Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
Volume 16, Issue 11
Displaying 1-29 of 29 articles from this issue
  • Yoshinori NAKAGAWA, Hitoshi HORI, Itaru YAMAMOTO, Hiroshi TERADA
    1993 Volume 16 Issue 11 Pages 1061-1064
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
    JOURNAL FREE ACCESS
    The bleaching of cyanine-type pentamethine tinuclear dyes by active oxygen species (AOS), superoxide and hydroxyl radical, were studied under the xanthine-xanthine oxidase system and the Fenton reaction, respectively. The gradual bleaching of dyes occurred as the result of superoxide produced in the xanthine-xanthine oxidase system. However, the bleaching of cyanine dyes by Fenton reagent varied depending on the reaction conditions. If a normal Fenton reaction of Fe(II) and hydrogen peroxide (H2O2) took place, a hydroxyl radical was generated instantly. A Fenton reaction with an excess of [H2O2] over [Fe(II)] resulted in a gradual bleaching of the dye initiated by the hydroxyl radical, Fe(III) and H2O2. In this reaction, cyanine dyes with shorter side chains were bleached faster than those with longer ones. We controlled the Fenton reaction condition (Fe(II) or Fe(III)/H2O2 at pH 3.5 in the dark) to generate a specific AOS such as a hydroxyl radical (·OH) or superoxide (·O-2). Studies using xanthine-xanthine oxidase (pH 7.8), the Fe(II)-dipyridyl complex and various scavengers such as superoxide dismutase (SOD) and hyaluronic acid, revealed that ·O-2 was the primary radical responsible for this controlled Fenton reaction. This finding shows that this controlled Fenton reaction would be an effective AOS generation method, and that cyanine dyes may be hopeful probes for the detection of AOS.
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  • Masayo SAKATA, Toru KAWAI, Kunio OHKUMA, Hirotaka IHARA, Chuichi HIRAY ...
    1993 Volume 16 Issue 11 Pages 1065-1068
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    We describe a method for the removal of endotoxins from various crude antigen solutions originating from gram-negative bacteria using aminated poly(γ-methyl L-glutamate) (PMLG) spherical particles. The aminated PMLG adsorbents showed high affinity for various purified endotoxins at an ionic strength of μ=0.1. The endotoxin-adsorbing capacity of the adsorbent increased with increase in the amino-group content of the adsorbent. The adsorbent (3.2 meq/g amino-group content) showed the highest affinity for endotoxin at ionic strengths ranging from μ=0.025-0.8. The adsorption of Bordetella pertussis antigen to the adsorbent decreased with increasing amino-group content of the adsorbent at an ionic strength of μ=0.2. The adsorption of B. bronchiseptica protein to the adsorbent increased with increasing amino-group content of the adsorbent, but decreased with increasing ionic strength. The adsorbent (3.2 meq/g of amino-group content) selectively reduced endotoxin in crude antigen solutions originating from gram-negative bacteria, B. pertussis, B. bronchiseptica and Pasteurella multocida, even at a high ionic strength (μ=0.2-0.4) without affecting the recovery of the protective antigens.
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  • Nobuyuki SUZUKI, Eriko FUJITA, Hiroyuki YASUI, Eiji AOYAMA, Hisashi TA ...
    1993 Volume 16 Issue 11 Pages 1069-1072
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    An improved flow injection analysis (FIA) method has been developed for the determination of trace selenium in biological samples, and this method has been applied to investigate the effect of captopril, an antihypertensive drug having a thiol group, on selenium concentrations in the rat blood, liver and urine. After oral administration of captopril, selenium levels in the blood decreased, while those in the liver increased significantly. However, no pronounced effect was observed on the urinary excretion rate. The glutathione peroxidase activities in the blood and the liver were comparable to the changes in the selenium levels.
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  • Kunio FUJIWARA, Tetsuya SAITA, Tsunehiro KITAGAWA
    1993 Volume 16 Issue 11 Pages 1073-1077
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    Luteinizing hormone-releasing hormone (LH-RH) was first labeled with an enzyme, β-D-galactosidase (β-Gal; EC 3.2.1.23), using N-[β-(4-diazophenyl)ethyl]maleimide (DPEM) as a heterobifunctional cross-linking agent. An antigen was similarly prepared by coupling LH-RH to mercaptosuccinylated bovine serum albumin with DPEM and was used for the immunization of rabbits for antibodies against LH-RH. A new, simple enzyme-linked immunosorbent assay (ELISA) for LH-RH was developed by using the principle of direct competition between LH-RH and β-Gal-labeled LH-RH for anti-LH-RH IgG which had been adsorbed to the plastic surface of microtiter plates. LH-RH concentrations lower than 50 pg/assay well were measurable reproducibly by the ELISA, the sensitivity of which was found to be about 6250 times greater than the corresponding high performance liquid chromatography (HPLC) procedure. The specificity of this ELISA seems to be primarily toward the C-terminal region of LH-RH, showing a cross-reaction with the LH-RH6-10 fragment to the same extent as with LH-RH, but no cross-reaction with the LH-RH1-3 and LH-RH4-6 fragments. Using this assay, LH-RH levels were easily measured in the blood and urine of rats following the administration of LH-RH in a single dose of 0.5 mg/kg i.v. The present, newly developed ELISA is nonradioactive, inexpensive and rapid method, and might be useful for elucidating experimental hypothalamic-pituitary-gonad interactions.
