Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
35 巻 , 10 号
選択された号の論文の37件中1~37を表示しています
Current Topics
  • Naoto Yamaguchi
    原稿種別: Current Topic
    2012 年 35 巻 10 号 p. 1621-
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
  • Minoru Fukuda
    原稿種別: Current Topic
    2012 年 35 巻 10 号 p. 1622-1625
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    Recent development in carbohydrate markers and functions are described. Identification of carbohydrate epitope for cancer-specific antibody is introduced. This novel approach involves the key glycosyltransferases that synthesize tumor-associated carbohydrate-antigens and elucidate the biosynthetic pathways. This is the true determination of carbohydrate ligands and glycan array is secondary to determine the epitope. Tumor suppressor activity of carbohydrate is described. Cell surface carbohydrate, which expressed in normal cells is diminished on cancer cells, function as a tumor suppressor. Glycans attached to α-dystroglycan function as laminin-binding glycans. In cancer cells, oncogene downregulates laminin-binding glycans and they do not bind to laminin in extracellular matrix, making cells to mobile. Thus, laminin-binding glycans function to suppress the cell mobility, thereby suppressing tumor formation in normal cells. This article summarizes the recent progress in the regulation of carbohydrate function in cancer cells. Since the review is short and not comprehensive, other several important topics may be missing.
  • Michiko N. Fukuda, Shingo Hatakeyama, Kazuhiro Sugihara
    原稿種別: Current Topic
    2012 年 35 巻 10 号 p. 1626-1632
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    Although numerous carbohydrates play significant roles in mammalian cells, development of carbohydrate-based reagents and therapeutics are hampered by the technical difficulty of chemically synthesizing complex carbohydrate structures. Use of carbohydrate mimetic peptides circumvents this difficulty, as short peptide can be easily synthesized and modified. We as well as others identified carbohydrate mimetic peptides by screening peptide displaying phage library using anti-carbohydrate antibodies and lectins. This review introduces our experiences with I-peptide that was used for identification of new carbohydrate binding receptor expressed in the lung endothelial cells, and those with IF7 peptide that can be used as a therapeutic against malignant tumors.
  • Shigeru Tsuboi
    原稿種別: Current Topic
    2012 年 35 巻 10 号 p. 1633-1636
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    During the process of hematogenous tumor metastasis, tumor cells that dissociated from the primary site enter the blood vessels and are exposed to innate immune systems in host blood circulation. In the innate immune systems, natural killer (NK) cells play a major role in rejecting tumors and suppressing metastasis. To establish metastasis, tumor cells therefore need to defend themselves against tumor rejection by NK cells. It has been recently discovered that some tumor cells develop defense systems against NK cell attack using certain types of cell-surface carbohydrates. The types of carbohydrates attached to cell-surface glycoproteins through serine or threonine residues contain a branch consisting of β-1,6-linkage of N-acetylglucosamine and N-acetylgalactosamine and are designated as core2 O-glycans. Tumor cells expressing core2 O-glycans evade NK cell-mediated tumor rejection, thereby surviving longer in host circulation and acquiring high-metastatic phenotypes. This review explains two types of tumor defense systems against NK cell immunity using core2 O-glycans.
  • Hiroto Kawashima
    原稿種別: Current Topic
    2012 年 35 巻 10 号 p. 1637-1641
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    MUC2 is the major gel-forming colonic mucin that forms the two mucus layers. Recent studies using gene-targeted mice have revealed the physiological functions of Muc2, the mouse counterpart of human MUC2, and its O-glycosylation in the colon. Muc2-deficient mice spontaneously developed colitis and colorectal cancer. As for the O-glycosylation of Muc2, conditional core 1-derived O-glycan-deficient mice in the intestines exhibited a breached inner mucus layer and spontaneously developed colitis. Similarly, core 3-derived O-glycan-deficient mice exhibited an increased susceptibility to colitis and colorectal cancer, suggesting that both core 1- and core 3-derived O-glycans on Muc2 are required for colonic protection. Mice deficient in core 2-branched O-glycans synthesized after the formation of core 1 O-glycans also exhibited increased experimental colitis. Furthermore, our recent studies using gene-targeted mice deficient in N-acetylglucosamine-6-O-sulfotransferase (GlcNAc6ST)-2 revealed that sulfation of the core 2-branched O-glycans of the colonic mucins by GlcNAc6ST-2 is required for the protection against experimental colitis. Taken together, these findings demonstrate the critical roles of the MUC2 mucin and its various O-glycans in the protection against colitis and colorectal cancer. Consistently, various alterations in the expression of mucins and their O-glycosylation have been noted in clinical samples of colorectal cancer. This review focuses on the roles of the MUC2 core protein and its O-glycosylation in health and disease.
  • Takao Taki
    原稿種別: Current Topic
    2012 年 35 巻 10 号 p. 1642-1647
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    We have established a new approach to glyco- and lipidomics using a thin layer chromatography (TLC)-Blot/matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) system. This new approach consists of a combination of a method for transferring lipids separated on a TLC-plate to a poly-vinylidene difluoride (PVDF) membrane and direct mass spectrometric analysis of the individual lipids on the membrane by ion trap-type MALDI-TOF MS. This technology was applied to the analysis of individual lipids from the human brain. Then, based on the results of this analysis, ganglioside molecular species in neural diseases were analyzed. The levels of gangliosides GD1b and GT1b were lower in the hippocampal gray matter of patients with Alzheimer’s disease than in the hippocampal gray matter of patients with Parkinson’s disease or the control patients. The molecular scanning of individual ganglioside molecular species showed a significant reduction of d20 : 1/C18 : 0 ceramide-containing gangliosides in patients with Alzheimer’s disease. These findings suggest that Alzheimer’s disease is a kind of ganglioside metabolic disease affecting the hippocampal area. A new approach to glycobiology by the TLC-Blot/MALDI-TOF MS system is proposed.
