Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
26 巻 , 8 号
選択された号の論文の34件中1~34を表示しています
Analytical Biochemistry
Regular Articles
  • Akiko Iwata, Koei Satoh, Mitsuhiro Murata, Mikio Hikata, Takao Hayakaw ...
    原稿種別: Regular Article
    専門分野: Analytical Biochemistry
    2003 年 26 巻 8 号 p. 1065-1069
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    To enhance the sensitivity of virus detection by polymerase chain reaction (PCR) and reverse-transcriptional (RT)-PCR, we developed a novel virus-concentration method using sulfonated (SO-) magnetic beads in the presence of divalent cations. In the presence of either Zn2+ or Cu2+ ions, we showed that SO-magnetic beads were able to concentrate non-enveloped model viruses, such as porcine parvovirus (PPV) and poliovirus, which were not concentrated by polyethyleneimine (PEI)-magnetic beads.1) Using the SO-magnetic beads, the sensitivity of virus genome detection by PCR or RT-PCR can be enhanced. Therefore, an efficient virus concentration method using either SO-magnetic beads or PEI-magnetic beads enhances the sensitivity of virus detection by PCR or RT-PCR.
Notes
  • Keiko Okimura, Kazuhiro Ohki, Sotoo Nagai, Naoki Sakura
    原稿種別: Note
    専門分野: Analytical Biochemistry
    2003 年 26 巻 8 号 p. 1166-1169
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    Acylation with long-chain fatty acids is a common modification at the N-terminal glycine residues of natural proteins. In this work, we performed HPLC analysis of myristoylglycine (Myr-Gly-OH), palmitoylglycine (Pal-Gly-OH) or lauroylglycine (Lau-Gly-OH), which were produced in the hydrolysates of synthetic Myr-Gly-, Pal-Gly-, or Lau-Gly-peptides, respectively, by means of a mild acid hydrolysis in methanesulfonic acid : dioxane : water (2 : 1 : 1) at 60 °C for 12 h. Myr-Gly-OH, Pal-Gly-OH and Lau-Gly-OH were quite stable under hydrolysis conditions. These fatty acyl-Gly-OH were conveniently detectable at a 20 nmol level by direct reversed-phase HPLC. Thus, mild acid hydrolysis, followed by HPLC analysis of the hydrolysate, provides a simple method of identification of the N-terminal structure of fatty acyl-Gly-peptides.
Biochemistry/Molecular Biology
Regular Articles
  • Risa Kurozumi, Sachiko Tokuzumi, Shuji Kojima
    原稿種別: Regular Article
    専門分野: Biochemistry/Molecular Biology
    2003 年 26 巻 8 号 p. 1070-1075
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    The mechanism underlying the elevation of intracellular glutathione (GSH) in RAW 264.7 cells exposed to low-level sodium nitroprusside (SNP) was investigated by measuring the expression of mRNA for γ-glutamylcysteine synthetase (γ-GCS), the rate-limiting enzyme of de novo GSH synthesis, and the GSH content. A significant elevation of expression of mRNA for γ-GCS was observed at 3 h after exposure of the cells to SNP at a concentration of 0.25 mM. 2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO), N-acetylcysteine (NAC), or ebselen (Ebs) significantly suppressed the elevations induced by SNP, suggesting that hydrogen peroxide or peroxynitrite (ONOO) is involved in this event as a triggering molecule. Hydrogen peroxide itself, however, did not induce the elevation of γ-GCS mRNA and glutathione. Chemiluminescenses induced by SIN-1, a chemical ONOO donor, and ONOO itself were completely blocked by Ebs. SIN-1 also significantly elevated the cellular glutathione level, and the elevation was absolutely blocked by Ebs. These results suggest that the elevation of intracellular GSH in RAW 264.7 cells exposed to low-level SNP occurs via the de novo GSH pathway through transcriptional up-regulation of the γ-GCS gene induced by peroxynitrite molecule.
  • Yoko Makuta, Yoshiko Sonoda, Daisuke Yamamoto, Megumi Funakoshi-Tago, ...
    原稿種別: Regular Article
    専門分野: Biochemistry/Molecular Biology
    2003 年 26 巻 8 号 p. 1076-1081
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    As an immunosuppressive and anti-inflammatory cytokine, IL-10 was recently reported to play roles in CCR5 expression in human monocytes. CCR5 promoter regions contain Oct-2, TCF-1α, GATA, and STAT binding sites. Here, we studied the signals involved in the CCR5 expression in IL-10-stimulated cells using the HL-60 cell line. HL-60 cells were stimulated with PMA and differentiated to macrophage-like cells, then stimulated with IL-10. IL-10 induced significant expression of CCR5 protein and CCR5 mRNA in these cells. The induction of CCR5 by IL-10 was inhibited by a MEK-1 inhibitor, PD98059. In addition, IL-10 induced tyrosine (Tyr) phosphorylation of Erk, as well as serine (Ser) and Tyr phosphorylation of STAT-3. Tyr phosphorylation of Erk and Ser phosphorylation of STAT-3 were inhibited by PD98059, while Tyr phosphorylation of STAT-3 was not inhibited by PD98059. DNA binding activity of STAT-3 was observed by the stimulation with IL-10, which was inhibited by PD98059. These results first indicate that Erk1/2 and STAT-3 regulate CCR5 expression, and that Erk-mediated phosphorylation of Ser is required for full stimulation of STAT-3 in CCR5 expression.
