Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
Volume 21 , Issue 12
Showing 1-35 articles out of 35 articles from the selected issue
  • Michael U. ADIKWU, Kenneth C. OFOKANSI, Anthony A. ATTAMA
    1998 Volume 21 Issue 12 Pages 1243-1246
    Published: December 15, 1998
    Released: April 10, 2008
    JOURNALS FREE ACCESS
    Spectrophotometric absorption studies geve evidence for the formation of strongly bonded charge-transfer complexes between moclobemide and promethazine hydrochloride with chloranilic acid in non-aqueous media comprising chloroform and 1, 4-dioxane. The transitions involved were detected at wavelengths longer than those of the single pure substances in the visible region. Equilibrium constants from the Scott equation could be measured together with other thermodynamic parameters and molar absorptivities at different temperatures. Theoretical arguments are presented as to the spontaneity of these interactions.
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  • Naoki NAKATSUBO, Hirotatsu KOJIMA, Kuniko SAKURAI, Kazuya KIKUCHI, Hir ...
    1998 Volume 21 Issue 12 Pages 1247-1250
    Published: December 15, 1998
    Released: April 10, 2008
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    A specific and sensitive detection method for nitric oxide (NO) in living cells and tissue culture systems is required in the search for novel NO synthase (NOS) inhibitors. We have improved a fluorometric determination with 2, 3-diaminonaphthalene (DAN) by the addition of 2-phenyl-4, 4, 5, 5-tetramehylimidazoline-3-oxide-1-oxyl (PTIO) as an oxidant to form NO2 from NO. This method is 3 times more sensitive than that without PTIO, and is suitable for examining the NOS-inhibitory activity of large numbers of test compounds using a 96-well micropolate reader. The improved method was applied to N-monomethyl-L-arginine (L-NMMA) as a known inhibitor and the derivatives of 2-phenyl-1, 2-benzisoselenazol-3(2H)-one as teat compounds in order to investigate the effect of these compounds on NO production from activated rat aortic smooth muscle cells. The results obtained indicate that this method is suitable for the rapid assay of large numbers of test compounds.
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  • Chuan LI, Masato HOMMA, Kitaro OKA
    1998 Volume 21 Issue 12 Pages 1251-1257
    Published: December 15, 1998
    Released: April 10, 2008
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    There has been little explanation of herbal medicines by modern medical sciences, including pharmacokinetics, whereas physicians follow empirical indications written in classical literature. Recent reports of herb-induced adverse reactions compelled us to proceed the investigation of a herbal medicine Shosaiko-to (TJ-9) from a pharmacokinetic point of view.To five healthy volunteers, a single 5g dose of TJ-9, consisting of 7 herbs, was administered. We conducted HPLC analysis of the timed-urine specimens to disclose the type and amount of compounds excreted. Excretion rate-time curves were analyzed individually.Four flavonoids, liquiritigenin, baicalein, wogonin and oroxylin A, were found both in the urine and TJ-9. The glycosides in TJ-9 were absorbed after microflora hydrolysis. Davidigenin, which was not found in TJ-9, was an intestinal metabolite of liquiritigenin. Also, two flavanones, S-dihydrowogonin and S-dihydrooroxylin A, were identified as the metabolites of wogonin and oroxylin A, respectively. Excretion rate-time curves of the flavonoids were divided into three types of structure-dependent absorption, i.e. (1) the fast absorption of herbal-origin aglycons, (2) the moderately-delayed absorption of aglycons derived from herbal glycosides, and (3) markedly-delayed absorption after the molecular transformation of herbal compounds.Individual excretion profiles seemed to depend on microflora activities. Two types of flavonones, S-dihydrowogonin and S-dihydrooroxylin A, were found in a half of the volunteers, suggesting there might be two kinds of volunteers, namely, rapid and poor metabolizers of flavonoids.
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  • Daisei MIYAMOTO, Naoko ENDO, Naoto OKU, Yaeno ARIMA, Takashi SUZUKI, Y ...
    1998 Volume 21 Issue 12 Pages 1258-1262
    Published: December 15, 1998
    Released: April 10, 2008
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    The cytotoxic effects of β-thujaplicin and five kinds of metal chelates were examined on mouse melanoma B16BL6 cells by cell viability and lactate dehydrogenase (LDH) release assay. β-Thujaplicin-zinc chalate and β-thujaplicin-copper chelate had higher cytotoxic effects than β-thujaplicin, and the 50% effective does (ED50) of these metal chelates were 12.5 and 25 μM, respectively. In addition, the zinc chelate induced DNA ladder formation in B16BL6 cells, as shown by the DNA fragmentation assay, suggesting that cell death induced by the zinc chelate is apoptosis. The zinc chelate also had a cytotoxic effect and induced DNA fragmentation on other tumor cell lines : HeLa, Meth A, and B16F1 cells, but not on normal human diploid fibroblasts FS-4. These results suggest that β-thujaplicin-zinc chelate induces apoptotic cell death in various tumor cell lines and is a potent antitumor agent for tumor cells including malignant melanomas.
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  • Yumi YOKOYAMA, Kohfuku KOHDA, Masaji OKAMOTO
    1998 Volume 21 Issue 12 Pages 1263-1266
    Published: December 15, 1998
    Released: April 10, 2008
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    CytA protein (27 kDa) is produced by Bacillus thuringiensis subsp. israelensis (BTI) and is contained in its inclusion bodies. We previously reported the isolation of 25 kDa portion of CytA protein (p25-CytA protein) and its strong cytotoxic activity to mammalian cells. When p25-CytA protein was applied to an anion-exchange column for further purification, three fractions (M1, M2 and M3) were separted. M1 and M2 fractions were both shown to be 25 kDa protein, while M3 was a high molecular weight complex composed of 25 kDa protein and DNA. Purification and amino acid sequence analysis showed that M1 and M2 fractions were proteins lacking 29 and 31 N-terminal amino acids from CytA protein, respectively, and M3 was M1 protein associated with DNA. DNA was detected in BTI cells co-localizing with inclusion bodies. Both M1 and M2 proteins could bind to double-stranded DNA of BTI genome in vitro; DNA binding ability of M1 protein was higher than that of M2 protein. These results suggested that CytA protein has DNA binding ability and is associated with DNA in the mother cell.
