Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
20 巻 , 11 号
選択された号の論文の21件中1~21を表示しています
  • Irina BLANGE, Victor DRVOTA, Paul M. YEN, Chister SYLVEN
    1997 年 20 巻 11 号 p. 1123-1126
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    Little is known about the cardiac expression of different thyroid hormone receptor (TR) isoforms. The aim of the study was to investigate such patterns of TR expression at the protein level in different species and in some human tissues. Western blot analysis with specific polyclonal rabbit antibodies to each TR isoform was performed with samples from myocardium of the left ventricle from man, dog, guinea pig, rat and mouse, as well as with samples from several human tissues such as heart, skeletal muscle, brain, liver and thyroid. The TRα1 isoform was present in all of the species examined. The TRα2 was recognized in human, dog and guinea pig heart, while no such band was recognized in rat and mouse hearts. TRβ1 was not detected in the human heart but in the other species. Simularly to TRα1, TRβ2 was detected in all of the species examined. In the human tissues studied, TRα1 was detected in heart and skeletal muscle, whereas TRα2 was found only in the heart. TRβ1 was not detected in any of the examined human tissues, while TRβ2 was found in all of them. These results revealed unique distributions of TR variants and they demonstrate common epitopes in TR in the different species. For the first time, the presence of a TRβ2 isoform has been shown in human tissues. TR isoforms may have a tissue and species specific role in the regulation of gene expression and may in part explain variable tissue effects of thyroid hormones.
  • Ken HASHIMOTO, Takashi TOBE, Jun-ichi SUMIYA, Ken-ichi SAGUCHI, Yoshih ...
    1997 年 20 巻 11 号 p. 1127-1130
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    The cDNA which encodes the mouse counterpart of human plasma hyaluronan-binding protein (PHBP) was isolated and characterized. The clone contained an insert of 2153bp, which contained the 1674-bp open reading frame coding for a polypeptide of 558 amino acid residues. The amino sequence of mouse PHBP predicted from the nucleotide sequence of cDNA shows reasonable homology to that of human PHBP. Like human PHBP, the amino acid sequence predicted from the nucleotide sequence of mouse PHBP cDNA exhibited significant homology to that of human hepatocyte growth factor activator (HGFA).
  • Shinji SOEDA, Yoshiaki SHIBATA, Hiroshi SHIMENO
    1997 年 20 巻 11 号 p. 1131-1135
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    Fucoidan is a sulfated poly(L-fucopyranose) present in brown marine algae. In this study, we examined the effect of native and chemically oversulfated fucoidans (NF and OSF) on the tube structure formation by human umbilical vein endothelial cells (HUVEC) on the basement membrane preparation, Matrigel. Unlike NF, OSF significantly decreased the tube formation : maximal inhibition (50% of control) was obtained with 25 μg/ml. The OSF effect was mediated, at least in part, through the inhibition of HUVEC migration, as determined by the ability to block chemotaxis in a Transwell chamber assay. Quantitative immunoreactive assays for tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) in the culture media indicated that OSF (25 μg/ml) increased the accumulation of PAI-1 antigen, but not of t-PA antigen, 2.7-fold compared with control. The release of both antigens by HUVEC was slightly affected by the addition of NF. Determination of the media levels of type IV collagenase activity and tissue inhibitor of metalloproteinase-1 (TIMP-1) antigen showed that OSF (25 μg/ml) decreased the collagenolytic activity by 50% compared to the control, without alteration of the TIMP antigen level. However, the collagenase inhibition by OSF was not observed in an assay system using purified enzyme. NF had no effect on collagenase activity or TIMP-1 antigen levels. These results indicate that the introduction of sulfate groups into NF enables it to effectively inhibit the formation of capillary-like structures by HUVEC on Matrigel by reducing the basement memberane destruction and cell migration. It is involved as at least one of the mechanisms by which the OSF-induced increase in HUVEC PAI-1 decreases plasmin formation and suppresses the following pro-collagenase activation.
  • Masafumi GOTO, Tatsuya TAKAHASHI, Fumiko YAMASHITA, Ako KOREEDA, Hirom ...
    1997 年 20 巻 11 号 p. 1136-1140
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    Low molecular weight thiol compounds have been found to be strong inhibitors of metallo-β-lactamase (IMP-1) produced by Serratia marcescens TN9106, which was expressed by Echerichia coli JM109 cells. Mercaptoacetic acid and 2-mercaptopropionic acid strongly and competitively inhibited IMP-1 with Ki of 0.23 and 0.19 μM, respectively. 2-Mercaptoethanol reversibly inhibited IMP-1 but did not show simple competitive inhibition.
