Faculty Profiles - NAMBA Hikaru

MISC - NAMBA Hikaru

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High-resolution immuno-scanning electron microscopy using a non-coating method: study of herpes simplex virus glycoproteins on the surface of virus particles and infected cells. Reviewed International journal

J A Padilla, F Uno, M Yamada, H Namba, S Nii

Journal of electron microscopy 46 ( 2 ) 171 - 80 1997

Language：English Publishing type：Article, review, commentary, editorial, etc. (scientific journal) Publisher：OXFORD UNIV PRESS

The expression of two glycoproteins, i.e. glycoprotein C (gC) and glycoprotein D (gD), of herpes simplex virus type 1 (HSV-1) on the surface of extracellular particles of this virus was examined by immuno-scanning electron microscopy. Scanning electron microscopy specimens of infected cells immuno-labelled against the glycoproteins with colloidal gold particles were prepared by a conventional coating and a non-coating method. Surface ultrastructure of infected cells and gold particles were observed more clearly with specimens prepared by the non-coating method. The appearance of virus particles in association with glycoprotein expression on these particles and on the surface of infected cells was then studied. Progeny virus particles began to appear 6 h after infection, increased in number as the infection proceeded, and covered most of the cell surface by 16 h. Six to 24 h after the infection, the labelling density for each glycoprotein on virus particles remained constant. The labelling density for gD was always higher than that for gC. The patch-like distribution of gold-labelling against gD was often detected on infected cell monolayers at the exponential and late stage of one cycle of virus growth. The labelling density for gD on virus particles was the highest on these produced in Vero and L-929 cells, moderate in MRC-5, BHK-21 and FL cells, and the lowest in HEp-2 cells.

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BDNF gene can Be activated by Ca2+ signals without involvement of de novo AP-1 synthesis. International journal

K Sano, H Nanba, A Tabuchi, T Tsuchiya, M Tsuda

Biochemical and biophysical research communications 229 ( 3 ) 788 - 93 1996.12

Language：English Publisher：ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS

Although stimulation of N-methyl-D-aspartate receptors or voltage-dependent calcium channels induces both the activation of c-fos and brain-derived neurotrophic factor (BDNF) genes, it is not certain how the activation of these genes is related. Using primary cultures of rat hippocampal neurons, we found that exposing the cells to cycloheximide allowed subsequent activation of BDNF mRNA expression, although activation of AP-1 DNA-binding activity resulting from the c-fos induction was abolished. Super-induction of BDNF gene was also caused by cycloheximide. The estimated half-life of BDNF mRNA was approximately 2.5 hrs, which was almost identical to that of c-fos mRNA. These results indicate that nascent AP-1 is not required for the activation of BDNF gene, leading to the notion that the BDNF gene can be activated by Ca2+ signals as an immediate early gene.

DOI： 10.1006/bbrc.1996.1881

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インフルエンザウイルスの細胞侵入に対するセシウム添加の影響(in vitro)

