157 results on '"Norio Ohashi"'
Search Results
2. Co-infection of tick-borne bacterial pathogens in ticks in Inner Mongolia, China.
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Dan Liu, Wulantuya, Hongxia Fan, Xiaona Li, Fangchao Li, Ting Gao, Xuhong Yin, Zitong Zhang, Minzhi Cao, Hiroki Kawabata, Kozue Sato, Norio Ohashi, Shuji Ando, and Gaowa
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Arctic medicine. Tropical medicine ,RC955-962 ,Public aspects of medicine ,RA1-1270 - Abstract
Tick-borne infectious diseases pose a serious health threat in certain regions of the world. Emerging infectious diseases caused by novel tick-borne pathogens have been reported that are causing particular concern. Several tick-borne diseases often coexist in the same foci, and a single vector tick can transmit two or more pathogens at the same time, which greatly increases the probability of co-infection in host animals and humans and can lead to an epidemic of tick-borne disease. The lack of epidemiological data and information on the specific clinical symptoms related to co-infection with tick-borne pathogens means that it is not currently possible to accurately and rapidly distinguish between a single pathogen infection and co-infection with multiple pathogens, which can have serious consequences. Inner Mongolia in the north of China is endemic for tick-borne infectious diseases, especially in the eastern forest region. Previous studies have found that more than 10% of co-infections were in host-seeking ticks. However, the lack of data on the specific types of co-infection with pathogens makes clinical treatment difficult. In our study, we present data on the co-infection types and the differences in co-infection among different ecological regions through genetic analysis of tick samples collected throughout Inner Mongolia. Our findings may aid clinicians in the diagnosis of concomitant tick-borne infectious diseases.
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- 2023
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3. Surveillance of Borrelia miyamotoi-carrying ticks and genomic analysis of isolates in Inner Mongolia, China
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Gaowa, Wulantuya, Kozue Sato, Dan Liu, Yunhong Cui, Xuhong Yin, Lihua Zhang, Hong Li, Tingfu Wang, Rongxin Liu, Lijing Wu, Saixia Lu, Ting Gao, Zitong Zhang, Minzhi Cao, Guodong Wang, Chunpu Li, Dacheng Yan, Norio Ohashi, Shuji Ando, and Hiroki Kawabata
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Ixodes persulcatus ,Borrelia miyamotoi ,MLSA ,Inner Mongolia ,Infectious and parasitic diseases ,RC109-216 - Abstract
Abstract Background Borrelia miyamotoi is a newly described relapsing fever spirochete transmitted by ixodid tick species. Little is known about the prevalence of B. miyamotoi infections in humans and ticks in Inner Mongolia, China. Therefore, we investigated the prevalence of B. miyamotoi in Ixodes persulcatus ticks, and we aimed to isolateB. miyamotoi from I. persulcatus from four regions of Greater Khingan, Inner Mongolia, China. Methods From May to June each year during the period 2016–2019, host-seeking adult I. persulcatus ticks were collected from vegetation. Genomic DNA was prepared from half of each tick body for PCR template, and the remaining half was used to cultivate B. miyamotoi in BSK-M medium. We employed quantitative real-time PCR (qPCR) to detect Borrelia DNA in the ticks and to calculate the prevalence of B. miyamotoi and infections with other borreliae. For characterization of the isolated B. miyamotoi, we performed draft genome sequencing and multilocus sequencing analysis (MLSA). Results A total of 2656 adult I. persulcatus ticks were collected. The overall prevalence of relapsing fever (RF) borreliae in ticks was 5.0% (134/2656) and that of Lyme disease (LD) borreliae was 43.8% (1164/2656). Co-infection with RF and LD borreliae was observed in 63 ticks (2.4%). Ticks that were positive for RF borreliae by qPCR were subjected to glycerophosphodiester diester phosphodiesterase gene (glpQ) PCR amplification and sequencing, through which we identified the RF borrelia specimens as B. miyamotoi. Furthermore, the B. miyamotoi strain Hetao-1 was isolated from I. persulcatus, and a draft genome sequence was obtained from the isolate. Sequencing determined the strain Hetao-1 genome to be approximately 906.1 kbp in length (28.9% average GC content), and MLSA identified the strain as ST633, which has previously been reported in Japan and Mongolia. Conclusion We detected B. miyamotoi from I. persulcatus ticks collected in Inner Mongolia, and successfully isolated a B. miyamotoi strain. To our knowledge, this is the first study to culture a B. miyamotoi isolate from China. The data on the prevalence of B. miyamotoi and other borreliae in I. persulcatus ticks will be fundamental for future epidemiological studies of B. miyamotoi disease in Inner Mongolia.