  • Akihiro Michihara, Kenji Akasaki, Yukio Yamori, Hiroshi Tsuji
    原稿種別: Regular Article
    専門分野: Biochemistry/Molecular Biology
    2003 年 26 巻 8 号 p. 1082-1085
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    We previously reported that treatment of rats with a diet containing 0.1% pravastatin and 5% cholestyramine markedly increased mevalonate pyrophosphate decarboxylase (MPD) activity in liver crude extracts compared with nontreated rats. In this study, we examined the change in the protein level of MPD in the tissues of mice administered pravastatin. When MPD content in the tissues of nontreated mice was analyzed by quantitative immunoblotting, a single protein band with an apparent molecular weight of 46 kDa was detected in all tissues and the specific protein content of MPD in liver and kidney was markedly higher than that in other tissues. When MPD content in the tissues of pravastatin-treated mice was analyzed by immunoblotting, MPD was markedly increased (9-fold) only in the liver compared with nontreated mice. Next, when MPD activity was measured in the liver between nontreated and pravastatin-treated mice, MPD activity as well as protein levels were markedly increased (11-fold) in the liver of pravastatin-treated mice compared with nontreated mice. These data suggest that a marked induction of MPD in the liver by pravastatin is responsible for the tissue-specific effect of pravastatin.
  • Won Chung Lim, Jeong Bae Park, Young Joo Lee
    原稿種別: Regular Article
    専門分野: Biochemistry/Molecular Biology
    2003 年 26 巻 8 号 p. 1086-1088
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    The renin-angiotensin cascade plays an important role in blood pressure control and sodium homeostasis. This study investigated whether cyclooxygenase-2 expression is regulated in the kidney, in an angiotensin II- and aldosterone-induced hypertension model. For this purpose, we treated male Sprague-Dawley rats (n=8 per group) with angiotensin II (9 mg/h, subcutaneously) for 14 d and aldosterone (0.75 mg/h, subcutaneously) for 42 d. Systolic blood pressure was significantly increased by angiotensin II (p<0.001) and by aldosterone (p<0.001). We found that angiotensin II downregulated cyclooxygenase-2 protein in the kidney cortex, whereas aldosterone showed no effect. These results indicate that angiotensin II may act directly to inhibit kidney cortex cyclooxygenase-2 protein expression, rather than acting via stimulation of aldosterone.
Notes
  • Ho-Young Shin, Ji-Hyun Lee, Jang-Yeon Lee, Yeo-Ock Han, Myung Joo Han, ...
    原稿種別: Note
    専門分野: Biochemistry/Molecular Biology
    2003 年 26 巻 8 号 p. 1170-1173
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    β-D-Xylosidase (EC 3.2.1.37) has been purified from ginsenoside Ra-metabolizing Bifidobacterium breve K-110, which was isolated from human intestinal microflora. β-D-Xylosidase was purified to apparent homogeneity by a combination of ammonium sulfate precipitation, QAE-cellulose, butyl-toyopearl, hydroxyapatit and Q-Sepharose column chromatographies with the final specific activity of 51.8 μmol/min/mg. Molecular weight of β-D-xylosidase is 49 kDa by SDS-PAGE and gel filtration, which consisted of a single subunit. β-D-Xylosidase showed optimal activity at pH 5.0 and 37 °C. The purified enzyme was potently inhibited by PCMS. β-D-Xylosidase acted to the greatest extent on p-nitrophenyl-β-D-xylopyranoside, followed by ginsenoside Ra1 and ginsenoside Ra2. This enzyme hydrolyzed xylan to xylose, but did not act on p-nitrophenyl-β-glucopyranoside, p-nitrophenyl-β-galactopyranoside or p-nitrophenyl-β-D-fucopyranoside. These findings suggest that this is the first reported purification of ginsenoside-hydrolyzing β-D-xylosidase from an anaerobic Bifidobacterium sp.
  • Atsushi Tsuji, Yuya Akaza, Shogo Nakamura, Ken-ichi Kodaira, Hiro Yasu ...
    原稿種別: Note
    専門分野: Biochemistry/Molecular Biology
    2003 年 26 巻 8 号 p. 1174-1177
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    The cellular slime mold Dictyostelium discoideum expresses three genes (sodA, sodB and sodC) encoding the extracellular Cu/Zn superoxide dismutases. Following H2O2 treatment, the expression of sodA and sodB increased while that of sodC decreased. The sodC null strain formed multinucleate cells in a shaking culture. These results suggest that sodC plays a unique role in Dictyostelium discoideum.
Pharmacology
Regular Articles
  • Jieyun Jiang, Chunxiang Zhou, Qiang Xu
    原稿種別: Regular Article
    専門分野: Pharmacology
    2003 年 26 巻 8 号 p. 1089-1094
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    The present study aims at examining the effects of Si-Ni-San, a prescription usually used for treating hepatitis in Traditional Chinese Medicine (TCM), on various experimental liver injury models and its mechanisms. The prescription showed significant hepatoprotection against CCl4-induced hepatic damage, both in vivo and in vitro. To the liver injury induced by Bacillus Calmette-Guerin (BCG) with lipopolysaccharide (LPS), Si-Ni-San also provided significant alleviation through enhancing nitric oxide (NO) release by macrophages. Against the liver injury induced by a delayed-type hypersensitivity reaction to picryl chloride (PCl-DTH), Si-Ni-San alleviated it remarkably when administered during either the induction or effector phase. A significant reduction of in-vitro hepatotoxicity, as measured by the inhibition of serum transaminase evaluation, was observed in nonparenchymal cells from liver-injured mice treated with Si-Ni-San. Si-Ni-San facilitated apoptosis in nonparenchymal cells from liver-injured mice, as well as in spleen cells activated by PCl in vivo or by Con A in vitro. These results suggest that Si-Ni-San provides alleviating effects against liver injury through multiple mechanisms, including protection of the hepatocyte membrane, enhancement of NO release, and dysfunction of liver-infiltrating cells mainly through causing their apoptosis.
