated rats showed significant decreases in the kidney glutathione (GSH) content and superoxide dismustase (SOD) activity and a significant increase in malondialdehyde (MDA) production in comparison to the values at 0 h (p<0.05). Seven days after Cap plus cisplatin treatments, the renal damage induced by cisplatin recovered to a significant statistically level. In addition, Cap prevented the rise of MDA and the reduction of SOD activities. These results suggest that Cap has protective effects against cisplatin-induced nephrotoxicity and lipid peroxidation in rats.
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  • Hyun-Sun Lee, Nam Hee Won, Kyoung Heon Kim, Hojoung Lee, Woojin Jun, K ...
    2005 Volume 28 Issue 9 Pages 1639-1644
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The ripe fruit of Terminalia chebula RETZIUS (T. chebula RETZ) (Combretsceae), which is a native plant in India and Southeast Asia, has traditionally been used as a popular folk medicine for homeostatic, antitussive, laxative, diuretic, and cardiotonic treatments. The objective of this study was to evaluate the protective effects of an aqueous extract of fruit of T. chebula on the tert-butyl hydroperoxide (t-BHP)-induced oxidative injury observed in cultured rat primary hepatocytes and rat liver. Both treatment and pretreatment of the hepatocytes with the T. chebula extract (TCE) significantly reversed the t-BHP-induced cell cytotoxicity and lactate dehydrogenase leakage. In addition, TCE exhibited in vitro ferric-reducing antioxidant activity and 2,2-diphenyl-1-picryhydrazyl free radical-scavenging activities. The in vivo study showed that pretreatment with TCE (500 or 1000 mg/kg) by gavage for 5 d before a single dose of t-BHP (0.1 mmol/kg i.p.) significantly lowered the serum levels of the hepatic enzyme markers aspartate aminotransferase and alanine aminotransferase and reduced the indicators of oxidative stress in the liver, such as the glutathine disulfide content and lipid peroxidation, in a dose-dependent manner. Histopathologic examination of the rat livers showed that TCE reduced the incidence of liver lesions, including hepatocyte swelling and neutrophilic infiltration, and repaired necrosis induced by t-BHP. Based on the results described above, we speculate that TCE has the potential to play a role in the hepatic prevention of oxidative damage in living systems.
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  • Anthony Siu Lung Chan, Eric Chun-Hung Yip, Lisa Ying Yung, Haihong Pan ...
    2005 Volume 28 Issue 9 Pages 1645-1650
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    CKBM is an herbal formula composed of five Chinese medicinal herbs (Panax ginseng, Schisandra chinensis, Fructus crataegi, Ziziphus jujube and Glycine Max) supplemented with processed Saccharomyces cerevisiae. Previous studies have demonstrated that CKBM is capable of triggering the release of IL-6 and TNFα from human peripheral blood mononuclear cells, and its anti-tumorigenic activity has been demonstrated in nude mice with gastric cancer. In this report, we utilized the THP-1 monocytic cell line as a cellular model to investigate how CKBM regulates the intracellular signaling of monocytes and the subsequent release of the produced cytokines. In terms of mitogen-activated protein kinase (MAPK) cascades, CKBM (20%) had no significant effect on ERK, but was linked to an inhibitory effect on JNK and a stimulatory effect on p38 MAPK. The differential responsiveness of JNK and p38 was dependent on the duration of treatment, as well as on the dosage of CKBM. Treatment of CKBM alone induced the release of IL-10 and IFNγ, but not IL-1β, IL-4, IL-6 and TNFβ, while increase of intracellular Ca2+ concentration by A23187 triggered the release of IL-10 only. Interestingly, A23187 synergized with the activities of CKBM-treated THP-1 cells in terms of IL-1β and IFNγ production, while the IL-10 production showed no synergistic relationship between CKBM and A23187. This A23187-induced synergism was associated with a dose-dependent character towards CKBM administration. In view of the intracellular Ca2+ elevation during monocyte activation, our results suggest that CKBM can serve as a promoting agent for modulating the functions of monocytes.
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  • Takayuki Seki, Mong-Heng Wang, Noriyuki Miyata, Michal Laniado-Schwart ...
    2005 Volume 28 Issue 9 Pages 1651-1654
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    We examined the effect of N-hydroxy-N′-(4-butyl-2-methylphenyl)-formamidine) (HET0016), an inhibitor of 20-hydroxy-5,8,11,14-eicosatetraenoic acid (20-HETE) synthesis on the ω-hydroxylation and epoxidation of arachidonic acid (AA) catalyzed by recombinant cytochrome P450 4A1 (CYP4A1), CYP4A2 and CYP4A3, and characterized the enzyme inhibitory profile of HET0016. The IC50 values of HET0016 for recombinant CYP4A1-, CYP4A2- and CYP4A3-catalyzed 20-HETE synthesis averaged 17.7 nM, 12.1 nM and 20.6 nM, respectively. The IC50 value for production of 11,12-epoxy-5,8,14-eicosatrienoic acid (11,12-EET) by CYP4A2 and 4A3 averaged 12.7 nM and 22.0 nM, respectively. The IC50 value for CYP2C11 activity was 611 nM which was much greater than that for CYP4As. The initial velocity study showed the Ki value of HET0016 for CYP4A1 was 19.5 nM and a plot of Vmax versus amount of recombinant CYP4A1 added shows HET0016 is an irreversible non-competitive inhibitor. These results indicate that HET0016 is a selective, non-competitive and irreversible inhibitor of CYP4A.