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- 2021
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4. Diversity unearthed by the estimated molecular phylogeny and ecologically quantitative characteristics of uncultured Ehrlichia bacteria in Haemaphysalis ticks, Japan
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Hongru Su, Eri Onoda, Hitoshi Tai, Hiromi Fujita, Shigetoshi Sakabe, Kentaro Azuma, Shigehiro Akachi, Saori Oishi, Fuyuki Abe, Shuji Ando, and Norio Ohashi
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Medicine ,Science - Abstract
Abstract Ehrlichia species are obligatory intracellular bacteria transmitted by arthropods, and some of these species cause febrile diseases in humans and livestock. Genome sequencing has only been performed with cultured Ehrlichia species, and the taxonomic status of such ehrlichiae has been estimated by core genome-based phylogenetic analysis. However, many uncultured ehrlichiae exist in nature throughout the world, including Japan. This study aimed to conduct a molecular-based taxonomic and ecological characterization of uncultured Ehrlichia species or genotypes from ticks in Japan. We first surveyed 616 Haemaphysalis ticks by p28-PCR screening and analyzed five additional housekeeping genes (16S rRNA, groEL, gltA, ftsZ, and rpoB) from 11 p28-PCR-positive ticks. Phylogenetic analyses of the respective genes showed similar trees but with some differences. Furthermore, we found that V1 in the V1–V9 regions of Ehrlichia 16S rRNA exhibited the greatest variability. From an ecological viewpoint, the amounts of ehrlichiae in a single tick were found to equal approx. 6.3E+3 to 2.0E+6. Subsequently, core-partial-RGGFR-based phylogenetic analysis based on the concatenated sequences of the five housekeeping loci revealed six Ehrlichia genotypes, which included potentially new Ehrlichia species. Thus, our approach contributes to the taxonomic profiling and ecological quantitative analysis of uncultured or unidentified Ehrlichia species or genotypes worldwide.
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- 2021
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5. Tomato saponin supplementation ameliorates the development of experimental arthritis by regulating inflammatory responses
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Yuko Yoshikawa, Yuki Katayanagi, Manatsu Kamiya, Yuri Yamamoto, Ryuta Fukutomi, Shinjiro Imai, Noriyuki Miyoshi, and Norio Ohashi
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Collagen-induced arthritis ,Esculeogenin A ,Inflammatory cytokines ,Tomato saponin ,Nutrition. Foods and food supply ,TX341-641 - Abstract
We evaluated the effect of tomato saponin, containing esculeoside A, a major saponin found in Japanese pink-colored tomato, on collagen-induced arthritis DBA/1J mice. Arthritis scores in collagen-injected mice were markedly lower by tomato saponin supplementation than that of the control throughout the 15-days observation period. Notably, there was a significant negative correlation between the arthritis scores and serum concentrations of esculeogenin A, an aglycon of esculeoside A (R2 = 0.84, p
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- 2018
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6. Spotted Fever Group Rickettsiae in Inner Mongolia, China, 2015–2016
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Gaowa, Wulantuya, Xuhong Yin, Shengchun Guo, Chunlian Ding, Minzhi Cao, Hiroki Kawabata, Kozue Sato, Shuji Ando, Hiromi Fujita, Fumihiko Kawamori, Hongru Su, Masahiko Shimada, Yuko Shimamura, Shuichi Masuda, and Norio Ohashi
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Rickettsia ,Rickettsia raoultii ,Rickettsia aeschlimannii ,Hyalomma asiaticum ,Dermacentor nuttalli ,Hyalomma marginatum ,Medicine ,Infectious and parasitic diseases ,RC109-216 - Abstract
We found Rickettsia raoultii infection in 6/261 brucellosis-negative patients with fever of unknown origin in brucellosis-endemic Inner Mongolia, China. We further identified Hyalomma asiaticum ticks associated with R. raoultii, H. marginatum ticks associated with R. aeschlimannii, and Dermacentor nuttalli ticks associated with both rickettsiae species in the autonomous region.
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- 2018
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7. Effect of (−)-Epigallocatechin Gallate to Staphylococcal Enterotoxin A on Toxin Activity
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Yuko Shimamura, Mio Utsumi, Chikako Hirai, Ami Kurokawa, Toshiyuki Kan, Norio Ohashi, and Shuichi Masuda
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catechin ,(−)-epigallocatechin gallate ,staphylococcal enterotoxin A ,binding ,Organic chemistry ,QD241-441 - Abstract
Staphylococcal enterotoxin A (SEA) functions both as superantigens that stimulate non-specific T cell proliferation as well as potent gastrointestinal toxins. We previously reported that (−)-epigallocatechin gallate (EGCG) binds to SEA. Therefore, the ability of EGCG to inhibit SEA toxin activity was examined. As a result, EGCG significantly decreased SEA-induced expression and production of interferon gamma (IFN-γ). In addition, EGCG inhibited SEA-induced spleen cell proliferation. To investigate the role of the galloyl group in EGCG on SEA cytotoxicity in more detail, the effect of the binding of a hydroxyl group at position 3 of the galloyl group in EGCG to SEA on SEA cytotoxicity was examined using two methylated EGCG. SEA cytotoxicity was significantly controlled in both (−)-3′′-Me-EGCG and (−)-4′′-Me-EGCG. These results suggest that EGCG inhibits toxic activity via direct interaction with SEA or without any interaction with SEA. The binding affinity between SEA and EGCG under in vivo conditions was examined using a model solution. Although after treatment under acidic and alkaline conditions, the presence of protein and the digestive tract model solution, EGCG still interacted with SEA. Our studies are the first to demonstrate the effect of the binding of EGCG to SEA on toxin activity.