  • Kunio OHYAMA, Tatsuo SHINOZUKA, Junichi YANAGIDA, Toshio YAMAKAWA
    1997 年 20 巻 11 号 p. 1141-1146
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    Three sialoglycoprotein fractions having reactivities against influenza A and B viruses (IV-A and IV-B) and influenza virus-hemagglutinin (IV-HA), were obtained by gel filtration using a Sephacryl S-300 column and an Asahipak GS-710HPLC column from water-soluble fraction prepared from human meconium (ME-WSF). Molecular weights of sialoglycoproteins in these fractions were estimated as 746 kDa (P-Fr. 1-3), 4470 kDa (P-Fr. 1-2) and a more than 10000 kDa (P-Fr. 1-1), respectively. The other in P-Fr. 2, with molecular weight estimated as 92 kDa, revealed positive but weak reactivity against IV-HA; however, the fraction did not show positive reactivity against either IVs-A or -B. All these fractions contained sialoglycoproteins with a high content of carbohydrates (31.0-59.3%, w/w). From their carbohydrate compositions, it can be suspected that sialoglycoproteins having O-glycosidically linked carbohydrate chains were predominating in the three fractions. In conclusion, it is suggested that at least three sialoglycoproteins with influenza virus-receptor activity showing different chemical properties to each other exist in human meconium.
  • Yuko TAKAOKA, Hiroichi NAGAI, Hiroshi MORI, Masahiko TANAHASHI
    1997 年 20 巻 11 号 p. 1147-1150
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    TRK-530 is a newly synthesized diphosphonate derivative. We investigated the effect of TRK-530 on type II collagen-induced arthritis (CIA) in mice in comparison to that of prednisolone and indomethacin. TRK-530 at a dose of 25 mg/kg showed a tendency to inhibit CIA. TRK-530 at a dose of 50 mg/kg inhibited the development of the CIA in terms of the progression of footpad swelling, bone damage and histopathological changes. TRK-530 at a dose of 50 mg/kg also significantly inhibited the delayed type hypersensitivity (DTH) response to type II collagen, but not the production of anti-type II collagen IgG antibody in arthritic mice.To investigate the inhibitory mechanism of TRK-530, the type of effect of TRK-530 on the production of IL-1β in vitro was studied. TRK-530 at a concentration of 10-4M inhibited LPS-induced IL-1β production from J774.1 cells.In conclusion, TRK-530 inhibited CIA in mice. The inhibition of the DTH reaction to type II collagen and the inhibition of IL-1β production may partly participate the anti-rheumatoid action of TRK-530.
  • Juvenia Bezerra FONTENELE, Glaucia Bezerra ARAUJO, Jose Wilson de ALEN ...
    1997 年 20 巻 11 号 p. 1151-1154
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    The present work shows an antinociceptive and dose-dependent effect of shark cartilage hydrosoluble fraction (HF) on writhing and formalin tests in mice. The effect was not altered by thalidomide, a known inhibitor of tumor necrosis factor-alfa (TNF-alfa) synthesis. Similarly, the antinosiceptive effect did not change in the presence of naloxone, indicating that the opioid system is not involved. However, the effect observed was blocked by L-arginine, a NO synthesis substrate, and it was potentiated by L-NAME, suggesting a role of the NO system in the shark cartilage antinociceptive effect. Effects similar to those seen with the HF were detected with peak II from gel filtration chromatography. The increase in vascular permeability induced by serotonin in rats was significantly abolished by the HF at the dose of 2 mg/kg, p.o., and again it was not potentiated by thalidomide. The observed blockade in the vascular permeability increase induced by histamine was detected only with a higher dose (10 mg/kg, p.o.).
  • Yoshio KASE, Mitsutoshi YUZURIHARA, Susumu IIZUKA, Atsusi ISHIGE, Yasu ...
