Biological and Pharmaceutical Bulletin
Online ISSN : 1347-5215
Print ISSN : 0918-6158
ISSN-L : 0918-6158
39 巻 , 11 号
選択された号の論文の23件中1~23を表示しています
Reviews
  • Atsuhiko Ichimura, Hiroshi Takeshima
    2016 年 39 巻 11 号 p. 1743-1747
    発行日: 2016/11/01
    公開日: 2016/11/01
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    Trimeric intracellular cation (TRIC) channel subtypes, namely TRIC-A and TRIC-B, are expressed in the endoplasmic/sarcoplasmic reticulum and nuclear envelope, and likely function as monovalent cation channels in various cell types. Our studies using knockout mice so far suggest that TRIC subtypes support Ca2+ release from intracellular stores by mediating counter-cationic fluxes. Several genetic mutations within the TRIC-B locus were recently identified in autosomal recessive osteogenesis imperfecta (OI) patients. However, the molecular mechanism by which the mutations cause human disease is not fully addressed. We found that Tric-b-knockout mice exhibit poor bone ossification and thus serve as an OI-model animal. Studies on Tric-b-knockout bones and cultured cell lines derived from the patients currently reveal the main part of the pathophysiological mechanism involved in the TRIC-B-mutated OI form. This mini-review focuses on the essential role of TRIC-B channels in bone ossification.

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  • Masahiro Hiratsuka
    2016 年 39 巻 11 号 p. 1748-1759
    発行日: 2016/11/01
    公開日: 2016/11/01
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    Genetic variations in CYP 2C (CYP2C) subfamily, CYP2C8, CYP2C9, and CYP2C19 contribute to interindividual variability in the metabolism of clinically used drugs. Changes in the drug metabolizing activity of CYP2C members may cause unexpected and serious adverse drug reactions and inadequate therapeutic effects. Therefore, CYP2C gene polymorphism is used as a genome biomarker for predicting responsiveness to administered drugs. The most direct method for understanding the extent of the effects of CYP2C gene polymorphism on drug pharmacokinetics is by evaluating the blood and urine concentrations of the drug in subjects. However, in vivo tests are highly invasive, and considering the risk of adverse drug reactions, the burden on the patient may be significant. In addition, examining the functions of rare variant enzymes with an allele frequency of ≤1% requires at least several hundred subjects. Furthermore, it is extremely difficult to evaluate the functions of all variant enzymes in an in vivo test. On the other hand, in vitro enzyme activity can be evaluated using a heterologous expression system to avoid the aforementioned problems. In vitro tests are extremely important as they complement in vivo information. This review focuses on recent findings of in vitro studies on 3 highly polymorphic CYP2C members: CYP2C8, CYP2C9, and CYP2C19.

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Regular Articles
  • Xiaomei Zhang, Na Li, Yuanyuan Yao, Xuming Liang, Xianyou Qu, Xiang Li ...
    2016 年 39 巻 11 号 p. 1760-1766
    発行日: 2016/11/01
    公開日: 2016/11/01
    [早期公開] 公開日: 2016/09/06
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    Species of genus Tripterygium (Celastraceae) have attracted much attention owing to their excellent effect on treating autoimmune and inflammatory diseases. However, due to high market demand causing overexploitation, natural populations of genus Tripterygium have rapidly declined. Tripterygium medicinal materials are mainly collected from the wild, making the quality of medicinal materials unstable. Additionally, identification of herbal materials from Tripterygium species and their adulterants is difficult based on morphological characters. Therefore, an accurate, convenient, and stability method is urgently needed. In this wok, we developed a DNA barcoding technique to distinguish T. wilfordii HOOK. f., T. hypoglaucum (LÉVL.) HUTCH, and T. regelii SPRAGUE et TAKEDA and their adulterants based on four uniform and standard DNA regions (internal transcribed spacer 2 (ITS2), matK, rbcL, and psbA–trnH). DNA was extracted from 26 locations of fresh leaves. Phylogenetic tree was constructed with Neighbor–Joining (NJ) method, while barcoding gap was analyzed to assess identification efficiency. Compared with the other DNA barcodes applied individually or in combination, ITS2+psbA–trnH was demonstrated as the optimal barcode. T. hypoglaucum and T. wilfordii can be considered as conspecific, while T. regelii was recognized as a separate species. Furthermore, identification of commercial Tripterygium samples was conducted using BLAST against GenBank and Species Identification System for Traditional Chinese Medicine. Our results indicated that DNA barcoding is a convenient, effective, and stability method to identify and distinguish Tripterygium and its adulterants, and could be applied as the quality control for Tripterygium medicinal preparations and monitoring of the medicinal herb trade in markets.