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  • Mayumi SUZUKI, Tadahide FURUNO, Reiko TESHIMA, Jun-ichi SAWADA, Mamoru ...
    1998 Volume 21 Issue 12 Pages 1267-1270
    Published: December 15, 1998
    Released: April 10, 2008
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    Culture media from rat basophilic leukemia cells (RBL-2H3) induced the neurite outgrowth of rat pheochromocytoma PC12 cells, a model system for neuronal differentiation. The extension of the neurite outgrowth was dependent on the culture time of RBL-2H3 cells in the DMEM medium. The DMEM medium conditioned by RBL-2H3 cells for 48 h induced neurite outgrowth of PC12 cells significantly. The neurite extension was much higher than that by medium containing 1 ng/ml nerve growth factor (NGF) but was rather lower than that by medium containing 10 or 50 ng/ml NGF. The neurite extension by 50 ng/ml NGF was completely suppressed by excess anti-NGF antibody (1-1.5 μg/ml), while the extension by culture medium conditioned by RBL-2H3 cells for 48 h was not completely suppressed in the presence of the same amount of anti-NGF antibody. The neurite extension by the culture medium of RBL-2H3 cells was also suppressed by anti-interleukin (IL)-6 antibody (1 μg/ml), although IL-6 itself (20 units) could scarcely induce the neurite outgrowth of PC12 cells. This suggests that IL-6 in the culture medium of RBL-2H3 cells could be effective in inducing the neurite extension in cooperation with NGF. In the presence of an excess of both anti-NGF and anti-IL-6 antibodies, the culture medium of RBL-2H3 cells induced the neurite extension of PC12 cells. This suggests that the action of the various factors from RBL-2H3 cells may be synergistic as far as the neurite outgrowth of PC12 cells is concerned.
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  • Akira SATO, Min-Zhao HUANG, Shiro WATANABE, Harumi OKUYAMA, Hideko NAK ...
    1998 Volume 21 Issue 12 Pages 1271-1276
    Published: December 15, 1998
    Released: April 10, 2008
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    We examined in vivo the effect of dietary fats and oils with different peroxidizability on protein carbonyl content, the presumed index of free radical-mediated protein oxidation. For 15.6 months, SHRSP (stroke-prone spontaneously hypertensive strain) rats were fed a diet supplemented with lard, safflower oil, perilla oil or fish oil/soybean oil, the peroxidizability of which increases in this order. The peroxidizability of tissue lipids was positively correlated with the protein carbonyl content in skeletal muscle, but not in the brain, heart or liver. The protein carbonyl content in the lard group was higher in the brain and liver compared to the other dietary groups. These results contradict the concept that long-term feeding of easily autoxidizable fatty acids allows the accumulation of lipid peroxides to accelerate the development of the free radical diseases, and suggest that tissue protein carbonyl content is not a simple reflection of autoxidizability-related lipid peroxidation but is also influenced by other biochemical processes.
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  • Yoshiko KASE, Kazuko SAITOH, Atsushi ISHIGE, Yasuhiro KOMATSU
    1998 Volume 21 Issue 12 Pages 1277-1281
    Published: December 15, 1998
    Released: April 10, 2008
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    To determine the mechanisms by which Hange-shashin-to (TJ-14) reduces prostaglandin E2 (PGE2) levels, the effects on blood corticosterone levels were examined in vivo and the effects on cyclooxygenase (COX) activity in virto assessed. TJ-14, orally administered to rats at dose levels between 125 and 1000 mg/kg, caused a dose-dependent increase in blood corticosterone levels. We also showed that Glycyrrhizae Radix and Ginseng Radix, constituents of TJ-14, are involved in the increase in blood corticosterone. The activity of COX-1 was not inhibited by TJ-14 even at a dose of 1000 μg/ml, while COX-2 was inhibited at dose levels between 10 and 1000 μg/ml. The constituents Scutellariae Radix, Glycyrrhizae Radix and Coptidis Rhizoma were believed to be involved in COX-2 inhibition.These results suggest that the effect of TJ-14 in decreasing PGE2 is partially mediated by corticosterone and inhibition of COX-2.
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  • Shigeyoshi HARADA, Toshiro MAEKAWA, Eiji HANEDA, Yuko MORIKAWA, Nobuyu ...
    1998 Volume 21 Issue 12 Pages 1282-1285
    Published: December 15, 1998
    Released: April 10, 2008
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    By means of successive Mono Q and glycyrrhizin (GL)-affinity column chromatography (HPLC), recombinant HIV-1 RT (rRT) was purified to apparent homogeneity from the Superdex 200 pg fraction of the crude protein extract of E. coli BL21 transfected with pET 21a(+)/HIV-1 PR-RT. It was found that (i) rRT functioned as an effective phosphate acceptor for recombinant human casein kinase II (rhCK-II) in vitro; (ii) this phosphorylation was inhibited by anti-HIV-1 substances [a glycyrrhetinic acid derivative (oGA) and quercetin] and a high dose (100 μM) of GL; (iii) RNA-dependent DNA polymerase (RDDP) activity was stimulated about 2.5-fold after full phosphorylatioin of rRT by rhCK-II; and (iv) oGA as well as NCS-chromophore effectively prevented the CKI-II-mediated stimulation of RDDP activity. These results suggest that the anti-HIV-1 effect of oGA may be involved in the selective inhibition of the CK-II-mediated stimulation of HIV-1 RT at the cellular level.