  • Qin Zhou, Dewi Kenneth Rowlands, Yu Lin Gou, Lai Ling Tsang, Yiu Wa Ch ...
    原稿種別: Regular Article
    専門分野: Pharmacology
    2003 年 26 巻 8 号 p. 1095-1099
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    The cardiovascular protective effects of the traditional Chinese medicine Bak Foong Pills (BFP) were investigated. Spontaneously hypertensive rats (SHR) were treated (3 g/kg) over a 5-month period and blood pressure measurements periodically tested with a plethysmographic tail cuff. Following treatment, blood samples were analysed for serum electrolyte levels and lipid levels and brain tissue subjected to micro-array analysis. In vitro experiments were also conducted to identify possible direct vasorelaxatory effect. The results showed that BFP was able to significantly reduce both systolic and diastolic blood pressure by about 30 mmHg in SHR following 5 months of treatment, when compared to untreated animals. Investigation for possible mechanisms of actions revealed that BFP treated rats had elevated blood serum K+ levels, and also demonstrated decreased serum triglyceride levels. Micro-array analysis of brain tissue showed altered expression of acetylcholine and lysosphingolipid receptor genes that are known to regulate blood pressure. In vitro experiments also showed that BFP caused a concentration-dependant vasorelaxation of isolated rat aortae when contracted with phenylepherine, which was partially inhibited by nitric oxide synthase inhibitor L-NAME (100 μM). These data suggest that BFP is able to significantly reduce hypertension in SHR through mechanisms probably involving a combination of increased serum K+, vasorelaxatory action, reduced serum triglyceride and altered gene regulation in the higher centres.
  • Hyean-Woo Lee, Yoon-Sun Park, Jong-Whan Choi, Sang-yeop Yi, Woon-Seob ...
    原稿種別: Regular Article
    専門分野: Pharmacology
    2003 年 26 巻 8 号 p. 1100-1103
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    The antidiabetic effect of chitosan oligosaccharide (COS) was investigated in neonatal streptozotocin (STZ)-induced noninsulin-dependent diabetes mellitus rats. The fasting glucose level was reduced by about 19% in diabetic rats after treatment with 0.3% COS. Glucose tolerance was lower in the diabetic group compared with the normal group. After diabetic rats had been treated with 0.3% COS for 4 weeks, glucose tolerance increased significantly versus the diabetic control group, and glucose-inducible insulin expression increased significantly. In addition, fed-triglyceride (TG) levels in diabetic rats drinking 0.3% COS were reduced by 49% compared with those in diabetic control rats. The cholesterol levels of animals treated with COS were reduced by about 10% in fed or fasting conditions versus the corresponding controls, although the difference was not statistically significant. It was found that COS has a TG-lowering effect in diabetic rats, and that COS reduces signs of diabetic cardiomyopathy such as vacuolation of mitochondria and the separation and degeneration of myofibrils. In conclusion, these results indicate that COS can be used as an antidiabetic agent because it increases glucose tolerance and insulin secretion and decreases TG.
  • Takafumi Naito, Hiroki Itoh, Masaharu Takeyama
    原稿種別: Regular Article
    専門分野: Pharmacology
    2003 年 26 巻 8 号 p. 1104-1107
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    Regarding the gastroprotective function as a neural emergency system, sensory afferent neurons in the gastrointestinal mucosa regulate neuropeptide (calcitonin gene-related peptide (CGRP), substance P, etc.) levels, and those peptides play various physiological roles. To determine whether the pharmacological effects of Hange-shashin-to and Rikkunshi-to on the gastrointestine are due to changes in gastrointestinal mucosa regulatory peptides levels, we investigated the levels of CGRP-like immunoreactive substances (IS) and substance P-IS in plasma from healthy subjects. A single oral administration of Hange-shashin-to caused significant increases in CGRP-IS (40—60 min) and substance P-IS (60—180 min) levels in the plasma compared with the levels induced by a placebo. Rikkunshi-to and a 5.0 g Pinelliae tuber extract had no significant effect on CGRP-IS and substance P-IS levels. Extract of a 2.5 g Zingiberis rhizoma significantly caused increases in CGRP-IS at 40 min and in substance P-IS at 60 min. These results, in comparison with Kampo medicines, might indicate that the pharmacological actions of Hange-shashin-to closely are related to changes in CGRP-IS and substance P-IS levels, while Zingiberis rhizoma partially might participate in those effects of Hange-shashin-to.
  • Narumi Sugihara, Arisa Kaneko, Koji Furuno
    原稿種別: Regular Article
    専門分野: Pharmacology
    2003 年 26 巻 8 号 p. 1108-1114
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    Sixteen flavonoids including quercetin and kaempferol and their relatives were examined for their ability to promote DNA degradation induced by the bleomycin (BLM)–Fe complex. Three hydroxyl groups in the flavonoidal nucleus were proposed as a crucial structural requirement for effectively promoting DNA degradation: 1) the C7-hydroxyl substitution in the A-ring; 2) the C4′-hydroxyl substitution in the B-ring; and 3) the C3-hydroxyl substitution in the C-ring. Flavonoids, which lack even one of these hydroxyl substitutions, showed remarkably diminished activity. There was a good correlation (r=0.920, p<0.001) between activity to promote DNA degradation and oxidizability, which was measured following the Fe(III)-induced oxidation of flavonoids themselves, among the 16 flavonoids. The oxidizability of flavonoids which have the crucial hydroxyl substitutions, was remarkably enhanced in the presence compared with the absence of BLM. On the other hand, the extent of oxidation of flavonoids lacking these substitutions was enhanced little or not at all by BLM. No correlation between the Fe(III)-reducing activity and DNA degradation-promoting activity was found among flavonoids satisfying the crucial structural requirements. Furthermore, the correlation between the extent of oxidation of flavonoids and the Fe(III)-reducing activity was not confirmed among these flavonoids. Therefore, it was suggested that Fe(III)-reducing activity was not the only factor determining DNA degradation-promoting activity in flavonoids having the three hydroxyl groups necessary for effectively promoting DNA degradation induced by BLM–Fe complex.