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  • Keita Shibata, Arisa Matsumoto, Ayumi Nakagawa, Keiko Akagawa, Akihiro ...
    2016 年 39 巻 11 号 p. 1767-1773
    発行日: 2016/11/01
    公開日: 2016/11/01
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    Community pharmacies in Japan have long been advocated as effective sources of nonprescription medicines and health-related advice. Consumers sometimes self-treat symptoms of minor illnesses without consulting a pharmacist because the benefits of such consultations are not adequately recognized. The aim of this study was to investigate the use and impact of pharmacist consultations before purchase of nonprescription laxatives. An online survey was conducted July 14–22, 2012 with 500 respondents (250 men, 250 women), ranging 20–60 years old. All participants had purchased nonprescription laxatives for constipation within the past year. Stratified analysis was used to compare responses in groups that had and had not consulted a pharmacist before purchase. Consulting a pharmacist appears to improve consumers’ awareness and makes them more likely to use appropriate medication. Those who consulted a pharmacist were better able to identify side effects and take appropriate action than the group that did not consult the pharmacist. Those who consulted a pharmacist were also significantly more likely to say that they would consult a pharmacist in the future. These results indicate that it is important for consumers to be able to consult with pharmacists, to improve consumers’ awareness of side effects and to self-medicate appropriately, and hence improve their quality of life. Pharmacists in community pharmacy could be more active in health promotion campaigns, such as drug safety, campaigns, to raise their public profile. Increased public awareness of what pharmacists in community pharmacy do will make it easier for patients to consult with them.

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  • Khalil Al-Ali, Hala Salah Abdel Fatah, Yaser Abdel-Moemen El-Badry
    2016 年 39 巻 11 号 p. 1774-1780
    発行日: 2016/11/01
    公開日: 2016/11/01
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    Ultrasound-assisted extraction of curcumin from Curcuma longa was performed in an ultrasonic bath at 30°C using ethanol for 40 min. A successful attempt has been made to prepare curcumin–zinc (Zn) complex using a simple chemical procedure. The complex formation and its stoichiometry were characterized using elemental analysis, Fourier transform (FT)-IR and UV spectroscopy which revealed the interaction of Zn(II) ion (M) with curcumin (ligand, L) to proceed via (ML) complex type formation. Oral administration of curcumin–Zn complex at a concentration of 150 mg/kg body weight/rat/d for 45 d in streptozotocin-induced diabetic rats in comparison to curcumin and/or Zn administration exerted a hypoglycemic effect. A significant reduction in blood glucose, glycosylated hemoglobin (Hb)A1c, and lipid profile parameters with an excellent improvement in plasma insulin levels have been attained. Also, the reduced activities of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea, and creatinine in the diabetic rats treated with the complex exhibited the non-toxic nature of the curcumin–Zn complex. Finally, the larger extent of the complex in hyperglycemic improvement in comparison to curcumin and/or Zn supplementation was interpreted by its dual action on glucose and insulin maintenance.

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  • Hiroaki Hayashi, Shinya Tamura, Ren Chiba, Isao Fujii, Nobuji Yoshikaw ...
    2016 年 39 巻 11 号 p. 1781-1786
    発行日: 2016/11/01
    公開日: 2016/11/01
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    The characteristics of 2 Glycyrrhiza plants, G. glabra and G. bucharica (=Meristotropis bucharica), were investigated in Tajikistan. The glycyrrhizin content in the underground parts of G. glabra varied from 2.56 to 9.29% of the dry weight, and the content of glabridin, a species-specific flavonoid of G. glabra, varied from 0.09 to 0.92% of the dry weight. Seeds of G. glabra plants from Tajikistan were cultivated for 3 years in Japan, and the glycyrrhizin content of the harvested roots ranged from 0.75 to 1.82% of the dry weight. In addition, HPLC analysis of leaf extracts indicated that the G. glabra plants collected in Tajikistan could be divided into various types, according to the flavonoid contents of the leaves. The endemic G. bucharica was also collected. A phylogenetic tree of rbcL nucleotide sequences from various Glycyrrhiza plants indicated that G. bucharica was closely related to the three glycyrrhizin-producing Glycyrrhiza spp. (G. uralensis, G. inflata, and G. glabra), even though G. bucharica does not produce glycyrrhizin.

