Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
17 巻 , 1 号
選択された号の論文の36件中1~36を表示しています
  • 平岡 厚, 三浦 勇夫, 服部 宗和, 冨永 格, 串田 克彦, 前田 昌徹
    1994 年 17 巻 1 号 p. 1-4
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    The proton magnetic resonance (1H-NMR) spectra were obtained from 38 cerebrospinal fluid (CSF) samples from 34 patients with various central nervous system (CNS) diseases and the H2O signals were reduced by the presaturation method. A doublet signal for CH3 protons of lactate, as well as a singlet one for α-CH proton of glucose, were clearly detected in all the spectra obtained. The relative lactate concentration values were calculated on the basis of the glucose concentrations which had been measured by a routine laboratory method in the hospital and the ratios of the peak heights (lactate CH3/glucose α-CH). In the CSF samples examined, the relative lactate values semi-quantitatively determined in this way were clearly elevated in cerebral infarction and bacterial meningitis, but not in other disorders including viral meningitis. On the other hand, one CSF component whose enhancement was found only in a sample from a hepatic encephalitis patient was identified as glutamine from the double quantum filtered-shift correlation spectrum. These results suggest that 1H-NMR spectroscopy of CSF can become a powerful aid in biochemical diagnosis of CNS diseases.
  • 池川 繁男, 村井 毅, 松井 陽, 藤間 貞彦
    1994 年 17 巻 1 号 p. 5-8
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    Radioimmunological measurement of conjugated 1β-hydroxycholic acid in dried blood spots obtained from newborns at 4-5 d after birth has been carried out to investigate a diagnostic utility for congenital biliary atresia. The method allowed the determination of this bile acid, with a detection limit of 50 pg in one disc (3 mm i.d.), and showed good reproducibility with inter- and intra-assay coefficients with variations of 3.5-7.7% and 6.3-8.8%. With the analysis, a relatively high concentration of this substance was observed in the patients, as compared to normal subjects, averaging 1.63 ng/disc in the former and 0.97 ng/disc in the latter.
  • 菊川 清見, 加藤 哲太, 八巻 真司, 葛西 洋芳
    1994 年 17 巻 1 号 p. 9-15
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    Extraction methods for measuring blue fluorescence generated in rat liver and brain in situ were examined. Great care should be taken when the traditional chloroform/methanol extraction method is employed because artificial blue fluorescence is readily produced by subsequent light irradiation, or by passage through a silicic acid column. In contrast, sodium dodecylsulfate (SDS) extraction were found to be suitable since no artificial blue fluorescence was produced by subsequent light irradiation. The levels of blue fluorescence in rat tissues were re-evaluated following SDS extraction. The levels of blue fluorescence in rat liver increased with age, whereas those in rat brain did not. Accumulation of blue fluorescence in the tissues was only slightly enhanced by vitamin E-deficiency. Blue fluorescence in the extracts did not seem to be derived from autofluorescent yellow lipofuscin in situ. The results were not always consistent with earlier observations obtained using the chloroform/methanol extraction.
  • 岡部 亘雄, 橋爪 規子
    1994 年 17 巻 1 号 p. 16-21
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    Binding properties of Sudlow's site-specific drugs to glycosylated human serum albumin (G-HSA) were investigated using fluorescence and circular dichroism (CD). Dansylamide, phenylbutazone and warfarin were used as site I-specific drugs, and dansylproline, ibuprofen and flufenamic acid were used as site II-specific ones. Similar changes in the fluorescence intensity of dansylamide occured in the presence of both G-HSA and intact human serum albumin (HSA), while the fluorescence enhancement of dansylproline caused by G-HSA was extremely weakened in comparison with that by HSA. These results suggest that the glycosylation of HSA inhibits the binding of the site II-specific drug, dansylproline, to HSA, while it does not influence the binding of the site I specific drug, dansylamide. The induced ellipticities of the complexes of ibuprofen, flufenamic acid and phenyl butazone with G-HSA were diminished in comparison with those with HSA. With the complexes of warfarin, the induced ellipticity was enhanced. These CD results suggest that the glycosylation of HSA induces microenvironmental changes in the binding sites for the above site-specific drugs which influence the drug binding ability of HSA.
  • 白神 歳文, 岩崎 一秀, 野崎 和吉, 田村 敏昭, 山添 康, 加藤 隆一, 高仲 正
    1994 年 17 巻 1 号 p. 22-28
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    Four different forms of cytochrome P450 (P450) were purified from liver microsomes of untreated or phenobarbital (PB)-treated male beagle dogs using HPLC techniques, and designated as DUT-1, DPB-1, DPB-2 and DPB-3, respectively. Specific contents of the purified DUT-1, DPB-1, DPB-2 and DPB-3 were 13.3, 9.6, 15.6 and 12.2 nmol/mg protein, respectively. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the monomeric molecular weights of DUT-1, DPB-1, DPB-2 and DPB-3 were estimated to be 57.5, 50.0, 47.0 and 50.0 kDa, respectively. The absolute spectra of the oxidized forms indicated that they exist in the low-spin state of heme in their oxidized forms. The NH2-terminal amino acid sequence of DUT-1 was unique and different from that of any other P450 so far reported. DUT-1 was active in the ω-hydroxylation of lauric acid. The amino-terminal sequences of DPB-1, DPB-2 and DPB-3 suggested that they belong to the P450 3A, 2C and 2B gene families, respectively. DPB-3 was a major form of P450 in PB-treated dog liver microsomes. Purified DPB-1 catalyzed nifedipine and (+)- and (-)-nilvadipine oxidations, as well as testosterone 6β-hydroxylation in the reconstituted system. These activities were enhanced 3- to 5-fold by the addition of cytochrome b5. DPB-2 and DPB-3 catalyzed aminopyrine N-demethylation, 7-ethoxycoumarin O-deethylation, biphenyl 4-hydroxylation and testosterone 16α-hydroxylation. We believe that DUT-1 is a new form not purified previously.