    1997 年 20 巻 11 号 p. 1155-1159
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    The effects of "Hange-shashin-to (TJ-14)" on gastric function were examined in comparison with "Sho-saiko-to (TJ-9)". Oral treatment with TJ-14 (125-500 mg/kg) caused dose-dependent suppression of ethanol-induced gastric injury, while it did not suppress gastric lesions induced by water-immersion stress. TJ-9 (125-500 mg/kg, p.o.) suppressed both water-immersion stress-induced gastric lesions and ethanol-induced gastric injury in a dose-dependent manner. Intraduodenal administration of TJ-14 even at 500 mg/kg did not affect gastric juice secretion, while TJ-9 at 125 to 500 mg/kg dose-dependently suppressed gastric juice secretion. TJ-14 (125-500 mg/kg, p.o.) accelerated gastric emptying in normal rats and improved the delayed gastric emptying induced by BaCl2 in a dose-dependent manner, whereas such effect was not noted with TJ-9. Oral treatment with TJ-14 at 500 mg/kg significantly suppressed apomorphine-induced vomiting, but it did not affect copper sulfate-induced vomiting. These results suggest that TJ-14 exhibits an anti-ulcer action (probably based on its ability to protect the gastric mucosa), improvement of gastric emptying and an anti-emetic action. TJ-9 also showed anti-ulcer effects, probably based on its ability to suppress gastric secretion and to protect the gastric mucosa.Thus, the present study demonstrated the effectiveness of TJ-14 and TJ-9 against disease, and provided basic data which explain the differences in clinical application between these two kampo medicines.
  • Yu-Jen CHEN, Mang-Lin TU, Hsiang-Chi KUO, Kuo-Hua CHANG, Yuen-Liang LA ...
    1997 年 20 巻 11 号 p. 1160-1164
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    Tetrandrine, an alkaloid isolated from the plant Stephania tetrandra, at low concentration (2 μg/ml) was shown to protect normal human mononuclear cells in vitro against damage due to a single high-dose of ionizing irradiation (10 Gy). The cell survival rate increased from 58.3±2.2% in the irradiated group to 78.0±2.6% in the tetrandrine-pretreated group, and similarly, the percentage of necrotic cells declined from 20.7±2.5% to 10.7±1.9%, respectively. This protective effect of tetrandrine for cell surviving fraction increased in a dose-dependent manner. Tetrandrine was also found to inhibit inflammatory responses induced by irradiation including the release of superoxide (NBT [nitroblue tetrazolium] reduction decreased from 21.3±2.3% to 10.2±2.5%) and phagocytic activity (decreased from 80.7±3.8% to 50.7±2.3%, the same range level as that of the control group). However, the alkaloid demonstrated no effect on the production of nitric oxide. In terms of cell morphology, only two types were observed-normal or necrotic cells, and there were no characteristics of programmed cell death. These results indicate that tetrandrine Possesses radioprotective activity against 10 Gy of ionizing irradiation and could suppress irradiation-induced inflammatory processes.
  • Hideaki MATSUDA, Yue DAI, Yasuko IDO, Masayuki YOSHIKAWA, Michinori KU ...
    1997 年 20 巻 11 号 p. 1165-1170
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    The 70% ethanol extract (KS-ext) from Kochiae Fructus (dried fruits of Kochia scoparia L.) has been screened for activity in experimental models of type I-IV allergy. In type I allergic models, KS-ext at doses of 200 and 500 mg/kg, p.o. exhibited an inhibitory effect on 48-h homologous passive cutaneous anaphylaxis (PCA) in rats, which is related to IgE, and 1.5-h heterologous PCA in mice, which is related to IgG. In a type III allergic model, KS-ext showed an inhibitory effect on direct passive arthus reaction (DPAR) in rats, while it had no inhibitory effect on reversed cutaneous anaphylaxis (RCA) in a type II allergic model. Furthermore, in a type IV allergic model, KS-ext had an inhibitory effect on the effector phase in picryl chloride-induced contact dermatitis (PC-CD). Also, its anti-pruritogenic component, momordin Ic (oleanane saponin) exhibited inhibitory effects on 48-h homologous PCA and PC-CD.These results indicate that Kochiae Fructus not only inhibits humoral immunity but also influences cellular immunity, and should be recognized as a material for anti-allergic reactions. Also, the mode of its anti-pruritogenic activity may be mediated by anti-allergic action, and its active component may be partially attributed to momordin Ic.
  • Keiko KAWASHIMA, Ken-ichi SAITO, Akiyoshi YAMADA, Shin-ichi OBARA, Tak ...