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  • Hajimu KURUMATANI, Kazuya KIKUCHI, Tetsuo NAGANO, Masaaki HIROBE, Jun ...
    1998 Volume 21 Issue 12 Pages 1286-1289
    Published: December 15, 1998
    Released: April 10, 2008
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    Direct detection of nitric oxide (NO) is essential for understanding the precise mechanism of its production from endothelial cells. Previously, we developed an NO detection system based on the chemiluminescence reaction between NO and luminol-H2O2. Here, we have applied this system to cultured endothelial cells for the direct and on-time measurement of NO. The perfusate from cultured endothelial cells was continuously mixed with luminol-H2O2. NG-monomethyl-L-arginine (L-NMMA) (10-4M) decreased the chemiluminescence signal of NO, suggesting the existence of basal NO release. Bradykinin (10-8-10-6M) increased the NO signal (10-6M;5.1±0.4 fmol/min, corresponding to 1.7 pM in the perfusate), and this was inhibited by 10-4M L-NMMA (1.8±0.3 fmol/min). These results corresponded to the changes in cGMP levels in RFL-6 cells, which provide an No bioassay system. We conclude that the luminol-H2O2 system is useful for the direct and continuous measurement of NO from coltured endothelial cells.
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  • Naotsugu HORIKAWA, Kazuyoshi KATAHA, Nobue WATANABE, Kunio ISHII, Nobo ...
    1998 Volume 21 Issue 12 Pages 1290-1293
    Published: December 15, 1998
    Released: April 10, 2008
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    The effects of helodermin, a basic 35-amino acid peptide isolated from the venom of a lizard salivary gland, on arterial blood pressure and heart rate were examined in the rat, focusing on the possibility that activation of ATP sensitive K+ (KATP channels is involved in the responses. The results were also compared with those of vasoactive intestinal polypeptide (VIP). Helodermin produced hypotension in a dose-dependent manner with approximately similar potency and duration to VIP. Hypotension induced by both peptides was significantly attenuated by glibenclamide, which abolished a levcromakalim-produced decrease in arterial blood pressure. Oxyhemoglobin did not affect helodermin-induced hypotension, whereas it shortened the duration of acetylcholine (ACh)-produced hypotension. These findings suggest that helodermin-produced hypotension is partly attributable to the activation of glibenclamide-sensitive K+ channels (KATP channels), which presumably exist on arterial smooth muscle cells. EDRF (endothelium-derived relaxing factor)/nitric oxide does not seem to play an important role in the peptide-produced hypotension.
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  • Ryo-ichi YASUNO, Tamami OGUMA, Yasusuke MASUDA
    1998 Volume 21 Issue 12 Pages 1294-1299
    Published: December 15, 1998
    Released: April 10, 2008
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    Oxethazaine (OXZ), a potent topical anesthetic, was found to induce red blood cell (RBC) lysis in vitro, depending upon concentrations of OXZ, RBC and Ca2+. In a 2% RBC suspension, 100 μM OXZ caused almost complete hemolysis in the presence of 1.3 mM Ca2+ with only a minimal effect in its absence, while higher concentrations of OXZ (400 μM<) produced hemolysis without Ca2+. The hemolysis induced by OXZ plus Ca2+ was poreceded by a rapid increase in 45Ca2+ uptake by RBCs, with both the hemolysis and Ca2+ uptake being inhibited by 1 mM CoCl2, NiCl2, and quinine. Together with the Ca2+ influx, rapid influx of Na+ and efflux of K+ occurred, and an increasing external K+/Na+ concentration ratio inhibited both hemolysis and Ca2+ influx. Morphologically, OXZ plus Ca2+ caused rapid transformation to spheroechinocytes, the formation of blebs and the pinching-off of blebs, whereas OXZ alone produced membrane invagination. SDS-PAGE analysis of the ghosts prepared from the RBCs treated with OXZ plus Ca2+ revealed derangement of cytoskeletal components. OXZ is a rate drug that exhibits a Ca2+ ionophore-like action, increasing the Ca2+ permeability of plasma membranes.
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  • Toshio OKANO, Kimie NAKAGAWA, Naoko TSUGAWA, Keiichi OZONO, Noboru KUB ...
    1998 Volume 21 Issue 12 Pages 1300-1305
    Published: December 15, 1998
    Released: April 10, 2008
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    1α, 25-Dihydroxsyvitamin D3 [1α, 25(OH)2D3] mediates its biological activities through specific binding to the nuclear vitamin D receptor (VDR). The VDR, bound to 1α, 25(OH)2D3, forms a heterodimer with a nuclear accessory factor, retinoid X receptor (RXR), and the complex subsequently binds to specific nucleotide sequences or a vitamin D-responsive element (VDRE) to induce gene transcriptions. Thus, an ideal analogue of 1α, 25(OH)2D3 for therapeutic applications has been considered to be one which has a high binding affinity for VDR, thus forming a stable VDR/RXR complex, and binding strongly to VDRE. By contrast, we report here evidence contrary to this hypothesis. Several singly dehydroxylated A-ring analogues of 19-nor-1α, 25-dihydroxyvitamin D3 and 19-nor-22-oxa-1α, 25-dihydroxyvitamin D3, all of which have an extremely low binding affinity for VDR, and some of which lack the 1α-hydroxyl group that is considered to be essential for VDR-mediated gene expression, have greater or equivalent potencies to 1α, 25(OH)2D3 for inhibiting the proliferation and inducing the differentiation of HL-60 cells, as well as inducing the transactivation of a luciferase reporter gene combining a rat 25-hydroxyvitamin D3 24-hydroxylase gene promoter containing two VDREs. The present findings open interesting possibilities as to the role of the VDR in the genomic action of 1α, 25(OH)2D3 and the development of new 19-nor-analogues of 1α, 25(OH)2D3.