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  • Takeshi OHYA
    1993 Volume 16 Issue 11 Pages 1078-1082
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    The reactivity of formaldehyde (FA), acetaldehyde (AA) and propanal (PA) towards malondialdehyde (MDA) under physiological conditions was investigated. These alkanals revealed a reactivity towards MDA. Especially, FA showed high reactivity; for instance, a mixture of MDA (1 mM) and FA (6 mM) incubated for 4 h, or a mixture of MDA (1 mM) and FA (1 mM) incubated for 24 h, resulted in almost a 100% decrease of MDA. The amount of free MDA decreased as the alkanal concentration increased and in the reaction of MDA with AA or PA, the loss of MDA was in approximate agreement with the yield of 2, 4-dihydroxymethylene-3-alkylglutaraldehyde (1b or 1c). In the reaction of MDA with FA, the yield of 1a decreased with an excess amount of FA. In a thiobarbituric acid (TBA) test, 1 gave a positive reaction. Compounds 1b and 1c, which contain 2 molecules of MDA, liberated 1 molecule of MDA during the TBA reaction. The results showed that the MDA recovery of 1b and 1c with the TBA test was 50%. In the case of 1a, MDA recovery was 32%. In the MDA/p-nitrophenylhydrazine (NPH)-HPLC method, the MDA recovery of 1a, 1b and 1c ranged from 3-7%.
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  • Seiji TSUBOI, Emiko FUJIWARA, Kazumi OGATA, Akihiro SAKAUE, Taiji NAKA ...
    1993 Volume 16 Issue 11 Pages 1083-1086
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    S-(1, 2-Dicarboxyethyl)glutathione (DCE-GS) in addition to being present in the liver, lens, and heart, also inhibited platelet aggregation. To clarify these inhibitory effects, the role of DCE-GS in the release of ATP and serotonin from platelets was studied, as was thromboxane A2 formation, cyclic AMP level and adenylate cyclase activity in human platelets. The results are as follows : DCE-GS at a concentration of 1.3 mM inhibited ATP and serotonin release from platelets induced by collagen, by 77.4±4.3 and 78.7±6.3%, respectively. At 1.5 mM DCE-GS also inhibited the formation of thromboxane B2 by 79.6±4.1%. Incubation of human platelet rich plasma with 2 mM of DCE-GS for 10 min increased the cyclic AMP level and the activity of adenylate cyclase by 204±28 and 211±11.7%, respectively. These results suggest that the inhibitory effect of DCE-GS on the platelet aggregation induced by collagen is due to an increase in the cyclic AMP level in plateletds, which in turn may be due to enhancement of the activity of adenylate cyclase.
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  • Masashi TOMODA, Keiko HIRABAYASHI, Noriko SHIMIZU, Ryoko GONDA, Naoko ...
    1993 Volume 16 Issue 11 Pages 1087-1090
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    Two acidic polysaccharides, called ginsenan-S-IA and ginsenan S-IIA, were isolated from the root of Panax ginseng C. A. MEYER. They were homogeneous on electrophoresis and gel chromatography, and their molecular masses were estimated to be 5.6×104 and 1.0×105, respectively. Ginsenan S-IA is composed of L-arabinose : D-galactose : D-galacturonic acid in the molar ratio of 8 : 8 : 1, and ginsenan S-IIA is composed of L-arabinose : D-galactose : D-glucose : D-galacturonic acid in the molar ratio of 15 : 10 : 2 : 5, in addition to small amounts of O-acetyl groups. About a half (ginsenan S-IA) and about a quarter (ginsenan S-IIA) of the hexuronic acid residues exist as methyl esters. Reduction of carboxyl groups, methylation analysis, nuclear magnetic resonance and periodate oxidation studies indicated that their structural features include mainly α-1, 5-linked L-arabino-β-3, 6-branched D-galactan type structural units. Both polysaccharides showed remarkable reticuloendothelial system-potentiating activity in a carbon clearance test and pronounced anti-complementary activity. These substances are the first examples having a relatively high content of both α-3, 5-branched L-arabinose and β-1, 4-linked D-galactose units among the acidic arabinogalactans with activities on phagocytosis and anti-complement.