Regular Articles
  • Noriyasu Fukuoka, Osamu Imataki, Hiroaki Tanaka, Kumiko Tani, Hiroaki ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1648-1654
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    Tacrolimus is commonly used in stem-cell transplants (SCT) for prophylaxis of graft-versus-host disease and is continuously administered throughout transplantation. The dose of tacrolimus is frequently decreased to maintain a desired concentration during the recovery of hemocytes after engraftment. If parameters which affect tacrolimus clearance are identified, it is of clinical use to estimate concentrations and aid dosing. The objective of this study was to identify which hematologic parameters affect tacrolimus clearance. Seventeen consecutive Japanese patients with hematological malignancies who received allogeneic SCT between March 2004 and January 2007 were enrolled in this study. Their steady-state concentrations were routinely measured and standardized as the concentration/dose (C/D) ratio ((ng/mL)/(mg/kg/d)). Multivariate analysis was performed to identify which hemocyte parameters affected the C/D ratio. Of the 13 patients, gradual dose reduction was required to combat elevated tacrolimus concentrations. The mean post-engraftment C/D ratio was higher than the pre-engraftment C/D ratio in each patient. The mean C/D ratio for all patients after engraftment was 1.56-fold higher (p=0.00004, range: 1.04–3.03) than that before engraftment. The variation ratio was calculated by dividing the C/D ratio by that on the engraftment day. Multivariate analysis revealed that the reticulocyte (RET) level (×103 count/µL) was the sole parameter influencing this ratio, and both parameters were expressed as: Variation ratio=0.004×RET+1.0. RET recovery of patients could influence the C/D ratio and tacrolimus clearance was affected by recipient original red blood cells, but not that of transfused red blood cells.
  • Naoki Matsumoto, Tohru Aomori, Masafumi Kanamoto, Tadashi Usui, Tatsuy ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1655-1660
    発行日: 2012/10/01
    公開日: 2012/10/01
    [早期公開] 公開日: 2012/08/06
    ジャーナル フリー
    Although landiolol is useful in the emergency management of atrial fibrillation, atrial flutter, and tachycardia, as well as in perioperative arrhythmia control, the influence of hemodynamic changes on the pharmacokinetics of landiolol is unknown. We investigated the influence of hemodynamic variation and the following hepatocirculatory changes after systemic heparinization on the pharmacokinetics of landiolol in patients undergoing cardiovascular surgery under cardiopulmonary bypass. Cardiac output and cardiac index (CI) were continuously monitored in 19 patients using an arterial pressure-based cardiac output monitor. The middle and right hepatic venous blood flow indexes (mHVBFI and rHVBFI) were measured by transesophageal echocardiography, and hemodynamic data were collected at points (T1–T3) as follows: T1, before administration of heparin and after sternotomy; T2, just before systemic heparinization (300 U/kg); T3, 10 min after T2. The plasma concentration of landiolol was measured by HPLC at the same point. After administration of heparin, mean arterial blood pressure, CI, mHVBFI, and rHVBFI were significantly decreased (<0.05). Heart rate was not significantly changed. After systemic heparinization, the landiolol concentration was significantly decreased from 0.407±0.251 µg·mL−1 to 0.232±0.207 µg·mL−1 (<0.01). There was no significant difference between T1 and T2 (=0.88). In conclusion, the plasma concentration of landiolol was decreased by diminished CI due to systemic heparinization, but not affected by the change of hepatic blood flow.
  • Kouichi Tanonaka, Kanataka Motegi, Toru Arino, Tetsuro Marunouchi, Nor ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1661-1668
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    Previous studies showed that myocardial Na+ overload during ischemia directly induced mitochondrial damage. The pathway for Na+ flux into mitochondria remains unclear. We examined possible routes for Na+ flux into mitochondria in the ischemic heart. Isolated perfused rat hearts were subjected to 15- to 35-min ischemia followed by 60-min reperfusion and then Na+ content and respiratory function in mitochondria of the ischemic heart were determined. The mitochondrial Na+ content of the ischemic heart was ischemic duration-dependently increased, associated with a reduction in mitochondrial respiratory function. To mimic induction of mitochondrial Na+ overload in vitro, isolated mitochondria were incubated with 6.25 to 50 mM NaCl or sodium lactate, a metabolite of anaerobic glycolysis, in the presence and absence of a mitochondrial Na+/Ca2+ exchanger inhibitor CGP37157 and a monocarboxylate transporter (MCT) inhibitor α-cyano-4-hydroxy cinnamic acid (CHCA). Incubation of mitochondria with NaCl or sodium lactate increased the mitochondrial Na+ concentration. This increase in mitochondrial Na+ was partially attenuated by the presence of either inhibitor. Combined treatment of mitochondria with both inhibitors attenuated sodium lactate-induced increase in Na+ content to a greater degree than that treated with either agent. These results suggest that mitochondrial Na+/Ca2+ exchanger and MCT inhibitor-sensitive Na+ transporter are possible pathways for the mitochondrial Na+ overload in the ischemic myocardium.
  • Kyung-Eun Lee, Sukyeong Mun, Hee-Bong Pyun, Myung-Suk Kim, Jae-Kwan Hw ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1669-1675
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    Exposure to ultraviolet (UV) light causes premature skin aging that is associated with upregulated matrix metalloproteinases (MMPs) and decreased collagen synthesis. Macelignan, a natural lignan compound isolated from Myristica fragrans HOUTT. (nutmeg), has been reported to possess antioxidant and antiinflammatory activities. This study assessed the effects of macelignan on photoaging and investigated its mechanisms of action in UV-irradiated human skin fibroblasts (Hs68) by reverse transcription-polymerase chain reaction, Western blot analysis, 2′,7′-dichlorofluorescein diacetate assay, and enzyme-linked immunosorbent assay. Our results show that macelignan attenuated UV-induced MMP-1 expression by suppressing phosphorylation of mitogen-activated protein kinases (MAPKs) induced by reactive oxygen species. Macelignan also increased type I procollagen expression and secretion through transforming growth factor β (TGF-β)/Smad signaling. These findings indicate that macelignan regulates the expression of MMP-1 and type I procollagen in UV-irradiated human skin fibroblasts by modulating MAPK and TGF-β/Smad signaling, suggesting its potential as an efficacious antiphotoaging agent.