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  • Yanmei LI, Takahiro SATO, Koichi METORI, Katsuya KOIKE, Qing-ming CHE, ...
    1998 Volume 21 Issue 12 Pages 1306-1310
    Published: December 15, 1998
    Released: April 10, 2008
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    We have reported that collagen synthesis was stimulated by the administration of a hot water extract from the leaves of Eucommia ulmoides OLIVER, Eucommiaceae (Du-Zhong leaves) in false aged model rats. In this paper, we set out to examine the compounds in Du-Zhong leaves that stimulated collagen synthesis in false aged model rats. In experiment 1, a methanol extract of Du-Zhong leaves also stimulated collagen synthesis in aged model rats. An acetone fraction was derived from the methanol extract by silica gel chromatography in experiment 2. The acetone fraction mainly contained iridoides mono-glycosides such as geniposidic acid and aucubin.The administration of geniposidic acid or aucubin stimulated collagen synthesis in aged model rats in experiments 3 and 4 (significance (p<0.05)). The reported pharmacological effects of Du-Zhong leaves, including healing organs and strengthening bone and muscle, are closely related to collagen metabolism. It appears that geniposidic acid and aucubin are the actual compounds in Du-Zhong which caused the effect in our experiments.
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  • Chun Zhen WU, Makoto INOUE, Yukio OGIHARA
    1998 Volume 21 Issue 12 Pages 1311-1316
    Published: December 15, 1998
    Released: April 10, 2008
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    The effect of Ogi-Keishi-Gomotsu-To-Ka-Kojin (OKGK), a traditional Chinese herbal medicione (Kampo medicine), on cholesterol metabolism was studied in male Sprague-Dawley rats. Intake of OKGK at doses of 1.38 g/kg for 4 weeks significantly reduced total cholesterol levels in the surum and liver of hypercholesterolemia rats fed a cholesterol-enriched diet. OKGK suppressed cholesterol absorption through the intestine and stimulated excretion of cholesterol into feces as bile acids. Biochemical study indicated that OKGK treatment enhanced cholesterol 7α-hydroxylase activity the rate limiting enzyme of cholic acid synthesis, in the liver without any effect on the rate limiting enzyme of cholesterol synthesis, 3-hydroxy-3-methylglutaryl coezyme A (HMG-CoA) reductase. Further, cholesterol-enriched diet containing cholic acid suppressed cholesterol 7α-hydroxylase activity, whereas OKGK administration reversed the suppression. In conclusion, these results supported the idea that OKGK may be an effective agent for treatment of patients with hypercholesterolemia.
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  • Hideaki MATSUDA, Norimichi TOMOHIRO, Masayuki YOSHIKAWA, Michinori KUB ...
    1998 Volume 21 Issue 12 Pages 1317-1321
    Published: December 15, 1998
    Released: April 10, 2008
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    A methanol extract (TMe-ext) from the dried rhizome of Alisma orientale was screened for anti-complemenetary activity in experimental models. In the animal models, it was found that TMe-ext inhibits zymosan-induced hind paw edema in rats and zymosan-activated rat serum (ZAS)-induced vascular permeability in mice. TMe-ext showed an inhibitory effect on complement-induced hemolysis through both the classical pathway and the alternative pathway. And TMe-ext inhibited hypotonic shock-induced hemolysis, but this effect was weak compared with the anti-complementary activities of TMe-ext. Four triterpenes (alisol A, alisol A monoacetate, alisol B and alisol B monoacetate) isolated from the rhizome also inhibited the complement-induced hemolysis through the classical pathway, but two sesquiterpenes (alismol and alismoxide) were ineffective. These results indicate that Alismatis Rhizoma shows anti-complementary activity, and its anti-complementary components are partially attributable to the terpene components mentioned above.
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  • Huiying LI, Tatsuro MIYAHARA, Yasuhiro TEZUKA, Tsuneo NAMBA, Nobuo NEM ...
    1998 Volume 21 Issue 12 Pages 1322-1326
    Published: December 15, 1998
    Released: April 10, 2008
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    Four water extracts of Kampo formulae (Yi-kkan-sen, Dai-ho-in-gan, Ni-chi-gan, Tsu-kan-gan) were screened for their inhibitory activities on bone resorption induced by parathyroid hormone (PTH) in organ culture using neonatal mouse parietal bones. Among the Kampo formulae, Tsu-kan-gan (TKG) showed the most potent inhibitory activity. We further fractionated the TKG water extract by monitoring the inhibitory activity on bone resorption stimulated by PTH in virto. The MeOH fraction of the water extract inhibited PTH-stimulated bone resorption, and its inhibitory activity was more potent than those of other fractions. The MeOH fraction was then subjected to Sephadex LH-20 column chromatography to give fractions I, II and III, which were examined for bone resorption activity. Fraction I inhibited PTH-stimulated bone resorption, and its inhibitory activity was more potent than those of the other fractions. Upon oral administration of the three fractions (100 mg/kg/d) to ovariectomized (OVX) mice, fractions I and III prevented the decrease of bone mineral density (BMD) of the lumbar vertebra. Eleven compounds isolated from the MeOH fraction were examined for their inhibitory effect on PTH-stimulated bone resorption. Among them, berberine (1), syringin (3), limonin (4) and mangiferin (10) showed a significant inhibitory effect on bone resorption. In the formation assay of osteoclast-like cells, these compounds decreased the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells (MNCs). The inhibitory effect of TKG on bone resorption may be at least partly due to the inhibitory action of these compounds.