  • 小野寺 敏
    1994 年 17 巻 1 号 p. 29-33
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    This paper presents a sensitive assay for sialidase activity based on the specific binding of lecting to N-acetyllactosamine. The substrate used for sialidase assay is fetuin (30-100 ng/50 μl) with sialilated oligo-saccharides, which was then coated on a 96-well microtiterplate. After removing sialic acids from the terminal positions of the glycoconjugate glycans by sialidase, it was subjected to biotin-labeled lectin (Ricinus communis agglutinin 120), which binds specifically to N-acetyllactosamine. This was followed by the addition of a peroxidase conjugated avidin-biotin complex. The amount of bound peroxidase was determined by a colorimetric assay. The sensitivity was enhanced 1000- to 10000-fold compared to the colorimetric assay using a synthetic substrate such as 2-O-(p-nitrophenyl)-N-acetyl-α-D-neuraminic acid (PNPN). In the established method, only very small amounts of substrate and sialidase were required; therefore, it can be applied to the quantitative assay of some sialidases from Vibrio cholerae, streptococcus, the influenza virus and rat liver.
  • 新田 淳美, 古川 美子, 林 恭三, 長谷川 高明, 鍋島 俊隆
    1994 年 17 巻 1 号 p. 34-38
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    We attempted to measure the change in the nerve growth factor (NGF) content in the hippocampus and parietal cortex following basal-forebrain lesions induced by ibotenic acid and electrolysis. The NGF content of the parietal cortex and hippocampus increased transiently on days 3 to 7, and then returned to the control level on day 14 after the lesion of the basal forebrain induced by ibotenic acid. Ibotenic acid decreased both the choline acetyltransferase (ChAT) activity in the parietal cortex and the dopamine content in the striatum. Electrolytic lesions of the basal forebrain decreased the dopamine content in the striatum, but did not affect the NGF content and ChAT activity in any of the brain regions examined. These results suggest that the mechanism of NGF synthesis is related to cholinergic, but not to dopaminergic, neurons in the basal forebrain.
  • 別府 正敏, 洞 昌千代, 菊川 清見
    1994 年 17 巻 1 号 p. 39-46
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    A simple method is described for the assessment of the binding of macrophage scavenger receptor and its ligands such as oxidized low density lipoprotein (ox-LDL) and maleylated bovine serum albumin (m-BSA). In this method, the binding of ox-LDL or m-BSA to macrophages is observed as the adherence of erythrocytes precoated with ox-LDL or m-BSA under phase-contrast microscopy. Erythrocytes were coated with native LDL or ox-LDL simply by incubating mouse erythrocytes with LDL. The ox-LDL-coated erythrocytes attached to the monolayer of mouse peritoneal macrophages without phagocytosis at 37°C, while native LDL-coated erythrocytes did not. The extent of the adherence of the ox-LDL-coated erythrocytes to the macrophages was conveniently expressed as the proportion of the macrophages binding the coated erythrocytes. An optimal concentration of ox-LDL for erythrocyte coating giving maximum erythrocyte adherence to macrophages was 10 μg/ml as protein at an erythrocyte concentration of 2%. The binding of the ox-LDL-coated erythrocytes to the macrophages could be inhibited by ligands for scavenger receptor, and reflected the extent of LDL oxidation. Mouse erythrocytes were successfully coated with m-BSA using chromium(III) ion as adsorbent. The m-BSA-coated erythrocytes attached to the macrophages, while native BSA-coated erythrocytes did not. The adherence was inhibitable with ligands for scavenger receptor, and reflected the extent of maleylation. The method would be useful particularly for measurement of the ligand-binding activity of the receptor, and the receptor-binding activity of the ligands.
  • 中谷 良人, 原 俊太郎, 村上 誠, 工藤 一郎, 井上 圭三
    1994 年 17 巻 1 号 p. 47-50
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    We previously reported that cultured mast cells expressed three discrete phospholipases A2 (PLA2s), one of which showed a remarkable preference for phospholipids bearing an arachidonoyl residue at the sn-2 position [M. Murakami, et al., J. Biochem., 111, 175 (1992)]. In the present study, we have purified and characterized this enzyme using rat mastocytoma RBL-2H3 as an enzyme source. The elution profiles of the arachidonoyl-preferential PLA2 activity of rat mastocytoma RBL-2H3 cells on several column chromatographies were indistinguishable from those of 85-kDa cytosolic PLA2 (cPLA2) characterized so far. The molecular mass of the partially purified PLA2 was estimated to be about 90 kDa by gel filtration and it hydrolyzed arachidonate-containing phospholipids preferentially in the presence of submicromolar Ca2+ concentrations. Furthermore, it was immunoprecipitated with an anti-rabbit cPLA2 antibody almost completely. From these observations, we have concluded that the arachidonoyl-preferential PLA2 in mast cells belongs to the "cPLA2" family.