  • Yoshiyuki Kagawa, Jun-ichi Sawada, Shizuo Yamada, Hiroaki Matsuda, Shi ...
    原稿種別: Regular Article
    専門分野: Pharmacology
    2003 年 26 巻 8 号 p. 1115-1119
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    This study was undertaken to investigate the relationship between blood concentration of cyclosporine A (CsA), administered intravenously by a 24-h continuous infusion, and drug-induced nephrotoxicity or hepatotoxicity. It was investigated retrospectively in 8 patients who had received an allogeneic bone marrow transplant (BMT). The correlation between daily doses and blood concentration of CsA was not significant. Then, the data of blood concentration of CsA and renal or liver function test result were divided into 5-d periods from the date of transplantation, and the mean value for each period was calculated. The maximum values of blood urea nitrogen (BUN) and serum creatinine (SCr) were consistently observed only after the period when the 5-d mean CsA concentration reached the peak level: the maximum BUN and SCr values were witnessed at Periods 2 to 10 and at Periods 1 to 9, respectively. On the other hand, no consistent correlation was found between the 5-d mean CsA concentrations and liver function test result. We also investigated the relationship between renal function and the cumulative dose or AUC of CsA. The parameters of renal function tests reached peak levels at the cumulative dose of 4000 to 10000 mg and at the cumulative AUC of 280000 to 660000 ng/ml·h. These results suggest that: 1) a deterioration of renal function occurs usually after the peak blood concentration of CsA is attained, and 2) the monitoring of the blood concentration of CsA is useful in predicting renal dysfunction in post-BMT patients.
  • Shinya Yodogawa, Takayuki Arakawa, Narumi Sugihara, Koji Furuno
    原稿種別: Regular Article
    専門分野: Pharmacology
    2003 年 26 巻 8 号 p. 1120-1124
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    Glucurono- and sulfo-conjugation of kaempferol in rat liver preparations and cultured hepatocytes were studied using high-performance liquid chromatography (HPLC) with two distinctly different elution solvents. Kaempferol glucuronides and sulfates were produced by treating kaempferol with microsomes plus UDPGA or with cytosol plus PAPS, respectively. HPLC analysis of the conjugates revealed one major and three minor glucuronides with solvent A and one sulfate with solvent B. Kaempferol metabolites produced by cultured hepatocytes also consisted of four glucuronides and one minor sulfate, all of which corresponded to their respective in vitro-produced conjugates in the liver subcellular preparations. The relative proportion of kaempferol sulfate accounted for about 9% of the total conjugates in the cultured hepatocytes. The kinetic data on glucurono- and sulfo-conjugation of kaempferol by the liver subcellular preparations correlated well with the preferential production of kaempferol glucuronides in the cultured hepatocytes. Glucurono- and sulfo-conjugation of 3-, 5- and 7-OH flavones in the liver subcellular preparations were also kinetically characterized. 7-OH flavone was predominantly conjugated to form a glucuronide compared to 3- and 5-OH flavones. These data suggest that glucuronidation at the 7-OH position on the A-ring is a major metabolic pathway of kaempferol in hepatic cells.
  • Kenji Ohishi, Haruji Sawada, Yasuto Yoshida, Wakae Yokoi, Hiroshi Hata ...
    原稿種別: Regular Article
    専門分野: Pharmacology
    2003 年 26 巻 8 号 p. 1125-1128
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    The effects of an acyl-CoA:cholesterol O-acyltransferase (ACAT) inhibitor, N-(3,5-dimethoxy-4-n-octyloxycinnamoyl)-N′-(3,4-dimethylphenyl)piperazine (YIC-C8-434), on cholesterol esterification in the intestine and liver were investigated in vitro and in vivo. YIC-C8-434 inhibited the formation of cholesteryl [3H]oleate from [3H]oleic acid and cholesterol both in human colon adenocarcinoma Caco2 cells and in human hepatoma HepG2 cells with IC50 values of 0.38 and 0.49 μM, respectively. However, it did not influence the incorporation of [3H]oleic acid into triacylglycerols and phospholipids. Oral administration of YIC-C8-434 at a dose of 8.3 mg/kg/d inhibited [14C]cholesterol absorption by 17% (p<0.01) in rats. YIC-C8-434 also significantly reduced the secretion of very low-density lipoprotein (VLDL) cholesterol from the liver into the plasma at an oral dose of 100 mg/kg/d after an intravenous injection of Triton WR-1339. These results suggest that oral administration of YIC-C8-434 reduces intestinal cholesterol absorption and hepatic VLDL cholesterol secretion by direct inhibition of ACAT in the intestinal epithelium and hepatocytes, respectively. However, the inhibitory action of YIC-C8-434 on cholesterol absorption rather than hepatic cholesterol secretion may play a more important role in its hypocholesterolemic activity, because the effective dose for the former was 12-fold lower than that for the latter.
Notes
  • Kenji Honda, Namiko Murao, Takae Ibuki, Hiro-o Kamiya, Yukio Takano
    原稿種別: Note
    専門分野: Pharmacology
    2003 年 26 巻 8 号 p. 1178-1180
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    We have examined the effects of intrathecal (i.t.) injection of the muscarinic acetylcholine receptor antagonist atropine on the clonidine-induced nociceptive effect in formalin-induced nociception in rats. The injection of 5% formalin into the hind paw caused biphasic nociceptive responses, and i.t. injection of clonidine inhibited both phases of the nociceptive response in a dose-dependent manner. Pretreatment with atropine (i.t.) only partially inhibited the nociceptive effect of clonidine. These results suggest that the nociceptive effect of clonidine in the rat formalin model may be at least partly mediated by muscarinic acetylcholine receptors in the spinal cord.