  • Linlin Peng, Jiayi Yang, Chen Ning, Jing Zhang, Xiangcheng Xiao, Dan H ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1676-1685
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    Studies have found overexpressed integrin-linked kinase (ILK) and disturbed matrix metalloproteinase-9/tissue inhibitor of metalloproteinase-1 (MMP-9/TIMP-1) ratio in diabetic nephropathy epithelial-mesenchymal transition (EMT). However, the underlying mechanisms of EMT and the inhibiting effect of rhein need further understanding. The aim of this study was to investigate the possible regulating effects of ILK towards MMP-9/TIMP-1 ratio in EMT and the inhibiting effect of rhein. The characteristic epithelial marker and mesenchymal marker of EMT were examined by cytoimmunostaining, real-time reverse transcription polymerase chain reaction (real-time RT-PCR) and Western blot. To observe the EMT inhibiting effects of rhein, specific ILK-small interfering RNA (ILK-siRNA) was used as a positive control. The results showed that in high glucose conditions, overexpression of ILK and an abnormal changing of MMP-9/TIMP-1 ratio occurred; ILK inhibition by siRNA could adjust MMP-9/TIMP-1 ratio to near normal. Meanwhile, rhein inhibited the overexpressing ILK and inhibits high glucose-induced EMT; the effect was similar to that of ILK-siRNA. The decreased expression of ILK regulated by rhein contributed to the adjustment of the MMP-9/TIMP-1 ratio. Our data indicates that rhein inhibits high glucose-induced-EMT partially through the inhibition of ILK expression and regulates the MMP-9/TIMP-1 ratio in HK-2 cells. This mechanism may be associated with rhein’s effect of ILK suppression.
  • Fei Li, Qihai Gong, Lina Wang, Jingshan Shi
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1686-1690
    発行日: 2012/10/01
    公開日: 2012/10/01
    [早期公開] 公開日: 2012/08/08
    ジャーナル フリー
    Osthole, a main active constituent from Cnidium monnieri (L.) CUSSON, has been considered therapeutic agent in the treatment of ischemic stroke. This study was designed to investigate the effect of osthole on permanent middle cerebral artery occlusion (MCAO) in rats. Osthole was administrated by gavage to the normal and the MCAO rats. Rats were assessed for neurological deficit after 24 h following MCAO, then their brains were evaluated to determine the infarct area, and the mRNA and protein levels of some inflammatory factors were detected. It was found that MCAO animals pre-treated with osthole for 7 d showed significant improvement in all neurological tests compared with vehicle-treated MCAO groups. In addition, there was a significant decrease in infarct volume 24 h after occlusion in animals pre-treated with osthole versus the vehicle-treated MCAO group. MCAO also dramatically caused some inflammatory factors increase. However, pretreatment with osthole restored the mRNA and protein levels of these factors, including tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS) of ischemic penumbra cortices, suggesting that osthole possessed the function of preventing brain against ischemic damage, while no significant difference was found in any of normal groups with or without osthole. The present study demonstrated that osthole may be a novel neuroprotective therapy in the treatment of focal ischemic stroke.
  • Shinji Koizumi, Shotaro Ohno, Fuminori Otsuka
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1691-1696
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    Gene expression processes are now recognized as important targets of the toxic effects exerted by industrial chemicals. The transient transfection assay is a powerful tool to evaluate such effects. Thus, we developed a versatile assay system by constructing a basic reporter plasmid in which the regulatory DNA sequence to be studied can easily be substituted. To verify the performance of this system, reporter plasmids carrying any of the three distinct regulatory sequences, estrogen responsive element (ERE), glucocorticoid responsive element (GRE) and xenobiotic responsive element (XRE) were constructed. After transfection of human cells, these plasmids successfully expressed the relevant reporter genes in response to specific inducers, β-estradiol, dexamethasone and 3-methylcholanthrene, respectively. Several industrial chemicals were assayed using these reporter plasmids, and the ability of p-dimethylaminoazobenzene to elevate GRE- and XRE-mediated transcription was detected. α-Naphthylamine and o-tolidine were also observed to increase the XRE-mediated response. The transfection assay system established here will be useful to evaluate the effects of a wide variety of industrial chemicals.
  • Shoko Sato, Osamu Nakagawasai, Takafumi Hayashi, Atsuko Oikawa, Fukie ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1697-1702
    発行日: 2012/10/01
    公開日: 2012/10/01
    [早期公開] 公開日: 2012/07/25
    ジャーナル フリー
    We investigated whether postweaning protein malnutrition (PM) affects serotonergic systems. Mice were fed a PM diet or normal protein (control) diet from weaning (21 d of age). Twenty days later, we tested for behavioral effects of the selective serotonin (5-HT)1A receptor agonist 8-hydroxy-N,N-dipropyl-2-aminoteralin (8-OH-DPAT) and the 5-HT releaser d-fenfluramine. The number of head weaving responses induced by 8-OH-DPAT or d-fenfluramine in the PM mice was significantly increased compared with the control diet group. The effects of 8-OH-DPAT and d-fenfluramine were blocked by pretreatment with the selective 5-HT1A receptor antagonist WAY-100635 (0.01 mg/kg). However, postpubertal (56 d of age) mice fed with the PM diet did not show an enhancement of the 8-OH-DPAT-induced head weaving response. These results indicate the occurence of a supersensitivity of postsynaptic 5-HT1A receptor in the postweaning PM group. Moreover, they highlight the postweaning stage as a vulnerable period to malnutrition-induced alterations in central serotonergic systems.