  • / 別府 正敏, 大石 憲司, 菊川 清見, Kiyomi KIKUGAWA
    1994 年 17 巻 1 号 p. 51-57
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    It is known that macrophages recongnize and take up oxidized low density lipoprotein (LDL) and this macrophage recognition has been suggested to be due to modification of the lysine (Lys) residues of apoprotein B (apo B). In order to determine whether such modification is involved in recognition, delipidated apo B was modified with lipid peroxidation products and the macrophage recognition of the modified apo B was examined. When delipidated apo B was treated with linoleic acid 13-mono-hydroperoxide (LOOH) and trans-2-octenal (octenal), apo B became fluorescent and its Lys and histidine (His) residues were decreased. When delipidated apo B, partially methylated at the ε-amino groups of Lys residues, was treated with LOOH and octenal, fluorescence was not produced and the free Lys residues were not decreased. LOOH- and octenal-modified delipidated apo B were recognized by mouse peritoneal macrophages. The macrophage recognition of the modified apo B was prevented by maleyl bovine serum albumin, indicating that the scavenger receptor was involved in recognition. Neither methyl apo B nor methyl apo B, modified with LOOH or octenal, was recognized by macrophages. Neutralization of positively charged Lys residues of apo B by modification with LOOH and octenal may be requisite for recognition. Bovine serum albumin modified with LOOH and octenal prevented the recognition of the modified apo B, indicating that none of the intrinsic structure of apo B is required for recognition.
  • 渡辺 孝寿, 林 治久, 森 陽
    1994 年 17 巻 1 号 p. 58-61
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    We investigated the effects of PAL (protocatechualdehyde), a metabolite of ACP (3, 4-diacetoxy benzylidene diacetate) which is a candidate as a novel antirheumatic agent, on the production of a tissue inhibitor of metalloproteinases (TIMP) using a primary chondrocyte culture from bovine nasal septum cartilage.The addition of human recombinant interleukin-1α (hrIL-1α) induced proteoglycan (PG) depletion from a chondrocyte matrix. PAL significantly reduced the induction of PG depletion. HrIL-1α increased the casein-degrading activity, matrix metalloproteinase (MMP) activity, and the level of TIMP secretion in the culture media. PAL significantly reduced the casein-degrading activity and further increased TIMP secretion under hrIL-1α stimulation. Phorbol myristate acetate (PMA) also increased the level of TIMP secretion, and staurosporine, a specific inhibitor of protein kinase C (PKC), reduced the TIMP secretion to the control level under hrIL-1α or PMA stimulation. Furthermore, PAL showed a significant increase following treatment with a low concentration of PMA which alone did not increase TIMP production. These findings suggest that the activation of the PKC pathway plays an important role in TIMP production and that PAL increases the level of TIMP production through an additive or synergistic effect with the activation of the PKC pathway.In conclusion, these findings demonstrate that the inhibition of the MMP activity and the increase of TIMP production by PAL contribute to the inhibition of PG depletion in a primary chondrocyte culture from bovine nasal septum cartilage.
  • 尾崎 昌宣, 宮本 賀之, 岸岡 史郎, 増田 康輔, 山本 博之
    1994 年 17 巻 1 号 p. 62-69
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    Met-enkephalin concentration-dependently and transiently inhibited the ileal twitch contraction and this inhibition gradually recovered with time. Bacitracin, phosphoramidon, thiorphan and captoril did not influence the twitch inhibition of met-enkephalin, but bestatin increased the twitch inhibitory potency of met-enkephalin and terminated it in a manner which almost paralleled that of untreated tissue. Transient inhibition of twitch contraction after tetanic stimulation (post-tetanic twitch inhibition) was obtained. Bestatin increased the potency of met-enkephalin and this was terminated within 2 min. Phosphoramidon tended to increase the potency and delayed the termination of post-tetanic twitch inhibition. Bacitracin, thiorphan and captopril did not influence either the potency or the termination of post-tetanic twitch inhibition. Morphine-induced twitch inhibition was not influenced by bacitracin, bestatin or phosphoramidon. These results suggest that bestatin-sensitive aminopeptidase and phosphoramidon-sensitive enkephalinase take part in post-tetanic twitch inhibition, acting in a different mode of action, and have an important role in the termination of the pharmacological action of endogenous opioids (post-tetanic twitch inhibition) in MPLM. This different mode of response of bestatin and phosphoramidon upon post-tetanic twitch inhibition may underlie that aminopeptidase is a more soluble enzyme and enkephalinase is membrane-bound in myenteric plexus-longitudinal muscle (MPLM).
  • 中村 俊哉, 金子 周司, 佐藤 公道
    1994 年 17 巻 1 号 p. 70-73
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    Effect of idebenone on the α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA)-selective glutamate receptor was evaluated using Xenopus oocytes injected with RNAs encoding mouse α1 and α2 AMPA receptors. Concanavalin A augmented current responses of the RNA-injected oocytes to glutamate, kainate, and AMPA and these responses were further potentiated by 100 μM idebenone. The minimum concentration of idebenone that gave a significant potentiation was 10 μM for glutamate. These results suggest that idebenone acts on AMPA-selective glutamate receptor channels composed of α1 and α2 subunits.