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  • Mamoru Ohkita, Atsushi Nakajima, Kyoko Ueda, Masanori Takaoka, Yoshino ...
    2005 Volume 28 Issue 9 Pages 1655-1657
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The effects of flavangenol on ischemia/reperfusion-induced acute renal failure (ARF) in rats were examined. Ischemic ARF was induced by occlusion of the left renal artery and vein for 45 min followed by reperfusion, 2 weeks after contralateral nephrectomy. Renal functional parameters such as blood urea nitrogen, plasma creatinine, creatinine clearance, urine flow, urinary osmolality and fractional excretion of sodium were measured. Renal function in ARF rats markedly decreased at 1 d after reperfusion. Pre-ischemic treatment with flavangenol (3—30 mg/kg, i.v.) attenuated the ischemia/reperfusion-induced renal dysfunction. Histopathological examination of the kidney of ARF rats revealed severe renal damage, such as tubular necrosis and proteinaceous casts in tubuli, which were also significantly suppressed by the administration of flavangenol. These findings suggest that flavangenol supplementation may be a promising candidate for treatments to improve the ischemia/reperfusion-induced renal injury.
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  • Takashi Shirakura, Feng Han, Norifumi Shiota, Shigeki Moriguchi, Jiro ...
    2005 Volume 28 Issue 9 Pages 1658-1661
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    Microsphere embolism (ME)-induced ischemia model in rat resembles to multiple brain embolism in human with several clinical features. We here tested whether nitric oxide (NO) production contributes to the neuronal injury in the ME model. A novel calmodulin antagonist, DY-9760e, having a potent inhibitory effect on neuronal nitric oxide synthase (nNOS), reduced brain infarct size in the ME-induced brain ischemia. Consistent with our previous observation with gerbil ischemia/reperfusion model, DY-9760e completely inhibited NO production immediately after and 24 or 48 h after ME. Unlike the gerbil ischemia/reperfusion model, protein tyrosine nitration markedly increased 6—48 h after ME. DY-9760e treatment completely inhibited the marked increase in the protein tyrosine nitration at 24 h after ME. These results suggest that the inhibition of NO production and protein tyrosine nitration by DY-9760e contribute to its neuroprotective action in the ME-induced brain damage.
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Notes
  • Hiroki Itoh, Fumihiko Katagiri, Kazuro Ikawa, Masaharu Takeyama
    2005 Volume 28 Issue 9 Pages 1752-1756
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    Domperidone, an upper gastrointestinal function regulatory medicine, has recently been evaluated for its clinical usefulness in the treatment of stress and depression. We examined the effects of domperidone on the plasma levels of motilin-immunoreactive substance (IS), somatostatin-IS, gastrin-IS, adrenocorticotropic hormone (ACTH)-IS, and cortisol under stress conditions by repetitive blood sampling. After a single oral administration of domperidone (30 mg), the plasma domperidone level was highest (58.6±6.4 ng/ml) in the sample taken 40 min after administration, after which the plasma level fell. Peak plasma motilin-IS levels (23.1±1.4 pg/ml) were achieved 40 min after administration of domperidone (p<0.01 vs. placebo), and returned to baseline levels within a further 40 min. Plasma somatostatin-IS levels (13.0±1.2 pg/ml) increased 60 min after administration of domperidone (p<0.01 vs. placebo). Plasma gastrin-IS levels did not change significantly. These results suggest that the pharmacological effects of domperidone on gastrointestinal functions are closely related to changes in motilin-IS and somatostatin-IS levels. Domperidone significantly suppressed increases in plasma ACTH-IS and cortisol levels compared with the response to a placebo. These modulatory effects might be beneficial in stress-related diseases and suggest that this medicine has clinical pharmacological activity.
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  • Yuhki Sato, Fumihiko Katagiri, Hiroki Itoh, Masaharu Takeyama
    2005 Volume 28 Issue 9 Pages 1757-1761
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    Some traditional Chinese herbal (Kampo) medicines have recently been evaluated for their clinical usefulness in stress and depression. These medicines have modulatory effects on the hypothalamic-pituitary-adrenal axis and sympathetic nervous system (SNS). We examined the effects of Rikkunshi-to, Hange-shashin-to, Hange-koboku-to, and Ninjin-to on plasma levels of neuropeptide Y (NPY), which is the representative neurotransmitter of the SNS, under venipuncture stress. Rikkunshi-to and Hange-shashin-to suppressed increases in plasma NPY-immunoreactive substance levels compared with the response to a placebo. In this study, Rikkunshi-to and Hange-shashin-to altered plasma levels of NPY under venipuncture stress. These effects might be benefical in stress-related diseases and our results suggest that these medicines have clinical pharmacologic activity.
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  • Nobuaki Matsui, Hiroshi Nakashima, Yuki Ushio, Toshimasa Tada, Akiko S ...