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  • Chao Yu, Shanjun Tan, Chunyu Zhou, Cuilin Zhu, Xin Kang, Shuai Liu, Sh ...
    2016 年 39 巻 11 号 p. 1787-1792
    発行日: 2016/11/01
    公開日: 2016/11/01
    [早期公開] 公開日: 2016/08/09
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    Berberine is one of the main active constituents of Rhizoma coptidis, a traditional Chinese medicine, and has long been used for the treatment of gastrointestinal disorders. The present study was designed to investigate the effects of berberine on the intestinal mucosal barrier damage in a rat uremia model induced by the 5/6 kidney resection. Beginning at postoperative week 4, the uremia rats were treated with daily 150 mg/kg berberine by oral gavage for 6 weeks. To assess the intestinal mucosal barrier changes, blood samples were collected for measuring the serum D-lactate level, and terminal ileum tissue samples were used for analyses of intestinal permeability, myeloperoxidase activity, histopathology, malondialdehyde (MDA) level, and superoxide dismutase (SOD) activity. Berberine treatment resulted in significant decreases in the serum D-lactate level, intestinal permeability, intestinal myeloperoxidase activity, and intestinal mucosal and submucosal edema and inflammation, and the Chiu’s scores assessed for intestinal mucosal injury. The intestinal MDA level was reduced and the intestinal SOD activity was increased following berberine treatment. In conclusion, berberine reduces intestinal mucosal barrier damage induced by uremia, which is most likely due to its anti-oxidative activity. It may be developed as a potential treatment for preserving intestinal mucosal barrier function in patients with uremia.

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  • Gerlane Coelho Bernardo Guerra, Marília Stefani Souza de Menezes, Auri ...
    2016 年 39 巻 11 号 p. 1793-1801
    発行日: 2016/11/01
    公開日: 2016/11/01
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    The objective of this study was to study the effect of olmesartan medoxomil (OLM), an antihypertensive drug, on intra-articular inflammation induced by zymosan (Zy) in Wistar rats. Intra-articular inflammation was induced in the right knees of rats by 1 mg Zy dissolved in saline. The animals were divided into the following groups: saline only (oral saline and intra-articular saline); Zy only (intra-articular Zy and oral saline), and intra-articular Zy and oral OLM (5, 15, or 30 mg/kg) or diclofenac sodium (SD; 100 mg/kg). Twenty-four hours after Zy injection, synovial fluid was collected for total leukocyte counts, blood was collected for biochemical measurements, and synovial tissue was collected for histopathology, immunohistochemistry, immunofluorescence and myeloperoxidase (MPO), malonaldehyde (MDA), and non-protein sulphydryl (NPSH) assays. OLM doses of 15 and 30 mg/kg had protective effects, as evidenced by improved histopathological parameters of synovium, reduced total leukocyte counts, reduced MPO and MDA levels, and increased NPSH group levels compared with the Zy group. OLM reduced immunostaining for cyclooxygenase 2, tumour necrosis factor and interleukin 17 and increased immunostaining for superoxide dismutase and glutathione peroxidase. SD produced similar results. The drugs studied caused no change in biochemical parameters of the animals. OLM showed protective effects in this model of Zy-induced intra-articular inflammation.

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  • Mi-Hee Han, Sung-Won Park, Hyun-Jin Do, Hak-Jae Chung, Hyuk Song, Jin- ...
    2016 年 39 巻 11 号 p. 1802-1808
    発行日: 2016/11/01
    公開日: 2016/11/01
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    Growth and differentiation factor 3 (GDF3), a mammalian-specific transforming growth factor β ligand, and OCT4, one of key stem cell transcription factors, are expressed in testicular germ cell tumors (TGCTs) as well as pluripotent stem cells. To understand the molecular mechanism by which OCT4 and GDF3 function in tumorigenesis as well as stemness, we investigated the transcriptional regulation of GDF3 mediated by OCT4 in human embryonic carcinoma (EC) NCCIT cells, which are pluripotent stem cells of TGCTs. GDF3 and OCT4 was highly expressed in undifferentiated NCCIT cells and then significantly decreased upon retinoic acid-induced differentiation in a time-dependent manner. Moreover, GDF3 expression was reduced by short hairpin RNA-mediated knockdown of OCT4 and increased by OCT4 overexpression, suggesting that GDF3 and OCT4 have a functional relationship in pluripotent stem cells. A promoter-reporter assay revealed that the GDF3 promoter (−1721-Luc) activity was significantly activated by OCT4 in a dose-dependent manner. Moreover, the minimal promoter (−183-Luc) was sufficient for OCT4-mediated transcriptional activation and provided a potential binding site for the direct interaction with OCT4. Collectively, this study provides the evidence about the regulatory mechanism of GDF3 mediated by OCT4 in pluripotent EC cells.