  • 岡 美智子, 根岸 学, 山本 貴昭, 佐藤 公彦, 廣橋 敏之, 市川 厚
    1994 年 17 巻 1 号 p. 74-77
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    The prostacyclin I1 (PGI1) analogue, SM-10906 and its methyl ester, SM-10902, have been compared with the PGI2 analogue, iloprost, with respect to binding to the PGI2 receptor, stimulation of adenylate cyclase activity and inhibition of thrombin-induced Ca2+ mobilization in mastocytoma P-815 cells. SM-10906 displaced [3H]iloprost binding to the membrane fraction, the IC50 value being 20 nM, but showed very low affinity for the prostaglandin E (PGE) receptor. SM-10906 dose-dependently stimulated GTP-dependent adenylate cyclase activity in the membrane fraction, the EC50 value being 35 nM. Furthermore, SM-10906 prevented a thrombin-induced increase in the intracellular Ca2+ concentration, the IC50 value being 100 nM. These IC50 and EC50 values are much lower than those of SM-10902. These results demonstrate that SM-10906, a stable PGI1 derivative, is an agonist for the [3H]iloprost-binding (PGI2) receptor, and that it prevents thrombin-induced Ca2+ mobilization through stimulation of the adenylate cyclase system in mastocytoma cells. On the other hand, a methyl ester derivative of PGI1, SM-10902, was inactive in the binding assay, but it seems to be a partial agonist for adenylate cyclase activity.
  • 兼松 雅博, 高木 邦明, 祐田 泰延
    1994 年 17 巻 1 号 p. 78-81
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    Nitrogen oxide, which is produced by activated macrophages, has been demonstrated to possess anti-tumor activity. We report herein the synergistic effect of sodium nitrite (NO-2) and/or sodium nitroprusside (SNP) on lysophospholipid (LysoPL)-mediated cytolysis. The incubation of 51Cr-labeled mouse melanoma (B16) cells with NO-2 alone for 3 h at 37°C did not induce cytolysis. On the other hand, NO-2 significantly enhanced the cytolysis of B16 cells in the presence of lysophosphatidylcholine (LysoPC; 2.0 μM). A similar effect of NO-2 on B16-cytolysis was also observed in the presence of 1-O-alkyl-sn-glycero-3-phosphocholine (LysoPAF). In addition, SNP (0.05-0.5 mM) synergistically enhanced B16-cytolysis in the presence of LysoPC. However, nitrate had no effect on the cytolysis of B16 cells treated with LysoPC. Furthermore, NO-2 synergistically enhanced the hemolysis of sheep erythrocytes in the presence of LysoPC, but not in the presence of an anti-sheep erythrocyte antibody and complement. These findings suggest that NO-2 directly affects membrane damage in the presence of LysoPL.
  • 鈴木 俊英, 根津 京子, 佐々木 晴代, 宮澤 陽夫, 礒野 秀夫
    1994 年 17 巻 1 号 p. 82-86
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    The effects of 11 chlorinated hydrocarbons on toxicological parameters and lipid peroxidation were examined in cultured rat hepatocytes. The cytotoxicity was evaluated by measuring the lactate dehydrogenase (LDH) released and the protein amount in cells attached to culture dishes. Phosphatidylcholine hydroperoxide (PCOOH) and phosphatidylethanolamine hydroperoxide (PEOOH) levels were determined by HPLC with chemiluminescence detection (CL-HPLC). Severe cytotoxicity was observed under the presence of the carbon tetrachloride, 1, 1, 1-trichloroethane, tetrachloroethylene and 1, 3-dichloropropene in a concentration of 10 mM. The cellular PCOOH and PEOOH levels were remarkably increased (more than 4 times) by adding these 4 chlorinated hydrocarbons. The other 7 chlorinated hydrocarbons examined did not cause any change on the cellular phospholipid hydroperoxide levels, and showed lower or no cytotoxicity at a concentration of 10 mM. The peroxidative degradation of membrane phospholipids may play an important role in causing the cytotoxicity of some chlorinated hydrocarbons.
  • 王 澤川, 掛川 寿夫, 松本 仁, 佐藤 利夫
    1994 年 17 巻 1 号 p. 87-92
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    The influence of mannose, glucose, galactose and their corresponding methyl and phenyl glycopyranosides, as well as a series of alkylphenyl α-D-mannopyranosides, on histamine release from rat peritoneal mast cells induced by concanavalin A was examined. Among the compounds tested, the inhibitory effects of compounds bearing a 2, 6-dimethyl substituent were stronger than the others. The results suggest that the binding ability of phenyl α-D-mannopyranoside may be markedly enchanced by the introduction of a 2, 6-dimethyl substituent into the benzene nuclei of aglycones.
  • 王 澤川, 掛川 寿夫, 松本 仁, 佐藤 利夫
    1994 年 17 巻 1 号 p. 93-96
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    The inhibitory effects of alkylphenyl α-D-mannopyranosides on histamine release from rat peritoneal mast cells induced by an antigen-antibody reaction were examined. Among the compounds tested, 2, 4, 6-trimethylphenyl α-D-mannopyranoside exhibited the strongest inhibitory effect. Furthermore, the 2, 4, 6-trimethylphenyl α-D-man-nopyranoside suppressed the Schultz-Dale reaction and 48 h homologous passive cutaneous anaphylaxis (PCA), suggesting that this compound may be a useful lead compound in the development of novel anti-allergy drugs.
