Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
Volume 16 , Issue 10
Showing 1-25 articles out of 25 articles from the selected issue
  • Atsushi HIRAOKA, Isao MIURA, Munekazu HATTORI, Itaru TOMINAGA, Sachiko ...
    1993 Volume 16 Issue 10 Pages 949-952
    Published: October 15, 1993
    Released: April 10, 2008
    JOURNALS FREE ACCESS
    Analysis systems for protein and amino acid components in cerebrospinal fluid (CSF) from patients with a variety of central nervous system (CNS) diseases were established employing capillary-zone electrophoresis (CZE). Under the conditions employed, the major proteins in CSF and the concurrent sera were separated into deveral respective fractions, and in the obtained electropherograms of serum proteins, the % values of areas of each major protein fraction (from γ-globulin to albumin) were in close accord with those in the densitograms of stained protein bands after electrophoresis on cellulose-acetate strips which had been done as a routine laboratory test in the hospital. Some unusual migration patterns of major proteins were observed in various CNS diseases, and an increase of glutamine in hepatic encephalitis was also detected in the electropherograms. A peak responsible for a minor protein component with the molecular weight of 10000-30000, which was tentatively identified as β-trace protein, was found to be contained in all the CSF samples examined, and its concentration levels were higher in some patients with cerebral infarction and multiple sclerosis. These results suggest that CZE can become a powerful aid in analyses of the protein and amino acid components in CSF for biochemical diagnosis of CNS diseases.
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  • Tohru MIZUSHIMA, Shunji NATORI, Kazuhisa SEKIMIZU
    1993 Volume 16 Issue 10 Pages 953-955
    Published: October 15, 1993
    Released: April 10, 2008
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    We examined the influence of chlorpromazine, a phenothiazine derivative, on DNA synthesis in Meth A cells. Pulse-labelling experiments with [3H]thymidine showed that chlorpromazine inhibited DNA synthesis in cells cultured in vitro. The drug also inhibited DNA synthesis in isolated nuclei. Observation by fluorescence microscopy of fibroblastic cells stained with chlorpromazine indicated that the drug was localized in the cytoplasm and nuclear membranes, suggesting that it inhibited DNA synthesis in a manner dependent on the interaction of replication proteins with nuclear membranes. Meth A sarcomas growing in the endoderm of BALB/c mice regressed on intra-tumor injection of chlorpromazine, indicating that the drug has an anticancer action.
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  • Takayuki SHIRAISHI, Yutaka UDA
    1993 Volume 16 Issue 10 Pages 956-959
    Published: October 15, 1993
    Released: April 10, 2008
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    Lignoceroyl-coenzyme A (CoA) ligase activity was detected in microsomal fractions from chicken liver in the presence of α-cyclodextrin as a solubilizing agent of lignoceric acid. Heptakis(2, 6-di-O-methyl)-β-cyclodextrin (dimethyl-β-cyclodextrin) and hexakis(2, 6-di-O-methyl)-α-cyclodextrin (dimethyl-α-cyclodextrin), among the cyclodextrins tested, were more effective than α-cyclodextrin as solubilizing agents. We have characterized lignoceroyl-CoA ligase activity in comparison with palmitoyl-CoA ligase activity. Lignoceroyl-CoA and palmitoyl-CoA ligase activities showed a similar dependency on CoA concentration. However, lignoceroyl-CoA ligase activity exhibited responses to the Mg2+ ion, adenosine triphosphate (ATP), ATP analogues (adenosine monophosphate (AMP) and adenosine diphosphate (ADP)), and heat treatment, which were distinctly different from the responses of palmitoyl-CoA ligase activity. These results are consistent with the idea that lignoceroyl-CoA and palmitoyl-CoA are synthesized by two different enzymes.
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  • Yasuki KATO, Kyoko WATANABE, Toshihito HOSOKAWA, Eiji HAYAKAWA, Kunio ...
    1993 Volume 16 Issue 10 Pages 960-964
    Published: October 15, 1993
    Released: April 10, 2008
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    The influence of HCO60 on the blood clearance and tissue distribution of soybean phosphatidylcholine (PC) liposomes, encapsulating α-tocopherol as a marker, was studied in rats. The liposomes were prepared by the hydration method from a lipid film containing different amounts of HCO60, and by extrusion through a 0.1 μm polycarbonate membrane filter. The blood clearance and liver uptake of α-tocopherol after i.v. administration increased with increasing the amount of HCO60 the liposome contained. With 80 wt% HCO60 liposomes, the accumulation of α-tocopherol in the liver was approximately three-fold that of the 100% PC liposomes. The uptake by lungs, spleen and kidneys did not change with the addition of HCO60. The findings obtained in a gel-filtration study suggested that α-tocopherol is not removed from the liposomes, with or without HCO60, by rat plasma proteins and the HCO60 micelle may form a complex with rat plasma proteins. Our findings suggest that liposomes containing large amounts of HCO60 (more than 60 wt%) will be useful for delivering drug to the liver.