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  • Mitsumasa Kaneta, Wataru Ochiai, Marina Nagae, Wataru Suto, Mika Hanag ...
    2016 年 39 巻 11 号 p. 1809-1814
    発行日: 2016/11/01
    公開日: 2016/11/01
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    Approximately 30% of patients with cancer pain experience concurrent neuropathic pain. Since these patients are not sufficiently responsive to morphine, the development of an effective method of pain relief is urgently needed. Decreased function of the μ opioid receptor, which binds to the active metabolite of morphine M-6-G in the brain, has been proposed as a mechanism for morphine resistance. Previously, we pharmacokinetically examined morphine resistance in mice with neuropathic pain, and demonstrated that the brain morphine concentration was decreased, expression level of P-glycoprotein (P-gp) in the small intestine was increased, and expression level and activity of uridine diphosphate glucuronosyltransferase (UGT)2B in the liver were increased. In order to clarify the mechanism of the increased expression of UGT2B, we examined the phase of neuropathic pain during which UGT2B expression in the liver begins to increase, and whether this increased expression is nuclear receptor-mediated. The results of this study revealed that the increased expression of UGT2B in the liver occurred during the maintenance phase of neuropathic pain, suggesting that it may be caused by transcriptional regulation which was not accompanied by increased nuclear import of pregnane X receptor (PXR).

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  • Teerapol Srichana, Chaveewan Ratanajamit, Siwasak Juthong, Tan Suwande ...
    2016 年 39 巻 11 号 p. 1815-1822
    発行日: 2016/11/01
    公開日: 2016/11/01
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    Inhalation therapy is a promising drug delivery approach for tuberculosis treatment. However, there is always concern about the safety of the dosage form by inhalation as it may induce inflammation. Developing a new dosage form for inhalation must include tests for its safety especially for the tumor necrosis factor (TNF)-α and interleukine (IL)-1β. The safety of four anti-tuberculosis (anti-TB) drugs administered via inhalation was assessed in healthy volunteers. Four anti-TB drugs; isoniazid, rifampicin, pyrazinamide and levofloxacin were prepared as dry powder and evaluated for uniformity of delivered dose and in vitro drug deposition. These four anti-TB dry powder formulations for inhalation met the criteria of uniformity of delivered dose and exhibited suitable size for lung delivery. Forty healthy volunteers were recruited and each was sequentially challenged with isoniazid, rifampicin, pyrazinamide and levofloxacin in different orders. Safety was monitored by measuring the pro-inflammatory cytokines in their sputum, lung function test, blood chemistry and adverse events. This study proves that all four anti-TB dry powders did not provoke inflammatory cytokines and are safe to healthy volunteers.

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  • Keiichi Motoyama, Rena Nishiyama, Yuki Maeda, Taishi Higashi, Yoshimas ...
    2016 年 39 巻 11 号 p. 1823-1829
    発行日: 2016/11/01
    公開日: 2016/11/01
    [早期公開] 公開日: 2016/09/06
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    Niemann–Pick disease type C (NPC) is an autosomal recessive lysosomal storage disorder, which is an inherited disease characterized by the accumulation of unesterified cholesterol in endolysosomes. Recently, 2-hydroxypropyl-β-cyclodextrin (HP-β-CyD) has been used for the treatment of NPC, and ameliorated a hepatosplenomegaly in the patients. However, to obtain the treatment efficacy, a high dose of HP-β-CyD was necessary. Therefore, the decrease in dose by using active intracellular delivery system of β-CyD to NPC cells is expected. In this study, to efficiently deliver β-CyD to NPC-like cells, we newly synthesized octaarginine (R8)-appended β-CyD with a spacer of γ-aminobutyric acid (R8-β-CyD) and evaluated its cytotoxicity, intracellular distribution, endocytosis pathway and cholesterol-lowering effect in Npc1-trap-Chinese hamster ovary (CHO) cells, cholesterol-accumulated cells through the impairment of NPC1 function. R8-β-CyD did not show cytotoxicity in the cells. In addition, Alexa568-labeled R8-β-CyD was actively internalized into Npc1-trap-CHO cells, possibly through micropinocytosis. Notably, R8-β-CyD significantly decreased intracellular cholesterol content compared with HP-β-CyD. These results suggest that R8-β-CyD may be a promising therapeutic agent for ameliorating cholesterol accumulation in NPC.