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- 2020
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8. Human Granulocytic Anaplasmosis, Japan
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Norio Ohashi, Gaowa, Wuritu, Fumihiko Kawamori, Dongxing Wu, Yuko Yoshikawa, Seizou Chiya, Kazutoshi Fukunaga, Toyohiko Funato, Masaaki Shiojiri, Hideki Nakajima, Yoshiji Hamauzu, Ai Takano, Hiroki Kawabata, Shuji Ando, and Toshio Kishimoto
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Anaplasma phagocytophilum ,anaplasmosis ,Rickettsia japonica ,rickettsiosis ,p44/msp2 ,16S rDNA ,Medicine ,Infectious and parasitic diseases ,RC109-216 - Abstract
We retrospectively confirmed 2 cases of human Anaplasma phagocytophilum infection. Patient blood samples contained unique p44/msp2 for the pathogen, and antibodies bound to A. phagocytophilum antigens propagated in THP-1 rather than HL60 cells. Unless both cell lines are used for serodiagnosis of rickettsiosis-like infections, cases of human granulocytic anaplasmosis could go undetected.
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- 2013
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9. Binding of Catechins to Staphylococcal Enterotoxin A
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Yuko Shimamura, Mio Utsumi, Chikako Hirai, Shogo Nakano, Sohei Ito, Ai Tsuji, Takeshi Ishii, Takahiro Hosoya, Toshiyuki Kan, Norio Ohashi, and Shuichi Masuda
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staphylococcal enterotoxin A ,catechins ,(−)-epigallocatechin gallate ,molecular docking ,Organic chemistry ,QD241-441 - Abstract
Staphylococcal enterotoxin A (SEA) is a toxin protein, and is the most common cause of staphylococcal food poisoning. Polyphenols, such as catechins, are known to interact with proteins. In this study, we investigated the binding of catechins to SEA using SPR (Biacore), Fourier transform infrared spectroscopy (FT-IR), isothermal titration calorimetry (ITC), and protein-ligand docking. We found that (−)-epigallocatechin gallate (EGCG) could strongly bind to SEA. According to thermodynamic parameters, a negative ΔG indicated that the interaction between EGCG and SEA was spontaneous, and the electrostatic force accompanied by hydrophobic binding forces may play a major role in the binding. Data from Western blot analysis and docking simulation suggest that the hydroxyl group at position 3 of the galloyl group in the catechin structure was responsible for binding affinity with the Y91 of the A-6 region of SEA active sites. Our results provide further understanding of the binding interactions between catechins and SEA, and the inhibition of toxin activities by catechins.
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- 2018
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10. Anaplasma phagocytophilum–infected Ticks, Japan
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Norio Ohashi, Megumi Inayoshi, Kayoko Kitamura, Fumihiko Kawamori, Daizoh Kawaguchi, Yuusaku Nishimura, Hirotaka Naitou, Midori Hiroi, and Toshiyuki Masuzawa
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Anaplasma phagocytophilum ,anaplasmosis ,Ixodes ,Japan ,p44 protein ,16S ribosomal RNA ,Medicine ,Infectious and parasitic diseases ,RC109-216 - Abstract
We report Anaplasma phagocytophilum infection of Ixodes persulcatus and I. ovatus ticks in Japan. Unique p44/msp2 paralogs (and/or 16S rRNA genes) were detected in tick tissues, salivary glands, and spleens of experimentally infected mice. These findings indicate the public health threat of anaplasmosis in Japan.
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- 2005
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11. Rickettsiae in Ticks, Japan, 2007–2011
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Gaowa, Norio Ohashi, Minami Aochi, Wuritu, Dongxing Wu, Yuko Yoshikawa, Fumihiko Kawamori, Toshiro Honda, Hiromi Fujita, Nobuhiro Takada, Yosaburo Oikawa, Hiroki Kawabata, Shuji Ando, and Toshio Kishimoto
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Rickettsiales ,Rickettsia ,rickettsiae ,Anaplasma phagocytophilum ,Ehrlichia ,tick-borne infections ,Medicine ,Infectious and parasitic diseases ,RC109-216 - Published
- 2013
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12. Interaction between Various Apple Procyanidin and Staphylococcal Enterotoxin A and Their Inhibitory Effects on Toxin Activity
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Yuko Shimamura, Chikako Hirai, Yuka Sugiyama, Mio Utsumi, Akio Yanagida, Masatsune Murata, Norio Ohashi, and Shuichi Masuda
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staphylococcal enterotoxin A ,Staphylococcus aureus ,procyanidin ,polymerization ,apple ,Medicine - Abstract
In this study, we investigated the interaction between apple polyphenols (AP; mainly consisting of procyanidin (PC) from an apple) and staphylococcal enterotoxin A (SEA), and the inhibitory effects of AP on SEA activity. According to the degree of polymerization, in particularly highly polymerized PC (more than pentamer) strongly interacted with SEA. The binding affinity of AP with SEA molecules was determined using Biacore analysis. AP reacted with SEA immobilized on a Biacore sensor chip. After treatment with pepsin and pancreatin, to examine the changes of binding affinity of AP in intragastric conditions, AP maintained interaction with SEA. We examined whether AP inhibits the proliferation and interferon-γ (IFN-γ) production induced by SEA in mouse spleen cells. AP strongly inactivated the proliferation and IFN-γ production induced by SEA. These results suggest that AP, which has a higher degree of polymerization, inactivates stronger biological activity of SEA through interaction with SEA. Our studies are the first to demonstrate the relationship between the degree of polymerization of AP and the inhibitory effects on SEA activities.