  • Sarka Studena, Jirina Martinkova, Dasa Slizova, Otakar Krs, Marian Sen ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1703-1710
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    A rat model of early sepsis induced by lipopolysaccharide (LPS) combined with interleukin-2 (IL-2) was developed. The primary aim was to assess the pharmacokinetics of gentamicin and sepsis-induced pathophysiological changes. Moreover, the effects on the glomerular filtration rate and tubular function were studied in septic and control rats. First, an intravenous (i.v.) bolus of LPSIL-2 (1 mg/kg-Pseudomonas aeruginosa, 15 µg/kg IL-2) or saline (controls, C) was administred. The Wistar rats were treated 30 min after LPSIL-2 with gentamicin as a 3 mg/kg i.v. bolus followed 10 min later by an i.v. 170-min infusion (GE, 0.09 mg/kg·min−1). The monitoring of vital functions, biochemistry and GE concentrations was performed. Creatinine clearance was 2–3 times lower and fractional urea excretion was 3–4 times less in septic rats as compared to controls(p<0.05), although urine flow was comparable. Capillary leakage caused a 55% elevation in the volume of distribution (Vc) in the LPSIL+GE group vs. C+GE (p<0.05). The renal CLge was less (2.2±0.59 vs. 3.8±0.53 mL/min·kg−1, p<0.05), while the total CLge was comparable (5.9±1.5 vs. 6.7±1.1 mL/min·kg−1; p=0.30). In the LPSIL+GE group relative to C+GE, the half-life (t1/2) was 79% higher (p<0.05) and GE concentrations detected at the end of the study in the plasma and kidney were elevated 2.5-fold (p=0.09) and 2.2-fold (p<0.05), respectively. The model reproduced several consequences of early sepsis like in patients such as capillary leak, a decreased glomerular filtration rate (GFR) and the changes in pharmacokinetics of GE (increased values of Vc and t1/2 and a drop in renal CLge proportional to that of CLcr). Nonrenal routes which, for the most part, compensate the reduced renal CLge in septic rats deserve further study.
  • Govinda Bhattarai, Young-Hee Lee, Nan-Hee Lee, In-Kyoung Lee, Bong-Sik ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1711-1719
    発行日: 2012/10/01
    公開日: 2012/10/01
    [早期公開] 公開日: 2012/08/02
    ジャーナル フリー
    In this study, a new lanostane triterpene glycoside (fomitoside-K) having biologically active molecules was isolated from a mushroom Fomitopsis nigra to test its anticancer activity on human oral squamous cell carcinomas (YD-10B). We focused on the effect of fomitoside-K on apoptosis, the mitochondria-mediated death pathway and the accumulation of reactive oxygen species (ROS) in YD-10B cells. Fomitoside-K could induce a dose and time-dependent apoptosis in YD-10B cells as characterized by cell morphology, cell cycle arrest, inhibition of survivin, activation of poly(ADP-ribose) polymerase (PARP), caspase-3, -9 and an increased expression ratio of Bax/Bcl-2. The mitochondria membrane potential loss and cytochrome c (Cyt C) release from mitochondria to cytosol were observed during the induction. Moreover, fomitoside-K caused dose-dependent elevation of intracellular ROS level and increase phosphorylation of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) in YD-10B cells. To further investigate the mechanisms, we examined the effects of ROS scavenger N-acetyl-L-cysteine (NAC) and selective inhibitors for mitogen activated protein kinase (MAPK) pathways on the cell death. The fomitoside-K induced cell death by ROS was significantly inhibited by NAC, ERK (PD98059) and JNK inhibitor (SP600125). In addition, fomitoside-K has a synergistic effect with adriamycin in suppressing the growth of YD-10B cells. These data suggest that fomitoside-K induces apoptosis in YD-10B cells through the ROS-dependent mitochondrial dysfunction pathway and provides a mechanistic framework for further exploring the use of fomitoside-K against the proliferation of human oral cancer.
  • Sureewan Duangjit, Yasuko Obata, Hiromu Sano, Shingo Kikuchi, Yoshinor ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1720-1728
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    Menthosomes, novel deformable carriers for the enhancement of transdermal delivery are introduced in this study. Meloxicam (MX)-loaded menthosomes were formulated, and their physicochemical characteristics and skin permeability were evaluated. A two-factor spherical and second-order composite experimental design was used to prepare the formulation of the menthosomes. Ten formulations of menthosomes composed of a phospholipid as the lipid bilayer carrier, cholesterol (Chol) as a stabilizer and cetylpyridinium chloride (CPC) and L-menthol as penetration enhancers were prepared. The amounts of Chol and CPC were selected as causal factors. Physicochemical characteristics (particle size, size distribution, zeta potential, elasticity and drug content) and an in vitro skin-permeation study of meloxicam-loaded menthosomes were evaluated. The concentrations of MX that permeated the skin at 2–12 h and the flux were selected as response variables. The optimal formulation was estimated using a nonlinear response-surface method incorporating thin-plate spline interpolation. The experimental values were very close to the values predicted by the computer programs in this study. A Bayesian network analysis was applied to gain a mechanistic understanding of the relationships between causal factors and response variables.
  • Sumiko Hyuga, Masumi Shiraishi, Atsushi Hori, Masashi Hyuga, Toshihiko ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1729-1739
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    Paclitaxel-resistant HuH-7 (HuH-7/PTX) cells were established by one-week exposure of HuH-7 cells to paclitaxel to analyze the effects of Kampo medicines on MDR-1-mediated multidrug resistance. HuH-7/PTX cells expressed high levels of MDR-1 and efficiently exported calcein-acetoxymethylester (calcein-AM), which is a substrate of MDR-1, suggesting that HuH-7/PTX cells resist paclitaxel by overexpressing MDR-1. We assessed the effects of 26 kinds of Kampo medicine on MDR-1 by calcein-AM efflux assay using HuH-7/PTX cells, and the results revealed that takushato and goreisan are potential inhibitors of drug efflux by MDR-1. Additionally, the sensitivity of HuH-7/PTX cells to paclitaxel was increased in combination with these Kampo medicines, indicating that takushato and goreisan overcame paclitaxel resistance in the cells by suppressing drug export by MDR-1. We further clarified that Alismatis Rhizoma contained in both takushato and goreisan reversed paclitaxel resistance by preventing drug efflux by MDR-1 without affecting the expression levels of MDR-1. Moreover, the principal components of Alismatis Rhizoma, Alisol A, Alisol B, and Alisol B acetate, were found to increase the sensitivity to paclitaxel in HuH-7/PTX by inhibiting drug export by MDR-1 without affecting the expression levels of MDR-1. These results suggested that the reversal effects of takushato and goreisan on paclitaxel resistance are derived from these principal components in Alismatis Rhizoma. Accordingly, Kampo medicines containing Alismatis Rhizoma such as takushato and goreisan may be useful as MDR-1 inhibitors.