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  • Suho Seo, Kyuhwa Seo, Sung Hwan Ki, Sang Mi Shin
    2016 年 39 巻 11 号 p. 1830-1838
    発行日: 2016/11/01
    公開日: 2016/11/01
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    Isorhamnetin is a flavonoid metabolite of quercetin and isolated from water dropwort (Oenanthe javanica, Umbelliferae). It has been reported that isorhamnetin exerts beneficial effects including antioxidant, anti-inflammatory, and anti-proliferative activities. The present study investigated whether the antioxidant activity of isorhamnetin is correlated with its anti-cancer effects on colorectal cancer cells. Isorhamnetin significantly repressed cobalt chloride (CoCl2)- or hypoxia-induced hypoxia inducible factor-1α (HIF-1α) accumulation in HCT116 and HT29 cells. When compared with quercetin, isorhamnetin showed potent inhibition of HIF-1α. Moreover, it inhibited CoCl2-induced activity of hypoxia response element reporter gene and HIF-1α-dependent transcription of genes such as glucose transporter 1, lactate dehydrogenase A, carbonic anhydrase-IX, and pyruvate dehydrogenase kinase 1. Isorhamnetin also blocked hydrogen peroxide (H2O2)-induced HIF-1α accumulation. The antioxidant effects of isorhamnetin were confirmed by observation of CoCl2- or H2O2-induced reactive oxygen species (ROS) production. Consistently, overexpressed HIF-1α was decreased by isorhamnetin or N-acetyl-L-cysteine in HEK293 cells. In vitro migration and invasion assay further confirmed the inhibitory effects of isorhamnetin on cancer cells. Collectively, these results demonstrate that isorhamnetin inhibits ROS-mediated HIF-1α accumulation, which contributes to its anti-metastatic efficacy.

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  • Pollyanna Francielli de Oliveira, Jaqueline Lopes Damasceno, Heloiza D ...
    2016 年 39 巻 11 号 p. 1839-1845
    発行日: 2016/11/01
    公開日: 2016/11/01
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    This study evaluated the influence of Styrax camporum stems hydroalcoholic extract (SCHE) and of chemical markers of the genus, egonol (EG) and homoegonol (HE), on DNA damage induced in V79 cells by mutagens with different mechanisms of action. These natural products were combined with different mutagens [methyl methanesulfonate (MMS), hydrogen peroxide (H2O2), (S)-(+)-camptothecin (CPT), and etoposide (VP-16)] to evaluate the modulatory effect on DNA damage. The results showed that SCHE was genotoxic at the highest concentration tested (60 µg/mL). Treatment with EG or HE alone induced no genotoxic effect, while genotoxic activity was observed when the two compounds were combined. The SCHE extract was able to reduce the frequency of micronuclei induced by H2O2 and VP-16. Similar results were observed when the cell cultures were treated with EG and/or HE plus VP-16. In contrast, the highest concentration (40 µg/mL) SCHE potentiated DNA damage induced by VP-16. Neolignan EG alone or combined with HE also potentiated H2O2-induced genotoxicity. However, these natural products did not influence the frequency of DNA damage induced by MMS or CPT. Therefore, the influence of SCHE and of chemical markers (EG and HE) of the genus on the induction of DNA damage depends on the concentration tested and on the mutagen used.