    2005 Volume 28 Issue 9 Pages 1762-1765
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    Magnolol has neurotrophic effects in primary cultured rat cortical neurons, which are expressed as the promotion of neurite outgrowth and neuronal survival. In this study, we investigated the protective effect of magnolol against age-related neuronal loss in the hippocampus using senescence-accelerated mouse (SAMP1). Magnolol (5, 10 mg/kg) was orally administered once a day for 14 d to 2- or 4-month-old mice, and evaluation was carried out when the mice were 4 or 6 months old. The density of neurofibrils decreased with aging in the stratum radiatum of the CA1 region in the hippocampus of SAMP1, not SAMR1. Treatment with magnolol significantly prevented the decrease of neurofibrils in the CA1, when it was administered in 2-month-olds. However, administration at 4 months of age did not result in a preventive effect. These findings suggest that the administration of magnolol before the initiation of neuronal loss may result in a protective effect in the hippocampus.
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  • Satoshi Uchiyama, Masayoshi Yamaguchi
    2005 Volume 28 Issue 9 Pages 1766-1769
    Published: 2005
    Released: September 01, 2005
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    The effects of β-cryptoxanthin, a carotenoid, on bone components in the femoral-diaphyseal and -metaphyseal tissues of streptozotocin (STZ)-diabetic rats was investigated. Rats received a single subcutaneous administration of STZ (6.0 mg/100 g body weight), and then the animal were orally administered β-cryptoxanthin (5 or 10 μg/100 g body weight) once daily for 7 or 14 d. The administration of STZ caused a significant decrease in body weight and a significant increase in serum glucose, triglyceride, and calcium levels, indicating a diabetic state. These alterations were significantly prevented by the administration of β-cryptoxanthin (5 or 10 μg/100 g) for 14 d. The administration of β-cryptoxanthin (5 or 10 μg/100 g) to normal rats for 14 d did not have a significant effect on body weight or on serum glucose, triglyceride, and calcium levels. Calcium content, alkaline phosphatase activity, and DNA content in the femoral-diaphyseal and -metaphyseal tissues were significantly decreased in STZ-diabetic rats. These decreases were significantly prevented by the administration of β-cryptoxanthin (5 or 10 μg/100 g) for 14 d. The administration of β-cryptoxanthin to normal rats for 14 d caused a significant increase in calcium content, alkaline phosphatase activity, and DNA content in the femoral-diaphyseal and -metaphyseal tissues. This study demonstrates that the intake of β-cryptoxanthin has a preventive effect on bone loss in STZ-diabetic rats.
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  • Mayumi Tomioka, Toshihiko Murayama, Makoto Kashiwayanagi
    2005 Volume 28 Issue 9 Pages 1770-1772
    Published: 2005
    Released: September 01, 2005
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    Plasma progesterone concentrations in female Wistar rats after exposure to urine preparations with and without protease-treatment were measured to explore the effects of protease-sensitive pheromones on the endocrine state. Exposure to crude urine excreted from male rats induced an increase in the plasma progesterone concentration in female rats. The progesterone concentration of oestrous females increased with an increase in the protein concentration in urine samples. Exposure of females in the oestrous state to urine preparations treated with protease did not induce increases in plasma progesterone. These results suggest that the presence of a protease-sensitive component in male urine exerts an influence on the endocrine state of oestrous females.
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Medicinal Chemistry
Notes
  • Masahiro Ogata, Takao Kunikane, Masako Seki, Kentaro Oka, Shiro Urano, ...
    2005 Volume 28 Issue 9 Pages 1773-1775
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The mode of action of dipropofol and its antibacterial activity in combination with other antibiotics against Gram-positive and -negative bacteria were investigated. Dipropofol showed a bactericidal effect against Staphylococcus aureus 209P, and inhibited the incorporations of 3H-glutamate and 3H-leucine into S. aureus 209P and Bacillus subtilis PCI219 cells. These results indicated that the mechanism of action of dipropofol was mediated by the inhibition of protein synthesis or amino acid incorporation. A synergistic effect was performed by checkerboard titration with Muller–Hinton agar plates containing dipropofol (0.39 μg/ml, 1/4×MIC) and various concentrations of nine other antibiotics in vitro. The synergism against vancomycin resistant Enterococcus faecium was confirmed in the combination of dipropofol with rifampicin. The MIC of rifampicin was decreased from 0.39 μg/ml to <0.005 μg/ml by the addition of dipropofol. This combinational effect in reversing vancomycin resistance of enterococci highlights novel drug targets and has importance in the design of new therapeutic regimens against resistant pathogens.
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  • Yu-Ming Chi, Motoyuki Nakamura, Toyokichi Yoshizawa, Xi-Ying Zhao, Wen ...
    2005 Volume 28 Issue 9 Pages 1776-1778
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The anti-inflammatory activities of α-truxillic acid (1) and 4,4′-dihydroxy-α-truxillic acid (2) as well as their monomer components (E)-cinnamic acid (3) and (E)-p-coumaric acid (4) were evaluated in the formalin test. α-Truxillic acid (1) and its derivative 4,4′-dihydroxy-α-truxillic acid (2) exhibited significant activity against inflammatory pain response, while their monomer components (E)-cinnamic acid (3) and (E)-p-coumaric acid (4) did not show any activity against either neurogenic or inflammatory pain responses induced by formalin in mice. These results suggested that the dimeric structure might play an important role for the expression of anti-inflammatory activity. Furthermore, in order to gain information on their potencies, their anti-inflammatory activities were compared with that of incarvillateine (5) which contains the same dimeric structure and showed more potent antinociceptive activity than morphine in the formalin test. The activities of α-truxillic acid (1) and 4,4′-dihydroxy-α-truxillic acid (2) at the dose of 40 mg/kg against inflammatory pain response were equal to that of incarvillateine at doses of 20 mg/kg.