    1997 年 20 巻 11 号 p. 1171-1174
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    We made a trial of searching for the bioactive substances from Sandohnin( ?? ?? ?? ). The blood and bile of rats after the oral administration of extracts of Sansohnin were analyzed by three-dimensional high-performance liquid chromatography (3D-HPLC). In blood, spinosin and feruloyl spinosin were found after the oral administration of a butanol extract of Sansohnin. In bile, spinosin and feruloyl spinosin were also identified after the oral administration of a water extract of Sansohnin. Spinosin and feruloyl spinosin induced the prolongation of hexobarbital sleeping time in mice at doses of 50 and 100 mg/kg, respectively. We concluded that spinosin and feruloyl spinosin were the bioactive constituents of Sansohnin. It was assumed that spinosin and feruloyl spinosin circulated through the intestine and liver, therefore, these results will provide support for the sedative and hypnotic use of this crude drug in Oriental medicine.
  • Jun-ichiro SAKUSHIMA, Mitsuhiko NOSE, Yukio OGIHARA
    1997 年 20 巻 11 号 p. 1175-1177
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    We investigated here the effect of Hachimi-jio-gan (HJ), a Japanese and Chinese herbal medicine, on immunoglobulin A (IgA) producing cells in Peyer's patch. The oral administration of HJ (0.1, 0.2, 1.0 g/kg for 2 d) enhanced the IgA producing cells almost 2-fold compared with the control group on days 4 and 5 after the administration. Furthermore, HJ augmented antigen-specific IgA (anti-SRBC IgA) production. These results suggested that HJ acted as a polyclonal B cell activator. To characterize its active ingredients, HJ was fractionated by ethanol precipitation. The crude polysaccharide fraction (EP fraction) showed the ability to augment IgA production, but low molecular weight fraction (ES fraction) did not. These results suggest that the crude polysaccharide fraction might be responsible for the augmentation of IgA production by HJ.
  • Keiji KAJIMURA, Yasuhiro TAKAGI, Keiichi MIYANO, Yoshiyuki SAWABE, May ...
    1997 年 20 巻 11 号 p. 1178-1182
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    The effect of Astragali Radix (AR) on IgM antibody production in mice of various ages (10 weeks, 36 weeks and 60 weeks) was examined. The antibody production levels in the 36- and 60-week-old mice were significantly decreased to about 70 and 60% of that in the 10-week-old mice. The enhancement effect of a crude polysaccharide AR fraction on the antibody production was nil in the 10-week-old mice, but significant enhancement effects were observed in the 36- and 60-week-old, mice, compared to the age-matched control.Two polysaccharides active in the enhancement of the IgM antibody production in the aged mice were isolated from the high molecular weight fraction of AR by cetavlon precipitation, ion-exchange and gel permeation chromatography. The molecular masses of these polysaccharides were calculated by HPLC in salt solution. Only one major peak was observed for each, and their molecular masses were estimated to be 1.2×104 and 2.2×104.The major components of these polysaccharides were neutral carbohydrates (89.3 and 95.5%), followed by uronic acid and protein / glucose was the predominant sugar component.
  • Jiro FUJISAKI, Yuji TOKUNAGA, Toshiya TAKAHASHI, Sumihisa KIMURA, Fumi ...
    1997 年 20 巻 11 号 p. 1183-1187
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    An osteotropic drug delivery system (ODDS) based on a bisphosphonic prodrug has been developed for 17β-estradiol (E2) to improve patient compliance in estrogen replacement therapy of postmenopausal osteoporosis. The biological disposition and the targeting efficiency of a bisphosphonic prodrug of E2, disodium [17β-(3'-hydroxy-1', 3', 5'-estratrienyloxy)carbonylpropyl carboxamidomethylene]bisphosphonate (E2-BP), was investigated in ovariectomized rats. After intravenous injection, E2-BP was rapidly taken up into the bone and subsequently cleared from the bone at a half-life of 13.5d. The bone concentration of regenerated E2 was maintained throughout 28d. In contrast, E2 injected intravenously showed extremely low bone distribution and rapid clearance from the bone, and E2 administered orally showed even lower bone distribution. Therapeutic availability (TA) and drug targeting index (DTI), which were calculated on the basis of the AUCs for E2 in the bone and plasma after injection of E2-BP and E2, were 64.6 and 451, respectively. These results suggest that ODDS has a potential to improve not only the apparent potency but also the therapeutic index of E2. As compared with the conventional estrogenic products, E2-BP should improve patient compliance with lower adverse effects and less frequent medication in long-term estrogen replacement therapy.