  • Shinichiro Yamachika, Chika Sugihara, Hayato Tsuji, Yasunori Muramatsu ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1740-1744
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    In order to find new anti-Pseudomonas agents, we carried out whole-cell based P. aeruginosa growth assay, and identified 1,2,3,4-tetrahydro-1,3,5-triazine (Compound A). This compound showed anti-Pseudomonas activity against wild as well as pumpless strain equally at a same concentration. Also, this compound was structurally very similar to A22, which is known to inhibit the bacterial actin-like protein MreB. By the analysis of resistant strains, the primary target of this compound in P. aeruginosa was definitely confirmed to be MreB. In addition, these compounds showed a bacteriostatic effect, and induced the morphology changes in P. aeruginosa from rod shape to sphere shape, which leads to be clinically favorable in terms of susceptibility to phagocytosis and release of endotoxin. These results display that Compound A is a very attractive compound which shows anti-P. aeruginosa activity based on inhibition of MreB without being affected by efflux pumps, and could provide a new step toward development of new promising anti-Pseudomonas agents, MreB inhibitors.
  • Tetsuya Endo, Yasuhiko Minoshima, Yohsuke Hisamichi, Osamu Kimura, Mor ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1745-1751
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    We analyzed the levels of total mercury (T-Hg), methylmercury (M-Hg) and Cd in the muscle and liver of kidako moray eels (Gymnothorax kidako) of different body lengths caught off Kochi Prefecture in southern Japan. Furthermore, we analyzed the levels of organohalogen compounds such as polychlorinated biphenyls (PCBs), p,p′-1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (p,p′-DDE), trans-nonachlor and 2,3,3′,4,4′,5,5′-heptachloro-1′-methyl-1-2′-bipyrrole (Q1) and stable isotope ratios of carbon (δ13C) and nitrogen (δ15N) in the muscle of eels. The concentrations of T-Hg and M-Hg in the muscle (edible part) were 0.31±0.08 µg/wet g and 0.25±0.06 µg/wet g (n=26), respectively, and those in large eels exceeded the Japanese legislated levels of T-Hg (0.4 µg/wet g) and M-Hg (0.3 µg/wet g) in fish and shellfish, respectively. The T-Hg and M-Hg concentrations in the liver were markedly higher than those in the muscle, respectively. The ratios of M-Hg to T-Hg in the muscle and liver were about 80 and 60%, respectively, and those ratios tended to decrease with increased body length. The Cd concentrations in the liver tended to increase proportionally with body length, while that in the muscle was trace (around or below 0.03 µg/wet g). The concentrations of PCBs, p,p′-DDE, trans-nonachlor in the muscle tended to increase proportionally with body length, while that of Q1 did not. The δ13C and δ15N values in the kidako moray eel were markedly higher than those in offshore habit predators reported elsewhere, which may reflect the inshore habitat of this eels.
  • Ya Li, Su-Yun Li, Jian-Sheng Li, Li Deng, Yan-Ge Tian, Su-Li Jiang, Yi ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1752-1760
    発行日: 2012/10/01
    公開日: 2012/10/01
    [早期公開] 公開日: 2012/08/03
    ジャーナル フリー
    To develop a stable chronic obstructive pulmonary disease (COPD) model in rats. Sprague-Dawley rats were treated with cigarette-smoke inhalation (CSI) for 12 weeks, repetitive bacterial infection (RBI) for 8 weeks, or the combination of the two (CCR) for 12 weeks and followed up for the additional 20 weeks. Tidal volume (VT), peak expiratory flow (PEF) and 50% VT expiratory flow (EF50), histological changes in the lungs, and levels of the cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-8, and IL-10 in serum and bronchial alveolar lavage fluid (BALF) were examined at intervals during the 32 week study period. The right ventricular hypertrophy index (RVHI) was also determined at the same times. VT, PEF, and EF50 were decreased in rats with COPD compared to the control. The expression of TNF-α, IL-8 and IL-10 increased in both serum and BALF with a similar trend. Bronchiole and arteriole wall thickness and the degree of bronchiole stenosis and alveolar size increased in COPD rats. RVHI was reduced gradually following the treatment. All of these changes were more pronounced in the CCR-treatment group than in the other groups. Our results have shown that CSI or RBI alone can induce COPD in rats, but that the combination of CSI with RBI induces a stable COPD that has more similarity to complications seen in patients with COPD. This combination may therefore provide a more appropriate model for study of human COPD.
  • Claudio Manuel Lezama-Dávila, Angelica Patricia Isaac-Má ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1761-1764
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    Here we studied ability of two naphthoquinones to inhibit Leishmania growth (2,3-dichloro-5,8-dihydroxy-1,4-naphthoquinone (TR 001) and 2,3-dibromo-1,4-naphthoquinone (TR 002). TR 001 was more efficient than TR 002 in inducing killing of promastigotes and intracellular amastigotes. These values compare well to those obtained with the standard first-line antileishmanial agent sodium stibogluconate (SSG). TR 001 also induced significantly more nitric oxide (NO) production than TR 002 or SSG. Taken together, these data show that TR 001 and TR 002 could be promising new drugs for treatment of visceral leishmaniasis.
  • Tatsuo Takahashi, Yusuke Tonami, Mami Tachibana, Masaaki Nomura, Tsuto ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1765-1774
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    One of the mediators of osteoclast differentiation is receptor activator of nuclear factor κB ligand (RANKL), which is produced by osteoblasts. Binding of RANKL to its receptor, RANK, activates several signaling pathways, including those involving mitogen-activated protein kinases (MAPKs), nuclear factor κB (NF-κB), nuclear factor of activated T cells c1 (NFATc1) and Ca2+-calcineurin. In the present study, we found that tetrandrine, a bisbenzylisoquinoline alkaloid extracted from the root of Stephania tetrandra S. MOORE, significantly ameliorated the decrease of bone mass in sciatic-neurectomized osteoporosis model mice. It appears that tetrandrine acts directly on osteoclast precursors, since tetrandrine inhibited osteoclast differentiation not only in mouse bone marrow cells, but also in monocultures of murine macrophage RAW 264.7 cells without osteoblasts. Tetrandrine suppressed RANKL-induced amplification of NFATc1, a master regulator of osteoclast differentiation. However, it did not affect other signaling molecules such as MAPKs and NF-κB. These results suggest that tetrandrine is a candidate for the treatment of bone-destructive diseases, or at least a suitable lead compound for further development.