  • Libin Zhan, Toru Hosoi, Yasunobu Okuma, Yasuyuki Nomura
    原稿種別: Note
    専門分野: Pharmacology
    2003 年 26 巻 8 号 p. 1181-1183
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    Increasing evidence has suggested that cholecystokinin (CCK) is involved in immune-to-brain communication. The afferent vagus nerve is an important component for transmitting peripheral immune signals to the brain, such as those determining interleukin (IL)-1β expression in the brain and anorexia. In the present study, we investigated whether the anorexic effect of CCK, which also activates the afferent vagus nerve, is mediated via IL-1β expression in the brain. CCK-8 dose-dependently (8—320 μg/kg, i.p.) inhibited food intake in mice. However, IL-1β transcripts in the hypothalamus, the hippocampus and the brainstem were not significantly increased after the administration of CCK-8, even at the larger dose of 320 μg/kg. These findings suggest that the CCK-induced inhibition of food intake may be independent of IL-1β production in the brain, and indicate the diverse role of CCK in the regulation of the neuro-immune interaction.
  • Mamunur Rashid, Habib Abul Muntasir, Masatomo Watanabe, Mikio Nakazawa ...
    原稿種別: Note
    専門分野: Pharmacology
    2003 年 26 巻 8 号 p. 1184-1187
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    The aim of the present study was to investigate the binding affinities and dissociation potencies of several 5-HT2 antagonists in M2 muscarinic receptor of rat heart membranes using [3H]QNB as a radioligand. The 5-HT2 antagonists used in this study were sarpogrelate, ketanserin and cyproheptadine. The results showed that sarpogrelate and ketanserin had very weak binding affinities to M2 muscarinic receptor, whereas cyproheptadine had higher binding affinity to this receptor than the muscarinic receptor antagonist, atropine. All of these three 5-HT2 antagonists as well as muscarinic receptor antagonists (atropine and pirenzepine) were readily dissociated from M2 muscarinic receptor in rat heart membranes after washing. Therefore, the findings of the present investigation suggest that the dissociation potencies of neither 5-HT2 antagonists nor muscarinic antagonists used correlate with their binding affinities to M2 muscarinic receptors in rat heart.
  • Masayoshi Yamaguchi, Satoshi Uchiyama
    原稿種別: Note
    専門分野: Pharmacology
    2003 年 26 巻 8 号 p. 1188-1191
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    The effects of various carotenoids and rutin on calcium content and alkaline phosphatase activity in the femoral-diaphyseal and femoral-metaphyseal tissues of young rats in vitro were investigated. Bone tissues were cultured for 48 h in serum-free Dulbecco's modified Eagle's medium containing either vehicle or a compound (10−8—10−6 M). The presence of β-cryptoxanthin (10−7 or 10−6 M) caused a significant increase in calcium content and alkaline phosphatase activity in the femoral-diaphyseal and femoral-metaphyseal tissues. Also, the lowest concentration (10−8 M) of β-cryptoxanthin used caused a significant increase in diaphyseal and metaphyseal calcium content. Lutein (10−8—10−6 M) had no effect on diaphyseal and metaphyseal calcium contents and diaphyseal alkaline phosphatase activity, while 10−7 and 10−6 M lutein significantly decreased metaphyseal alkaline phosphatase activity. Lycopene (10−8—10−6 M) or rutin (10−8—10−6 M) did not have a significant effect on bone calcium content and alkaline phosphatase activity. The present study suggests that the carotenoid β-cryptoxanthin has a unique anabolic effect on bone calcification in vitro.
  • Yoshio Tanaka, Takao Okamoto, Toshiyasu Imai, Takahiro Horinouchi, Hik ...
    原稿種別: Note
    専門分野: Pharmacology
    2003 年 26 巻 8 号 p. 1192-1194
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    Urinary bladder smooth muscle (UBSM) exhibits spontaneous rhythmic contraction. This spontaneous mechanical activity is generated in the presence of neuronal blockade and thus is myogenic in origin. The spontaneous myogenic contraction of UBSM may be the fundamental determinant of the physiological functions of the urinary bladder to store and excrete urine. Although the mechanisms by which UBSM generates spontaneous contraction have not been completely ascertained, its induction has been suggested to be intimately associated with smooth muscle cell action potentials to enhance extracellular Ca2+ influx through voltage-gated L-type Ca2+ channels. However, the alteration of membrane electrical activity does not seem to be the exclusive trigger mechanism for the generation of the spontaneous contraction. In the present study, we show that spontaneous mechanical activity of guinea pig UBSM is substantially diminished by an inhibitor of phospholipase C (PLC), U-73122, but is not affected by its inactive form, U-73343. Significant attenuation of the mechanical activity can be also obtained with another PLC inhibitor 2-nitro-4-carboxyphenyl-N,N-diphenylcarbamate. Our present findings suggest a significant role for the activation of PLC and subsequent inositol 1,4,5-trisphosphate-induced Ca2+ release mechanism as an alternative triggering system for inducing spontaneous mechanical activity of UBSM. The present results support the idea that the action potential is not the sole pacemaker mechanism by which spontaneous contraction is induced in UBSM.
Toxicology
Regular Articles
  • Kazuyuki Hiramoto, Yoshinobu Yasuhara, Ken-ichi Sako, Kazuhiro Aoki, K ...