  • 竹村 泰隆, 藤林 靖久, 土手 宣人, 谷内 英之, 飯島 直子, 小西 淳二, 横山 陽
    1994 年 17 巻 1 号 p. 97-101
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    We combined 2-hydroxy-3-methylbenzoylhydrazide (HMBH) with glycosides as a novel method for the radioiodination of physiologically active glycosides. This method was tested using digoxin, which is one of the cardiac glycosides. A digoxin-HMBH conjugate was synthesized by periodate cleavage of the third sugar ring, and was readily radiolabelled with Na[125I] by the chloramine-T method. 125I labelled digoxin-HMBH conjugate retained Na+, K+-ATPase binding in vivo and in vitro, and also retained immunoreactivity to an anti-digoxin antibody. Thus, this 125I labelled digoxin-HMBH conjugate represents a potential radiopharmaceutical for Na+, K+-ATPase imaging, as well as for the radioimmunoassay of digoxin.
  • 藤林 靖久, 脇 厚生, 和田 耕一, 上野 正樹, 間賀田 泰寛, 米倉 義晴, 小西 淳二, 竹田 俊男, 横山 陽
    1994 年 17 巻 1 号 p. 102-105
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    Using an accelerated senescence-prone model mouse strain, SAMP8, with spontaneously occurring age-related deficits in learning and memory, cerebral glucose and amino acid accumulation were investigated to study the metabolic abnormalities in relation to age. The findings were compared with those in an accelerated senescence-resistant mouse strain, SAMR1, without deterioration of ability in learning and memory.[14C]-2-Deoxyglucose accumulation in the SAMP8 brain was normal at 1 month of age but decreased from 2-3 months of age onwards. In contrast, tyrosine accumulation was unchanged from 1 to 5 months of age. The impairment of memory in the SAMP8 at 2-3 months of age corresponded with the decrease in [<14>C]-2-deoxyglucose accumulation, but was not related to Tyr. This animal model may help provide new information on the metabolic changes in aging.
  • 金 洛瑞, 梅島 啓之, 宮田 香織, 大久保 真弓, 後藤 茂, 加藤 哲雄, 三上 幸雄, 鷺谷 広道, 井上 春行
    1994 年 17 巻 1 号 p. 106-111
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    The pharmacokinetics of RK-28, a new hypoxic cell radiosensitizer, was studied in rats following its intravenous, intraportal, oral, and rectal administration. The pharmaceutical design of an RK-28 suppository form was also examined. After intravenous injection, RK-28 was rapidly removed from the plasma (biological half-life of 17 min) and its area under the curve (AUC) was proportional to the amount of RK-28 administered. The absolute bioavailability of RK-28 was 59.7% for intraportal administration and 36.9% for oral administration. Following oral administration of RK-28, it seems likely that specific acid-catalyzed decomposition of RK-28 takes place in the stomach and then the absorbed RK-28 undergoes first-pass metabolism in the liver. When "solidified RK-28 suppository, " made by solidifying molten RK-28, was inserted into the rectum, hepatic first-pass metabolism could be avoided substantially. The resulting absolute bioavailability of RK-28 increased to 75%. Furthermore, "RK-28 emulsion suppository, " prepared by emulsifying RK-28 with 1-hexadecanol and hydrogenated castor oil (HCO 60) at 80°C, showed a plasma concentration-time curve very suitable for radiation therapy; the maximum plasma concentration was attained 30 min after rectal administration and then decreased within a short period. Administration of "RK-28 emulsion suppository, " resulted in an absolute bioavailability of 76%.
  • 小木曽 太郎, 岩城 正宏, 谷野 公俊, 川淵 利津子, 畠 澄子
    1994 年 17 巻 1 号 p. 112-116
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    To better define the mechanism of the increased plasma concentration of propranolol (PL) after meals, the effect of the dietary constitution of a meal on the kinetics of PL and β-naphthoxylactic acid (NLA), a main metabolite, after administration of the drug, was investigated in rats. Additionally, the hepatic uptake of PL and cytochrome P-450 (P-450) content and uridin 5'-diphosphoglucuronyltransferase (UDPGT) activity in liver were measured after glucose intake. As a result, protein (skim milk) intake slightly, but not significantly, increased the area under the plasma concentration-time curve (AUC) and bioavailability of PL, with a slight increase (16%) in hepatic blood flow, and enhanced PL metabolism to NLA. Soybean oil and fatty acid intake significantly decreased the bioavailability of PL, while glucose intake dramatically decreased the hepatic uptake of PL and P-450 content at high glucose levels, resulting in a decrease in the plasma PL concentration at the initial time period and in the inhibition of a metabolic conversion to NLA. Thus, a possible mechanism involved in the effect of food on PL bioavailability could have been due largely to the decreased microsomal P-450 content and hepatic uptake of PL after glucose intake, but only partly to the increased hepatic blood flow after protein intake.