  • Naoki Ito, Atsushi Hori, Takeshi Yabe, Takayuki Nagai, Tetsuro Oikawa, ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1775-1783
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    Neuropeptide Y (NPY) and Orexin-A (OX-A), well-known neuropeptides associated with feeding and arousal, show antidepressant-like properties via hippocampal cell proliferation. Previous studies have revealed that kososan, a Kampo (Japanese herbal) medicine, has an antidepressant-like effect in behavioral animal models of depression; the mechanisms underlying this effect may involve the orexinergic system and subsequent upregulation of hippocampal cell proliferation. However, the roles of NPY in kososan’s antidepressant-like effect remain unclear. Here we investigated whether the regulation of the NPY system could play crucial roles in this effect in the stress-induced depression-like model mice. The antidepressant-like effect of kososan administered orally (1.0 g/kg) for 28 d was abolished by a continuous intracerebroventricular injection of BIBO3304, a neuropeptide Y1 receptor antagonist, for 7 d. Likewise, BIBO3304 injection blocked the kososan-induced increases in hippocampal cell proliferation and cluster formation of neural progenitor cells. On the other hand, BIBO3304 injection did not affect kososan-induced increases in hypothalamic OX-A-producing cells or in serum OX-A levels. These results suggest that the control of the NPY system in the brain plays an essential role in kososan’s antidepressant-like effect and facilitates hippocampal cell proliferation, both of which could be attributed, at least in part, to the control of the NPY system subsequent to the control of the OX-A system.
  • Shigeo Miyata, Nao Yamada, Tomie Kawada
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1784-1793
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    The aim of this study was to clarify the hypothalamic neuropeptides that are associated with hyperphagic feeding in Tsumura Suzuki Obese Diabetes (TSOD) mice, a model of type 2 diabetes with polygenic abnormalities. TSOD mice showed an increase in body weight and hyperleptinemia from 1 month of age and hyperphagic feeding, hyperglycemia, hyperlipidemia and hyperinsulinemia from 3 to 12 months of age compared with age-matched non-diabetic control Tsumura Suzuki Non Obesity (TSNO) mice. The mRNA level of nucleobindin-2 (NUCB2), the precursor of the anorexigenic neuropeptide nesfatin-1, was significantly decreased in the hypothalamus of TSOD mice compared with that in TSNO mice from 3 to 12 months of age. The protein level of NUCB2 was significantly decreased in the hypothalamus of TSOD mice compared with that in TSNO mice at 3 months of age. The mRNA levels of galanin, melanin-concentrating hormone, neuropeptide Y, and pro-opiomelanocortin were significantly changed in the hypothalamus in TSOD mice at several time points. Another model of type 2 diabetes, db/db mice, which is a mutant mouse that lacks a functional leptin receptor, showed hyperphagic feeding but no change in hypothalamic NUCB2 mRNA compared with non-diabetic control db/+ mice. The results suggest that the disrupted control of hypothalamic NUCB2-mediated signaling may contribute to hyperphagic feeding in TSOD mice. In addition, the mechanism for the development of hyperphagic feeding in TSOD mice is different than that in db/db mice.
  • Wenqiang Chang, Ying Li, Li Zhang, Aixia Cheng, Yongqing Liu, Hongxian ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1794-1801
    発行日: 2012/10/01
    公開日: 2012/10/01
    [早期公開] 公開日: 2012/08/03
    ジャーナル フリー
    Candida albicans is one of the most prevalent human opportunistic pathogens. C. albicans undergoes a yeast-to-hyphal transition that has been identified as a virulence factor as well as a critical element for mature biofilm formation. A previous study in our lab showed retigeric acid B (RAB), a lichen derived pentacyclic triterpenoid, displayed synergistic antifungal activity with azoles. We now showed that this combination also proved to be adequate in combating the formation of hyphae in vitro. In vivo tests with mice demonstrated RAB could markedly enhance the efficacy of fluconazole to promote the host’s longevity through inhibiting hyphae formation and adherence to host cells. It was also observed that RAB and azoles interacted synergistically to block the formation of biofilm. Our data suggested the attenuated yeast-to-hyphal switch contributed to the defect of mature biofilm formation. Moreover, quantitative real-time polymerase chain reaction (qPCR) analysis showed RAB could reduce the transcript level of MDR1, a multidrug efflux pump, and caused a slight transcriptional reduction for another drug pump related gene CDR1. Taken together, our work provides a potential application to combat candidiasis using the combination of RAB and azoles.
  • Lang Zhuo, Ming Liao, Li Zheng, Min He, Quanfang Huang, Ling Wei, Renb ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1802-1810
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    This study was to investigate the possibility of enhancing the anti-fibrotic effect by using a combination therapy with taurine, epigallocatechin gallate and genistein in a rat liver fibrosis model induced by alcohol, and to explore its underlying mechanism. Hepatic fibrosis was induced by intragastric administration with various amount of alcohol (5.0–9.5 g/kg) within 24 weeks in rats. The model group received alcohol only, and treatment groups received the corresponding drugs plus alcohol respectively, while the normal control group received an equal volume of saline. The antifibrotic effects of combination therapy were assessed directly by hepatic histology, and indirectly by measurement of serum biochemical markers, the fibrosis markers and related key cytokines/proteins. The results showed that combination therapy could significantly improve the liver function, as indicated by decreasing levels of alanine transaminase, aspartate transaminase, alkaline phosphatase, γ-glutamyltransferase, interleukin-6 and tumor necrosis factor-α. Moreover, combination therapy could effectively suppress the serum levels of fibrosis markers and hepatic hydroxyproline content, inhibit collagen deposition and reduce the pathological tissue damage. Research on mechanism showed that combination therapy was able to markedly reduce lipid peroxidation and recruit the anti-oxidative defense system, and inhibit the expression of B-cell lymphoma 2, α-smooth muscle actin, transforming growth factor β1 and small mothers against decapentaplegic homolog 3 proteins. Our results showed that combination therapy is effective in attenuating hepatic injury and fibrosis in the alcohol-induced rat model. The improved efficacy of the combination therapy with its good safety profile could represent a new protective approach for liver fibrosis.