    原稿種別: Regular Article
    専門分野: Toxicology
    2003 年 26 巻 8 号 p. 1129-1134
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    The results of the present study have shown that unoxidized linoleic acid (LA) and low density lipoprotein (LDL) suppressed free radical-induced supercoiled plasmid DNA strand breaks. Unoxidized LA suppressed DNA strand breaks induced by free radicals generated from hydrogen peroxide/Fe(II) ion, 2′-azobis(2-amidinopropane)hydrochloride (AAPH), and 4-(hydroxymethyl)benzene diazonium salt. Thiobarbituric acid reactive substances (TBARS) of LA were increased on treatment with the radical generators. The intensities of the electron spin resonance (ESR) signals of the spin adducts of the radicals were reduced by unoxidized LA. Although LA hydroperoxide caused DNA strand breaks as has already been shown, its strand breaking activity was observed only at the higher concentrations. Unoxidized LDL inhibited ascorbic acid/Cu(II) ion-, ascorbic acid/Fe(II) ion-, peroxynitrite- and AAPH-induced DNA strand breaks. The TBARS of LDL were increased by treatment with the agents. LDL oxidized with Cu(II) ion did not cause DNA strand breaks. The results indicate that the potency of the free radicals to cause DNA strand breaks was attenuated by the fatty acid and the lipoprotein through lipid peroxidation.
Notes
  • Ki-Young Moon, Young Jong Lee, Yeong Shik Kim
    原稿種別: Note
    専門分野: Toxicology
    2003 年 26 巻 8 号 p. 1195-1197
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    Effect of alkylating carcinogens, i.e., N-nitroso-N-methylurea (NMU) and N-nitroso-N-ethylurea (NEU), as well as the simpler alkylating agents, methyl iodide and ethyl iodide, on the activation of NF-κB was evaluated in human epidermal squamous cell carcinoma (SCC-13) keratinocytes in order to investigate the possible correlation of cellular NF-κB activity with chemical carcinogenesis. The activities of NF-κB induced by chemical carcinogens were determined in human SCC-13 keratinocytes transfected with pNF-κB-SEAP-NPT plasmid, permitting expression of the secretory alkaline phosphatase (SEAP) reporter gene in response to the NF-κB activity and contains the neomycin phosphotransferase (NPT) gene conferring resistance to the geneticin. In this cell-based assay system, all alkylating carcinogens significantly upregulated the cellular NF-κB activations in a time- and dose-dependent manner until 72 h, at concentrations of 0.5—5 μM. These results suggest that carcinogenicity by alkylating chemicals may be associated with the modulation of cellular NF-κB activity in human skin cells.
Pharmacognosy
Regular Articles
  • Yohei Sasaki, Hirozo Goto, Chihiro Tohda, Fumiyuki Hatanaka, Naotoshi ...
    原稿種別: Regular Article
    専門分野: Pharmacognosy
    2003 年 26 巻 8 号 p. 1135-1143
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    The effectiveness of Curcuma drugs against “Oketsu” and the differences in their efficacy were evaluated by examining their vasomotional effects as one index. Since nitric oxide (NO) is the relaxation factor of vascular smooth muscle and also an inhibitor of platelet aggregation in blood vessels, substances showing NO-dependent relaxation are thought to be effective in improving Oketsu. In this study, five Curcuma drugs derived from Curcuma longa, C. kwangsiensis, C. phaeocaulis, C. wenyujin, and C. zedoaria were used. Methanol extracts exhibited intense effects on relaxation in rings precontracted by prostaglandin F (PGF) despite pretreatment with and without NG-nitro-l-arginine methyl ester (L-NAME) as an inhibitor of NO synthesis. The maximal activities were approximately 80% at 10−3 g/ml. From these methanol extracts, curcumin and eight sesquiterpenes were isolated. Since all these compounds showed NO-independent relaxation effects with almost the same intensities, the relaxation effects of Curcuma drugs can be estimated by the total amounts of curcumin and sesquiterpenes. Polysaccharides, the main constituents of methanol-insoluble compounds of water extracts, in contrast, showed contraction effects; only polysaccharides in C. zedoaria showed NO-dependent relaxation as well as contraction. All water extracts showed relaxation effects as sum of the methanol-soluble compounds-induced relaxation and polysaccharides-induced contraction. Therefore, all Curcuma drugs tested in the present study can be effective for vasodilation. Moreover, the drug derived from C. zedoaria has potential to cure Oketsu with its various acting points.
  • Yutaka Yamamoto, Takami Majima, Ikuo Saiki, Tadato Tani
    原稿種別: Regular Article
    専門分野: Pharmacognosy
    2003 年 26 巻 8 号 p. 1144-1149
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    To clarify the feasibility of medicinal use of the cultivated Glycyrrhiza resources, the equivalency between the G. uralensis roots cultivated in eastern Nei-Meng-Gu of China and medicinal licorice (Glycyrrhizae Radix, Gancao in Chinese and Kanzo in Japanese) was examined. The HPLC fingerprint including glycyrrhizin (GL) of the cultivated roots was similar to that of medicinal Gancao, but different from that of non-medicinal Xinjiang-Gancao (Shinkyo Kanzo in Japanese). Similarity between the cultivated roots and two medicinal Gancao was confirmed quantitatively by hierarchical cluster analysis on the basis of HPLC-7-peak-area data. Moreover, the 4-year-old adventitious roots conformed to the five standards described in the Japanese Pharmacopoeia XIV (JP XIV). The 4-year-old adventitious roots had similar pharmaceutical properties to those of medicinal Dongbei-Gancao (Tohoku Kanzo in Japanese) as determined by examining IgE-mediated triphasic skin reaction in mice and pharmacokinetic profile of glycyrrhetic acid, an anti-allergic metabolite of GL. The present pharmaceutical study suggests that the 4-year-old adventitious roots of G. uralensis cultivated in eastern Nei-Meng-Gu of China are comparable to medicinal Gancao conforming to the JP XIV, and may be a potential medicinal source to compensate for the insufficiency of wild Glycyrrhiza plants caused by collection restriction in China.