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- 2017
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13. Anaplasma phagocytophilum Antibodies in Humans, Japan, 2010–2011
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Yuko Yoshikawa, Norio Ohashi, Dongxing Wu, Fumihiko Kawamori, Asaka Ikegaya, Takuya Watanabe, Kazuhito Saitoh, Daisuke Takechi, Yoichi Murakami, Daisuke Shichi, Katsumi Aso, and Shuji Ando
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Anaplasma phagocytophilum ,anaplasmosis ,serodiagnosis ,p44/msp2 ,P44 immunodominant protein ,Japan ,Medicine ,Infectious and parasitic diseases ,RC109-216 - Published
- 2014
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14. Proteomic analysis of growth phase-dependent expression of Legionella pneumophila proteins which involves regulation of bacterial virulence traits.
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Tsuyoshi Hayashi, Masahiro Nakamichi, Hirotaka Naitou, Norio Ohashi, Yasuyuki Imai, and Masaki Miyake
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Medicine ,Science - Abstract
Legionella pneumophila, which is a causative pathogen of Legionnaires' disease, expresses its virulent traits in response to growth conditions. In particular, it is known to become virulent at a post-exponential phase in vitro culture. In this study, we performed a proteomic analysis of differences in expression between the exponential phase and post-exponential phase to identify candidates associated with L. pneumophila virulence using 2-Dimentional Fluorescence Difference Gel Electrophoresis (2D-DIGE) combined with Matrix-Assisted Laser Desorption/Ionization-Mass Spectrometry (MALDI-TOF-MS). Of 68 identified proteins that significantly differed in expression between the two growth phases, 64 were up-regulated at a post-exponential phase. The up-regulated proteins included enzymes related to glycolysis, ketone body biogenesis and poly-3-hydroxybutyrate (PHB) biogenesis, suggesting that L. pneumophila may utilize sugars and lipids as energy sources, when amino acids become scarce. Proteins related to motility (flagella components and twitching motility-associated proteins) were also up-regulated, predicting that they enhance infectivity of the bacteria in host cells under certain conditions. Furthermore, 9 up-regulated proteins of unknown function were found. Two of them were identified as novel bacterial factors associated with hemolysis of sheep red blood cells (SRBCs). Another 2 were found to be translocated into macrophages via the Icm/Dot type IV secretion apparatus as effector candidates in a reporter assay with Bordetella pertussis adenylate cyclase. The study will be helpful for virulent analysis of L. pneumophila from the viewpoint of physiological or metabolic modulation dependent on growth phase.
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- 2010
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15. Correction: Comparative Genomics of Emerging Human Ehrlichiosis Agents.
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Julie C Dunning Hotopp, Mingqun Lin, Ramana Madupu, Jonathan Crabtree, Samuel V Angiuoli, Jonathan A Eisen, Rekha Seshadri, Qinghu Ren, Martin Wu, Teresa R Utterback, Shannon Smith, Matthew Lewis, Hoda Khouri, Chunbin Zhang, Hua Niu, Quan Lin, Norio Ohashi, Ning Zhi, William Nelson, Lauren M Brinkac, Robert J Dodson, M. J Rosovitz, Jaideep Sundaram, Sean C Daugherty, Tanja Davidsen, Anthony S Durkin, Michelle Gwinn, Daniel H Haft, Jeremy D Selengut, Steven A Sullivan, Nikhat Zafar, Liwei Zhou, Faiza Benahmed, Heather Forberger, Rebecca Halpin, Stephanie Mulligan, Jeffrey Robinson, Owen White, Yasuko Rikihisa, and Hervé Tettelin
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Genetics ,QH426-470 - Published
- 2006
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16. Comparative genomics of emerging human ehrlichiosis agents.