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  • Akio HIRATA, Wataru ITOH, Makiko KOMODA, Kengo TABATA, Shinji ITOYAMA, ...
    1993 Volume 16 Issue 11 Pages 1091-1093
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    In order to detect immunoreactive schizophyllan (SPG), one of the β-1, 3-glucans in solution, a murine antischizophyllan monoclonal antibody, SPG1-HS, was used as a primary antibody for solid-phase enzyme-linked immunosorbent assay (ELISA). Polysaccharide-bovine serum albumin (BSA) and polysaccharide-bovine hemoglobin (Hb) conjugate were prepared as an antigen. The absorbance at 490 nm in ELISA was directly proportional to the SPG concentration in SPG-BSA or SPG-Hb conjugates, allowing the accurate measurement of SPG at concentrations of more than 1.0 μg/ml. Also, this ELISA detected SPG in SPG-BSA and SPG-Hb conjugates in human serum in a similar manner. These results suggest that this ELISA is applicable for the determination of SPG concentration in SPG-BSA and SPG-Hb conjugates in human tissue and blood.
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  • Hideto KAWANO, Kentaro KOGURE, Kenji FUKUZAWA, Hiroshi TERADA
    1993 Volume 16 Issue 11 Pages 1094-1098
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    The effect of inorganic orthophosphate (Pi) on iron-induced lipid peroxidation in non-respiring rat liver mitochondria was studied and compared with that of adenosine 5'-diphosphate (ADP) at pH 7.4. Under the experimental conditions used, lipid peroxidation was induced by Fe2+ in conjunction with Pi and/or ADP, but not by Fe2+ alone. The characteristics of Fe2+-induced lipid peroxidation supported by Pi and by ADP were different : ADP-supported peroxidation took place after a lag-time and proceeded until all the O2 in the incubation medium was consumed, whereas the Pi-supported peroxidation exhibited a longer lag-time and a higher rate, but stopped when half the O2 was consumed. These features were ascribed to the fact that Pi forms active iron-oxygen complexes more rapidly than ADP, but the lifetimes of these complexes are less than those of the complexes with ADP. Furthermore, the ability of Pi to form active iron-oxygen complexes was found to be less than that of ADP. Fe2+, in conjunction with Pi and/or ADP, was found to induce lipid peroxidation in respiring mitochondria, although to a lesser extent than in non-respiring mitochondria.
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  • Hiroichi NAGAI, Fumio TSUJI, Shouichi GOTO, Akihide KODA
    1993 Volume 16 Issue 11 Pages 1099-1103
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    The effects of OKY-046 (thromboxane A2 (TXA2) synthetase inhibitor) and ONO-3708 (TXA2 receptor antagonist) on antigen-induced airway hyperreactivity in guinea pigs were investigated. Ketotifen was used as a reference drug. Seven inhalations of an antigen into actively sensitized animals resulted in an increase in airway reactivity to acetylcholine. Twenty-four hours after the final inhalation, the number of leukocytes (macrophages, neutrophils, eosinophils and lymphocytes) and the quantity of mediators (thromboxane B2, leukotriene D4 and histamine) in bronchoalveolar lavage fluid increased.All examined drugs inhibited the antigen-induced airway hyperreactivity to acetylcholine. Whereas ketotifen inhibited an accumulation of inflammatory cells (eosinophils and neutrophils) in bronchoalveolar lavage fluid, OKY-046 and ONO-3708 had no effect on the accumulation of inflammatory cells. OKY-046, but not ketotifen and ONO-3708, inhibited an increase of thromboxane B2 in the bronchoalveolar lavage fluid after antigen provocation.These results suggest the participation of TXA2 in the onset of antigen-induced airway hyperresponsiveness in guinea pigs, and the efficacy of TXA2 inhibitors, without affecting the accumulation of inflammatory cells in bronchoalveolar lavage fluid.
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  • Masaaki ISHIKAWA, Yoshio TAKAYANAGI, Ken-ichi SASAKI
    1993 Volume 16 Issue 11 Pages 1104-1107
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    We investigated the possibility that chlorpromazine (CPZ), an antiemetic frequently used to control nausea and vomiting in cancer patients receiving chemotherapy, might modify the progression of the renal toxicity and lethality of cisplatin (CDDP). In mice the preadministration of CPZ (i.p.) 1 h prior to CDDP (i.p.) injection efficiently reduced not only the lethal toxicity, but also the renal (indicated by increased blood urea nitrogen values) and intestinal toxicity (indicated by the incidence of diarrhea) which are usually observed in mice treated with CDDP alone. To further study the apparent protective activity of CPZ against CDDP nephrotoxicity we chose rats a species more commonly used as a model for nephrotoxicity. In F344 rats, CPZ ameliorated CDDP-induced increases in blood urea nitrogen (BUN), urine glucose, protein and lactate dehydrogenase (LDH). The preadministration of CPZ had no observed effect on the antitumor activity of CDDP in mice inoculated i.p. with Sarcoma 180, EL-4 lymphoma, or P-388 leukemia cells. The present study suggests that CPZ may be of therapeutic benefit when used with CDDP. This study also provides a rational basis for the selection of antiemetic therapy.