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Pharmacognosy
Regular Articles
  • Dae Gill Kang, Eun Jin Sohn, Mi Kyoung Moon, Yun Mi Lee, Ho Sub Lee
    2005 Volume 28 Issue 9 Pages 1662-1667
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The present study was designed to examine whether aqueous extract of steamed root of Rehmannia glutinose (ARR) has an ameliorative effect on renal functional parameters in association with the expressions of aquaporin 2 (AQP 2), Na,K-ATPase, and heme oxygenase-1 (HO-1) in the ischemia-reperfusion induced acute renal failure (ARF) rats. Polyuria caused by down-regulation of renal AQP 2 in the ischemia-induced ARF rats was markedly restored by administration of ARR (200 mg/kg, p.o.) with restoring expression of AQP 2 in the kidney. The expressions of Na,K-ATPase α1 and β1 subunits in the renal medullar and cortex of the ARF rats were also restored in the ARF rats by administration of ARR. On the other hand, administration of ARR lowered the renal expression of HO-1 up-regulated in rats with ischemia-induced ARF. The renal functional parameters including creatinine clearance, urinary sodium excretion, urinary osmolality, and solute-free reabsorption were also markedly restored in ischemia-ARF rats by administration of ARR. Taken together, these data indicate that RSR ameliorates renal defects in rats with ischemia-induced ARF.
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  • Hyun-Ju Jung, Sei-Gun Kim, Jung-Hwan Nam, Kwang-Kyun Park, Won-Yun Chu ...
    2005 Volume 28 Issue 9 Pages 1668-1671
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    As an attempt to search for bioactive natural products exerting antiinflammatory activity, we have isolated two saponins were isolated from the aerial portion of Pleurospermum kamtschaticum (Umbelliferae) by nitrite assay activity-directed chromatographic fractionation. They were identified as saikogenin F 3-O-{β-D-glucopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→3)]-β-D-fucopyranoside} (buddlejasaponin IV, 1) and 3β,16β,23,28-tetrahydroxy-11α-methoxyolean-12-ene 3-O-{β-D-glucopyranosyl(1→2)-[β-D-glucopyranosyl(1→3)]-β-D-fucopyranoside} (buddlejasaponin IVa, 2). Compound 1 significantly inhibited nitric oxide (NO) production, and it also significantly decreased prostaglandin E2 (PGE2) and tumor necrosis factor-α (TNF-α) release in the lipopolysaccharide (LPS)-activated macrophage Raw 264.7 cells whereas compound 2 was much less active. Saikogenin A (3) and –H (4) were obtained by hydrolyzing 1 and 2. Although these sapogenin showed strong NO inhibition, these effects were caused by the cytotoxic effect on Raw 264.7 cells. These results supported the notion that buddlejasaponin IV is a major inhibitors of NO, PGE2 and TNF-α production in P. kamtschaticum.
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  • Kanjana Sanugul, Teruaki Akao, Yan Li, Nobuko Kakiuchi, Norio Nakamura ...
    2005 Volume 28 Issue 9 Pages 1672-1678
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The C-glucosyl bond of C-glucosides generally tolerates acid and enzymatic hydrolysis. Many C-glucosides are cleaved by human intestinal bacteria. We isolated the specific bacterium involved in the metabolism of mangiferin (2-β-D-glucopyranosyl-1,3,6,7-tetrahydroxyxanthone), C-glucosyl xanthone, from a mixture of human fecal bacteria. The anaerobic Bacteroides species named MANG, transformed mangiferin to the aglycone, norathyriol, suggesting cleavage of a C-glucosyl bond. However, B. sp. MANG cleaved C-glucosyl in a dose- and time-dependent manner only when cultivated in the presence of mangiferin. Cleavage was abolished by inhibitors of RNA and protein syntheses, such as rifampicin and chloramphenicol, respectively, indicating that the enzyme that cleaves C-glucosyl is induced by mangiferin. In contrast, mangiferin did not affect bacterial α- and β-glucosidase activities under any conditions. The C-glucosyl-cleavage in cell-free extracts was not altered by potent glucosidase inhibitors such as 1-deoxynojirimycin and gluconolactone. Therefore, the C-glucosyl-cleaving enzyme substantially differs from known glucosidases that cleave O-glucosides. This is the first description of a specific intestinal bacterium that is involved in the metabolism of mangiferin and which produces a novel and inducible C-glucosyl-cleaving enzyme.
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Notes
  • Tsuyoshi Ikeda, Kazumi Yokomizo, Masafumi Okawa, Ryota Tsuchihashi, Ju ...