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  • Kalpana SRIVASTAVA, Tomomi HATANAKA, Kazunori KATAYAMA, Tamotsu KOIZUM ...
    1998 Volume 21 Issue 12 Pages 1327-1333
    Published: December 15, 1998
    Released: April 10, 2008
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    The purpose of this work was to investigate the disposition characteristics and pharmacodynamics of a barbiturate, thiopental, in renal dysfunction rats. Normal and renal dysfunction rats were infused with 40 and 20 mg/kg of thiopental, respectively. The quantitative electroencephalographic (EEG) method was used as the surrogate measure of pharmacological response. Signals from two electrodes fitted on the skull of rats were continuously measured, recorded and subjected to off-line analysis. Total amplitude (0.5-29.9 Hz) from aperiodic analysis was taken as the EEG parameter. Thiopental concentration in plasma and cerebrospinal fluid (CSF) was assayed by an HPLC method. Steady-state volume of distribution was increased in renal dysfunction rats due to decreased plasma protein binding, while no change in clearance or volume of distribution based on the plasma unbound concentration was observed. Amplitude changes induced by thiopental in both normal and renal dysfunction rats were characterized by the sigmoidal Emax model. The unbound plasma concertration at half maximal effect was lowered by 30% in renal dysfunction rats as compared to the normal rats. In addition to considerable alteration in the pharmacokinetics of thiopental, it was also evident that renal impairment is associated with an increase in apparent pharmacological sensitivity, which is related to affinity.
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  • Katsuji OHTA, Yoshiki FUKASAWA, Jun-ichi YAMAGUCHI, Yoshiro KOHNO, Kiy ...
    1998 Volume 21 Issue 12 Pages 1334-1337
    Published: December 15, 1998
    Released: April 10, 2008
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    Formation of a nucleophilic 4(5H)(or 5(4H))-imidazolone structure has been postulated from in vitro studies to be one of the causative elements involved in the retention of drugs with imidazole moiety in connective tissue.To confirm this, we searched for the imidazolone-related metabolite in rats after intravenous dosing of 2-methylimidazole (2MI; 14C-labeled and unlabeled form, 3 and 300 μmol/kg body weight) as a model compound.The excreted urine, the major route of elimination of the compound, was collected and analyzed using the HPLC/MS system with a counterion effect for metabolite separation. 2-Methyl-4(5H)(or 5(4H))-imidazolone (2MIone) was identified as a urinary metabolite by chromatographic and mass-spectral inspection with the corresponding authentic standard. Pretreatment of rats with either SKF-525A (50 mg/kg, i.p.) or cimetidine (200 mg/kg, i.p.) significantly increased the excreted amount of 2MIone in urine and the irreversible binding of 2MI equivalents in the aortic tissue, whereas both factors were reduced by pretreatment with triethylenetetramine dihydrochloride (150 mg/kg/d for 5 d, s.c.). These results support the aforementioned deduction, and also raise the possibility that a cytochrome P450-independent, copper-related metabolic reaction might be involved in the imidazolone formation in vivo.
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  • Kazuya FUKUMURA, Kiyoshi YAMAOKA, Mitsuo HIGASHIMORI, Terumichi NAKAGA ...
    1998 Volume 21 Issue 12 Pages 1338-1343
    Published: December 15, 1998
    Released: April 10, 2008
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    A pharmacokinetic analysis program based on a tank-in-series model, MULTI(TIS), was developed for the evaluation of dose-dependency in the local disposition of a drug. The program written in FORTRAN was constructed by expanding MULTI(RUNGE). The reliability of MULTI(TIS) was verified by analyzing the experimental data based on linear and nonlinear tank-in-series models. Linear one- and two-compartment tank-in-series models were adopted to analyze outflow time profiles in single-pass hepatic perfusion following a pulse input of 5'-deoxy-5-fluorouridine (DFUR). The estimated parameters agreed well with those by MULTI(FILT) which is widely used for linear kinetic analysis. The nonlinear models adopted were one-compartment model with Michaelis-Menten elimination and two-compartment models with Michaelis-Menten elimination from central and peripheral compartments. Oxacillin was used as a model drug, because time courses of oxacillin show a capacity-limited hepatic disposition following a pulse input in high doses to the liver (300, 1000, 3000 and 5000 μg). The hepatic recovery ratio (FH) of oxacillin increased with dose, whereas the mean transit time (t^-H) was almost constant. The maximum elimination rate constant (Vmax) and Michaelis constant (Km) of oxacillin were estimated to be 1980 μg/ml/min and 54.1 μg/ml, respectively. Thus, the reliability of MULTI(TIS) was demonstrated for the analysis of nonlinear local disposition, especially, capacity-limited elimination in the liver.
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  • Shigeyuki KITAMURA, Akira TERADA, Naoko INOUE, Hiroshi KAMIO, Shigeru ...