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  • Takeshi MINAMI, Hirofumi NAKAGAWA, Shigeru YOSHIMOTO, Ikuo ASANO, Yuko ...
    1993 Volume 16 Issue 11 Pages 1108-1110
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    pancreatic metallothionein (MT) and zinc levels were measured after the administration of 4-aminopyrazolo(3, 4-d)pyrimidine (4APP) to mice to determine the role of pancreatic MT. 4APP was found to increase the pancreatic MT level dose-dependently; the pancreatic MT level in mice administered 175 mg/kg of 4APP was 3.9 fold higher than that in control mice. When 40 mg/kg or less of 4APP was administered, the zinc concentration of the pancreatic cytosol fraction did not differ from that in control mice, while after 175 mg/kg of 4APP it was 1.7-fold higher. There was no difference in plasma glucose and insulin levels between the controls and mice administered 175 mg/kg of 4APP, although the plasma α-amylase activity was reduced in the latter. Moreover, a linear negative relationship was observed between pancreatic MT level and plasma α-amylase activity (r=-0.795, p<0.01).These findings suggest that pancreatic MT is affected by the disruption of the exocrine function in 4APP-administered mouse pancreas.
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  • Teruo TSUCHIYA, Takamasa YOSHIDA, Atsumune IMAEDA, Miho SAKUSHIMA, Tad ...
    1993 Volume 16 Issue 11 Pages 1111-1113
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    The effect of carbohydrate sulfate salts on the absorption and excretion of the herbicide, paraquat dichloride (PQ), was investigated in rats. A mixture of PQ and sodium dextran sulfate (DS) or sodium glucose sulfate (GS) was given orally to rats and then feces and urine were collected daily for 14 d. The cumulative amount of PQ excreted in the feces of rats given the mixture PQ and DS, or PQ and GS was 82.6% and 76.7% respectively. When PQ alone was administered, 56.7% was excreted in feces. Most (nearly 90%) of the PQ detected in feces was excreted within the first 24 h of the 14 d period in each case. The cumulative amount of PQ excreted in the urine over the 14 d period was 1.2% for the mixture PQ and DS and 0.7% for the mixture PQ and GS. When PQ was given alone the corresponding figure was 7.2%.In addition, the effect of DS and GS on PQ absorption from the rat small intestine was investigated using an in situ reciculating perfusion method. During 180 min period of recirculating perfusion, using PQ alone in normal saline, 13.4% of the herbicide was absorbed from the intestine. However, when the mixture of PQ and DS (or PQ and GS) in saline was used, the absorption was 3.9% for PQ-DS, of 4.8% for PQ-GS. Furthermore, an association of PQ with DS, or GS, was observed using gel filtration of the mixture of PQ-DS and PQ-GS in saline solution.The results indicate that the efficiency of the carbohydrate sulfate, i.e., DS or GS, counteracting the acute toxicity of PQ is due to inhibition of PQ absorption from the intestine. The carbohydrate sulfate inhibits PQ absorption from the intestine by associating with the herbicide, and increasing the rate of transit of PQ through the intestine.
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  • Shinya SHINOZAWA, Yutaka GOMITA, Yasunori ARAKI
    1993 Volume 16 Issue 11 Pages 1114-1117
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    The protective effects of clinically used drugs on the toxicity and microsomal lipid peroxidation induced by doxorubicin (adriamycin, ADM), an anthracycline type antitumor agent, were studied in mice and rats. Regarding the effects of anthracyclines (aclarubicin, ACL; daunorubicin, DAU; ADM; epirubicin, EPI; pirarubicin, PIR) on rat liver microsomal lipid peroxidation, ACL had the smallest effect, and effectiveness increased in the order of PIR, ADM, DAU and EPI. The increasing effect of lipid peroxidation induced by these drugs was closely correlated with the decrease in the body weight of mice administered intraperitoneally at a dose of 20 mg/kg and in rats at LD50 of the drugs. The survival times of ADM-administered mice (which were injected 15 mg/kg of ADM twice) treated with the following drugs, expressed as a percent of that of the control group, were 236% for adenosine triphosphate, 224% for coenzyme Q10 (Co Q), 235% for dextran sulfate (DS), 123% for dipyridamole, 121% for flavin adenine dinucleotide, 213% for reduced glutathion, 155% for inositol nicotinate, 157% for nicardipin and 297% for nicomol. The rat heart microsomal lipid peroxidation levels in vivo may be one of the indications of ADM-induced toxicity. The levels treated with DS correlated well with the development of ADM-induced toxicity : mouse survival time, change of body weight and tissue wet weight loss. Another type of drug, such as Co Q, may improve the myocardiac mitochondrial functions compared to those of ADM-administered mice.