    2005 Volume 28 Issue 9 Pages 1779-1781
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The anti-herpes simplex virus type 1 (HSV-1) activity of 15 oleanan-type triterpenoides including glycyrrhizin and its sapogenol was examined and their structure–activity relationships were discussed. Although glycyrrhizin which exhibited in vivo efficacy against HSV-1 replication showed moderate in vitro anti-HSV-1 activity, its sapogenol, glycyrrhetic acid, showed 10 times greater action than glycyrrhizin. Therefore, the in vivo anti-HSV-1 activity of glycyrrhizin administered orally could be reasonably attributed to glycyrrhetic acid generated by hydrolysis by intestinal bacteria. Since the activity of soyasapogenol A was less than 1/20 of that of soyasapogenol B, the hydroxylation at C-21 seemed to reduce anti-HSV-1 activity. Since kudzusapogenol A, abrisapogenols B and C lacked the activity, the C-29 hydroxy group would eliminate anti-HSV-1 activity. On the other hand, since the methylesters of kudzusapogenol B and glycyrrhetic acid exhibited greater action, a methoxy carboxy group at C-20 might enhance activity.
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  • Kyung-Ah Jung, Tae-Chul Song, Daeseok Han, In-Ho Kim, Young-Eon Kim, C ...
    2005 Volume 28 Issue 9 Pages 1782-1785
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    It is currently accepted that the consumption of fruit-derived antioxidants such as vitamin C, carotenoids, and flavonoids provides a preventive effect against cardiovascular disease. The purpose of the present study was to investigate potential cardiovascular protective properties of aqueous and 70% ethanol extracts from kiwifruit by analyzing the antioxidative, antihypertensive, hypocholesterolemic, and fibrinolytic activities in vitro. Aqueous and 70% ethanol extracts at 50 mg/ml showed DPPH-radical scavenging activities of 72.31% and 70.75%, respectively. Total antioxidant activity in linoleic acid emulsion was 85—88% at 10 mg/ml and 96—98% at 50 mg/ml of kiwifruit extract. Inhibitory activities against angiogensin I-converting enzyme of kiwifruit extracts were 21—26% at 10 mg/ml and 46—49% at 50 mg/ml, and inhibitory activities on HMG-CoA reductase were 13—14% at 10 mg/ml and 19—30% at 50 mg/ml. Fibrinolytic activity of kiwifruit was also observed at a high concentration of 100 mg/ml in both aqueous and 70% EtOH extracts. Based on our results, kiwifruit have potential cardiovascular protective properties in vitro.
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  • Yoshimi Ueda, Hisae Oku, Munekazu Iinuma, Kyoko Ishiguro
    2005 Volume 28 Issue 9 Pages 1786-1790
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The anti-anaphylactic and anti-pruritic activities of a 35% EtOH extract (IT) of the flowers of Impatiens textori MIQ. were investigated by in vivo assay. IT and apigenin (1), apigenin 7-glucoside (2) and luteolin (3), principal compounds from IT, inhibited compound 48/80 (COM)-induced by blood pressure (BP) decrease, which was an immunoglobulin (Ig)E-independent anaphylaxis-like response. Compounds 1—3 all inhibited BP decrease induced by IgE-dependent anaphylaxis. Furthermore, IT also inhibited the blood flow (BF) decrease induced by antigen-induced anaphylaxis in actively sensitized mice. IT showed a significant inhibitory effect on scratching behavior induced by COM without a central depressant. IT also significantly inhibited platelet activating factor (PAF)- and serotonin (5-HT)-induced scratching behavior and mitigated protease (PA)-induced scratching behavior. These findings showed that the flowers of I. textori can be utilized as an anti-anaphylactic and anti-pruritic agent in addition to the traditional applications of this plant.
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  • Filomena Conforti, Monica Rosa Loizzo, Giancarlo Antonio Statti, Franc ...
    2005 Volume 28 Issue 9 Pages 1791-1794
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The yield of methanolic extract and total phenol and non polar content of flowered parts from Achillea ligustica ALL. are reported. GC-MS analysis of the non polar fraction showed that the triterpene moretenol was the major constituent (17.228%) followed by stigmast-6-en-3β-ol, veridiflorol and β-amyrin (7.524%, 5.078% 4.470%, respectively). The antioxidant activities of the methanolic extract and its fractions from A. ligustica were carried out using two different in vitro assays, 2,2-diphenyl-1-picrylhydrazyl (DPPH) test and lipid peroxidation of liposomes assay. Methanolic extract showed higher radical scavenging activity on DPPH (IC50 of 50 μg/ml). This activity is probably due to the phenolic fraction which shown an IC50 value of 22 μg/ml. A different result was obtained from the methanolic extract on the lipid peroxidation of liposomes (IC50 of 416 μg/ml). The α-amylase inhibition assay was applied to evaluate antidiabetic activity. The methanolic extract showed weak activity (28.18% at 1 mg/ml) while the n-hexane fraction showed 74.96% inhibition at 250 μg/ml.
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  • Yu Mi Han, Hyuncheol Oh, MinKyun Na, Beom Seok Kim, Won Keun Oh, Bo Ye ...