山下信子, 小川寛人, 難波ひかる, 山田雅夫

臨床とウイルス 48 ( 3 ) S91 - S91 2020.9

唾液からのHHV6とHHV7のDNA検出率の前方視的検討保育園入園半年と非就園半年の比較

鳥越貞義, 渡辺正博, 難波ひかる, 山下信子, 小川寛人, 山田雅夫, 菅秀, 根来麻奈美

臨床とウイルス 47 ( 2 ) S84 - S84 2019.4

セシウムがインフルエンザウイルス増殖に及ぼす影響

山下信子, 小川寛人, 難波ひかる, 山田雅夫

日本ウイルス学会学術集会プログラム・予稿集(Web) 67th 2019

岡山大学自然生命科学研究支援センターに搬入される家畜ブタのE型肝炎ウイルス感染状況について

小川寛人, 平山晴子, 田中爽暉, 矢田範夫, 難波ひかる, 山下信子, 米満研三, 前田健, 樅木勝巳, 山田雅夫

岡山実験動物研究会報 ( 35 ) 2019

ヒトヘルペスウイルス6Bのテグメント蛋白質間の相互作用解析

難波ひかる

日本分子生物学会年会プログラム・要旨集(Web) 42nd 2019

HHV6Bテグメント蛋白質間相互作用の網羅的解析

難波ひかる, 小川寛人, 山下信子, 山田雅夫

日本ウイルス学会学術集会プログラム・予稿集(Web) 67th 2019

養豚農場から大学動物実験施設に搬入される家畜ブタのE型肝炎ウイルス感染状況について

小川寛人, 平山晴子, 田中爽暉, 矢田範夫, 難波ひかる, 山下信子, 米満研三, 前田健, 樅木勝巳, 山田雅夫

日本獣医学会学術集会講演要旨集 161回 475 - 475 2018.8

動物実験施設に搬入される家畜ブタにおけるE型肝炎ウイルスの遺伝子検出および血清学的解析

小川寛人, 難波ひかる, 山下信子, 山田雅夫, 田中爽暉, 矢田範夫, 平山晴子, 樅木勝巳, 米満研三, 前田健

臨床とウイルス 46 ( 2 ) S58 - S58 2018.4

抗GFP-Nanobody共免疫沈降法によるウイルステグメント蛋白質の相互作用解析

難波ひかる

生命科学系学会合同年次大会 2017年度 [3LBA - 114] 2017.12

Metabolic pathway catalyzed by Vanin-1 pantetheinase plays a suppressive role in influenza virus replication in human alveolar epithelial A549 cells. International journal

Nobuko Yamashita, Masato Yashiro, Hirohito Ogawa, Hikaru Namba, Nobuyuki Nosaka, Yousuke Fujii, Tsuneo Morishima, Hirokazu Tsukahara, Masao Yamada

Biochemical and biophysical research communications 489 ( 4 ) 466 - 471 2017.8

Language：English Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE

Our previous analysis of gene expression profiles in the peripheral blood from patients with influenza A (H1N1) pdm09 pneumonia revealed elevated transcription levels of the vanin-1 (vascular non-inflammatory molecule 1, VNN1) gene, which encodes an epithelial ectoenzyme with pantetheinase activity involved in recycling coenzyme A. Here, to elucidate the role of VNN1 in influenza A virus (IAV) H1N1 infection, we investigated the change of VNN1 expression in the context of IAV infection and the effects of its related substances, i.e., its direct substrate pantetheine and its two metabolites pantothenic acid and cysteamine on the replication of IAV in the human alveolar epithelial carcinoma cell line A549. The messenger RNA expression of VNN1 in A549 cells was significantly increased (by 4.9-fold) after IAV infection under an elevated concentration of pantetheine. Moreover, VNN1 mRNA levels were elevated by > 100-fold in response to pro-inflammatory cytokines, especially TNF-α and IL-1β. Pantetheine significantly reduced the IAV replication and IAV Matrix 1 (M1) mRNA levels when it was administered prior to and during infection. In addition, cysteamine treatment during IAV infection significantly reduced the viral replication and IAV M1 mRNA levels, whereas pantothenic acid did not. These findings suggest that the metabolic pathway catalyzed by VNN1 pantetheinase plays a suppressive role in IAV infection in the respiratory tract, especially in severe conditions under hypercytokinemia.

DOI： 10.1016/j.bbrc.2017.05.172

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抗GFP-Nanobody共免疫沈降法によるウイルステグメント蛋白質の相互作用解析

難波ひかる

日本生化学会大会(Web) 90th [3LBA - 114] 2017

集団保育児における唾液中のHHV-6とHHV-7DNAの排泄

難波ひかる, 渡辺正博, 鳥越貞義, 山下信子, 山田雅夫

臨床とウイルス 45 ( 2 ) S76 - S76 2017

突発性発疹の感染様式に関する研究

鳥越貞義, 難波ひかる, 渡辺正博, 宮崎裕樹, 森島恒雄, 山田雅夫

臨床とウイルス 43 ( 2 ) S111 - S111 2015

Vanin-1がインフルエンザウイルス感染に果たす役割

山下信子, 難波ひかる, 山田雅夫, 森島恒雄

臨床とウイルス 41 ( 2 ) S97 - S97 2013.5

インフルエンザウイルス増殖に対する酸化ストレス関連遺伝子Vanin-1の影響

山下信子, 難波ひかる, 山田雅夫, 森島恒雄

日本ウイルス学会学術集会プログラム・抄録集 61st 2013

HHV-6Bテグメント蛋白質の単独発現系による細胞内局在解析

難波ひかる, 山下信子, 山田雅夫

日本ウイルス学会学術集会プログラム・抄録集 60th 2012

Vanin-1がインフルエンザウイルス感染に果たす役割

山下信子, 山下信子, 難波ひかる, 山田雅夫, 森島恒雄

日本ウイルス学会学術集会プログラム・抄録集 60th 2012

Enhancive effects of D-glucose and its analogs on expression of d-glucose-unrelated transgenes in mammalian cells. Reviewed