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Julie C Dunning Hotopp, Mingqun Lin, Ramana Madupu, Jonathan Crabtree, Samuel V Angiuoli, Jonathan A Eisen, Rekha Seshadri, Qinghu Ren, Martin Wu, Teresa R Utterback, Shannon Smith, Matthew Lewis, Hoda Khouri, Chunbin Zhang, Hua Niu, Quan Lin, Norio Ohashi, Ning Zhi, William Nelson, Lauren M Brinkac, Robert J Dodson, M J Rosovitz, Jaideep Sundaram, Sean C Daugherty, Tanja Davidsen, Anthony S Durkin, Michelle Gwinn, Daniel H Haft, Jeremy D Selengut, Steven A Sullivan, Nikhat Zafar, Liwei Zhou, Faiza Benahmed, Heather Forberger, Rebecca Halpin, Stephanie Mulligan, Jeffrey Robinson, Owen White, Yasuko Rikihisa, and Hervé Tettelin
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Genetics ,QH426-470 - Abstract
Anaplasma (formerly Ehrlichia) phagocytophilum, Ehrlichia chaffeensis, and Neorickettsia (formerly Ehrlichia) sennetsu are intracellular vector-borne pathogens that cause human ehrlichiosis, an emerging infectious disease. We present the complete genome sequences of these organisms along with comparisons to other organisms in the Rickettsiales order. Ehrlichia spp. and Anaplasma spp. display a unique large expansion of immunodominant outer membrane proteins facilitating antigenic variation. All Rickettsiales have a diminished ability to synthesize amino acids compared to their closest free-living relatives. Unlike members of the Rickettsiaceae family, these pathogenic Anaplasmataceae are capable of making all major vitamins, cofactors, and nucleotides, which could confer a beneficial role in the invertebrate vector or the vertebrate host. Further analysis identified proteins potentially involved in vacuole confinement of the Anaplasmataceae, a life cycle involving a hematophagous vector, vertebrate pathogenesis, human pathogenesis, and lack of transovarial transmission. These discoveries provide significant insights into the biology of these obligate intracellular pathogens.
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- 2006
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17. A Case of Human Granulocytic Anaplasmosis Suspected to be Complicated by Cholangitis
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Manri KAWAKAMI, Fusao IKEDA, Shinichi FUJIOKA, Hiromi FUJITA, Koji KIDA, and Norio OHASHI
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General Medicine - Published
- 2022
18. Surveillance of Borrelia miyamotoi-carrying ticks and genomic analysis of isolates in Inner Mongolia, China
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Dacheng Yan, Lijing Wu, Chunpu Li, Lihua Zhang, Tingfu Wang, Dan Liu, Minzhi Cao, Gaowa, Rongxin Liu, Shuji Ando, Ting Gao, Saixia Lu, Hong Li, Zitong Zhang, Xuhong Yin, Kozue Sato, Yunhong Cui, Guodong Wang, Hiroki Kawabata, Norio Ohashi, and Wulantuya
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0301 basic medicine ,MLSA ,China ,relapsing fever ,030231 tropical medicine ,Borrelia miyamotoi ,Ixodes persulcatus ,Infectious and parasitic diseases ,RC109-216 ,Tick ,Real-Time Polymerase Chain Reaction ,law.invention ,03 medical and health sciences ,0302 clinical medicine ,Lyme disease ,law ,Borrelia ,medicine ,Animals ,Humans ,Polymerase chain reaction ,Ixodes ,biology ,Research ,Relapsing Fever ,Genomics ,biology.organism_classification ,medicine.disease ,bacterial infections and mycoses ,Virology ,Bacterial Typing Techniques ,Inner Mongolia ,030104 developmental biology ,Infectious Diseases ,Parasitology ,Epidemiological Monitoring ,Multilocus Sequence Typing - Abstract
Background Borrelia miyamotoi is a newly described relapsing fever spirochete transmitted by ixodid tick species. Little is known about the prevalence of B. miyamotoi infections in humans and ticks in Inner Mongolia, China. Therefore, we investigated the prevalence of B. miyamotoi in Ixodes persulcatus ticks, and we aimed to isolateB. miyamotoi from I. persulcatus from four regions of Greater Khingan, Inner Mongolia, China. Methods From May to June each year during the period 2016–2019, host-seeking adult I. persulcatus ticks were collected from vegetation. Genomic DNA was prepared from half of each tick body for PCR template, and the remaining half was used to cultivate B. miyamotoi in BSK-M medium. We employed quantitative real-time PCR (qPCR) to detect Borrelia DNA in the ticks and to calculate the prevalence of B. miyamotoi and infections with other borreliae. For characterization of the isolated B. miyamotoi, we performed draft genome sequencing and multilocus sequencing analysis (MLSA). Results A total of 2656 adult I. persulcatus ticks were collected. The overall prevalence of relapsing fever (RF) borreliae in ticks was 5.0% (134/2656) and that of Lyme disease (LD) borreliae was 43.8% (1164/2656). Co-infection with RF and LD borreliae was observed in 63 ticks (2.4%). Ticks that were positive for RF borreliae by qPCR were subjected to glycerophosphodiester diester phosphodiesterase gene (glpQ) PCR amplification and sequencing, through which we identified the RF borrelia specimens as B. miyamotoi. Furthermore, the B. miyamotoi strain Hetao-1 was isolated from I. persulcatus, and a draft genome sequence was obtained from the isolate. Sequencing determined the strain Hetao-1 genome to be approximately 906.1 kbp in length (28.9% average GC content), and MLSA identified the strain as ST633, which has previously been reported in Japan and Mongolia. Conclusion We detected B. miyamotoi from I. persulcatus ticks collected in Inner Mongolia, and successfully isolated a B. miyamotoi strain. To our knowledge, this is the first study to culture a B. miyamotoi isolate from China. The data on the prevalence of B. miyamotoi and other borreliae in I. persulcatus ticks will be fundamental for future epidemiological studies of B. miyamotoi disease in Inner Mongolia.