    2005 Volume 28 Issue 9 Pages 1795-1797
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    Protein tyrosine phosphatase 1B (PTP1B) acts as a negative regulator of insulin signaling, and selective inhibition of PTP1B has served as a potential drug target for the treatment of type 2 diabetes. In the course of screening for PTP1B inhibitory natural products, the MeOH extract of the dried root of Salvia miltiorrhiza BUNGE (Labiatae) was found to exhibit significant inhibitory effect. Bioassay-guided fractionation and purification afforded three related abietane-type diterpene metabolites 1—3. Compounds 1—3 were identified as isotanshinone IIA (1), dihydroisotanshinone I (2), and isocryptotanshinone (3) mainly by analysis of NMR and MS data. Compounds 1—3 non-competitively inhibited PTP1B activity with 50% inhibitory concentration values of 11.4±0.6 μM, 22.4±0.6 μM and 56.1±6.3 μM, respectively.
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  • Sachiko Tsukamoto, Takeshi Wakana, Keiichirou Koimaru, Takushi Yoshida ...
    2005 Volume 28 Issue 9 Pages 1798-1800
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The extract of Anemarrhenae Rhizoma (rhizomes of Anemarrhena asphodeloides BUNGE) showed neurotrophic activity toward rat pheochromocytoma (PC-12) cells. Bioassay-guided purification afforded four compounds, 2,6,4′-trihydroxy-4-methoxybenzophenone (1), 7-hydroxy-3-(4-hydroxybenzyl)chroman (2), broussonin B (3), and cis-hinokiresinol (4). Compounds 1—3 induced neurite outgrowth in PC-12 cells at concentration of 50 μg/ml, while 4 was less active. In addition, compounds 2—4 showed moderate inhibitory activities against a chymotrypsin-like activity of the proteasome.
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Biopharmacy
Regular Articles
  • Yotaro Arakawa, Shigeru Kawakami, Fumiyoshi Yamashita, Mitsuru Hashida
    2005 Volume 28 Issue 9 Pages 1679-1683
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    We investigated the effect of low-molecular-weight β-cyclodextrin (β-CyD) polymer on in vitro release of two drugs with different lipophilicities (i.e., lidocaine and ketoprofen) from mucoadhesive buccal film dosage forms. When β-CyD polymer was added to hydroxypropylcellulose (HPC) or polyvinylalcohol (PVA) film dosage forms, the release of lidocaine into artificial saliva (pH 5.7) was reduced by 40% of the control. In contrast, the release of ketoprofen from the polymer film was enhanced by addition of β-CyD polymer to the vehicle. When lidocaine and ketoprofen was incubated with β-CyD polymer in the artificial saliva, concentration of free lidocaine molecules decreased in a β-CyD polymer concentration-dependent manner. The association constant with β-CyD polymer was 6.9±0.6 and 520±90 M−1 for lidocaine and ketoprofen, respectively. Retarded release of the hydrophilic lidocaine by β-CyD polymer might be due to the decrease in thermodynamic activity by inclusion complex formation, whereas enhanced release of the lipophilic ketoprofen by the β-CyD polymer might be due to prevention of recrystallization occurring after contacting the film with aqueous solution. Thus, effects of low-molecular-weight β-CyD polymer to the drug release rate from film dosage forms would vary according to the strength of interaction with and the solubility of active ingredient.
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  • Usanee Kumprakob, Junichi Kawakami, Isao Adachi
    2005 Volume 28 Issue 9 Pages 1684-1688
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    Permeation enhancement of ketoprofen (KP) from supersaturated systems and the effects of antinucleant polymers on both stability and permeation of supersaturated KP were investigated using silicone membrane as a skin model. The supersaturation was prepared by the cosolvent technique with water and propylene glycol (PG). Saturated solubility of KP in water/PG cosolvent increased markedly with an increase in PG percentage. The time-profiles of the cumulative amount of released KP from supersaturated solutions through the membrane increased linearly, and this KP flux had a significant correlation with the degree of saturation (DS) in 80 : 20, 60 : 40, 50 : 50, and 40 : 60 (v/v) water/PG cosolvent systems. The influence of 1% solutions of antinucleant polymers, hydroxypropylmethylcellulose (HPMC), polyvinylpyrrolidone (PVP), and sodium carboxymethylcellulose (SCMC) on the DS and the stability of supersaturated KP was examined in 60 : 40 (v/v) water/PG cosolvent. The remaining DS for 24 h after mixing the solvents increased in the presence of HPMC and SCMC but not PVP. In the presence of SCMC, the physical stability of supersaturated KP was higher, however, the KP flux was lower than that in the control and in the presence of the other polymers. In conclusion, the supersaturation system can be applied to achieve higher transmembrane permeation of KP, and appropriate antinucleant polymers such as HPMC can optimize the physical stability and permeability of KP.