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  • Yasuki KATO, Toshihito HOSOKAWA, Yuichi OKUBO, Eiji HAYAKAWA, Kunio IT ...
    1993 Volume 16 Issue 10 Pages 965-969
    Published: October 15, 1993
    Released: April 10, 2008
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    The efficiency of encapsulation of doxorubicin (DOX) into liposomes containing HCO60 was studied. Empty egg phosphatidylcholine (PC) liposomes containing various amounts of HCO60 were passed through a polycarbonate membrane filter (0.1 μm). Closed vesicles were formed even with liposomes containing 90 wt% HCO60. DOX was encapsulated efficiently (more than 1 mg DOX/8 mg lipids) using a pH-gradient method. HCO60 accelerated the leakage of DOX from liposomes into phosphate buffer at temperatures above 25°C. HCO60 suppressed the leakage of DOX from egg PC/cholesterol (7/3, w/w) liposomes into phosphate buffer containing 50% fetal bovine serum (FBS). Studies on the interaction between liposomes and serum components using zeta potential and turbidity suggest that HCO60 prevents the liposomes from interacting with serum components.
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  • Tomomi NAKAMURA, Mizuho OGASAWARA, Ikuo KOYAMA, Masami NEMOTO, Tsuguch ...
    1993 Volume 16 Issue 10 Pages 970-972
    Published: October 15, 1993
    Released: April 10, 2008
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    The effect of taurine in protecting biomembrane against oxygen radicals was investigated using canine erythrocytes. 2, 2'-Azobis(2-amidinopropane) dihydrochloride (AAPH), a water-soluble azo-compound, was used as the oxygen-radical generator. Taurine suppressed erythrocyte hemolysis more effectively than α-alanine, used for comparison. To clarify the relationship to the lipid peroxidation, the amount of lipid peroxide was measured using liposomes prepared with egg yolk lecithin. However, the peroxidation was not suppressed by taurine. When intact erythrocytes were subjected to hemolysis by hyposmotic solutions, taurine suppressed the osmotic hemolysis more effectively than α-alanine. These results suggest that taurine does not have an antioxidative effect like vitamin E, but interacts with the biomembrane, and helps to protect it against damage caused by AAPH.
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  • Yasuhiro KOHAMA, Kentaro IIDA, Keishi TANAKA, Taro SEMBA, Masahiko ITO ...
    1993 Volume 16 Issue 10 Pages 973-977
    Published: October 15, 1993
    Released: April 10, 2008
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    The biological effects of a cytotoxic substance (BS-1), isolated from Bacillus stearothermophilus UK563 and identified as bis(2-hydroxyethyl) trisulfide, on elicited mouse peritoneal macrophages induced by a casein injection, were investigated in vitro. BALB/c mouse macrophages treated or pretreated with BS-1 (1-10 μg/ml) showed cytotoxicity against syngeneic DBA/2 mouse P815 mastocytoma. BS-1 also showed weak cytotoxicity directly against P815 in the absence of macrophages. BS-1 significantly increased the glucose consumption of macrophages without producing cytotoxicity. This trisulfide compound increased nitric oxide formation, interleukin-1 production and prostaglandin E2 release in macrophages. It did not, however, increase the production of active oxygen species in macrophages, but it reduced cytochrome c in the presence of phagocytes. These results indicate that BS-1 activates macrophages to the cytolytic stage.