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  • Atsushi KATO, Toshihiro MIURA
    1993 Volume 16 Issue 11 Pages 1118-1120
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    The hypoglycemic effect of different dose of Polygonati Rhizoma, i.e., "Ousei", was investigated in both normal and streptozotocin-induced diabetic mice. The methanol extract of Polygonati Rhizoma (OM) (800 mg/kg) reduced the blood glucose of normal mice from 202±7 to 144±13 mg/100 ml 4 h after intraperitoneal administration (p<0.01), and also lowered significantly the blood glucose of streptozotocin-induced diabetic mice from 589±34 to 396±15 mg/100 ml under similar conditions (p<0.001). However, the hypoglycemic effects were not accompanied by any alteration in the serum insulin in these mice. OM also suppressed epinephrine-induced hyperglycemia in mice. These results support, therefore, the use of Polygonati Rhizoma in patients with diabetes and confirm its role as a traditional medicine. In addition, one of the active components of OM was identified as a spirostanol glycoside (PO-2).
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  • Ayako KAMIZONO, Nobuo INOTSUME, Syouji FUKUSHIMA, Masahiro NAKANO
    1993 Volume 16 Issue 11 Pages 1121-1123
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    Pharmacokinetics of sultopride enantiomers was examined following a single dose in a human, rabbits and rats. Pharmacokinetic profiles were similar between (+)- and (-)-enantiomers of sultopride in human, whereas the serum concentrations of (-)-sultopride were slightly higher than those of (+)-sultopride after i.v. administration of 50 mg/kg of racemic sultopride to rats and rabbits.
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  • Naomi YAGI, Harumi KENMOTSU, Yoshie SHIMODE, Katsuhiko ODA, Hitoshi SE ...
    1993 Volume 16 Issue 11 Pages 1124-1129
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    The bioavailability of bumetanide following the oral administration of tablets, or the rectal administration of either macrogol suppositories or suppositories with and without weak acids were evaluated in human subjects. The absorption of bumetanide from those suppositories containing bumetanide without weak acids (control suppositories) was extremely poor, while the absorption from those suppositories containing citric acid or tartaric acid was enhanced. The mean area under the plasma concentration-time curve (AUC) following the rectal administration of the suppositories containing citric acid and tartaric acid was 52 and 62%, respectively, of that following the oral administration. On the other hand, the absorption rate constant (ka) and the mean residence time (MRT) following the rectal administration of the suppositories containing weak acids increased significantly compared to those administered orally. The time (Tmax) required to achieve the maximum plasma concentration (Cmax) in the plasma following the rectal administration of the suppositories containing weak acids was significantly shortened compared to the time of those administered orally. These results indicated that the bumetanide contained in the suppositories containing weak acids might be absorbed rapidly after administration. The diuretic effect of bumetanide following the oral and rectal administration was also evaluated. Sufficient diuretic effects were obtained following the rectal administration of the suppositories containing weak acids.
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  • Nobuhito SHIBATA, Harumi SHIMAKAWA, Tokuzo MINOUCHI, Akira YAMAJI
    1993 Volume 16 Issue 11 Pages 1130-1135
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    We examine the pharamacokinetic profile of cyclosporin A (CyA) after intravenous administration to rats prepared as models of various disease states found in patients who receive organ transplantations. After intravenous bolus administration to normal rats, the total blood clearance (CLt) of CyA showed a dose-dependent increase. The CLt was reduced in anemic (ANE) rats prepared by venesection, in carbon tetrachloride-induced acute hepatic failure (AHF) rats, and in glycerol-induced acute renal failure (ARF) rats. On the other hand, the volume of distribution at a steady state (Vss) of CyA increased significantly in ANE and aged (AGE) rats. CyA distribution was tissue-specific, and the tissue CyA concentration was disease state-dependent. Linear relationships between the CyA concentration in whole blood and various tissues (liver, kidney and heart) were found in AGE, ANE and AHF animals. However, in ARF rats, tissue concentration was not increased to a great extent in comparison with the other disease models, even though the whole blood CyA concentration was increased. The tissue per blood concentration ratio (Kb), which represented the CyA tissue transfer from systemic circulation, was influenced by the disease state. In the liver, in particular, the Kb increased in the AHF and AGE groups, whereas it decreased in the ANE and ARF rats. The CLt of CyA was negativerly related to the erythrocyte per plasma concentration ratio (E/P), and the E/P exhibited disease state-dependent changes, suggesting that this ratio is a valuable indicator for predicting variations in CyA total blood clearance in organ transplant patients during episodes of anemia, nephrotoxicity and hepatotoxicity.