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  • Masataka Tajima, Yoshinori Kato, Jun Matsumoto, Iori Hirosawa, Mariko ...
    2016 年 39 巻 11 号 p. 1846-1851
    発行日: 2016/11/01
    公開日: 2016/11/01
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    Linezolid (LZD) is an antimicrobial that is commonly used for treatment of vancomycin-resistant Enterococci and methicillin-resistant Staphylococcus aureus infections. However, the development of thrombocytopenia, one of the most frequent adverse side effects of this antimicrobial, can lead to discontinuation of LZD treatment. While clinical studies indicate that risk factors for the development of LZD-induced thrombocytopenia include treatment for >14 consecutive days, renal dysfunction, and chronic liver disease, the fundamental mechanism governing the pathogenesis of this disorder remains unclear. In this study, we aimed to elucidate the mechanism of LZD-induced thrombocytopenia by investigating the impact of LZD treatment on platelet destruction and production using rat platelet-rich plasma (PRP) and human immortalized cell lines, respectively. Compared to the control population, an increase in lactate dehydrogenase release was not detected upon exposure of rat PRP to varying concentrations of LZD, indicating that this compound is not cytotoxic towards platelets. Meanwhile, LZD treatment resulted in a significant dose-dependent increase in the proliferation of HEL human erythroleukemia and MEG-01 human megakaryoblast cells in vitro, but did not influence the differentiation of these cell lines. Lastly, LZD treatment yielded elevated levels of phosphorylation of myosin light chain 2 (MLC2), which regulates platelet release, in MEG-01 cells. Based on these results, we speculate that LZD induces thrombocytopenia by promoting MLC2 phosphorylation and thereby suppressing the release of platelets from mature megakaryocytes. These findings provide the first insight into the mechanism of LZD-mediated thrombocytopenia and may facilitate the development of strategies to treat and/or prevent this disease.

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  • Naoki Mukoyama, Akira Yoshimi, Aya Goto, Haruka Kotani, Kazuhiro Ishik ...
    2016 年 39 巻 11 号 p. 1852-1858
    発行日: 2016/11/01
    公開日: 2016/11/01
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    There are individual differences in the frequency of chemotherapy-induced nausea and vomiting (CINV) in cancer patients. We investigated the individual variability in susceptibility to CINV with focus on both behavioral factors and genetic factors in Japanese cancer patients. We performed a prospective study to investigate the association between patient attributes (backgrounds and habits as well as gene polymorphisms) and anorexia, nausea, or vomiting in 55 Japanese cancer patients undergoing chemotherapy at Nagoya University Hospital. We found that gender (female), use of non-steroidal anti-inflammatory drugs, susceptibility to motion sickness, and anxiety were associated with the frequency of CINV. Gene polymorphisms of rs1076560 (dopamine D2 receptor gene), rs6766410 (serotonin 5-HT3C receptor gene) and rs4680 (catechol-O-methyltransferase gene) were also associated. Our data suggest that these attributes may thus be risk factors for CINV. Our results provide novel information that can be used to predict the incidence of CINV in Japanese patients undergoing chemotherapy; this can help provide a substantial improvement in supportive care for patients with different types of cancer.

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  • Yasuhiro Abe, Kumiko Sakai-Kato, Yukihiro Goda
    2016 年 39 巻 11 号 p. 1859-1867
    発行日: 2016/11/01
    公開日: 2016/11/01
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    RNA interference via small interfering RNA (siRNA) has many potential therapeutic applications, and liposomal-based systems are useful for improving the pharmacokinetics of siRNAs, including their intracellular release and distribution. However, for the successful translation of this technology into clinical applications, it is important to understand how liposomal encapsulation changes the cellular uptake and immunostimulatory adverse effects of siRNAs. Here we evaluated the cellular uptake and innate immune activation by an immunostimulatory siRNA encapsulated within a liposome carrier in commercially available human peripheral blood mononuclear cells (PBMCs). We found considerable lot-to-lot variation in cytokine production by the PBMCs. Flow cytometric analysis in conjunction with intracellular staining of tumor necrosis factor-α (TNF-α) revealed that after treating PBMCs with the liposomal siRNA, approximately 5% of the cells produced TNF-α and more than 90% of the TNF-α-producing cells were positive for CD14 expression. We also showed that peripheral blood CD14+ monocytes in the cytokine release assay had low inter-lot variabilities in TNF-α production, suggesting that the peripheral blood CD14+ monocyte-based cytokine release assay is a specific means of alleviating the lot-to-lot variability in the cytokine release profiles of commercially available PBMCs. Our results also show that the peripheral blood CD14+ monocyte-based cytokine release assay can be used to identify the siRNA recognition receptors that mediate individual cytokine production.