Miyuki Kimura, Hikaru Namba, Manabu Okubo, Mai Ezumi, Nao Susumu, Masao Yamada, Yujiro Arao

Journal of bioscience and bioengineering 112 ( 2 ) 194 - 201 2011.8

Language：English Publishing type：Article, review, commentary, editorial, etc. (scientific journal) Publisher：SOC BIOSCIENCE BIOENGINEERING JAPAN

We studied the effects of d-glucose on transgene expression in mammalian cells by a reporter gene assay using CV-1 cells and a CMV promoter-controlled EGFP gene. Treatment of CV-1 cells with 5% D-glucose unchanged the number of fluorescent cells in fluorescence microscopic observation but significantly intensified fluorescence in the fluorometric assay. Furthermore, EGFP itself and mRNA became more abundant in Western blot and quantitative RT-PCR analyses of 5% D-glucose-treated cells, respectively. These results indicate that elevated D-glucose can activate transgene expression via transcriptional stimulation, at least in part. The same concentrations of L-glucose led to only negligible increases in transgene expression, indicating that D-glucose's effect is different from its osmotic effect. The D-glucose-induced augmentation of fluorescence was observed not only in the experiment using the CMV promoter-controlled EGFP gene but also in experiments using the SV40 and RSV promoter-controlled ones, suggesting that elevated D-glucose can enhance transgene expression regulated by various promoters commonly used in transgene expression. The assessment of D-glucose analogs for their enhancive effects on transgene expression revealed that 1,6-anhydro-D-glucose and β-methyl-D-glucoside had stronger effects than D-glucose. From this result, we can expect to find more effective carbohydrates to enhance transgene expression. The α- and β-M-D-glucosides, which are slightly different from each other in three-dimensional structure, exerted largely distinct stimulative effects on transgene expression, suggesting that fundamental rules determine the enhancive effects of saccharides and that the modification of the saccharide by applying such rules will enable us to develop more powerful substances for transgene expression.

DOI： 10.1016/j.jbiosc.2011.04.010

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G型ボツリヌス毒素遺伝子をコードするプラスミドの解析

阪口義彦, 林哲也, 林哲也, 山本由弥子, 難波ひかる, 中山恵介, 馬少博, ZHANG Kai, 幸田知子, 小崎俊司, 小熊惠二

日本細菌学雑誌 65 ( 1 ) 162 - 162 2010

コレラ菌およびコレラ毒素Bサブユニットに対するニワトリ抗体(IgY)の有用性

平井一行, 衛藤友美, 大野佑子, 田村臣哉, 山本由弥子, 難波ひかる, 阪口義彦, 横田憲治, 小熊惠二

日本薬学会年会要旨集 130th ( 3 ) 84 - 84 2010

G型ボツリヌス毒素遺伝子をコードするプラスミドの解析

阪口義彦, 林哲也, 林哲也, 中山恵介, 山本由弥子, 難波ひかる, 細見晃司, 幸田知子, 向本雅郁, 小崎俊司, 小熊惠二

日本ゲノム微生物学会年会要旨集 4th 2010

C型とD型ボツリヌス菌ホスホリパーゼCの酵素活性と生物活性

阪口義彦, 小田真隆, 山本由弥子, 難波ひかる, MA Shaobo, ZHANG Kai, 田村臣哉, 唐澤忠宏, 小林敬子, 永浜政博, 櫻井純, 小熊恵二

日本分子生物学会年会講演要旨集 32nd ( Vol.2 ) 2009

哺乳動物培養細胞における糖応答シグナルの解析

難波ひかる, 荒尾雄二郎

日本分子生物学会年会講演要旨集 32nd ( Vol.1 ) 2009

ウイルスプロモーターからの遺伝子発現に対する糖の増大効果

大久保学, 難波ひかる, 木村美幸, 荒尾雄二郎

日本ウイルス学会学術集会プログラム・抄録集 56th 2008

ヒトサイトメガロウイルス前初期遺伝子プロモーターで制御された遺伝子発現に対する糖の刺激効果

大久保学, 難波ひかる, 木村美幸, 荒尾雄二郎

日本ウイルス学会学術集会プログラム・抄録集 55th 2007

ウイルス由来プロモーターで制御された遺伝子発現に対する糖の刺激効果

難波ひかる, 大久保学, 江角真依, 進七緒, 木村美幸, 荒尾雄二郎

生化学 80回・30回 4T11 - 4 2007

Characterization and usefulness of a monoclonal antibody against human herpesvirus-7 DNA polymerase processivity factor Reviewed