Notes
  • Tsuyoshi Ikeda, Hidetsugu Tsumagari, Takehiko Honbu, Toshihiro Nohara
    原稿種別: Note
    専門分野: Pharmacognosy
    2003 年 26 巻 8 号 p. 1198-1201
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    Since some Solanum-genera plants have traditionally been used as anti-cancer and anti-herpes agents from olden times, we examined the cytotoxic activity of typical steroidal glycosides with the framework of spirostane, furostane, spirosolane, and pregnane obtained from Solanum plants. Among these steroidal glycosides, the spirostanol glycosides having a β-lycotetraosyl moiety were the most effective against PC-12 and HCT-116 cell lines. The potency of activity was observed to be decreased in the order of spirostane, furostane, spirosolane, and pregnane type steroid glycosides. It was also suggested that the activity depend on the kind of oligosaccharide moiety and aglycone moiety.
  • Seung Hyun Kim, Young Pyo Jang, Sang Hyun Sung, Chang Jong Kim, Jin Wo ...
    原稿種別: Note
    専門分野: Pharmacognosy
    2003 年 26 巻 8 号 p. 1202-1205
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    Three dibenzylbutyrolactone lignans, (−)-arctigenin, (−)-traxillagenin, and (−)-4′-demethyltraxillagenin, isolated from the bark of Torreya nucifera SIEB. et ZUCC. (Taxaceae) showed significant hepatoprotective activity in primary cultures of rat hepatocytes injured by carbon tetrachloride (CCl4). These lignans reduced the release of glutamic pyruvic transaminase into the culture medium from the CCl4-injured primary cultures of rat hepatocytes. Further investigation revealed that the three lignans significantly preserved the level of glutathione (GSH) and activities of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase in the CCl4-injured rat hepatocytes. The lignans also ameliorated lipid peroxidation as demonstrated by a reduction in malondialdehyde-related products. Moreover, these lignans significantly attenuated the GSH reduction caused by diethylmaleate which depletes GSH through the formation of stable conjugates. However, these lignans showed no effect on the GSH synthesis inhibited by buthionine sulfoximine. From these results, it can be concluded that arctigenin, traxillagenin, and 4′-demethyltraxillagenin may protect hepatocytes from CCl4 injury by maintaining the GSH level.
Biopharmacy
Regular Articles
  • Cui Lili, Nobuhito Shibata, Yukako Yoshikawa, Kanji Takada
    原稿種別: Regular Article
    専門分野: Biopharmacy
    2003 年 26 巻 8 号 p. 1150-1154
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    In this study, the effect of glycerol-induced acute renal failure (ARF) on the pharmacokinetics of lidocaine after transdermal application was investigated in rats. Microdialysis method was applied in vitro and in vivo to the abdominal skin of rats. After topical application of 1% lidocaine, the cumulative amount of lidocaine permeated through the excised rat abdominal skin showed parallel effect between normal and ARF rats with no significant difference in the in vitro permeability coefficient of lidocaine between them, while area under the plasma concentration versus time curve of lidocaine in ARF rats increased significantly. The protein binding rate of lidocaine in ARF plasma and the blood vessel permeability to muscle tissues, assessed by β-D-glucopyranosyl fluorescein isothiocyanate-labeled (FITC) albumin, increased significantly. After intravenous infusion of 5 mg/h/kg lidocaine, both of the total body clearance and the volume of distribution of lidocaine in the ARF rats decreased significantly. These results suggested that renal dysfunction did not have any effect on the skin permeability of lidocaine, but might change the plasma protein binding of drug and blood vessel permeability which led to high plasma concentration of lidocaine.
  • Makiko Shimizu, Masahiro Tatsuno, Reiko Matsushita, Junko Totsuka, Yuk ...
    原稿種別: Regular Article
    専門分野: Biopharmacy
    2003 年 26 巻 8 号 p. 1155-1165
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    This study was conducted to explore the relationship between physicochemical property and toxic effectiveness using rat red blood cells (RBCs). The toxic effectiveness of acid nonsteroidal anti-inflammatory drugs (NSAIDs) was systemically examined by the depletion of intracorpuscular adenosine triphosphate (ATP), glutathione (GSH), and hemoglobin (Hb) at various doses, increased every 5 fmol/RBC. When the RBCs were incubated with NSAIDs, the drugs attained maximum levels within RBC, and the levels were then reduced. The ATP depletion seemed to be observed on the excretion of the drugs prior to the depletions of GSH and Hb. The physicochemical properties of NSAIDs were obtained from QMPRPlusTM, SMILES code, and CS ChemRaw Ultra. Correlation between their physicochemical properties and their doses for the depletions of ATP, GSH and Hb was performed in comparison with those of the membrane bound enzyme (MBE) inhibiting- and methemoglobin (MHb)-generating drugs. The ATP depletion by NSAIDs was correlated with the GSH depletion and intracorpuscular levels of the drugs, but not with the Hb depletion. The GSH depletion was correlated with the Hb depletion and participated in the lipophilicity of the drugs.
Notes
  • Refiye Yanardağ, Şehnaz Bolkent, Ayse Tabakoğlu-O ...