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  • Yasuo ODA, Yoshiyuki TATSUMI
    1993 Volume 16 Issue 10 Pages 978-981
    Published: October 15, 1993
    Released: April 10, 2008
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    Four isolectins were isolated from bulbs of Croccus sativum (saffron), using affinity chromatography on mannan-Sepharose 4B, gel filtration and ion-exchange column chromatography in the presence of 8 M urea. The relative molecular masses of these lectins were determined by gel filtration to be approximately 48 kDa. On polyacrylamide gel electrophoresis, relative molecular masses of 8 kDa were obtained, suggesting that the lectins are hexamers. No carbohydrates were detected in the lectins. The lectins agglutinated the yeasts of Saccharomyces cerevisiae at a minimum concentration of 30 μg/ml, however, they did not agglutinate animal erythrocytes (human, sheep, rabbit and mouse) even at a concentration of 1000 μg/ml. In inhibitory experiments, mannan from Saccharomyces cerevisiae was the most potent inhibitor of C. sativum lectins. While D-mannose showed no inhibition, manno-oligosaccharides with either (α 1-2, 1-3) or (α 1-6) linkages were potent inhibitors, suggesting that C. sativum lectins recognized manno-oligosaccharide units larger than mannobiose. Ovomucoid also exhibited potent inhibition, however, the other carbohydrates examined did not.
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  • Masaaki ISHIKAWA, Yoshio TAKAYANAGI, Ken-ichi SASAKI
    1993 Volume 16 Issue 10 Pages 982-988
    Published: October 15, 1993
    Released: April 10, 2008
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    We attempted to produce a model mouse with a liver injury resulting from an immunological mechanism in C57BL/6J mice, and the effect of hepatitis on the hepatic microsomal mixed-function oxidase system was studied. An experimental immunological liver injury model was caused by the intravenous injection of an anti-basic liver protein (BLP) antibody in mice which had been previously immunized with normal rabbit IgG (RGG) and complete Freund's adjuvant. C57BL/6J strain mice showed the highest susceptibility to the immunological liver injury. Typical histopathological changes in the liver included submassive hepatocellular necrosis and infiltration of lymphocytes into the portal tract and sinusoid area in a necrotic lesion. The liver injury in this model was markedly inhibited by the administration of prednisolone (20 mg/kg, p.o.), cyclophosphamide (15 mg/kg, i.p.), levamisole (10 mg/kg, p.o.), glycyrrhizin (50 mg/kg, i.p.) and cepharanthine (10 mg/kg, i.p.), which act on the immune system. Twenty-four hours after the injection of anti-BLP antibody, the activities of aminopyrine N-demethylase, aniline hydroxylase and NADPH-cytochrome c reductase and the content of cytochrome P-450 were mostly reduced, whereas cytochrome b5 and NADH-ferricyanide reductase were not. These results suggest that the experimental liver injury model in C57BL/6J mice is useful as a model of liver injury model, and its hepatitis was shown to inhibit the cytochrome P-450-dependent biotransformation of drugs in the mouse.
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  • Riyo ENOMOTO, Kiyokazu OGITA, Yukio YONEDA
    1993 Volume 16 Issue 10 Pages 989-996
    Published: October 15, 1993
    Released: April 10, 2008
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    Specific binding of [3H]1, 3-di-o-tolylguanidine (DTG) was found not only in synaptic membrane fractions but also in subcellular fractions enriched of microsomes, nuclei and mitochondria/myelins, with different sensitivities to displacement by the antipsychotic haloperidol. The highest binding was detected in microsomal fractions followed by, in order of decreasing binding, fractions enriched in nuclei, synaptic membranes, mitochondria/myelins and homogenates. [3H]DTG binding was completely abolished by prior treatment of the synaptic membranes with a low concentration of Triton X-100. [-3H]DTG binding reached a plateau within 30 min of the incubation at 2°C, whereas raising the incubation temperature to 30°C resulted in marked shortening of the time required to attain equilibrium, without altering the binding at equilibrium. The binding was inhibited by haloperidol in a concentration-dependent manner over a concentration range of 1 nM to 0.1 mM but with a potency more than 100 times weaker than the value reported in the literature, irrespective of the termination method employed and the external proton concentrations. [3H]DTG binding was markedly displaced by a variety of compounds including sigma ligands, benzomorphan opiates and noncompetitive antagonists at the N-methyl-D-aspartate (NMDA) receptor in synaptic membranes of the cortex, hippocampus and cerebellum. However, sigma ligands such as haloperidol, DTG and (+)-3-(3-hydroxyphenyl)-N-(1-propyl)piperidine were more potent in displacing [3H]DTG binding in cortical membranes than in hippocampal and cerebellar membranes, while the potencies of the NMDA antagonists were not significantly different from each other among these 3 different central structures. These results suggest that the rodent brain may contain multiple sigma sites with different low affinities for haloperidol in the micromolar range.
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  • Hiroyuki KOMAZAWA, Ikuo SAIKI, Miho AOKI, Hiroshi KITAGUCHI, Hideaki S ...