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- 2021
19. Growth Characteristics of Rickettsia Species LON Strains Closely Related to Rickettsia japonica Isolated from Haemaphysalis longicornis Ticks in Mouse Derived L929 and Human-Derived THP-1 Host Cell Lines
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Ai Takano, Hiromi Fujita, Hongru Su, Naoya Takamoto, Fuyuki Abe, Norio Ohashi, Saori Oishi, Shuji Ando, and Hitoshi Tai
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0301 basic medicine ,Microbiology (medical) ,Rickettsia japonica ,biology ,Host (biology) ,030106 microbiology ,General Medicine ,biology.organism_classification ,medicine.disease ,Japonica ,Microbiology ,03 medical and health sciences ,Open reading frame ,chemistry.chemical_compound ,0302 clinical medicine ,Infectious Diseases ,chemistry ,medicine ,bacteria ,Japanese spotted fever ,THP1 cell line ,030212 general & internal medicine ,Haemaphysalis longicornis ,DNA - Abstract
Non-pathogenic Rickettsia species LON strains closely related to an agent of Japanese spotted fever (JSF), R. japonica, were isolated in Japan from Haemaphysalis longicornis ticks in 2001. However, the biological properties of LONs in mammalian host cells are poorly understood. In this study, microscopic analysis showed that LONs in a mouse-derived L929 host cell line were rod shaped with sizes of 0.3-0.5 × 0.5-2.0 μm. Molecular analysis revealed the existence of a LON-specific disrupted open reading frame in R. japonica-related group-specific DNA regions. Growth kinetics of LON-2 and LON-13 strains analyzed by a quantitative real-time PCR showed 100-fold or more increment of LONs cultured in L929 host cells at 30°C and slightly less increment at 33°C, and 25-fold increment in human-derived THP-1 host cells at 35°C on day 7 (168 h) post infection. The generation times of the two LON strains cultured in L929 and THP-1 were estimated to be 9.4-12.9 h and 9.6-10.9 h, respectively. To our knowledge, this is the first report on the biological characteristics of Rickettsia sp. LON strains in mammalian cells, which may provide significant information for the experimental approaches for other rickettsiae.
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- 2021
20. Predominant Shift of Different P44-Expressing Anaplasma phagocytophilum in Infected HL-60, THP-1, NB4, and RF/6A Cell Lines
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Yuko Shimamura, Norio Ohashi, Naoya Takamoto, Masahiko Shimada, Haruka Sasahara, Hongru Su, and Shuji Ando
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0301 basic medicine ,Microbiology (medical) ,endocrine system ,Human granulocytic anaplasmosis ,030106 microbiology ,RNA-Seq ,environment and public health ,03 medical and health sciences ,0302 clinical medicine ,Antigen ,parasitic diseases ,medicine ,Intracellular bacterium ,THP1 cell line ,030212 general & internal medicine ,biology ,General Medicine ,biology.organism_classification ,medicine.disease ,Anaplasma phagocytophilum ,Molecular biology ,enzymes and coenzymes (carbohydrates) ,Infectious Diseases ,Cell culture ,biological phenomena, cell phenomena, and immunity ,Bacterial outer membrane - Abstract
Anaplasma phagocytophilum, an agent of human granulocytic anaplasmosis, is an obligatory intracellular bacterium that dominantly produces P44 outer membrane proteins encoded by the p44/msp2 multigene family, which are major antigens for serodiagnosis. However, A. phagocytophilum antigens from cultures with different cell lines seem to have varying reactivities with sera. In this study, we performed RNA-seq to investigate the P44 expression of A. phagocytophilum propagated in 4 cell lines. In infected HL-60 cells, the P44-2b transcript was predominant in the first RNA-seq analysis (HL-60.1). However, the P44-23 transcript was predominant in the second RNA-seq analysis at 1 month after additional passages (HL-60.2). We further analyzed the P44 expression of A. phagocytophilum cultured in THP-1, NB4, and RF/6A cells through consecutive passages in the same cell lines for 1 year after transferring A. phagocytophilum from infected HL-60 cells to the respective cell lines. In the long-term cultures, P44-18, P44-78, and P44-51 were predominantly transcribed in infected THP-1, NB4, and RF/6A cells, respectively. Therefore, the predominant shifts of different P44-expressing transcripts of A. phagocytophilum might occur during cell culture even in the same cell line at different time points of sample harvest (HL-60.1 and HL-60.2), which may be attributed to host cell adaptation/selection/interaction.