難波ひかる

岡山医学会雑誌 117 ( 3 ) 219 - 224 2006

単純ヘルペスウイルスの吸着・侵入による宿主細胞のエステラーゼ,並びに排出ポンプ活性の上昇

ミラー真理, 大久保学, 難波ひかる, 木村美幸, 荒尾雄二郎

日本ウイルス学会学術集会プログラム・抄録集 54th 2006

落葉状天疱瘡に生じHHV-6の再活性化を認めなかったDDSによるdrug-induced hypersensitivity syndromeの1例

濱田利久, 秋山尚範, 岩月啓氏, 山田潤, 藤本亘, 吉田まり子, 難波ひかる, 山田雅夫

西日本皮膚科 65 ( 3 ) 299 - 299 2003.6

ICE症候群におけるヘルペスウイルスの関与

永山幹夫, 松尾俊彦, 山口樹一郎, 田村直之, 楳田知子, 大月洋, 難波ひかる

緑内障 11 ( 臨増 ) 131 - 131 2001.8

Monitoring of human herpesvirus-6 and -7 genomes in saliva samples of healthy adults by competitive quantitative PCR Reviewed International journal

Nobukiyo Fujiwara, Hikaru Namba, Reiko Ohuchi, Hiroki Isomura, Fumio Uno, Mariko Yoshida, Shiro Nii, Masao Yamada

Journal of Medical Virology 61 ( 2 ) 208 - 213 2000.6

Language：English Publisher：Wiley

Human herpesviruses-6 and -7 (HHV-6 and HHV-7) are thought to be transmitted during early infancy through saliva. However, the kinetics of the virus shedding in saliva of healthy adults, from whom children are assumed to acquire the viruses, is mostly unknown. This study was conducted to determine how many copies of the genome are secreted in saliva of healthy adults and to clarify the relationship between viral DNA load and virus isolation of HHV-6 and HHV-7. Competitive PCR was performed using primer sets in the U42 gene of each viral genome. In saliva samples from 29 healthy adults, HHV-6 and HHV-7 DNA was detected in 41.4% and 89.7%, respectively. The average copy number of the HHV-7 genome in the positive samples was higher than that of the HHV-6 genome. Follow-up studies of six seropositive individuals for 3 months showed that the amount of HHV-7 DNA was constant in each individual and that "high producers" and "low producers" could be distinguished. By contrast, the amount of HHV-6 DNA varied drastically over time in each individual. Although HHV-6 was never isolated from the saliva of any of the six individuals during the follow-up period, HHV-7 was isolated from each individual several times. The amount of HHV-7 DNA tended to be higher at the times when the virus was isolated than at the times when the virus was not isolated. These data demonstrate a striking contrast between HHV-6 and HHV-7 in the kinetics of genome and virus shedding.

DOI： 10.1002/(sici)1096-9071(200006)61:2<208::aid-jmv6>3.0.co;2-1

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Suppressive effects of human herpesvirus-6 on thrombopoietin-inducible megakaryocytic colony formation in vitro. Reviewed International journal

H Isomura, M Yoshida, H Namba, N Fujiwara, R Ohuchi, F Uno, M Oda, Y Seino, M Yamada

The Journal of general virology 81 ( Pt 3 ) 663 - 73 2000.3

Language：English Publisher：SOC GENERAL MICROBIOLOGY

Two clinical observations, the association of human herpesvirus-6 (HHV-6) with delayed engraftment after stem cell transplantation and thrombocytopenia concomitant with exanthema subitum, prompted us to evaluate the suppressive effects of HHV-6 on thrombopoiesis in vitro. Different culture conditions for thrombopoietin (TPO)-inducible colonies in semi-solid matrices were examined. Using cord blood mononuclear cells as the source of haematopoietic progenitors, two types of colonies, megakaryocyte colony-forming units (CFU-Meg) and non-CFU-Meg colonies, were established. The former colonies were identified by the presence of cells with translucent cytoplasm and highly refractile cell membrane, most of which were positive for the CD41 antigen. Although the plating efficiency of both types was much higher under serum-containing conditions than under serum-free conditions, the proportion of CFU-Meg to non-CFU-Meg colonies was consistently higher under serum-free conditions. The plating efficiency of CFU-Meg colonies was doubled by adding stem cell factor to the serum-free matrix. The effects of two variants of HHV-6 (HHV-6A and 6B) and human herpesvirus-7 (HHV-7) on TPO-inducible colonies were then compared. HHV-6B inhibited both CFU-Meg and non-CFU-Meg colony formation under serum-free and serum-containing conditions. HHV-6A had similar inhibitory effects. In contrast, HHV-7 had no effect on TPO-inducible colony formation. Heat-inactivation and ultra-filtration of the virus sample completely abolished the suppressive effect. After infection of CD34(+) cells with HHV-6, the viral genome was consistently detected by in situ hybridization. These data suggest that the direct effect of HHV-6 on haematopoietic progenitors is one of the major causes of the suppression of thrombopoiesis.