  • 相良 一禎, 山田 一磨呂, 川添 康之, 水田 博彰, 柴田 正宏
    1994 年 17 巻 1 号 p. 117-120
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    The gastrointestinal (GI) physiology of beagle dogs was effectively regulated with a combined treatment using intramuscular pentagastrin (10 μg/kg×2) and intravenous atropine sulfate (0.02 mg/kg×1). The superiority of the GI physiology regulated-dogs over the intact dogs was confirmed by comparative bioavailability studies using two classes of preparations of poorly water-soluble 4-(2-chlorophenyl)-2-[2-(4-isobutylphenyl)ethyl]-6, 9-dimethyl-6H-thieno[3, 2-f][1, 2, 4]triazolo[4, 3-a][1, 4]diazepine (Y-24180). Both the fine granules and the tablets of Y-24180 exhibited similar absorption profiles in the intact dogs, whereas the latter preparations revealed a delayed plasma curve of the drug in the regulated-dogs. The absorption profiles of the two classes of Y-24180 preparations in the regulated-dogs simulated those in healthy volunteers. The combined-treatment of beagle dogs with pentagastrin and atropine sulfate was suggested to supply a useful animal model for predicting the absorption characteristics of poorly water-soluble drugs and their preparations in humans.
  • 滝野 十一, 小西 清志, 高倉 喜信, 橋田 充
    1994 年 17 巻 1 号 p. 121-125
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    With the aim of developing of emulsion carrier systems for lipophilic drugs with the potential for prolonged circulation in the blood or hepatic targeting, the in vivo disposition of four model compounds, i.e., [3H]prostaglandin E1, [3H]retinoic acid, [14C]cholesterol, and [14C]cholesteryl oleate with calculated log PCoct values of 2.15, 6.61, 9.46, and 18.3, respectively, injected with various emulsion formulations, were studied in mice. Small sized emulsions of about 100 nm in diameters, with compositions of egg phosphatidylcholine (PC) : soybean oil=1 : 1 (small PC emulsion) and PC : egg sphingomyelin (SM) : soybean oil=0.7 : 0.3 : 1 (small SM emulsion), and a conventional emulsion with a diameter of about 250 nm and a composition of PC : soybean oil=1 : 1 (large PC emulsion) were compared. Highly lipophilic [14C]cholesteryl oleate, a marker of emulsion particles, indicated diverse in vivo behaviors; i.e., the small SM emulsion produced prolonged circulation in the blood, and the small PC emulsion followed this, while the large PC emulsion was rapidly uptaken by the liver. Thus, a reduction in size and coating with SM on the surface of oil droplets resulted in avoidance of the reticuloendothelial system (RES). Disposition profiles of other test compounds differed, depending on their lipophilicities : [14C]cholesterol showed disposition patterns in all formulations similar to those of [14C]cholesteryl oleate, but moderately lipophilic [3H]prostaglandin E1 and [3H]retinoic acid showed common disposition profiles, regardless of emulsion types, suggesting their rapid release from the emulsion carriers. These results suggest that small SM emulsion and large PC emulsion can act respectively as long circulating and liver targeting carriers for highly lipophilic drugs with log PCoct larger than 9.
  • 木村 都, 飯島 史朗, 小林 きよ美, 古幡 博
    1994 年 17 巻 1 号 p. 126-130
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    Previous studies show that the combined method using thrombolytic drugs and ultrasonic irradiation enhances the effect of thrombolysis in in vitro and in vivo experiments.However, the mechanism of the clot lysis was unclear. In previous studies the evaluation of the clot lysis effect was determined by the rate of decrease in clot weight or the decrease in radioisotope-labeled fibrinogen in the clot. However, it was unclear whether or not the fibrinogen in the clot was degraded.We measured quantitatively the fibrin degradation product, D-dimer (FDP-DD), produced from clots subjected to recombinant tissue-type plasminogen activator (rt-PA, 2.0 μg/ml), with and without ultrasonic irradiation.Two levels of continuous ultrasound were used, 300 kHz (100 V, 0.07 W/cm2) and 1 MHz (20 V, 0.4 W/cm2). The quantity of FDP-DD produced was 1.74 times at 300 kHz and 1.80 times at 1 MHz, greater than that measured in rt-PA solution without ultrasonic irradiation; clot lysis was not observed in normal saline with ultrasonic irradiation. Our experiment clarifies that the mechanism of clot lysis when subjected to a combination of drug and ultrasound leads to degradation of fibrin, allowing a quantitative measurement of the enhancement of clot lysis. A high correlation was observed between the FDP-DD produced from the clots and the rate of decrease in clot weight.
  • 須田 昌宏, 大野 尚仁, 安達 禎之, 宿前 利郎
    1994 年 17 巻 1 号 p. 131-135
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    Distribution of 3H-labeled (1→3)-β-D-glucan([3H]SSG) obtained from the culture filtrate of Sclerotinia sclerotiorum IFO 9395, in various tissues in tumor-bearing mice was examined. [3H]SSG administered intra-peritoneally was mainly detected in liver, spleen, kidney and tumor masses. In contrast to i.p. administration, intra-lesionally administered [3H]SSG was not released from the tumor. Similarly, in a double grafted tumor system, [3H]SSG was located in the administered tumor and not distributed in the distant site tumor, in spite of the fact that significant antitumor effect was shown in both tumor sites in this system. Winn assay confirmed the activation of the systemic antitumor immunity. These results suggested that the distribution of glucans would be one important factor in determining their antitumor effects. However, this would not always be necessary if systemic immunity could be induced.