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- 2019
21. Molecular Detection and Characterization of p44/msp2 Multigene Family of Anaplasma phagocytophilum from Haemaphysalis longicornis in Mie Prefecture, Japan
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Shigehiro Akachi, Shuichi Masuda, Yuko Shimamura, Shigetoshi Sakabe, Eri Onoda, Takashi Kanda, Hongru Su, Norio Ohashi, Saori Oishi, Ayaka Sato, Fuyuki Abe, and Hiromi Fujita
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0301 basic medicine ,Microbiology (medical) ,biology ,Obligate ,Phylogenetic tree ,Human granulocytic anaplasmosis ,030106 microbiology ,General Medicine ,Amplicon ,Tick ,bacterial infections and mycoses ,biology.organism_classification ,medicine.disease ,Anaplasma phagocytophilum ,Virology ,03 medical and health sciences ,0302 clinical medicine ,Infectious Diseases ,parasitic diseases ,medicine ,030212 general & internal medicine ,Haemaphysalis longicornis ,Nymph - Abstract
Anaplasma phagocytophilum is an obligate intracellular bacterium that causes human granulocytic anaplasmosis (HGA), an emerging tick-borne infectious disease. This bacterium expresses various 44-kDa major outer membrane proteins encoded by the p44/msp2 multigene family to avoid the host immune system. We previously detected A. phagocytophilum p44/msp2 from the tick Haemaphysalis longicornis in Mie Prefecture, Japan in 2008. In this study, we further investigated a total of 483 H. longicornis ticks (220 adults and 263 nymphs) collected from the Mie Prefecture by PCR targeting p44/msp2 to characterize the p44/msp2 multigene family of A. phagocytophilum. Six of the 483 ticks tested were PCR-positive for A. phagocytophilum p44/msp2, and these positive individuals were at the nymph stage of the tick life cycle. Cloning, sequencing, and phylogenetic analyses of the amplicons revealed that the 11 p44/msp2 clones obtained from the positive ticks shared a 54.9%-99.3% amino acid sequence similarity with the 27 previously identified clones from HGA patients in Japan. In particular, 6 p44/msp2 clones displayed the highest similarities (97.2%-99.3%) with 3 previously identified clones (FJ417343, FJ417345, FJ417357). Thus, the data from this study provide important public health information regarding A. phagocytophilum infection transmitted by H. longicornis ticks, especially at the nymph stage.
- Published
- 2019
22. Surveillance of Borrelia Miyamotoi-Carrying Ticks and Genomic Analysis of Isolates in Inner Mongolia, China
- Author
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H Gaowa, W Wulantuya, Kozue Sato, Dan Liu, Yunhong Cui, Xuhong Yin, Lihua Zhang, Hong Li, Tingfu Wang, Rongxin Liu, Lijing Wu, Saixia Lu, Ting Gao, Zitong Zhang, Minzhi Cao, Guodong Wang, Chunpu Li, Dacheng Yan, Norio Ohashi, Shuji Ando, and Hiroki Kawabata
- Abstract
Background: Borrelia miyamotoi is a newly described relapsing fever spirochete transmitted by Ixodid tick species. Little is known about the prevalence of B. miyamotoi infections in humans and ticks in Inner Mongolia, China. Therefore, we investigated the prevalence of B. miyamotoi in Ixodes persulcatus, and we aimed to isolate B. miyamotoi from I. persulcatus ticks from four regions of Greater Hinggan, Inner Mongolia, China.Methods: During May to June of 2016-2019, host-seeking adult ticks of I. persulcatus were collected from vegetation. Genomic DNA was prepared from half of each tick body for PCR template, and the remaining half was used to cultivate B. miyamotoi in BSK-M medium. We employed quantitative real-time PCR (qPCR) to detect Borrelia DNA in the ticks and to calculate the prevalence of B. miyamotoi and other borreliae infections. For the characterization of isolated B. miyamotoi, we performed draft genome sequencing and multi-loci sequencing analysis (MLSA). Results: A total of 2,656 I. persulcatus adult ticks were collected. The over-all prevalence of relapsing fever (RF) borreliae in ticks was 5.0% (134/2,656) and that of Lyme disease (LD) borreliae was 43.8% (1,164/2,656). Co-infection by RF and LD borreliae was observed in 63 ticks (2.4%). Ticks that were positive for RF borreliae on qPCR were subjected to glycerophosphodiester diester phosphodiesterase gene (glpQ) PCR amplification and sequencing, through which we identified the RF borreliae specimens as B. miyamotoi. In this study, we successfully isolated the B. miyamotoi strain Hetao-1 from I. persulcatus, and a draft genome sequence was obtained for the isolate. Genomic sequencing revealed the strain Hetao-1 genome to be approximately 906.1 kbp in length (28.9% average GC content), and MLSA analysis identified the strain as ST633, which was previously reported in Japan and Mongolia.Conclusion: We detected B. miyamotoi from I. persulcatus ticks collected in Inner Mongolia, and successfully isolated a B. miyamotoi strain. To our knowledge, this is the first study to culture a B. miyamotoi isolate in China. The data on the prevalence of B. miyamotoi and other borreliae in I. persulcatus ticks will be fundamental for future epidemiological studies of B. miyamotoi disease in Inner Mongolia.