  • Hiroaki YUASA, Naomi SOGA, Yukie KIMURA, Jun WATANABE
    1997 年 20 巻 11 号 p. 1188-1192
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    The effect of aging on the intestinal transport of hydrophilic drugs (and probe compounds) was investigated in the rat small intestine.Passive transport was suggested to be unchanged with aging from 8 (young) to 54 (old) and further to 101 (very old) weeks old, as shown for D-xylose and urea in single-pass intestinal perfusion (under urethane anesthesia), where steady-state transport across the intestinal membrane into the blood stream was evaluated. The passive transports of cephradine, 5-fluorouracil (5-FU) and L-glucose were also unchanged, though they were compared only between the young and the old. Consistently, the passive uptake in the intestinal everted sacs, where the entry process into the membrane was evaluated for 5-FU, D-xylose, urea and polyethylene glycol (PEG) 900, was unchanged with aging from the young to the very old.The carrier-mediated tranport of cephradine was also unchanged with aging from the young to the old in perfusion under anesthesia, though that of D-glucose was declined by about 50% with aging from the young to the old and thereafter remained constant in the very old.In perfusion in unanesthetized rats, age independency in passive transport (examined for cephradine, L-glucose and D-xylose) and an age-dependent decline in D-glucose transport were also observed, suggesting that the findings under anesthesia are not qualitatively distorted.These results suggest that, although carrier-mediated transport may moderately decline with aging, the barrier function of the intestinal membrane to passive permeation of hydrophilic drugs (with molecular weight below 1000) may be unaffected by aging, supporting the suggestion from our previous in vivo studies that age-dependent increases in the orally absorbed fraction may be predicted for incompletely absorbed drugs because of delayed intestinal transit rather than increased intestinal transport (membrane permeability).
  • Toyofumi SUZUKI, Taiyo SUGANUMA, Junichi NISHINO, Manabu HANANO
    1997 年 20 巻 11 号 p. 1193-1198
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    In order to assess quantitatively the pharmacodynamic process of pentazocine (PTZ), time courses of its plasma concentration and of the occupation of specific opioid receptors in the brain were investigated after intravenous (i.v.) administration of PTZ to rats. The plasma concentration of PTZ was determined by HPLC and the pharmacokinetic parameters were analyzed using nonlinear least-squares analysis. Measurement of ex vivo receptor occupation was made by comparing the specific [3H]naloxone (opioid receptor antagonist) binding in vitro to the crude P2-synaptosomal fractions between vehicle-treated rats (control) and PTZ-treated rats. Following the i.v. administration of PTZ, the occupancy of specific opioid receptors decreased rapidly until 10 min, depending on the two pharmacological doses (2.5 and 10 mg/kg). The results strongly suggest the fast binding kinetics of PTZ in terms of its association with and dissociation from specific opioid receptor sites in the brain in addition to its fast rate of disappearance from the brain compartment. Furthermore, we demonstrated that the time profile of receptor occupancy correlated well (r=0.8650) with that of the unbound concentration in plasma until 120 min after the i.v. administration of PTZ to rats.
  • Shigeki MURAMATSU, Yuko KOMOKATA, Yorihisa TANAKA, Hidekuni TAKAHAGI
    1997 年 20 巻 11 号 p. 1199-1203
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    When incubated with isolated rat hepatocytes, pravastatin sodium (PS) yielded a small amount of a metabolite in addition to two major metabolites that have already been reported. The previously uncharacterized metabolite was found to be formed by at first being enzymatically dehydrogenated to 6'-keto intermediate (R-104), followed by decomposition to give the aromatized metabolite (R-195), through spontaneous deesterification with accompanying aromatization. The PS-6'β-hydroxydehydrogenase activity was localized in cytosolic fraction and required NADP, preferentially over NAD, as a cofactor. The formation of R-195 by rat liver cytosol was strongly inhibited by indomethacin, 3α-hydroxysteroids (but not 3β-isomers) and 3-ketosteroids. The results and high substrate specificity of purified PS-6'β-hydroxydehydrogenase toward 3α-hydroxysteroids suggested that the enzyme is identical to 3α-hydroxysteroid dehydrogenase.
  • Naganori NUMAO, Yukiko HIROTA, Akiyo IWAHORI, Masanori SASATSU, Isamu ...