  • 笹津 備規, 清水 慶一, 野口 雅久, 河野 恵
    1994 年 17 巻 1 号 p. 136-138
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    The relationship between an effective concentration and the duration of exposure to antiseptics was evaluated in strains of Staphylococcus aureus with a known genetic background, which include methicillin-resistant strains, using a modified version of the phenol coefficient method as part of an effort to investigate the antiseptic resistance of S. aureus. Chlorhexidine digluconate killed an antiseptic-sensitive strain within 1.5 min at 22°C at a standard concentration (0.1%), whereas resistant strains still survived after 1.5 min. Povidone-iodine killed the sensitive strain within 1.5 min at a concentration of 0.1%, whereas it took this agent 3.0 and 4.5 min to kill low- and high-level resistant strains, respectively, at a concentration of 0.8%. These results indicate that the modified phenol coefficient method used is suitable for the evaluation of the sensitivity of microorganisms to antiseptics. An antiseptic-resistant chain that was associated with the ebr gene exhibited cross-resistance to povidone-iodine.
  • 大附 映子, 鈴木 誠, 高柳 美行, 八代 有
    1994 年 17 巻 1 号 p. 139-141
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    A simple enzymatic method for the spectrophotometric determination of xanthurenic acid (XA) in urine is described. The method is based on the formation of a pigment derived from an oxidative coupling reaction with XA and N, N-diethyl-p-phenylenediamine (DE-PPD) in the presence of hydrogen peroxide and horseradish peroxidase. The color of the reaction mixture was developed with a peak absorbance at 750 nm. The recoveries against XA added to urine ranged from 91.0 to 96.5%. With this method, XA can be determined within 5 min, without a time-consuming sample preparation. The detection range of this method was between 5 and 150 μM, with a coefficient of variation of 2.7% or less. The correlation between this and the fluorometric method was good.
  • 後藤 勝博, 加藤 清治, 住谷 貞春, 上原 総一郎, 平山 亮夫, 坂本 亘, 徳光 幸子, 高田 昌彦
    1994 年 17 巻 1 号 p. 142-145
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    Urinary γ-carboxyglutamic acid (γ-Gla) levels were determined in healthy subjects of all ages. The urinary γ-Gla levels were highest in infants (0-1 years), then fell in an age-dependent manner, again in subjects reaching a minimum value in adults, then gradually increased over 60 years of age. Urinary γ-Gla levels therefore change markedly with aging. The relationships between the urinary γ-Gla excretion and plasma levels of prothrombin and protein C in patients with various hepatic diseases or diabetes mellitus were examined and compared with those in healthy adults. Both plasma prothrombin and protein C levels were decreased in all patients with liver disease compared with healthy adults. In patients with hepatitis and liver cirrhosis, the decrease did not, however, affect the γ-Gla excretion. In addition, in patients with hepatoma or carcinoma with liver metastases, the urinary γ-Gla levels were increased. In patients with diabetes mellitus, the urinary γ-Gla levels and plasma levels of prothrombin and protein C tended to increase, but this was not significant. The present results indicate that simultaneous measurement of the levels of urinary γ-Gla and plasma prothrombin and protein C is a useful tool for the diagnosis of liver diseases and diabetes mellitus.
  • 芳野 恭士, 原 征彦, 佐野 満昭, 富田 勲
    1994 年 17 巻 1 号 p. 146-149
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    The antioxidative activity of theaflavins (TFs) and thearubigin (TR) purified from the infusion of black tea leaves was examined using the tert-butyl hydroperoxide-induced lipid peroxidation of rat liver homogenates.The concentrations which produced 50% inhibition of lipid peroxidation (IC50) by theaflavin (TF), theaflavin monogallate-A (TFM-A), and TR were 4.88×10-4, 4.09×10-4, and 4.95×10-4% (w/v), respectively. The antioxidative activity of these compounds was higher than that of glutathione, L(+)-ascorbic acid, dl-α-tocopherol, butylated hydroxytoluene, butyl hydroxyanisole, etc., but was lower than the activity of (-)-epicatechin gallate, (-)-epigallocatechin, and (-)-epigallocatechin gallate. As to the IC50 in molarity, the antioxidative activity of TFM-A was the second highest among all the samples used in this study.The antioxidative activity of lyophilized tea infusions was compared. The activity of black tea was about as potent as that of green tea.These results suggest that black tea infusion containing TFs and TR could inhibit lipid peroxidation in biological conditions in the same way as green tea infusion containing epicatechins.
  • 池永 敏彦, 日迫 睦美, 田島 みよこ, 中島 憲一郎
    1994 年 17 巻 1 号 p. 150-151
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    Five Artemisia capillaris plants were selected at random from among a bed of twenty plants cultivated in a field for one year. Several branches were collected from each plant once a month, except in August and October, when two collections were made during budding and flowering. Each organ (leaf, stem, and capitulum) was separated from the branch and dried, and later analyzed for capillarisn and 6, 7-dimethylesculetin content by high-performance liquid chromatography (HPLC).The capillarisn and 6, 7-dimethylesculetin content reached maximum levels in the leaf just before the appearance of flower buds at end of July. About one month later, at the end of August during budding and flowering, capillarisin content in the capitulum reached a peak and then decreased. On the other hand, 6, 7-dimethylescuretin content reached a maximum at the beginning of September, two weeks after the capillarisn maximum.The results suggest that the most appropriate time to harvest A. capillaris for use as a crude drug is between the flower bud stage and early flower stage, from late August to early September.