    1998 Volume 21 Issue 12 Pages 1344-1347
    Published: December 15, 1998
    Released: April 10, 2008
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    Rat blood exhibited a significant quinone-dependent N-oxide reductase activity towards imipramine N-oxide. The reduction mediated by the blood proceeded in the presence of both NAD(P)H and menadione under anaerobic conditions. When menadione was replaced with 1, 4-naphthoquinone or 9, 10-phenanthrenequinone, similar results were obtained. The reduction was also mediated by the combination of rat erythrocytes and plasma. The reducing activity was inhibited by dicumarol and carbon monoxide. When boiled plasma was combined with untreated erythrocytes, the N-oxide reducing activity was abolished. In contrast, when boiled erythrocytes were combined with untreated plasma, the activity was unchanged. These results suggest that the activity is caused by the heme of hemoglobin in erythrocytes and quinone reductase in plasma. In fact, erythrocytes and hemoglobin have the ability to reduce the N-oxide when supplemented with DT-diaphorase purified from rat liver in the presence of both NAD(P)H and menadione. Hemoglobin also exhibits N-oxide reductase activity with reduced menadione (menadiol). Furthermore, hematin exhibits a significant reducing activity in the presence of menadiol. The reduction appears to proceed in two steps. The first step is enzymatic reduction of quinones to dihydroquinones by quinone reductse(s) with NADPH or NADH in plasma. The second step is nonenzymatic reduction of imipramine N-oxide to imipramine by the dihydroquinones, catalyzed by the heme group of hemoglobin in erythrocytes. Cyclobenzaprine N-oxide and brucine N-oxide are similarly transformed to the corresponding amines by the above reducing system in blood.These results suggest that blood plays an important role in the reduction of tertiary amine N-oxide to tertiary amines.
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  • Hiroyuki FUJIMORI, Ranko SATO, Masahiro YASUDA, Hidemitsu PAN-HOU
    1998 Volume 21 Issue 12 Pages 1348-1351
    Published: December 15, 1998
    Released: April 10, 2008
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    To study the physiological significance of adenosine 3'-monophosphate (3'-AMP), an intracellular P-site inhibitor of adenylate cyclase, in rat liver mitochondria, a specific, rapid and reliable assay method for determination of 3'-AMP and the activity of its forming enzyme is required. 3'-AMP in rat liver was determined to be ca. 23±7 nmol/g wet weight, but no 2-deoxy-3'-AMP, another P-site inhibitor of adenylate cyclase, was detected, even when using a reversed-phase HPLC column with a fluorescent-reaction, as established in this study.By using the optimized assay method developed here, 3'-AMP forming enzyme activity in rat crude mitochondrial extract was found to be enhanced by EDTA and inhibited by p-chloromercurybenzoate. The optimum pH was ca. 5.8 and no divalent cation was required for activity. From these results, 3'-AMP forming enzyme(s) in rat liver mitochondria could be classified as acid exoribonuclease, which mainly existed in an active form. The results obtained in this study will help to gain more insight into the physiological roles of 3'-AMP in living systems.
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  • Yoshio KOIDE, Akira IGARASHI, Nobuhiro SAKUMA, Kanako SATO, Shinobu SH ...
    1998 Volume 21 Issue 12 Pages 1352-1355
    Published: December 15, 1998
    Released: April 10, 2008
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    Stanniocalcin (STC) is a glycoprotein hormone that was first identified in fish, where it regulates the calcium level in the body fluid. The cDNA which encodes human STC has recently been reported but the function has not been completely elucidated. We have prepared a monoclonal antibody against human STC using an analogous peptide of the putative antigenic domain in human STC; it was conjugated with keyhole limpet hemocyanin (KLH). The monoclonal antibody specifically stained the distal convoluted tubules in human kidney which is a putative target organ of STC. The ELISA was established using the monoclonal antibody and recombinant human STC as a standard antigen. The monoclonal antibody prepared in this study provides a useful tool for clinical studies of STC in human.
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  • Shizuo NAKAJIN, Noriko MINAMIKAWA, Michael E. BAKER, Satoshi TOYOSHIMA
    1998 Volume 21 Issue 12 Pages 1356-1360
    Published: December 15, 1998
    Released: April 10, 2008
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    We have isolated an NADPH-dependent reductase from neonatal pig testes that metabolizes andorogens and a variety of xenobiotics. This enzyme is distinct from 3α/β, 20β-hydroxysteroid dehydrogenase or its homologue, carbonyl reductase, as judged by its immunological and molecular properties and its much narrower specificity for steroids. This reductase and 3α/β, 20β-hydroxysteroid dehydrogenase may be part of a mechanism for regulating androgen levels the neonatal pig testes. Interestingly, we could not find multiple isoforms of 3α/β, 20β-hydroxysteroid dehydrogenase/carbonyl reductase in pig testes unlike human and rat testes and other organs in which multiple isoforms are expressed.
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  • Xiuzhong WU, Yoko ONO, Atsuko NODA, Hiroshi NODA
    1998 Volume 21 Issue 12 Pages 1361-1363
    Published: December 15, 1998
    Released: April 10, 2008
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    The enantioselective N-acetylation of N-desisopropylpropranolol (NDP), one of the main metabolites of propranolol (PL), by rat liver acetyltransferase (AT), was investigated. R(+)-NDP or S(-)-NDP was used as a substrate at concentrations ranging from 10 to 200 μM. The cytosol fraction of a rat liver containing 3.93 mg protein/ml served as the source of AT. For 1-amino-3-(1-naphthyloxy)-2-propanol (AcNDP) formation from R(+)-NDP or S(-)-NDP in the presence of infinite AcCoA (250 μM), the Km value was calculated to be 67.5 or 62.4 μM, and the Vmax value was 0.462 or 0.205 nmol/min/mg protein. Based on these findings, the enantioselective N-acetylation of NDP was proved, i.e., AcNDP formation from R(+)-NDP was found to take place more easily than that from S(-)-NDP. Furthermore, AcNDP formation from NDP was competitively inhibited by the exogenous arylamine, p-aminobenzoic acid (PABA), which is well-known to be a typical substrate of AT. The presence of enantioselective inhibition for AcNDP formation was thus confirmed based on the Ki values, 440 μM in the case of R(+)-NDP and 250 μM in the case of S(-)-NDP, respectively, i.e. two-fold enantioselective inhibition was demonstrated based on the Ki values in S(-)-enantiomer incomparison with R(+)-enantiomer.