    1997 年 20 巻 11 号 p. 1204-1207
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    Magainin 2, isolated from the skin of the Xenopus laevis, is an antimicrobial peptide which reacts directly with the biological membrane to lyse various bacteria from negative and positive microorganisms. In a previous report, we showed that (±)1-(4-aminobutyl)-6-benzylindane (PM2), which mimicked the conformation of the side-chains of a complementary unit on the amino acid sequence of magainin 2 analogs, expressed the in vitro antibacterial activity not only against Helicobacter. pylori (ATCC43526, ATCC43579), but also against Escherichia coli (ATCC25922) and Staphylococcus aureus (ATCC25923). In addition, PM2 caused human blood red cells (RBCs) to lyse at the minimum inhibitory concentration (MIC) value.Based on the antibacterial activities of 9-phenylnonanoic acid (pC9c), we further synthesized (±)-6-benzyl-1-(3-carboxypropyl) indane (PM2c), which replaced a positive charge of PM2 with a negative one, and tested the biological activities. PM2c had the ability to inhibit the growth of H. pylori strains, but its activity to inhibit the growth of E. coli and S. aureus was not detected and weak, respectively. Moreover, PM2c showed non-hemolytic activity against RBCs at the MIC value. These results indicate the possibility that PM2c may be more useful than PM2 either alone or in combination with well-known therapeutic agents for the treatment of H. pylori infection.
  • Toshiyasu MATSUOKA
    1997 年 20 巻 11 号 p. 1208-1211
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    A sensitive and rapid colorimetry for determination of methemoglobin (MetHb) in hemolysate of the test blood was devised by measurement of absorbance at 563 nm, the isosbestic point of spectra of cyanomethemoglobin and oxyhemoglobin, at pH 6.8. MetHb was determined as the difference in absorption caused by cyanide in the absence of potassium ferricyanide divided by the difference in absorption caused by cyanide in the presence of the ferricyanide.Carboxyhemoglobin (COHb) in the blood was also estimated from the absorbance values of the hemolysates with or without potassium ferricyanide after the addition of cyanide. The method requires only 3 μl of test blood and 10 min for determinations of MetHb and COHb in blood. Results obtained by the method were in satisfactory agreement with theoretical results for mixuture of MetHb, COHb, and oxyhemoglobin. The method was compared with two other methods using 55 forensic blood samples containing various amounts of MetHb and COHb, and proved to be suitable for practical samples.
  • Michiko MIYAHARA, Akiko NAKAMURA, Katsutoshi MISE
    1997 年 20 巻 11 号 p. 1212-1214
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    Two restriction endonucleases (ENases) were found by screening 38 standard phage strains of Salmonella (S.) Enteritidis. An isoschizomer of SacII ENase that recognizes the sequence 5'-CCGC/GG-3' was identified in S. Enteritidis PT14b, and an isoschizomer of XmaIII ENase (5'-C/GGCCG-3') was found in S. Enteritidis PT16.It is of special interest that the recognition specificities of all known ENases in Salmonella, including those of the S. Enteritidis ENases, are very similar to each other.
  • Shizuo NAKAJIN, Fumihiro TAMURA, Noriko TAKASE, Satoshi TOYOSHIMA
    1997 年 20 巻 11 号 p. 1215-1218
    発行日: 1997/11/15
    公開日: 2008/04/10
    ジャーナル フリー
    The carbonyl reductase activity exhibited by pig testicular 20β-hydroxysteroid dehydrogenase (20β-HSD) was examined using a recombinant enzyme. Kinetic parameters were obtained for 48 carbonyl group-containing substrates, including aromatic aldehydes, aromatic ketones, cycloketones, quinones, aliphatic aldehydes and aliphatic ketones. 20β-HSD showed a high affinity towards quinones, such as 9, 10-phenanthrenequinone, α-naphthoquinone and menadione (Km values of 4, 2 and 5 μM, respectively), and the substrate utilization efficiency (Vmax/Km) of the enzyme against these quinones was very high. Cyclohexanone and 2-methylcyclohexanone were also reduced with a high Vmax/Km value, but not cyclopentanone or 2-methylcyclopentanone. Various aromatic aldehydes and ketones including benzaldehyde- and acetophenone-derivatives were reduced by 20β-HSD. Especially, 4-nitrobenzaldehyde and 4-nitroacetophenone were reduced with high Vmax/Km values in related compounds. The enzyme also reduced the pyridine-derivatives, 2-, 3-, and 4-benzoylpyridine, with the Vmax/Km value for 2-benzoylpyridine being the highest. 20β-HSD reduced aliphatic aldehydes and aliphatic ketones, but was more effective on the former. The correlation between the structure of carbonyl compounds and their substrate Vmax/Km is discussed.
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