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  • Hirokazu Okamoto, Takashi Sakai, Kazumi Danjo
    2005 Volume 28 Issue 9 Pages 1689-1694
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The effect of sucrose fatty acid esters on transdermal permeation of lidocaine (LC) and ketoprofen was examined. A drug solution was applied to excised hairless mouse skin pretreated with a sugar ester solution to examine the direct effects of the sugar esters on skin permeability. LC was applied with a pH 6 buffer solution (98.8% ionized), pH 10 buffer solution (99.2% unionized), or propylene glycol, while KP was applied with a pH 6 buffer solution (99.1% ionized), pH 2 buffer solution (98.9% unionized), or propylene glycol. Pretreatment with J-1216 (sucrose laurate, HLB=16) or J-1205 (sucrose laurate, HLB=5) significantly increased the permeation of LC from the pH 6 solution and KP from propylene glycol, respectively. The permeability coefficients of the ionized and unionized LC and KP were calculated from the permeability data. The ionized LC and KP permeated even through skin not pretreated with sugar esters, although the permeability coefficients were 24 times and 38 times less than those of the unionized LC and KP, respectively. J-1216 pretreatment increased the permeability of ionized LC from aqueous vehicle 2.7 fold. In the next series of experiments, we formulated 1.5% of J-1205 and J-1216 in various vehicles to examine their effect on the permeation of LC applied on the excised hairless mouse skin with no pretreatment. The results coincided with the results of the pretreatment experiment, and the effect of J-1205 in propylene glycol was more remarkable than that observed in the pretreatment study. When these sugar esters were dissolved in propylene glycol at 1.5%, J-1205 increased significantly the KP permeation rate as well as LC permeation rate, suggesting that the combination of J-1205 and propylene glycol would be a potent vehicle for transdermal formulations.
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  • Jia-You Fang, Yaw-Bin Huang, Hsin-Yuan Wang, Yi-Hung Tsai
    2005 Volume 28 Issue 9 Pages 1695-1701
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The objective of this study was to examine the transdermal delivery of sodium nonivamide acetate (SNA) using iontophoresis and electroporation with ultra high molecular weight polyethylene membranes (Solupor®) to achieve controlled transdermal drug delivery. A derivative of SNA, sodium nonivamide propionate (SNP), was also used as a model drug in this investigation. Iontophoresis increased the transdermal permeation of SNA as compared to passive diffusion. Most Solupor membranes were rate-limiting for the iontophoretic permeation of SNA except for Solupor 8P07, which showed negligible resistance to SNA delivery. The tortuosity (Gurley number), pore size, and the current density-induced attachments on the surface of the Solupor membranes may have been important for their rate-controlling effect. The trends for inhibiting or controlling SNA permeation were similar for both iontophoretic and electroporation applications. The higher molecular size and lower hydrophilicity of SNP compared to SNA resulted in lower permeation of SNP using electrically-assisted methods. Moreover, the various types of Solupor membranes showed similar trends for both SNA and SNP. This present study indicates that Solupor membranes act as rate-limiting membranes for controlling the release and skin permeation of both SNA and SNP by electrically-assisted methods.
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  • Yasuyuki Sadzuka, Chieko Inoue, Saho Hirooka, Tomomi Sugiyama, Keizo U ...
    2005 Volume 28 Issue 9 Pages 1702-1706
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    We previously showed that theanine, is a major amino acid in green tea, enhanced doxorubicin (DOX) induced antitumor activity. Besides, theanine induced the elevation of glutathione (GSH) level attributable to the increase of glutamate in the liver of mice, namely theanine would reduce the adverse reaction of DOX. Consequently, theanine was thought to be effective against the tissue changes with GSH level reduction. On the other hand, it is suggested excessive uptake of alcohol causes a production of free radicals, a decrease of GSH level, and an increase in the amount of lipid peroxide (LPO) in liver, and shifting to an alcoholic liver injury. Then, aiming at the prevention and medical treatment of a hepatic toxicity by the food components with little toxicity, we have studied the effect of theanine (i.p.) on ethanol metabolism and hepatic toxicity using ethanol (p.o.) single-administered mice. On the 1st hour after ethanol administration, the ethanol concentrations in blood of the theanine combined groups decreased compared with the ethanol-alone group. The alcohol dehydrogenase and aldehyde dehydrogenase activities in the liver increased by combined theanine. Since the elevation of cytochrome P450 (CYP) 2E1 activity was controlled in the theanine-combined groups, it was considered that these disorders attributable to CYP2E1 in ethanol long-term uptake might be avoidable by theanine. Although LPO increased in 3 h after by single-administration of ethanol, the increase was controlled by theanine-administration and was improved until the normal level. In conclusion, it was indicated that theanine was effective against alcoholic liver injury.
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  • Masaki Otsuka, Kentaro Goto, Seishi Tsuchiya, Yukihiko Aramaki
    2005 Volume 28 Issue 9 Pages 1707-1710
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    We explored the involvement of the phosphatidylserine (PS)-receptor in the production of TGF-β by macrophages treated with PS-liposomes. The binding of anti-PS-receptor antibody to macrophages was specifically inhibited by PS-liposomes. The antibody led to an increase in the production of TGF-β, and also activated ERK, a member of the MAP kinase. But no activations in p38 and JNK were observed. ERK inhibitor, U0126 completely prevented TGF-β production. On the addition of a TGF-β neutralizing antibody or U0126, the inhibitory effect of the anti-PS-receptor antibody on macrophage function, nitric oxide production, was restored. These findings suggested that TGF-β is one of factors produced by PS-liposomes, and the ERK signaling pathway via the PS-receptor is intimately involved in the production of TGF-β in macrophages.