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  • Kiyoshi Watanabe, Hidemasa Nakaminami, Chihiro Azuma, Ippei Tanaka, Ke ...
    2016 年 39 巻 11 号 p. 1868-1875
    発行日: 2016/11/01
    公開日: 2016/11/01
    [早期公開] 公開日: 2016/08/11
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    Staphylococcus epidermidis, a major skin flora on hands, acts as a reservoir of various antimicrobial resistance determinants including staphylococcal cassette chromosome mec (SCCmec) and contributes to multidrug resistance for S. aureus. The aim of this study was understanding the characteristics of commensal S. epidermidis on the hands of hospital workers and healthy individuals. A total of 23 hospital workers (physicians, nurses, and hospital pharmacists), 13 community pharmacists, and 24 healthy individuals (students) were studied. Commensal bacteria on hands were recovered using a glove-juice method. For methicillin-resistant S. epidermidis (MRSE), we performed SCCmec typing, pulsed-field gel electrophoresis (PFGE), and determined the antimicrobial susceptibility. The detection rates of MRSE in community pharmacists (92.3%) and students (87.5%) were higher than those in hospital workers (66.7 to 81.8%). SCCmec type IV strains were predominant in both hospital workers and students. PFGE analysis strongly suggested that the MRSE of hospital workers and students were normal inhabitants of each subject. The antimicrobial resistance rates and levels in MRSE of hospital workers were higher than those of students. Our findings showed that MRSE was frequently colonized on the hands of healthy individuals as well as hospital workers.

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    Editor’s picks

    In their report, Watanabe et al. described the prevalence of methicillin-resistant Staphylococcus epidermidis (MRSE), one of the major nosocomial pathogens, on the hands of healthy individuals and hospital workers. The detection rates of MRSE in community pharmacists (92.3%) and students (87.5%) were higher than those in hospital workers (66.7% to 81.8%). Pulsed-field gel electrophoresis (PFGE) analysis strongly suggested that the MRSE strains on the hands of hospital workers and students were normal inhabitants of each subject. Their results lead to a new finding that MRSE is commonly colonized on the hands of not only hospital workers but also of healthy individuals.



  • Osamu Nakajima, Tomoko Nishimaki-Mogami, Kazunari Kondo
    2016 年 39 巻 11 号 p. 1876-1880
    発行日: 2016/11/01
    公開日: 2016/11/01
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    Genome editing has undergone rapid development during the last three years. It is anticipated that genetically modified organisms (GMOs) for food purposes will be widely produced using the clustered regularly interspaced short palindromic repeat/Cas9 (CRISPR)/Cas9 system in the near future. However, the Cas9 gene may then enter the genomes of GMOs for food if the breeding process is not strictly managed, which could lead to the Cas9 protein or associated peptides being produced within these organisms. A variety of peptides could theoretically be produced from the Cas9 gene by using open reading frames different from that of Cas9 in the GMOs. In this study, Cas9 and the peptides potentially encoded by Cas9 genes were studied regarding their immunogenicity, in terms of the digestibility of Cas9 and the homology of the peptides to food allergens. First, the digestibility and thermal stability of Cas9 were studied. Digestibility was tested with natural or heat-denatured Cas9 in simulated gastric fluid in vitro. The two types of Cas9 were digested rapidly. Cas9 was also gradually degraded during heat treatment. Second, the peptides potentially encoded by Cas9 genes were examined for their homology to food allergens. Specifically, an 8-mer exact match search and a sliding 80-mer window search were performed using allergen databases. One of the peptides was found to have homology with a food allergen.

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  • Noriaki Nagai, Yu Mano, Yoshimasa Ito
    2016 年 39 巻 11 号 p. 1881-1887
    発行日: 2016/11/01
    公開日: 2016/11/01
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    Disulfiram (DSF) is a dimer of diethyldithiocarbamate (DDC) that we previously added to a solution of 2-hydroxypropyl-β-cyclodextrin (DSF solution). We found that the instillation of this DSF solution delayed lens opacification in a hereditary cataractous ICR/f rat. In this study, we attempted to design an ophthalmic formulation containing DSF nanoparticles for use as a lens targeted drug delivery system (nano-DSF suspension), and investigated the changes in drug content in the lens after the instillation of DSF solution or nano-DSF suspension. The nano-DSF suspension was prepared by a bead mill method to yield a mean particle size of nano-DSF of 181 nm. Following the instillation of 1.4% DSF solution or the nano-DSF suspension, DDC was detected only in the aqueous humor and lens; in both, the area under the curve (AUC) and mean residence time (MRT) for the nano-DSF suspension were higher than for the DSF solution. In addition, we found that the DDC residence time in the cortex and nucleus of the lens was higher than in the capsule-epithelium. Although DDC was not detected in the cortex and nucleus of lenses following the instillation of the 1.4% DSF solution, the instillation of a 1.4% nano-DSF suspension led to the accumulation of DDC in both areas. In conclusion, it is possible that the instillation of a nano-DSF suspension can supply more DDC into the aqueous humor and lens than a conventional formulation, and these findings provide information significant for the prevention of cataracts and the design of a lens targeted drug delivery system.