  • Yuji Ishii, Naoko Iida, Yuu Miyauchi, Peter I. Mackenzie, Hideyuki Yam ...
    原稿種別: Regular Article
    2012 年 35 巻 10 号 p. 1811-1817
    発行日: 2012/10/01
    公開日: 2012/10/01
    [早期公開] 公開日: 2012/08/08
    ジャーナル フリー
    Morphine is an important drug used to alleviate moderate to severe pain. This opiate is mainly metabolized by glucuronidation to a non-analgesic metabolite, morphine-3-glucuronide (M-3-G) and an active metabolite morphine-6-glucuronide (M-6-G). To understand the modulation of morphine glucuronidation activity by environmental factors, the effect of endogenous and food-derived compounds on morphine uridine 5′-diphosphate (UDP)-glucuronosyltransferase (UGT) in rat and human microsomes was evaluated examining the 50% inhibitory concentration (IC50). The liver microsomes from Sprague-Dawley rats (RLM) and humans (HLM, 150 donors, pooled microsomes) were used as enzyme sources. Of 27 compounds tested, monoterpenoid alcohols, such as borneol and iso-borneol, exhibited a strong inhibitory effect on morphine glucuronidation in rat liver microsomes (RLM), whereas we failed to detect any inhibitory effect of endogenous substances including amino acids and sugars. The substances which have the ability to inhibit the activity in RLM are also inhibitory toward morphine glucuronidation in HLM and UGT2B7 baculosomes. However, the difference was that while the strongest inhibitory effect was observed for iso-menthol in HLM, borneol was the strongest inhibitor of the activity mediated by RLM. Although zidovudine is a typical substrate of UGT2B7, the inhibition of morphine glucuronidation by zidovudine was far weaker than that of monoterpenoid alcohols. These results demonstrate that dietary and supplementary monoterpenoid alcohols modify the pharmacokinetics and pharmacodynamics of morphine through inhibition of UGT2B7.
Notes
  • Renata Grespan, Marcia Paludo, Henrique de Paula Lemos, Carmem Patr&ia ...
    原稿種別: Note
    2012 年 35 巻 10 号 p. 1818-1820
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    This study was designed to test the efficacy of eugenol, a compound obtained from the essential oil of cloves (Syzygium aromaticum) in collagen-induced arthritis (CIA), a well characterized murine model of rheumatoid arthritis. Macroscopic clinical evidence of CIA manifests first as periarticular erythema and edema in the hind paws. Treatment with eugenol starting at the onset of arthritis (day 25) ameliorated these clinical signs of CIA. Furthermore, eugenol inhibited mononuclear cell infiltration into the knee joints of arthritic mice and also lowered the levels of cytokines (tumor necrosis factor (TNF)-α, interferon (IFN)-γ and tumor growth factor (TGF)-β) within the ankle joints. Eugenol treatment did not affect the in vitro cell viability as assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Therefore, eugenol ameliorates experimental arthritis and could be useful as a beneficial supplement in treating human arthritis.
  • Kazuhiko Yamaji, Yoichi Kawasaki, Kei Yoshitome, Hisashi Matsunaga, To ...
    原稿種別: Note
    2012 年 35 巻 10 号 p. 1821-1825
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    In this study, levels of the photoinitiator 1-hydroxycyclohexyl phenyl ketone (1-HCHPK) in aqueous injection solutions were analyzed by GC-MS. In our previous studies, photoinitiators such as 2-methyl-4′-(methylthio)-2-morpholinopropiophenone (MTMP) were detected in intravenous (i.v.) injection bag solution, and they were found to be cytotoxic to human monocytes. Therefore, we hypothesized that 1-HCHPK might display similarly cytotoxicity. The purpose of this study was to quantitate the amount of contaminants from plastic containers such as those used for peripheral parenteral nutrition and to determine the cytotoxicity of such extracts on human monocytes. The sample extraction procedure for GC-MS analysis involved a liquid-phase extraction. The solvent was evaporated under a stream of nitrogen at 50°C to yield a residue, which was dissolved in n-hexane and injected into a GC-MS. Normal human peripheral blood mononuclear cells (PBMC), isolated from the buffy coat by centrifugation, were suspended in RPMI 1640 medium supplemented with 10% (v/v) heat-inactivated fetal calf serum. In the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell viability assay, cells (1×104) were treated with 1-HCHPK for 24 h or 48 h at 37°C. From the GC-MS analysis, 6.13–8.32 µg/mL of 1-HCHPK was found in 20 mL vials of water for injection solution. In the MTT assay, 1-HCHPK decreased cell viability for both the 24 h and 48 h incubation periods. In conclusion, our findings suggest that 1-HCHPK could promote adverse events in patients. Future studies will clarify the possible health risks of photoinitiator accumulation in human cells.
  • Dong Ho Jung, Young Sook Kim, Jin Sook Kim
    原稿種別: Note
    2012 年 35 巻 10 号 p. 1826-1830
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    Activation of the receptor for advanced glycation endproducts (RAGE) triggers cellular responses implicated in the pathogenesis of diabetic complications; blockade of RAGE has been shown to inhibit the development of diabetic complications. To develop a screening system to identify novel disruptors of advanced glycation endproducts (AGE)-RAGE binding, we used an AGE-RAGE binding system in RAGE-overexpressing cells; test compounds were screened using this system. To construct human RAGE-overexpressing cells, mouse mesangial cells (MMCs) were stably transfected with the pcDNA-human RAGE (hRAGE) vector and selected under 1 mg/mL gentamicin (G418). RAGE expression in hRAGE-overexpressing MMCs was analyzed by Western blotting with specific RAGE antibody. To identify novel disruptors of AGE-RAGE binding, 50 single compounds and AGE-bovine serum albumin (BSA)-Alexa 488 (AGE-BSA labeled with Alexa 488) were treated to the hRAGE-overexpressing MMCs. Nonbinding AGE-BSA-Alexa 488 was washed and fluorescence measured by microtiter plate reader (excitation wavelength, 485 nm; emission wavelength, 528 nm). In hRAGE-overexpressing cells, only treatment with AGE-BSA-Alexa 488 significantly increased fluorescence intensity in a dose-dependent manner. Of 50 compounds tested, genistein disrupted AGE-RAGE binding in a dose-dependent manner. This AGE-RAGE binding system using AGE-BSA-Alexa 488 in hRAGE-overexpressing cells was suitable for screening of agents that disrupt AGE-hRAGE binding.