    原稿種別: Note
    専門分野: Biopharmacy
    2003 年 26 巻 8 号 p. 1206-1210
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    This study investigated both morphologically and biochemically whether parsley (Petroselinum crispum), which is used as a folk remedy to decrease blood glucose, has any antidiabetic effect on pancreatic B cells of rats. Parsley extract was given to male diabetic rats. In the diabetic group given parsley extract, it was detected that the number of secretory granules and cells in islets and other morphologic changes were not different from the control diabetic group, while the blood glucose levels in the diabetic group given the plant extract were reduced in comparison to the diabetic group. In addition, a decrease was observed in the weight of the control diabetic group and the diabetic group given the plant extract. It is suggested that the plant therapy can provide blood glucose homeostasis and cannot regenerate B cells of the endocrine pancreas.
  • Maruthamuthu Balasubramanian, Sathiah Thennarasu, Thankiah Sudhakaran, ...
    原稿種別: Note
    専門分野: Biopharmacy
    2003 年 26 巻 8 号 p. 1211-1214
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    Ethyl acetoacetate and methyl acetoacetate have been tested for the determination of hiprex (methenamine hippurate). Both the esters react with hexamine and hiprex forming fluorescent yellow Hantzsch esters. The products show absorption maximum at 370 nm and a fluorescence emission maximum at 465 nm in aqueous n-propanol. Important parameters such as pH, temperature, mole ratio and time of reaction have been optimized. The co-efficient of variation for inter and intra-day precisions were less than 1%. The chromogens show linearity in the concentration ranges of 2.0—35.0 μg ml−1 and 0.6—40.0 μg ml−1 for the spectrometric and fluorimetric determinations of hiprex, respectively.
Miscellaneous
Notes
  • Takeshi Ohya, Masaru Niitsu
    原稿種別: Note
    専門分野: Miscellaneous
    2003 年 26 巻 8 号 p. 1215-1218
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    Relative reactivities of histamine and indoleamines such as tryptamine, 5-hydroxytryptamine and 5-methoxytryptamine with acetaldehyde (AA) under physiological conditions were investigated. AA was found to have much higher reactivity towards histamine than towards indoleamines. For example, when a reaction mixture of AA (1 mM) and histamine or tryptamine (5 mM) in 0.1 M phosphate buffer (pH 7.4) was incubated at 37 °C for 24 h, AA decreased by 11% in the case of tryptamine, while in the case of histamine, it decreased 88%. In addition, the reaction product of AA with histamine was investigated. Mixtures of a fixed amount of histamine (5 mM) and various amounts of AA (1—20 mM) in phosphate buffer (pH 7.4) were incubated for 5 h at 37 °C. In all cases, only one product, 4-methylspinaceamine (4-MSPA), was observed. The yield of 4-MSPA was in approximate agreement with the losses of histamine and AA, indicating that the loss of histamine caused by the reaction of AA was quantatively converted to 4-MSPA. These results show that the reaction of AA with histamine easily takes place to produce 4-MSPA in an aqueous medium close to physiological conditions.
  • Tomoko Okubo, Toshinari Suzuki, Yoshiko Yokoyama, Kazutaka Kano, Itsu ...
    原稿種別: Note
    専門分野: Miscellaneous
    2003 年 26 巻 8 号 p. 1219-1224
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    Phthalates are man-made chemicals abundantly found in the environment. Estrogenic activities of phthalate di and monoesters were studied by in vitro assay of human breast cancer MCF-7 cell proliferation. Since phthalate monoesters are formed from diesters by degradation and are found in the environment, we selected some phthalate monoesters in addition to diesters. Among 19 compounds tested, dicyclohexyl phthalate (DCHP), di(2-ethylhexyl) phthalate (DEHP) and butyl benzyl phthalate (BBP) were found to have estrogenic activities, all of which were completely suppressed by the addition of pure anti-estrogen ICI 182,780. DCHP stimulated cell proliferation with maximal cell yield at 5×10−5 M. Its estrogenic potency was approximately 1700000 times less than that of 17β-estradiol. DEHP and BBP stimulated cell proliferation only slightly at >10−3 M. No other phthalate diesters or monoesters tested were estrogenic. Anti-estrogenic activities were also examined by estimating the suppression of cell proliferation in the presence of 10−11 M 17β-estradiol. Mono-n-pentyl phthalate (MPP), monocyclohexyl phthalate (MCHP), monobenzyl phthalate (MBZP), monoisopropyl phthalate (MIPrP) and BBP were suggested to have anti-estrogenic activities at higher than 10−4 M. Among commonly used phthalate esters and those with related structures, some were found to be estrogenic and others were anti-estrogenic in vitro.
  • Toshie Harada, Noriko Nagi Miura, Yoshiyuki Adachi, Mitsuhiro Nakajima ...
    原稿種別: Note
    専門分野: Miscellaneous
    2003 年 26 巻 8 号 p. 1225-1228
    発行日: 2003年
    公開日: 2003/08/01
    ジャーナル フリー
    A branched β-glucan from Sparassis crispa (SCG) is a major 6-branched 1,3-β-D-glucan showing antitumor activity. In the present study, we examined the anti-SCG antibody in naive mice by ELISA. Using SCG coated plate, sera of naive DBA/1 and DBA/2 mice contained significantly higher titers of antibody than other strains of mice. Anti-SCG Ab titers of each DBA/1 and DBA/2 mice were significantly varied. Using various polysaccharide-coated plate, sera of DBA/2 mice also reacted with a β-glucan from Candida spp. (CSBG) having 1,3-β and 1,6-β-glucosidic linkages. The SCG specific immunoglobulin (Ig) M but G was detected in sera. The reactivity of sera to coated SCG was neutralized by adding soluble SCG and CSBG as competitor. These results suggested that DBA/1 and DBA/2 strains carry specific and unique immunological characteristics to branched 1,3-/1,6-β-glucan.
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