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  • Malik Suliman Mohamed, Toshihide Maki, Mohammad Monir Shah, Yoshio Ich ...
    2016 年 39 巻 11 号 p. 1888-1892
    発行日: 2016/11/01
    公開日: 2016/11/01
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    Two hydroquinone derivatives were prepared and their antimicrobial activity evaluated. Their minimum inhibitory concentrations (MICs) were determined using a broth dilution method. Gentamycin and ciprofloxacin were used as reference antibiotics. The antimicrobial activity of 4-(benzyloxy)phenol (monobenzone) was also evaluated based on its structural similarity to the new compounds; activity was comparable to that of 3,5-dimethyl-4-((4-nitrobenzyl)oxy)phenol (4a). 2,3,5-Trimethyl-4-((4-nitrobenzyl)oxy)phenol (4b) exhibited the best antibacterial activity against both clinical isolates and type strain of Moraxella catarrhalis (M. catarrhalis), with a MIC value of 11 µM, comparable to ciprofloxacin 9 µM.

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Notes
  • Yoko Idota, Yumi Kogure, Takako Kato, Kentaro Yano, Hiroshi Arakawa, C ...
    2016 年 39 巻 11 号 p. 1893-1896
    発行日: 2016/11/01
    公開日: 2016/11/01
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    We investigated the relationship between the physical parameters of various metal ions, including toxic metal ions, and the binding affinity of these metal ions for alginate (Alg). The binding constant, K, of Sr2+ was the highest among all tested metal ions. The order of K values was: Sr2+>Pb2+>Tb3+>Dy3+>Ca2+>Cd2+>Mg2+>Fe2+>Fe3+>Co2+>Al3+>Ni2+>Cs+>Cu2+>Ag+>Li+>K+. The metal ions showing the highest K values had ionic radii within the range of about 90–120 pm. Moreover, the K values of divalent or trivalent metal ions tended to be higher than those of monovalent ions. The number of binding sites per 1 mg of Alg (n) was highest for K+, followed by Pb2+ and Cs+. The order of affinity (calculated as the product of n and K) was Pb2+>Dy3+>Tb3+>Sr2+>Ca2+>Mg2+>Cd2+>Fe2+, Fe3+>Cs+>Al3+>Co2+>Ni2+>Cu2+>Ag+>K+>Li+. Our results support the idea that Alg would be effective as an excretion accelerator and/or absorption inhibitor for various toxic metal ions.

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  • Naoyoshi Maeda, Atsushi Furukawa, Kosuke Kakita, Masahiro Anada, Shuni ...
    2016 年 39 巻 11 号 p. 1897-1902
    発行日: 2016/11/01
    公開日: 2016/11/01
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    Herpes simplex virus type 1 (HSV-1) is a causative agent for a variety of diseases. Although antiherpetic drugs such as acyclovir have been developed to inhibit virus replication through interaction with DNA kinases, their continuous administration leads to an increase in the frequency of drug-resistant HSV-1, which is an important clinical issue that requires urgent solution. Recently, we reported that the sialylated O-linked sugar T antigen (sTn) and its attached peptide region (O-glycosylated sTn peptide) derived from the HSV-1 glycoprotein B (gB) protein inhibited HSV-1 infection by specifically targeting paired immunoglobulin-like type 2 receptor alpha (PILRα) in vitro. In this study, to further identify novel inhibitors of gB-mediated HSV-1 infection in vitro, we established a cell-based fusion assay for rapid drug screening. Chinese hamster ovary (CHO) cells were transfected with expression plasmids for HSV-1 gB, gD, gH, and gL, and T7 RNA polymerase, and were designated as the effector cells. The CHO-K1 cells stably expressing PILRα were transfected with the expression plasmid for firefly luciferase under the T7 promoter, and were designated as the target cells. The effector and target cells were co-cultured, and luminescence was measured when both cells were successfully fused. Importantly, we found that cell-to-cell fusion was specifically inhibited by O-glycosylated sTn peptide in a dose dependent manner. Our results suggested that this virus-free cell-based fusion assay system could be a useful and promising approach to identify novel inhibitors of gB-mediated HSV-1 infection, and will aid in the development of antiviral therapeutic strategies for HSV-1-associated diseases.

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