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  • Yutaka NISHIOKA, Syojiro KYOTANI, Masashi OKAMURA, Saburo OHNISHI, Yas ...
    1993 Volume 16 Issue 11 Pages 1136-1139
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    cis-Diamminedichloroplatinum(II) (CDDP) albumin microspheres were prepared with various chitin concentrations and microsphere CDDP contents, specific surface area, surface structure and other microsphere properties. CDDP release in vitro and the antitumor effect in VX2 tumor model rabbits were investigated. CDDP content was increased as the concentration of chitin increased; at a chitin concentration of 6.0% it was about 2 times that without chitin. Specific surface area also increased with chitin concentration. CDDP release rate from various microspheres in vitro was suppressed as chitin concentration increased.Thus, microsphere properties, especially surface structure, are affected by an increase in chitin concentration. In experiments in vivo, various microspheres were injected into the hepatic artery of VX2 hepatocellular carcinoma model rabbits, and the effects of the chitin concentration on the time course of blood platinum (Pt) level and the antitumor effect were assessed. The blood Pt concentration increased with increase in chitin concentration, with a maximum of 0.45 μg/ml at a concentration of 6.0%, even 7 d following microsphere administration. Tumor growth was suppressed when the chitin concentration was increased. Tissue Pt concentrations also increased in the presence of chitin. These findings suggest that increasing the chitin concentration might promote microsphere decomposition and hence CDDP release in vivo, thus improving immunopotentiating activity and resulting in enhanced CDDP antitumor effect. The detailed mechanism of the action, however remains to be studied.
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  • Toshikiro KIMURA, Seiya NAKAYAMA, Tadanao YAMAO, Yuji KUROSAKI, Taiji ...
    1993 Volume 16 Issue 11 Pages 1140-1145
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    The pharmacokinetics of indocyanine green (ICG) following intravenous administration was compared between normal rats and rats with experimentally induced acute hepatic diseases; namely D-galactosamine-intoxicated (GAL) rats, CCl4-intoxicated (CCl4) rats and rats with obstructive jaundice (OJ). The total body clearance of ICG was decreased in all rats with these hepatic diseases, especially in the rats with OJ. Hepatic plasma flow was also decreased in all rats with the three hepatic diseases. The profile of biliary excretion of ICG was characteristic of rats with each hepatic disease. The cumulative biliary excretion of ICG was markedly low in GAL rats, almost unchanged in CCl4 rats and the appearance of ICG was markedly delayed in rats with OJ. The pharmacokinetic analysis of these data showed that the decreased total plasma clearance of ICG in GAL and CCl4 rats results from both lowered influx across the sinusoidal plasma membrane of hepatocytes and the decreased hepatic plasma flow. In rats with OJ, the decreased total plasma clearance results only from a markedly lowered influx across the sinusoidal plasma membrane of hepatocytes without any relation to the decreased hepatic plasma flow. The decreased biliary recovery in GAL rats might relate to the decreased bile flow. The time lag of the appearance of ICG into the bile of rats with OJ was due to its slow efflux across the bile canalicular membrane, in addition to its slow influx across the sinusoidal membrane of the hepatocyte and its remarkably long period of stay in the hepatocyte.
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  • Yoshifumi KIYOHARA, Kohshi NISHIGUCHI, Fusao KOMADA, Seigo IWAKAWA, Mi ...
    1993 Volume 16 Issue 11 Pages 1146-1149
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    When epidermal growth factor (EGF) ointment containing a protease inhibitor, nafamostat (NM), was applied to burn sites in rats, the superoxide dismutase (SOD) enzyme activity and protein content increased 45% and 60%, respectively, at these sites 1 d after the burns compared with the control ointment. Following treatment with EGF plus NM (EGF+NM) ointment, messenger RNA for SOD also increased, to about 1.6 times that of the control at 1 d after the burn, indicating that this ointment stimulates SOD synthesis at burn sites in vivo. In contrast, following treatment with EGF+NM ointment, the content of heat shock protein (HSP 70) in the burned tissue decrease to about 70% of the control value 1 d after the burn. These findings suggest that EGF+NM ointment alleviated tissue damage at burn sites at an early stage, and that this was related to the stimulation of SOD synthesis and reduced HSP 70 levels. We also examined the effects of SOD ointment on wound healing at burn sites. A dose-dependent increase in the dry weight of granulation tissue at wound sites 3 d after the burn was observed following the application of this ointment. These results suggest that SOD may play an important role in wound healing after burns.