    1993 Volume 16 Issue 10 Pages 997-1003
    Published: October 15, 1993
    Released: April 10, 2008
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    Synthetic peptide analogues of the Arg-Gly-Asp-Ser(RGDS) sequence of fibronectin in which the amino acid of Gly was substituted with another one, named X, i.e. Arg-X-Asp-Ser (R-X-DS), and N-terminal modified R-X-DS have been synthesized to examine their antimetastatic effects in murine lung or liver metastasis models, as well as the inhibitory effect on tumor cell invasion, migration and adhesion in vitro. R-X-DS[X=Leu (L) or D-Leu (1)], as well as RGDS at a high dose of 3000 μg, significantly reduced the number of lung tumor colonies when they were co-injected with B16-BL6 melanoma. At a dose of 1000 μg/mouse, N-terminal modified R-X-DS, i.e. acetyl-D-R-X-DS [AcDR-X-DS : X=G, L or l], showed a more potent inhibitory effect on the lung or liver metastasis of B16-BL6 melanoma or L5178Y-ML25 lymphoma cells, respectively, as compared with RGDS or R-X-DS. AcDRLDS and AcDRlDS prevented the invasion of B16-BL6 cells into Matrigel/fibronectin- and Matrigel/laminin- coated filters, haptotactic migration, and the adhesion of the cells to both fibronectin- and laminin-coated substrates, whereas AcDRGDS inhibited only fibronectin-mediated cell functions. The intermittent i.v. administration of a water soluble vinylpolymer [poly(carboxyethylmethacrylamide), poly(CEMA)] containing R-X-DS (X=L or l) or RGDS, following the subcutaneous inoculation of B16-BL6 cells, significantly inhibited spontaneous lung metastasis as compared with multiple administrations of RGDS, R-X-DS or the untreated control. These results indicate that synthetic AcDR-X-DS (X=G, L or l) analogues and poly(CEMA-R-X-DS) conjugates may be useful for preventing cancer metastasis. It is of great interest that AcDR-X-DS (X=L or l) was able to regulate tumor cell adhesion, migration and invasion mediated by laminin as well as by fibronectin differently than AcDRGDS.
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  • Naoto INOUE, Kisa YAMADA, Kimie IMAI, Tachio AIMOTO
    1993 Volume 16 Issue 10 Pages 1004-1007
    Published: October 15, 1993
    Released: April 10, 2008
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    Sex-related differences in hepatic epoxide hydrolase (EH) activities towards 7-(2', 3'-epoxy)propoxycoumarin (7-glycidoxycoumarin, GOC) were investigated, mainly in mice but also in rats.Hepatic subcellular EH activities in the ddY mouse were higher in microsomes than in the soluble and mitochondrial fractions and sex-related differences were noted in all the subcellular fractions where males had significantly higher activities than females. Sex differences in the hepatic microsomal and soluble activities similar to those in the ddY strain were also observed in two other strains of mice, A/J and C3H/He, and in Wistar rats.In the ddY strain, castration of the males caused decreases in microsomal and soluble EH activities, while no alteration in the activities in those fractions was found following castration of females. Treatment of the male castrates with testosterone led to recovery of the activities in microsomal and soluble fractions while hormone treatment of female castrates caused a rise only in microsomal activity. Estradiol treatments of castrates of both sexes did not cause any changes in the hepatic subcellular activities.In intact ddY mice, testosterone treatment did not affect the male microsomal and soluble EH activities, but resulted in stimulation of both subcellular enzyme activities in females. In contrast, estradiol treatment showed a suppressive effect on both subcellular activities in males, but had no effect on female activities.These results show that hepatic EH activities towards GOC are mainly under androgenic stimulatory control in mice.
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  • Katsunori NAGAI, Kazuhito WATANABE, Shizuo NARIMATSU, Hiroshi GOHDA, T ...