  • 一川 暢宏, 直良 浩司, 岩本 喜久生
    1994 年 17 巻 1 号 p. 152-155
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    Pharmacokinetic behavior involved in the entry of four quinolone antibacterial agents, norfloxacin (NFLX), ciprofloxacin (CPFX), ofloxacin (OFLX) and nalidixic acid (NA), into cerebrospinal fluid (CSF) was comparatively investigated in rats.Periodically, after the bolus i.v. dose of each quinolone (10 mg/kg), aliquots of CSF were collected by cisternal puncture and blood samples were then withdrawn from the jugular vein. CSF and serum (total and unbound) levels of the drugs were determined by HPLC method. Transport parameters for three new quinolones (NFLX, CPFX, OFLX) into CSF were obtained by physiological model analysis.Serum levels of OFLX and NFLX declined bi-exponentially with time, whereas the serum levels of NA and CPFX declined in mono-exponential and tri-exponential fashion, respectively. Fractions of each quinolone unbound to serum protein (approximately 0.7 for NFLX, CPFX, and OFLX, 0.12 for NA) were almost the same at any point in time. The CSF levels of these quinolones rose quite rapidly after drug administration, and then declined, along with their serum levels. Both the CSF level and the ratio of CSF concentration to serum unbound concentration were the highest for NA, followed by OFLX, CPFX and NFLX. These values of the four quinolones were almost proportional to the apparent partition coefficient (Papp) between n-octanol and phosphate buffer (pH 7.0) values of each reported in a previous paper [Tsuji et al., Antimicrob. Agents Chemother., 32, 190 (1988)]. In the three new quinolones, OFLX had a larger value of apparent diffusional clearance between blood and CSF (PAc) than CPFX and NFLX. We found that the values of PAc for CPFX, OFLX and NFLX correlated well with the reported values of Papp for these quinolones.The present results suggest that the permeability of these quinolones between blood and CSF may depend predominantly on the lipophilicities of the drugs.
  • 五郎丸 毅, 松木 一浩, 松木 洋子
    1994 年 17 巻 1 号 p. 156-159
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    The effect of acarbose on the digestion of starch was examined by a stable isotope tracer technique. [U-13C]-Starch was administered orally to rats with or without acarbose. After the addition of [2H3]-D-glucose as the internal standard, the plasma samples were treated successively for defatting, deproteinizing and desalting. Glucose was converted to sorbitol by reduction with sodium borohydride. The cyclic butylboronate of sorbitol was injected into a gas chromatograph-mass spectrometer, and the concentration of labeled glucose was measured by selected monitoring of the quasi-molecular ion. The plasma concentration of labeled glucose was decreased significantly by the addition of acarbose. The effect of acarbose on the digestion of starch was clearly confirmed using [U-13C]starch.
  • 稲森 善彦, 室 親明, 船越 裕一郎, 宇佐美 吉英, 辻坊 裕, 沼田 敦
    1994 年 17 巻 1 号 p. 160-162
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    2-Thiophenecarboxylic acid (I) exhibited growth-inhibitory activity in five kinds of plants. In particular, I strongly inhibited the growth of the roots of Lactuca sativa L. var. longifolia LAM and Echinochloa utilis OHWI et YABUNO, even at the low concentration of 5.0×10-5 M. Furthermore, all of the I-related compounds (II-V and VII-X) except for VI, showed more or less obvious inhibitory activity on the seeds of Sesamum indicum L. Compounds VII-X, in which the carboxyl group of I was replaced by acetic acid, propionic acid, butyric acid and acrylic acid, and exhibited more potent phytogrowth-inhibitory activity than I. Among these compounds, 2-thiophenebutyric acid (IX) showed the strongest activity. Esterification of the carboxyl group in I increased the inhibitory activity relative to that of I, while amidation and reduction of this group markedly decreased its inhibitory activity. The radicles of the plants treated with each of the compounds except for VI showed negative geotropism, even though germination occurred.
  • 笹津 備規, 柴田 義之, 野口 雅久, 河野 恵
    1994 年 17 巻 1 号 p. 163-165
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    The range of substrates of antiseptic resistance associated with the ebr gene is very wide and includes antiseptics, DNA-intercalating drugs and preservatives which have no quaternary ammonium group in methicillin-resistant Staphylococcus aureus. Antiseptic resistance was inhibited by calcium channel blockers and calmodulin inhibitors similarly to the resistance to multiple antitumor agents encoded by the mdr gene in human cells. These results show that the common properties of the mdr gene and the ebr gene may be shared by the mechanisms that are involved in the removal of toxic substances from the cell.
  • 小野 敦央, 山口 雅章, 堀越 勇, 新谷 智美, 上野 雅晴
    1994 年 17 巻 1 号 p. 166-168
    発行日: 1994/01/15
    公開日: 2008/04/10
    ジャーナル フリー
    Characteristics of the calcein release from temperature-sensitive liposomes with or without stirring were studied. Calcein release from DPPC(dipalmitoylphosphatidylcholine) REV(reverse phase evaporation vesicle) with stirring occurred at a temperature lower by 8 degrees than that without stirring. DPPC-DSPC(distearoylphasphatidylcholine)-mixed REV showed the maximum release(%) for calcein, near 40 °C with stirring and 45 °C without stirring.
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