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  • Satoru YUI, De YANG, Masatoshi YAMAZAKI
    1993 Volume 16 Issue 11 Pages 1150-1155
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    The present study demonstrates the co-existence of heat stable interleukin-1 (IL-1) and IL-1-suppressing activity in murine inflammatory polymorphonuclear leukocytes (PMNs). When the IL-1 activity in casein-induced PMN lysate was examined by lymphocyte activating factor assay, it peaked at a concentration of 2.5×106 cells per ml, but this activity was reduced to nothing at higher concentrations, suggesting the co-existence of IL-1 and IL-1-suppressive factors in PMNs. Although IL-1 has been reported to be inactivated by heat treatment, approximately one-fourth of the IL-1 activity was restored after treatment at 100°C for 10 min. In gel chromatography, the IL-1 activity of the PMN-soluble fraction was eluted in two broad peaks with apparent molecular sizes of 17-23 kDa and 23-43 kDa, respectively. Since the heat stable IL-1 activity was mainly eluted in fractions corresponding to the latter peak, and was neutralized by IL-1 receptor antagonist (Il-1ra) or anti-IL-1α antiserum, the heat stable Il-1 is suggested to be precursor form IL-1α. A high concentration of soluble fraction from PMN lysate, in contrast, completely suppressed the activity of recombinant IL-1α or β, and the suppressive activity was heat labile. Since thymocyte mitogenesis by 5 μg/ml of concanavalin A (Con A) with or without IL-2 was also completely suppressed, the suppressive factor(s) may not be IL-1ra or transforming growth factor β which were reported to be present in PMNs.
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  • Fumio NANJO, Miwa HONDA, Kazuo OKUSHIO, Natsuki MATSUMOTO, Fumiko ISHI ...
    1993 Volume 16 Issue 11 Pages 1156-1159
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    The effects of dietary tea catechins on the levels of α-tocopherol and lipid peroxidation in both plasma and erythrocytes, as well as their effects on erythrocyte deformability, were examined in rats fed on high palm and perilla oil diets. The decrease in α-tocopherol concentration and the increase in lipid peroxidation level were much more pronounced in the perilla oil group than in the palm oil group. The addition of tea catechins to these diets significantly prevented the α-tocopherol concentration from decreasing. These results suggest that tea catechins may counteract a decrease in α-tocopherol by acting as an antioxidant in vivo. Furthermore, the lipid peroxidation in the plasma of rats fed perilla oil was slightly but significantly reduced by the supplemented tea catechins. However, no measurable differences were observed in the deformability of the erythrocytes in any of the groups. It is therefore likely that the erythrocytes are not severely enough affected by the lipid peroxidation to influence their deformability.
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  • Naganori NUMAO, Shun-ichi KIDOKORO
    1993 Volume 16 Issue 11 Pages 1160-1163
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    The discriminant analysis of complementary units and repeated sequences of amino acids in an initial sample of 48 different enzymes produces practically useful empirical-functions which allow catalytic sites to be distinguished from non-catalytic sites. The independent variables in the discriminant functions were almost all composed of complementary units of amino acids, that is amino acid sequences whose nucleotide coding sequences were complementary to each other. In order of evaluate the validity of the functions, we applied them to the amino acid sequences of 17 different kinds of enzymes as well as 30 non-enzymes such as receptors, oncoproteins, cytokines, hormones and so on. The functions proved to be effective in predicting not only the catalytic sites of enzymes but also the binding sites of the other proteins. The result suggests that complementary units are evolutionarily conserved as a signal around the active sites of various proteins.
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  • Sumihiro SHIRAISHI, Teruko IMAI, Masaki OTAGIRI
    1993 Volume 16 Issue 11 Pages 1164-1168
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    Alginate gel beads containing indomethacin (IMC) were prepared using a gelation of alginate with calcium cation. Though IMC was not released from the beads at pH 1.2 because of its extremely low solubility in aqueous media, it was released according to the swelling of alginate gel at pH 6.8. The release of IMC from the beads decreased with the decreasing ratio of mannuronic acid (M) to guluronic acid (G) in alginate, and with the larger molecular weight or alginate. The maximum concentration and the area under the time curve of plasma concentration of IMC after oral administration of alginate gel beads to beagle dogs were lower than those of IMC powder. The absorption rate of IMC was lower and more prolonged after administration of beads containing IMC than IMC powder alone. The high plasma concentration of IMC was maintained because absorption continued for a longer period when the bead consisting of the alginate with 0.65 M-block/G-block ratio and 66000 molecular weight was administered. The availability of alginate gel beads of IMC was further demonstrated by oral administration to human volunteers. A serum concentration which was comparable to maximum serum concentration was maintained from 2 to 6 h after administration, while an effective serum concentration was maintained even 12 h after administration of the beads. These data suggest that alginate is useful for a controlled-release formulation of IMC.
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  • Mohammed Habibur RAHMAN, Keishi YAMASAKI, Young-Hee SHIN, Chia Chiem L ...