    1993 Volume 16 Issue 10 Pages 1008-1013
    Published: October 15, 1993
    Released: April 10, 2008
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    The oxidative metabolism of cannabidiol (CBD) at the 8, 9-double bond was examined. 8R, 9-Epoxy-CBD was identified by GC-MS as a new metabolite of CBD produced by hepatic microsomal fractions of guinea pigs, rats and mice. The reaction required NADPH as a cofactor and molecular oxygen. The optimal pH for the reaction was 7.4-8.0. The 8R, 9-epoxy-CBD forming activity was highest in guinea pigs, followed by mice and rats in the presence of 3, 3, 3-trichloropropene-1, 2-oxide (TCPO), an inhibitor of epoxide hydrolase. The activity was significantly suppressed by SKF 525-A, α-naphthoflavone, metyrapone and carbon monoxide. 8R, 9-Epoxy-CBD was further converted to 6β-hydroxymethyl-Δ9-tetrahydrocannabinol (6β-CH2OH-Δ9-THC) and 8, 9-dihydro-8, 9-dihydroxy-CBD by hepatic microsomes of guinea pigs, rats and mice. Microsomal formation of 6β-CH2OH-Δ9-THC was markedly increased in the presence of TCPO with a concomitant decrease in the formation of 8, 9-dihydro-8, 9-dihydroxy-CBD in all animal species examined. Furthermore, 6β-CH2OH-Δ9-THC was also identified as a new metabolite of CBD produced by hapatic microsomes of guinea pigs. 6β-CH2OH-Δ9-THC exhibited THC-like pharmacological effects, catalepsy, analgesia, pentobarbital-induced sleep prolongation and hypothermia in mice, although these effects were less marked than those of Δ9-THC. This study presents the first example of the biotransformation of CBD to a Δ9-THC derivative which exhibits some pharmacological effects.
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  • Tomoko YAMASHITA, Junzo HIROSE, Masahide NOJI, Reiko SAITO, Hisao TOMI ...
    1993 Volume 16 Issue 10 Pages 1014-1018
    Published: October 15, 1993
    Released: April 10, 2008
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    Several Pt(IV) and Pt(II) complexes containing 1R, 2R-cyclohexanediamine (1R, 2R-dach) as a carrier ligand were synthesized. The cytotoxicities and the uptake of the platinum complexes by leukemia L1210 cells were compared in order to study the correlation between their structures and cytotoxicities.[Pt(II)Cl2(1R, 2R-dach)], [(Pt(II)(oxalato)(1R, 2R, -dach)], and [Pt(II)(malonato)(1R, 2R-dach)], which have excellent anticancer properties, exhibited very high cytotoxicities and were easily taken up by leukemia L1210 cells. [Pt(IV)Cl4(1R, 2R-dach)], trans(Cl)-[Pt(IV)Cl2(oxalato)(1R, 2R-dach)], and trans(Cl)-[Pt(IV)Cl2(malonato)(1R, 2R-dach)] also had high cytotoxicities. After a short incubation time, the uptake of [Pt(II)Cl2(1R, 2R-dach)], [Pt(II)(oxalato)(1R, 2R-dach)], and [Pt(II)(malonato)(1R, 2R-dach)] by leukemia L1210 cells were respectively very similar to those of [Pt(IV)Cl4(1R, 2R-dach)], trans(Cl)-[Pt(IV)Cl2(oxalato)(1R, 2R-dach)], and trans(Cl)-[Pt(IV)Cl2(malonato)(1R, 2R-dach)].In addition, trans(OH)-[Pt(IV)(OH)2Y2(1R, 2R-dach)] (Y2 : oxalato or malonato) did not exhibit cytotoxicity towards leukemia L1210 cells, whereas trans(Cl)-[Pt(IV)Cl2Y2(1R, 2R-dach)] (Y2 : oxalato or malonato) were highly cytotoxic. The accumulation of trans(OH)-[Pt(IV)(OH)2Y2(1R, 2R-dach)] in leukemia L1210 cells was much lower than that of trans(Cl)-[Pt(IV)Cl2Y2(1R, 2R-dach)]. Platinum(IV) complexes, in which leaving groups are replaced by hydroxide groups, have decreased cytotoxic activity, because the hydroxide groups of the platinum(IV) complex reduce the uptake of platinum by the cells. trans(OH), cis(Cl)-[Pt(IV)(OH)2Cl2(1R, 2R-dach)], which has hydroxide and chloride groups, was easily incorporated into the cells and exhibited the high cytotoxic activity. This behavior indicates that the chloride group apparently overcomes the ameliorating effect of the hydroxide group.
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  • Seiji MIYAUCHI, Yasufumi SAWADA, Tatsuji IGA, Manabu HANANO, Yuichi SU ...