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  • Akira TAKAGI, Yoshiyuki YABE, Mitsunobu YOSHIDA, Yoshinobu TAKAKURA, M ...
    1998 Volume 21 Issue 12 Pages 1364-1366
    Published: December 15, 1998
    Released: April 10, 2008
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    The hepatic disposition characteristics of recombinant human interleukin-11 (rhIL-11) were investigated in perfused rat liver to clarify the mechanism of hepatic clearance which is a major contributor to the rapid clearance of rhIL-11 in vivo. We analyzed the disposition characteristics of [111In]-labeled rhIL-11 using a single-pass constant infusion mode at different concentrations of rhIL-11. The venous outflow rapidly reached a steady-state condition at every concentration. Liver extraction ratio at steady-state (ESS) was decreased with increase in the concentration, suggesting that there is a saturable interaction between the liver cells and rhIL-11 molecule. Cellular localization experiments demonstrated that rhIL-11 was taken up by both liver parenchymal and non-parenchymal cells depending on their surface area, suggesting that this uptake was mediated by electrostatic interaction due to cationic charges in the cytokine.
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  • Hisahiro YOSHIDA, Akira KAMIYA
    1998 Volume 21 Issue 12 Pages 1367-1370
    Published: December 15, 1998
    Released: April 10, 2008
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    To establish a quicker preparation procedure for cryoprecipitate (Cryo) from a patient's autologous plasma, to be used as fibrin glue, we examined the effects of various conditions on the concentrations and yields of coagulation factors in Cryo. Human plasma from healthy volunteers was divided and treated under various freezing, skaking and defrosting conditions. The concentrations of fibrinogen, plasminogen, fibronectin, and factor XIII in Cryo were then measured. Results were as follows : (1) concentrations and yields of plasma components in Cryo obtained from plasma stored at -20°C were significantly higher than those in Cryo from plasma stored at -80°C; (2) shaking at 70 cycles/min during the freezing process had a favorable effect on the concentrations and yields of coagulation factors in the Cryo; (3) a shaking thaw process in a cold water bath was a rapid method for obtaining adequate yields of coagulation factors; (4) shaking in the defrosting process did not affect the yields of coagulation factors. These results indicated that Cryo containing high concentrations of coagulation factors could be prepared easily and rapidly from a patient's autologous plasma (within 4-5h).
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  • Hirotaka NAITOU, Jun-ichi MIMAYA, Yasuo HORIKOSHI, Yoshifumi TAKASHIMA ...
    1998 Volume 21 Issue 12 Pages 1371-1375
    Published: December 15, 1998
    Released: April 10, 2008
    JOURNALS FREE ACCESS
    The amount of human cytomegalovirus (CMV) DNA in sera is considered to be a direct marker for CMV infection. We established conditions for nested PCR that detected one copy of CMV DNA, and for competitive PCR, which detected five or more copies of CMV DNA quantitatively. We tested 50 μl each of 16 freeze-stored and 5 fresh sera from patients, for CMV DNA. In sera obtained from the same patient at different time points, small amounts of CMV DNA were detected before the onset of CMV pneumonia. In sera from certain CMV-in-fected patients who were treated with the anti-CMV agent, ganciclovir, CMV DNA was not detected. Quantitative PCR detection of CMV DNA seems to be suitable for predicting early recurrent CMV infection and monitoring the efficacy of antiviral therapy. The qualitative nested PCR examination of CMV DNA in 40 cord blood plasma samples was carried out for the purpose of preventing CMV infection by cord blood stem cell transplantation, and they were all negative for CMV DNA.
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  • Takafumi KOMORI, Kazuki SHIMOISHI, Hideyoshi HARASHIMA, Masaki OTAGIRI
    1998 Volume 21 Issue 12 Pages 1376-1378
    Published: December 15, 1998
    Released: April 10, 2008
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    The goal of the present work was to determine the effect of clarithromycin (CAM) administration on the pharmacokinetic properties of pindolol in rats. The binding of pindolol to serum components increases proportionally with increasing α1-acid glycoprotein (AGP) concentration, indicating that AGP might play a major role in the binding of pindolol. After intravenous administration of pindolol to rats, the CAM-treated group showed a decrease in the volume of distribution, an increase in AUC and no change in the half-life as compared to the control group. Treatment with CAM increased the AGP concentration only. The serum concentration of albumin and creatinine, as well as the metabolic activity of hepatic microsomes towards pindolol, were not altered. Good correlation was observed between the AUC of pindolol in rats and the AGP concentration in serum. Moreover, at 5 min after the administration of an intravenous bolus dose of pindolol to CAM-treated rats, the free concentration of pindolol was lower but the total concentration was higher, compared with the control rats. These results suggested that the influence of CAM on the pharmacokinetic properties of pindolol in CAM-treated rats can be explained by protein binding which, in turn, may be associated with variations in AGP concentration.
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  • Kohshi NISHIGUCHI, Fusao KOMADA, Yusuke TANIGAWARA, Yumiko SAKAI, Seig ...