  • Satoko Uchida, Shin Endo, Kenji Akita, Tsunetaka Ohta, Shigeharu Fukud ...
    原稿種別: Note
    2012 年 35 巻 10 号 p. 1831-1835
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    The aim of this study is to evaluate the effects of NK-4, a kind of cyanine dye, on cholinergic memory deficits in mice. We examined whether NK-4 could reverse scopolamine-induced amnesia in mice since NK-4 displays a potent and selective inhibitory effect on acetylcholinesterase (AChE) in vitro. Intraperitoneal administration of NK-4 significantly reversed scopolamine-induced cognitive impairments in mice in the Y maze and the passive avoidance tests, and NK-4 also improved spatial learning ability in the Morris water maze test. Despite NK-4 displaying remarkable AChE inhibitory activity in vitro, we could not detect a significant reduction of AChE activity in brain homogenates of NK-4-treated mice. Although the mechanism through which NK-4 reverses cognitive impairments in scopolamine-treated mice remains unclear, these data suggest that NK-4 may have potential as a therapeutic agent for the treatment of dementia.
  • Nobuyuki Yamagishi, Yoko Yamamoto, Chika Noda, Takumi Hatayama
    原稿種別: Note
    2012 年 35 巻 10 号 p. 1836-1840
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    Polyglutamine (polyQ) diseases are inherited neurodegenerative diseases characterized by the aggregation of proteins containing expanded polyQ tract. Recently, we have shown that GRP78, the endoplasmic reticulum (ER) chaperone, was significantly decreased in the cells expressing enhanced green fluorescent protein with a pathological-length polyQ tract (EGFP-polyQ97), but not with a non-pathological-length polyQ tract (EGFP-polyQ24), and the expression levels of GRP78 were inversely related to the aggregation of EGFP-polyQ97. In this study, we performed the screening for compounds that modulate the GRP78 expression in herbal medicines, and found that naringenin, one of the major constitutions of Kanzo (Glycyrrhizae Radix), induced the expression of GRP78 in several mammalian cells. Furthermore, naringenin suppressed the protein aggregation caused by EGFP-polyQ97 in mammalian cells. These findings suggested that naringenin seemed to be a new inducer of GRP78 in mammalian cells, and may be a potential therapeutic agent for diseases caused by ER stress such as polyQ diseases.
  • Masashi Muroi, Keisuke Shima, Masayuki Igarashi, Yasuyoshi Nakagawa, K ...
    原稿種別: Note
    2012 年 35 巻 10 号 p. 1841-1845
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    The ability of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) to discriminate laboratory-derived antibiotic-resistant bacterial strains of known genetic origin was examined. A computer-based cluster analysis of spectral data successfully discriminated the majority of single- as well as multiple-antibiotic-resistant Escherichia coli strains examined. Cluster analysis of Staphylococcus aureus strains with different levels of novobiocin resistance showed that as the degree of resistance increased similarity to the wild-type strain decreased. These results demonstrate that MALDI-TOF MS is capable of discriminating antibiotic-resistant bacterial strains and may have potential for differentiating bacterial strains with varying degrees of antibiotic-resistance.
  • Kouichi Kurose, Tomoko Koizumi, Jun Nishikawa, Keiko Maekawa, Yoshiro ...
    原稿種別: Note
    2012 年 35 巻 10 号 p. 1846-1848
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    High-density oligonucleotide microarrays are widely used in genome-wide association studies. The purpose of this study was to assess the influence of various factors during the preparation of DNA on genotype calling for the Affymetrix high-density oligonucleotide microarray 250K GeneChip. DNA was extracted from peripheral whole blood by solution-based and silica-membrane-based methods. Blood was stored at 4°C or 25°C for 4 or 24 h, followed by DNA extraction. To examine the effects of freeze-thaw cycles, blood and DNA were also subjected to 5 and 10 or 20 of freeze-thaw cycles, respectively. The suitability of variously DNA preparations for the array was assessed by the call rate resulting from genotyping. All DNA samples showed mean call rates of more than 0.99, which passed the quality criteria for genotyping (greater than 0.95). The results indicated that the solution-based method and the silica-membrane-based DNA extraction method could provide DNA of sufficient quality for genotyping. In addition, DNA quality suitable for high-density oligonucleotide microarrays is not strongly dependent on the preparation conditions under standard procedures.
  • Sayuri Hama, Yasuhiro Ishihara, Masatomo Watanabe, Sonoko Danjo, Yu Na ...
    原稿種別: Note
    2012 年 35 巻 10 号 p. 1849-1853
    発行日: 2012/10/01
    公開日: 2012/10/01
    ジャーナル フリー
    Treatment of intracerebral hemorrhage is often pointless, although considerable effort has been devoted to developing treatments for ischemic stroke. The purpose of this study was to determine the influence of drugs in improving neurological outcomes with pharmaceutical therapy after intracerebral hemorrhage. The free-radical hypothesis for intracerebral hemorrhage is based on the cytotoxicity triggered by blood components and its degradation products, such as heme and iron as a potent pro-oxidant atom. Sulfaphenazole (SPZ) has a different mechanism such as reactive oxygen species scavenging, in addition to the inhibition of superoxide production by cytochrome P450. The present study investigated the properties of SPZ in collagenase-induced intracerebral hemorrhage rat brain damage. The results show that systemic SPZ treatment after intracerebral hemorrhage reduces striatal dysfunction, the elevation of lipid peroxidation, and brain edema in the rat. These results suggest that SPZ is a potentially effective therapeutic approach for intracerebral hemorrhage as the effect of SPZ was initiated for either 1 h or 3 d post-intracerebral hemorrhage.
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