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  • Toshiro Niwa, Toshifumi Shiraga, Ikuko Ishii, Akira Kagayama, Akira Ta ...
    2005 Volume 28 Issue 9 Pages 1711-1716
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    The metabolic activities of six psychotropic drugs, diazepam, clotiazepam, tofisopam, etizolam, tandospirone, and imipramine, were determined for 14 isoforms of recombinant human hepatic cytochrome P450s (CYPs) and human liver microsomes by measuring the disappearance rate of parent compounds. In vitro kinetic studies revealed that Vmax/Km values in human liver microsomes were the highest for tofisopam, followed by tandospirone>clotiazepam>imipramine, diazepam, and etizolam. Among the recombinant CYPs, CYP3A4 exhibited the highest metabolic activities of all compounds except for clotiazepam and imipramine. The metabolism of clotiazepam was catalyzed by CYP2B6, CYP3A4, CYP2C18, and CYP2C19, and imipramine was metabolized by CYP2D6 most efficiently. In addition, the metabolic activities of diazepam, clotiazepam, and etizolam in human liver microsomes were inhibited by 2.5 μM ketoconazole, a CYP3A4 inhibitor, by 97.5%, 65.1%, and 83.5%, respectively, and the imipramine metabolism was not detected after the addition of 1 or 10 μM quinidine, a CYP2D6 inhibitor. These results suggest that the psychotropic drugs investigated are metabolized predominantly by CYP3A4, except that CYP2D6 catalyzes the metabolism of imipramine. In addition, this approach based on the disappearance rate appears to be useful for the identification of the responsible CYP isoform(s) of older drugs, for which metabolic profiles have not been reported.
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  • Atsushi Ishikado, Hiromichi Imanaka, Takashi Takeuchi, Etsumori Harada ...
    2005 Volume 28 Issue 9 Pages 1717-1721
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    It is known that lactoferrin is one of the functional proteins contained in mammalian milk and that it plays an important role in the immune system. In this study, we prepared multi-lamellar liposomal bovine lactoferrin composed of egg yolk phosphatidylcholine and phytosterol for oral delivery, and examined any resulting anti-inflammatory effects. Oral pretreatment of liposomal lactoferrin exhibited more suppressive effects than did non-liposomal lactoferrin on CCl4-induced hepatic injury in rats as well as on lipopolysaccharide-induced TNF-alpha production from mouse peripheral blood mononuclear leukocytes. Further investigation revealed that the liposomalization did not exert influence on the absorbability of lactoferrin to the venous blood or lymph following an intraduodenal administration in rats. Furthermore, there was no significant difference exhibited between the antigenicity of liposomal and non-liposomal lactoferrin, which was measured using the passive cutaneous anaphylaxis reaction following oral sensitization to them in guinea pigs. These results suggest that liposomal lactoferrin might act more effectively than conventional lactoferrin in the intestinal site, which is regarded as an active site of orally administered lactoferrin, although the biological mechanism is not fully understood yet. Consequently we propose that liposomal lactoferrin could be a novel active constituent useful for preventive and therapeutic treatment of inflammatory diseases.
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  • Sew-Fen Leu, Chi-Hsien Chien, Chi-Yu Tseng, Yu-Ming Kuo, Bu-Miin Huang
    2005 Volume 28 Issue 9 Pages 1722-1725
    Published: 2005
    Released: September 01, 2005
    JOURNALS FREE ACCESS
    Cordyceps sinensis (CS), an Ascomycetes fungus parasitic to Lepidoptera larvae, has been traditionally used as nutritious food for the enhancement in immuno-modulation in Chinese society for a long time. Previous report has demonstrated the CS water extract stimulates in vitro corticosterone production in rat primary adrenal cells. In the present studies, we determined the in vivo effects of CS and its fractions on plasma corticosterone production in mouse. Different concentrations of CS and CS fractions dissolved in water (0.02 and 0.2 mg/g body weight) were fed to immature and mature mice from 1, 3 or 7 d. The plasma levels of corticosterone were determined by radioimmunoassay (RIA), and the weight of adrenal gland and body weight were also evaluated. Results illustrated that plasma corticosterone levels were significantly induced by F2 at 0.02 mg/g body weight with 7 d feeding in immature mice, and by CS at 0.02 mg/g body weight with 3 d feeding and F3 at 0.02 mg/g body weight for 7 d feeding in mature mice, respectively (p<0.05). There were no differences of adrenal gland weight except there was significant stimulation by CS at 0.2 mg/g body weight with 3 d feeding in mature mice (p<0.05) and there were significant inhibitions by both dosages of F3 for 3 d feeding in immature mice and F2 for 7 d feeding in mature mice (p<0.05), respectively. Concerning body weight, the stimulatory effects were observed with CS feeding at 0.2 mg/g body weight for 7 d and F3 feeding at 0.02 mg/g body weight for 3 and 7 d in mature mice. Whereas, the inhibitory effect were observed in F2 feeding at 0.2 mg/g body weight for 7 d in immature mice and at both dosages for 7 d in mature mice, respectively. Taken together, these studies illustrate that CS and its fractions stimulated mouse in vivo corticosterone production. However, CS and its fractions didn't have constant stimulatory or inhibitory effects on the weights of body and adrenal glands.
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