    1993 Volume 16 Issue 11 Pages 1169-1174
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    A site-oriented study of nine non-steroidal anti-inflammatory drugs (NSAIDs), suprofen, ibuprofen, diclofenac sodium, pirprofen, flurbiprofen, ketoprofen, phenylbutazone, oxyphenbutazone, and ketophenylbutazone, on human serum albumin (HSA) was carried out at pH 7.4 by various direct and indirect methods to gain insignt into the high affinity binding sites of NSAIDs. The binding was determined by equilibrium dialysis, circular dichroism and fluorescence methods in order to strengthen the results. Irrespective of the method used, close agreement between the binding parameters obtained by the different methods was obtained. A site-oriented description, as revealed by the fluorescent probe displacement method, suggests that site II and site I probes bound to HSA were selectively displaced by NSAIDs with a carboxyl group and without a carboxyl group, respectively. The absence of a carboxyl group of the NSAIDs changed the binding site from site II to site I. Different binding models describing the competitive and independent binding of a NSAID and a site-specific probe bound simultaneously to HSA further describe the respective high affinity binding sites for NSAIDs, thus underscoring the necessity of a carboxyl group in order for a NSAID to bind to site II. The presence of tyrosine, lysine and histidine amino acid residues in the binding of carboxyl group-containing NSAIDs to HSA was evident, whereas tryptophan is believed to take part in the binding of non-carboxyl group-containing NSAIDs. The present findings support the proposal that two separate primary binding sites exist for different NSAIDs; hence, an attempt to correlate the present results with those in literature has been made.
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  • Yoshiharu KOBAYASHI, Kayoko SAIKI, Fukuko WATANABE
    1993 Volume 16 Issue 11 Pages 1175-1178
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
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    The characteristics of the mass spectra of sixty steroids were investigated using atmospheric pressure chemical ionization-mass spectrometry (APCI-MS). In APCI-MS, the drift voltage and nebulizer temperature affected the appearance of molecular ions. Solvent adducted ions [M+H+CH3CN]+ and [M+H+H2O]+ were decreased with an increase in drift voltage. The optimum drift voltage differed slightly for each steroid.These sixty steroids were divided into two groups according to their mass spectra profiles : one having a carbonyl group at position 3 together with a double bond at position 4 (group A) and the other bearing a hydroxyl group at the 3 position (group B). In group A, the predominant peak observed corresponded to the protonated molecular ion [M+H]+. The fragment ion corresponding to the elimination of CH2OH, COCH3 and/or COCH2OH from the steroid skeleton appeared as a base peak in some steroids of group A. In group B, predominantly [M+H-H2O]+ and/or [M+H-2H2O]+ ion(s), originating from the loss of water molecules, were observed. Other major ions in this group were the protonated molecular ions. Like other chemical ionization mass spectrometry, the mass spectrum of each steroid was very simple. These results suggest that APCI-MS is a suitable tool for the determination of the mass number of polar, nonvolatile and thermolabile steroids without derivatization.
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  • Masayoshi YAMAGUCHI, Seiko KISHI, Takeshi HOSHI
    1993 Volume 16 Issue 11 Pages 1179-1181
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
    JOURNAL FREE ACCESS
    The effect of insulin administration on bone formation was investigated in the femoral diaphysis of rats with skeletal unloading. When the femoral-diaphyseal tissues from normal rats were cultured in the presence of insulin (10-9 and 10-8 M) for 48 h, the hormone produced a significant increase of alkaline phosphatase activity, collagen, deoxyribonucleic acid (DNA) and calcium contents in the bone tissues, indicating that it has a direct stimulatory effect on bone formation. Moreover, when normal rats received a subcutaneous administration of insulin (5.0 IU/100 g body weight) 5 times at 24-h intervals, the hormone caused a significant increase of alkaline phosphatase activity, DNA and calcium contents in the femoral diaphysis. Those increases were not seen by insulin administration to the skeletal-unloading rats with hindlimb hang for 4 d. The present results indicate that the stimulatory effect of insulin on bone formation is lost by skeletal unloading. The finding further supports the view that insulin impairment plays an important role in the deterioration of bone formation by microgravity.
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  • Mami YAMAZAKI, Aya SATO, Kazuki SAITO, Isamu MURAKOSHI
    1993 Volume 16 Issue 11 Pages 1182-1184
    Published: November 15, 1993
    Released on J-STAGE: April 10, 2008
    JOURNAL FREE ACCESS
    The restriction fragment length polymorphisms (RFLPs) in genomic DNA were detected among six species of Lupinus plants using rice DNA coding for ribosomal RNA (rDNA) as a probe. Additionally, the fragment patterns were compared between alkaloid-rich 'better' forms and alkaloid-poor 'sweet' forms of L. albus and L. luteus. The hybridizing patterns for several enzymes were distinguished among these species and between bitter form and sweet form of L. albus. The phylogenic tree constructed from RFLP profiles was related with the pattern of alkaloid production, indicating the usefulness of RFLP for DNA characterization of medicinal plants.
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