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Development of a dot blot neutralizing assay for HHV-6 and HHV-7 using specific monoclonal antibodies Reviewed International journal

T Tsukazaki, M Yoshida, H Namba, M Yamada, N Shimizu, S Nii

JOURNAL OF VIROLOGICAL METHODS 73 ( 2 ) 141 - 149 1998.8

Language：English Publisher：ELSEVIER SCIENCE BV

To elucidate further immune responses to human herpesviruses 6 and 7 (HHV-6 and -7), a neutralizing antibody assay was established for these viruses using a dot blot method. Three monoclonal antibodies against HHV-6 and 12 monoclonal antibodies against HHV-7 were developed and characterized by radio-immunoprecipitation. One monoclonal antibody which recognizes the 135 kDa late polypeptide of HHV-6 and several which recognize the 125 kDa late polypeptide of HHV-7 were selected to monitor virus growth by a dot blot antigen-detection method. The dot blot method was then used for the assay of HHV-6 and -7 neutralizing antibodies in human serum samples. The neutralization endpoints determined by the dot blot were comparable to those determined by immunofluorescence (IF). The neutralizing antibody titers appeared to correlate with the antibody titers determined by the indirect IF antibody test. The dot blot neutralization assay is easy to perform, is highly reproducible and objective when compared with the conventional methods based on cytopathology or IF for determining neutralization endpoints. (C) 1998 Elsevier Science B.V. All rights reserved.

DOI： 10.1016/S0166-0934(98)00051-2

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High-resolution immuno-scanning electron microscopy using a non-coating method: study of herpes simplex virus glycoproteins on the surface of virus particles and infected cells Reviewed

J. A. Padilla, F. Uno, M. Yamada, H. Namba, S. Nii

Journal of Electron Microscopy 46 ( 2 ) 171 - 180 1997.1

Language：English Publisher：Oxford University Press (OUP)

The expression of two glycoproteins, i.e. glycoprotein C (gC) and glycoprotein D (gD). of herpes simplex virus type 1 (HSV-1) on the surface of extracellular particles of this virus was examined by immune-scanning electron microscopy. Scanning electron microscopy specimens of infected cells immuno-labelled against the glycoproteins with colloidal gold particles were prepared by a conventional coating and a non-coating method. Surface ultrastructure of infected cells and gold particles were observed more clearly with specimens prepared by the non-coating method. The appearance of virus particles in association with glycoprotein expression on these particles and on the surface of infected cells was then studied. Progeny virus particles began to appear 6 h after infection, increased in number as the infection proceeded, and covered most of the cell surface by 16 h. Six to 24 h after the infection, the labelling density for each glycoprotein on virus particles remained constant. The labelling density for gD was always higher than that for gc. The patch-like distribution of gold-labelling against go was often detected on infected cell monolayers al the exponential and late stage of one cycle of virus growth. The labelling density for go on virus particles was the highest on these produced in Vero and L-929 cells, moderate in MRC-5, BHK-21 and FL cells, and the lowest in HEp-2 cells.

DOI： 10.1093/oxfordjournals.jmicro.a023505

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Long PCR and multiple SSCP for mutation analysis of a whole gene.

難波ひかる, 花房裕子, 吉鷹知也, 松原長秀, 佃和憲, 谷野元彦, 清水憲二

日本分子生物学会年会プログラム・講演要旨集 18th 1995

グルタミン酸応答で活性化される遺伝子発現カスケードの解析

佐野訓明, 難波ひかる, 田岡章子, 市川大介, 田淵明子, 土屋友房, 津田正明

日本薬学会年会要旨集 115th ( Pt 3 ) 1995

Possible regulation of BDNF mRNA expression by AP-1 binding activity induced via glutamate receptors.

難波ひかる, 佐野訓明, 市川大介, 土屋友房, 津田正明

日本分子生物学会年会プログラム・講演要旨集 17th 1994

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