    1993 Volume 16 Issue 10 Pages 1019-1024
    Published: October 15, 1993
    Released: April 10, 2008
    JOURNALS FREE ACCESS
    Concentration-dependency in the hepatic elimination of l-propranolol (l-PR) was investigated over a wide range of concentrations from 60 to 2200 μM in an isolated rat liver perfusion system. Under the steady-state condition produced by unlabeled l-PR at various concentrations, 3H-l-PR and 14C-inulin were bolusly injected into the portal vein, and the outflow was collected at 0.5 s intervals over 30 s. Up to 300 μM, the instantaneous hepatic availability of l-PR was approximately 4%, while it abruptly increased when the perfusate concentration exceeded 300 μM. To determine which process (influx or efflux or sequestration process) caused the nonlinearity, we calculated the rate constants k1 (influx), k2 (efflux), and k3 (sequestration) based on the "distributed" model. With increasing l-PR concentration in the perfusate, k2 increased approximately two times, whereas k3 decreased to approximately one-half. In contrast, k1 was independent of the perfusate concentration. The concentration-dependency of k2 was explained by saturation of l-PR tissue binding, since the tissue unbound fraction of l-PR obtained with liver homogenates and isolated hepatocytes increased approximately two times. The efflux and sequestration clearances were then normalized by the unbound fractions in the liver. The efflux clearance for unbound l-PR was constant irrespective of the perfusate concentration, whereas the sequestration clearance for unbound l-PR (CLint) showed Michaelis-Menten type saturation (Km=28 μM, Vmax=2.8 μmol/min/g liver, α (nonspecific) = 20 ml/min/g liver). Based on the comparison of the kinetic parameters representing these processes, the rate-determining process for the hepatic clearance of l-PR is a combination of the membrane transport and sequestration processes at low concentrations, while at high concentration, the rate-determining process is the "sequestration" process. Such concentration dependent shift of the rate determining process can be attributed to the saturation of its tissue binding as well as to the sequestration process.
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  • Taro OGISO, Masahiro IWAKI, Tadatoshi TANINO, Osamu MURAOKA, Genzou TA ...
    1993 Volume 16 Issue 10 Pages 1025-1030
    Published: October 15, 1993
    Released: April 10, 2008
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    The derivatives of phenytoin (DPH) were synthesized by the reaction at 3 position of hydantoin ring with valproic acid and valeric acid, producing valproyl DPH (VPDPH) and valeroyl DPH (VADPH), respectively. These derivatives showed much higher lipid solubilities than that of DPH. Their distribution and elimination were compared to those of DPH. Additionally, the concentration profiles of the drugs in brain and plasma were analyzed with a modified 2-compartment model. DPH and its derivatives, without hydrolysis to DPH in blood, were found rapidly distributed into brain, although the distribution of derivatives was much less, probably due to the high protein binding capacities. The distribution of DPH and its derivatives into brain regions was similar to that into the cortex cerebri. VPDPH and VADPH were more rapidly eliminated from plasma and brain than DPH, giving smaller mean residence time (MRT) values (0.92 and 0.85 h) and much smaller cortex/plasma concentration ratio than those of DPH. The VPDPH and VADPH concentrations in the cerebrospinal fluid (CSF) were also much lower than that of DPH. The time course of plasma and brain concentrations of DPH and its derivatives after i.v. administration was successfully described by the modified 2-compartment models presented.
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  • Jun WATANABE, Masami HABA, Hiroya MURANISHI, Hiroaki YUASA
    1993 Volume 16 Issue 10 Pages 1031-1034
    Published: October 15, 1993
    Released: April 10, 2008
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    The dose-dependent uptake of fractionated 3H-heparin in the subpopulations of liver cells, parenchymal and non-parenchymal cells, was chatacterized in rats in vivo. Following the intravenous administration of fractionated 3H-heparin, the radioactivity in plasma was eliminated according to the first order kinetics at each dose. However, the elimination rate constant decreased with dose over the dose range of 0.3 to 100 U/kg, suggesting nonlinear elimination. In accordance with the delay in the plasma elimination, the uptake rate constant of radioactivity by parenchymal as well as non-parenchymal cells of liver, the major distribution organ, also decreased. Although heparin has long been considered to be taken up by a reticuloendothelial system (RES) such as non-parenchymal cells in the liver, the uptake of fractionated 3H-heparin by parenchymal cells was found to be comparable with that by non-parenchymal cells at the lowest dose of 0.3 U/kg, and even larger than that by non-parenchymal cells at the highest dose of 100 U/kg. The uptake clearances of fractionated 3H-heparin at the dose of 0.3 U/kg were 86.4 and 504 ml/108 cells/d, respectively, for parenchymal and non-parenchymal cells. These values were much larger than those reported for polyvinylpyrrolidone, which has been suggested to be taken up by fluid phase endocytosis. Thus, the present study revealed the significant contribution of parenchymal cells in the hepatic uptake of fractionated 3H-heparin. The dose-dependent uptake with high clearance values in both parenchymal and non-parenchymal cells provides an in vivo suggestion of the specialized transport of fractionated heparin in these two subpopulations of liver cells.