    1998 Volume 21 Issue 12 Pages 1379-1381
    Published: December 15, 1998
    Released: April 10, 2008
    JOURNALS FREE ACCESS
    the effects of Cu, Zn-superoxide disumutase (SOD) delivered by genetically modified skin fibroblasts on cold-induced skin edema were studied in rats. Cold-induced skin edema was induced on the dorsal skin following transplantation of ILSOD cells, genetically modified skin fibroblasts which release secretable SOD protein into the extracellular space. The degree of skin edema induced by cold injury was estimated by measuring the amounts of Evans' blue (FB) leaking into the injured skin following intravenously administration. The amounts of FB leakage were significantly reduced by transplantation of ILSOD cells relative to that observed following transplantation of host cells as a control. The degrees and durability of these effects of ILSOD cells were dependent on the number of cells transplanted. Also, the increases of lipid peroxidation following cold injury were significantly reduced by transplantation of ILSOD cells but not of host cells. These findings suggested that transplantation of ILSOD cells was a suitable delivery system for obtaining efficient and continuous effects of SOD. This strategy using genetically modified skin fibroblasts may also be useful as a drug delivery system for other therapeutic proteins.
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  • Shuji KITAGAWA, Miori HASEGAWA, Shinji SATO
    1998 Volume 21 Issue 12 Pages 1382-1384
    Published: December 15, 1998
    Released: April 10, 2008
    JOURNALS FREE ACCESS
    Characteristics of the transport of doxifluridine (5'-DFUR) in rabbit small intestinal brush-border membrane vesicles were examined. 5'-DFUR was rapidly incorporated and a linear relation was found between the concentration of 5'-DFUR and its uptake rate in the concentration range tested (0.1-10 mM). The uptake rate increased at an acidic pH, at which 5'-DFUR is present mostly in an undissociated form. However, the uptake rate was not change either by the extravesicular Na+ concentration or by the inside negative membrane potential. Nucleosides, uridine and adenosine only slightly inhibited the uptake, even at a fifty-fold concentration, and nucleobases, uracil and thymine had no effect. These results suggest the possibility of the major involvement of passive diffusion of the undissocated form of 5'-DFUR in the uptake of the drug through the brush-border membrane at a relatively high concentration.
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  • Yoshiki MINO, Hajime KITAGAKI, Masahiro SASAKI, Kazunari ISHII, Tetsuy ...
    1998 Volume 21 Issue 12 Pages 1385-1388
    Published: December 15, 1998
    Released: April 10, 2008
    JOURNALS FREE ACCESS
    To find new contrast agents for magnetic resonance imaging (MRI), the spin-lattice relaxation time (T1)-reducing activities of metal complexes of EDTA, N-hydroxyethyethylenediamine-N, N', N'-triacetic acid (HEDTA), diethylenetriamine-N, N, N', N", N&tprime;-pentaacetic acid (DTPA), deferoxamine, mugineic acid, and pectin with Fe(III) or Mn(II) were investigated. Strong activity was found in Fe(III)-deferoxamine, Fe(III)-mugineic acid, or Mn(II)-pectin. In the actual MRI tomogram, Fe(III)-deferoxamine exhibited a contrast-enhancing effect comparable with that of Gd(III)-DTPA, and a much stronger effect was observed for Mn(II)-pectin. Fe(III)-deferoxamine and the Mn(II)-pectin appear to be candidates, respectively, as a new intravenous contrast agent and an oral gastrointestinal one.
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  • Hiroyuki ICHIKI, Toshihiro MIURA, Masayoshi KUBO, Eriko ISHIHARA, Yasu ...
    1998 Volume 21 Issue 12 Pages 1389-1390
    Published: December 15, 1998
    Released: April 10, 2008
    JOURNALS FREE ACCESS
    Mangiferin (MF) and its glucosides (mangiferin-7-O-β-glucoside) (MG) isolated from Anemarrhena asphodeloides Bunge rhizome, were tested for their antidiabetic activity in KK-Ay mice, an animal model of non-insulin-dependent diabetes mellitus (NIDDM). MF and MG lowered the blood glucose level of KK-Ay mice after oral administration. However, no affect on the blood glucose level in normal mice was seen, indicating that MF and MG are useful in treating NIDDM. In addition, MF or MG improved hyperinsulinemia in KK-Ay mice. From these findings, it seems likely that MF and MG exert their its antidiabetic activity by increasing insulin sensitivity.
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  • Hiroyuki TANAKA, Yukihiro SHOYAMA
    1998 Volume 21 Issue 12 Pages 1391-1393
    Published: December 15, 1998
    Released: April 10, 2008
    JOURNALS FREE ACCESS
    The ratio of hapten and bovine serum albumin in an antigen conjugate was determined by matrix-assisted laser desorption/ionization-tof mass spectrometry. Hybridomas secreting monoclonal antibody against glycyrrhizin were produced by fusing splenocytes immunized with a glycyrrhizin-bovine serum albumin conjugate with the HAT-sensitive mouse myeloma cell line P3-X63-Ag8-653. A very small cross-reaction appeared with glycyrrhetic acid monoglucuronide and glycyrrhetic acid. The full measuring range of the assay extends from 20 ng ml-1 to 200 ng ml-1 of glycyrrhizin.
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  • Makoto INOUE, Mari UEDA, Yukio OGIHARA, Iclal SARACOGLU
    1998 Volume 21 Issue 12 Pages 1394-1395
    Published: December 15, 1998
    Released: April 10, 2008
    JOURNALS FREE ACCESS
    A phenylpropanoid glycoside acteoside was found to induce interleukin (IL)-1, IL-6, and tumor necrosis factor-α(TNF-α) in macrophage-like cell line J774.A1 at 1-100 ng/ml. In a addition, when the stimulatory action of acteoside was studied using the bovine glomerular endothelial cell line GEN-T, it stimualted IL-6 production. These stimulatory activities were not abolished by treatment with polymixin B, which inactivates lipopolysaccharide (LPS), indicating that the action was not a contamination of LPS.
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