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  • Jun WATANABE, Hiromu KONDO, Hiroya MURANISHI, Kimihiko URANO, Masami H ...
    1993 Volume 16 Issue 10 Pages 1035-1039
    Published: October 15, 1993
    Released: April 10, 2008
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    The concentration-dependent hepatic uptake of fractionated [3H]heparin, a macromolecular model drug, was kinetically characterized and the effect of plasma proteins, albumin and α-globulin, was evaluated in the perfused rat liver as part of an ongoing effort to elucidate the mechanism of interaction of macromolecular drugs with biological macromolecules and the role of this interaction in the drugs' distribution. In the absence of proteins, the uptake of fractionated [3H]heparin was saturable with the maximum uptake velocity (Vmax) of 7.6 pmol/min/g liver and the Michaelis constant (Km) of 32.2 nM, suggesting the involvement of a specialized transport. α-Globulin (8.0 mg/ml) reduced the uptake of fractionated [3H]heparin at lower heparin concentrations. However, albumin (40 mg/ml) did not affect the uptake of fractionated [3H]heparin, suggesting an insignificant interaction. Assuming that fractionated [3H]heparin bound to α-globulin cannot be uptaken and that the reduction in uptake was solely attributable to the saturable Scatchard-type binding of fractionated [3H]heparin to α-globulin, the dissociation constant (Kd) and the binding capacity (n) were estimated to be 2.1 nM and 0.002, respectively.In in vitro binding experiments by ultrafiltration, Kd and n were estimated as 168 nM and 0.5, respectively, for α-globulin and 1021 nM and 0.02, respectively, for albumin, suggesting lower affinity and higher capacity in vitro for each protein. The results of in vitro binding experiments did not agree with those of perfusion experiments in that the binding of fractionated [3H]heparin to albumin was significant and the unbound fraction in the presence of α-globulin was smaller than that estimated in the analysis of hepatic uptake. The apparent enhancement of dissociation of fractionated [3H]heparin in perfusion suggests the participation of "protein-mediated transport, " as reported for low molecular weight compounds in the transport of macromolecular compounds.Thus, the present study successfully characterized the hepatic uptake of fractionated [3H]heparin as a Michaelis-Menten type process in the perfused rat liver and confirmed the involvement of protein-mediated transport.
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  • Yasuko ISHII, Hisayuki TANIZAWA, Yoshio TAKINO
    1993 Volume 16 Issue 10 Pages 1040
    Published: October 15, 1993
    Released: April 10, 2008
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    As individual differences in sensitivity in the laxative activity of barbaloin were observed in rats, a selection test for barbaloin (31.1 mg/kg, p.o.) was carried out between 1989 and 1991. The annual percentages of rats exhibiting a reaction positive to barbaloin were almost constant (about 57%). We propose that rats positive to barbaloin should be selected for studies on the laxative effect of barbaloin.
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  • Kazuhito WATANABE, Yoko HIDA, Tamihide MATSUNAGA, Ikuo YAMAMOTO, Hidet ...
    1993 Volume 16 Issue 10 Pages 1041-1043
    Published: October 15, 1993
    Released: April 10, 2008
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    The hepatic microsomal metabolism of cocaine and the pharmacological effects of some metabolites were studied in experimental animals. Hepatic microsomes from mice, rats and guinea pigs catalyzed the oxidation of cocaine to m- and p-hydroxycocaines. Only trace amounts of m-hydroxycocaine were detected in the extract of the incubation mixture with rabbit hepatic microsomes. The total amount of the two hydroxycocaines was less than 12% that of norcocaine which was the most predominant microsomal metabolite in all the animal species examined. The administration of p-hydroxycocaine (20 mg/kg i.p.) to mice significantly increased locomotor activity (total distance and number of rearing movements). The effect of p-hydroxycocaine was more active or comparable with that of cocaine, indicating that this metabolite is an active metabolite of cocaine.
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  • Shuji WATANABE, Shigeaki MARUO, Hideya MUKAI, Akira MORINO
    1993 Volume 16 Issue 10 Pages 1044-1045
    Published: October 15, 1993
    Released: April 10, 2008
    JOURNALS FREE ACCESS
    The utilization of intravenously infused maltotriose, maltotetraose and maltopentaose was investigated in rabbits. Rabbits were infused with isotonic solutions of each maltooligosaccharide for 2 h and the utilization was assessed by the 24-h urinary loss of carbohydrates. The utilization of these maltooligosaccharides was good (≥87.5%) at a rate of 1 ml/kg/h (≤250 mg/kg/h) and poor (≤36.2%) at a rate of 5 ml/kg/h (>250 mg/kg/h). Furthermore, these maltooligosaccharides infused as 5% solution showed good utilization (≥86.2%) at a rate of 5 ml/kg/h (250 mg/kg/h). These results suggest the existence of a threshold dose in the utilization of intravenously infused maltooligosaccharides, and this threshold value in rabbits is estimated to be nearly 250 mg/kg/h.
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  • Jiro NEMOTO, Naohito OHNO, Kazuo SAITO, Yoshiyuki ADACHI, Toshiro YADO ...
    1993 Volume 16 Issue 10 Pages 1046-1050
    Published: October 15, 1993
    Released: April 10, 2008
    JOURNALS FREE ACCESS
    OL-2, a highly branched (1→3)-β-D-glucan, is an antitumor glucan showing strong hematopoietic activity with weaker adjuvant activity than schizophyllan (SPG), also an antitumor glucan and one which is clinically used. This paper deals with the gene expression of the interleukin 1 (IL-1) family in mice by OL-2 and SPG in order to characterize the immunopharmacological activity. Gene expression was examined by reverse transcriptase-polymerase chain reaction method. Intraperitoneal administration of OL-2 (250 μg/mouse) expressed all three genes of IL-1α, β, and IL-1 receptor antagonist (IL-1ra) in the peritoneal exudate cells, while SPG induced a strength of IL-1α mRNA comparable to that by OL-2 but a weaker level of IL-1β mRNA. SPG did not induce IL-1ra. Similar patterns were seen in spleen and liver by OL-2 or SPG administration. These findings suggest that the immunopharmacological characteristics of (1→3)-β-D-glucan are regulated under the gene expression of the IL-1 family.
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  • Yukihiro SHOYAMA, Tomohiro FUKADA, Takashi TANAKA, Akihiko KUSAI, Kazu ...
    1993 Volume 16 Issue 10 Pages 1051-1053
    Published: October 15, 1993
    Released: April 10, 2008
    JOURNALS FREE ACCESS
    Opium alkaloids (thebaine, codeine and morphine) have been conjugated with bovine serum albumin (BSA) to give individual antigen conjugates which are analyzed by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. It became clear that 9 molecules of thebaine were contained in a thebaine-BSA conjugate. Codeine and morphine contents in individual conjugates were determined to be 12 and 6 molecules, respectively.
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  • Dai SASAKI, Satoshi KOSUNAGO, Takeshi MIKAMI, Tatsuji MATSUMOTO, Masuk ...
    1993 Volume 16 Issue 10 Pages 1054-1056
    Published: October 15, 1993
    Released: April 10, 2008
    JOURNALS FREE ACCESS
    Two subclones (K562-L and -H) were previously isolated from K562 human leukemic cells according to hemoglobin production : K562-L was expressed in less than 5% and K562-H in more than 90% of dianisidine positive cells. 12-O-Tetradecanoylphorbol-13-acetate (TPA) suppressed the expression of the erythrocytic (glycophorin A) and myelocytic (CD11b) antigens in K562-L, but increased the expression of these antigens in K562-H. TPA increased the megakaryocytic (CD61) antigens in both cells. These findings suggest that there are distinct TPA responsible factors in K562-L and -H on the expression of the erythrocytic and myelocytic antigens.
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  • Eiji TAKAHARA, Osamu NAGATA, Hideo KATO, Shigeru OHTA, Masaaki HIROBE
    1993 Volume 16 Issue 10 Pages 1057-1059
    Published: October 15, 1993
    Released: April 10, 2008
    JOURNALS FREE ACCESS
    Inter-individual differences of drug plasma concentration were recognized in male Sprague-Dawley (SD) rats after a p.o. or i.v. administration of (+)-MPPB. Rats could be divided into two phenotypes, the rapid-metabolizing group (RM) and the slow-metabolizing group (SM). The hepatic clearance (CLliver) of RM was about 10 times larger than that of SM. Outbred SD(♂, ♀), and Wister(♂) rats were mixtures of RM and SM. On the other hand, all inbred Lewis(♂) rats were RM, and all inbred F344(♂) and ACl(♂) rats were SM.
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