- Published
- 2021
23. Growth Characteristics of Rickettsia Species LON Strains Closely Related to Rickettsia japonica Isolated from Haemaphysalis longicornis Ticks in Mouse Derived L929 and Human-Derived THP-1 Host Cell Lines
- Author
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Hitoshi, Tai, Hongru, Su, Naoya, Takamoto, Hiromi, Fujita, Ai, Takano, Saori, Oishi, Fuyuki, Abe, Shuji, Ando, and Norio, Ohashi
- Subjects
DNA, Bacterial ,Mice ,Ticks ,Ixodidae ,Japan ,THP-1 Cells ,Animals ,Humans ,Rickettsia ,Spotted Fever Group Rickettsiosis ,Real-Time Polymerase Chain Reaction ,Cell Line - Abstract
Non-pathogenic Rickettsia species LON strains closely related to an agent of Japanese spotted fever (JSF), R. japonica, were isolated in Japan from Haemaphysalis longicornis ticks in 2001. However, the biological properties of LONs in mammalian host cells are poorly understood. In this study, microscopic analysis showed that LONs in a mouse-derived L929 host cell line were rod shaped with sizes of 0.3-0.5 × 0.5-2.0 μm. Molecular analysis revealed the existence of a LON-specific disrupted open reading frame in R. japonica-related group-specific DNA regions. Growth kinetics of LON-2 and LON-13 strains analyzed by a quantitative real-time PCR showed 100-fold or more increment of LONs cultured in L929 host cells at 30°C and slightly less increment at 33°C, and 25-fold increment in human-derived THP-1 host cells at 35°C on day 7 (168 h) post infection. The generation times of the two LON strains cultured in L929 and THP-1 were estimated to be 9.4-12.9 h and 9.6-10.9 h, respectively. To our knowledge, this is the first report on the biological characteristics of Rickettsia sp. LON strains in mammalian cells, which may provide significant information for the experimental approaches for other rickettsiae.
- Published
- 2020
24. Diversity unearthed by the estimated molecular phylogeny and ecologically quantitative characteristics of uncultured Ehrlichia bacteria in Haemaphysalis ticks, Japan
- Author
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Hongru Su, Fuyuki Abe, Shigehiro Akachi, Norio Ohashi, Hitoshi Tai, Kentaro Azuma, Shuji Ando, Hiromi Fujita, Shigetoshi Sakabe, Saori Oishi, and Eri Onoda
- Subjects
0301 basic medicine ,DNA, Bacterial ,Science ,animal diseases ,030106 microbiology ,Ehrlichia ,Tick ,Article ,03 medical and health sciences ,Ticks ,Bacterial Proteins ,Japan ,parasitic diseases ,Animals ,Humans ,Phylogeny ,Genetics ,Infectious-disease epidemiology ,Multidisciplinary ,biology ,Phylogenetic tree ,Ehrlichiosis ,Biodiversity ,biology.organism_classification ,Haemaphysalis ,16S ribosomal RNA ,rpoB ,bacterial infections and mycoses ,Housekeeping gene ,030104 developmental biology ,Molecular phylogenetics ,Medicine ,bacteria ,Infectious diseases - Abstract
Ehrlichia species are obligatory intracellular bacteria transmitted by arthropods, and some of these species cause febrile diseases in humans and livestock. Genome sequencing has only been performed with cultured Ehrlichia species, and the taxonomic status of such ehrlichiae has been estimated by core genome-based phylogenetic analysis. However, many uncultured ehrlichiae exist in nature throughout the world, including Japan. This study aimed to conduct a molecular-based taxonomic and ecological characterization of uncultured Ehrlichia species or genotypes from ticks in Japan. We first surveyed 616 Haemaphysalis ticks by p28-PCR screening and analyzed five additional housekeeping genes (16S rRNA, groEL, gltA, ftsZ, and rpoB) from 11 p28-PCR-positive ticks. Phylogenetic analyses of the respective genes showed similar trees but with some differences. Furthermore, we found that V1 in the V1–V9 regions of Ehrlichia 16S rRNA exhibited the greatest variability. From an ecological viewpoint, the amounts of ehrlichiae in a single tick were found to equal approx. 6.3E+3 to 2.0E+6. Subsequently, core-partial-RGGFR-based phylogenetic analysis based on the concatenated sequences of the five housekeeping loci revealed six Ehrlichia genotypes, which included potentially new Ehrlichia species. Thus, our approach contributes to the taxonomic profiling and ecological quantitative analysis of uncultured or unidentified Ehrlichia species or genotypes worldwide.
- Published
- 2020
25. Effect of (-)-Epigallocatechin Gallate to Staphylococcal Enterotoxin A on Toxin Activity
- Author
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Mio Utsumi, Yuko Shimamura, Chikako Hirai, Toshiyuki Kan, Ami Kurokawa, Shuichi Masuda, and Norio Ohashi
- Subjects
binding ,(−)-epigallocatechin gallate ,Pharmaceutical Science ,Epigallocatechin gallate ,Catechin ,Analytical Chemistry ,chemistry.chemical_compound ,Enterotoxins ,Mice ,Drug Discovery ,Superantigen ,Interferon gamma ,heterocyclic compounds ,Drug Interactions ,Cytotoxicity ,0303 health sciences ,Molecular Structure ,food and beverages ,Gallate ,Hydrogen-Ion Concentration ,medicine.anatomical_structure ,Chemistry (miscellaneous) ,Molecular Medicine ,Cytokines ,Female ,medicine.drug ,Protein Binding ,T cell ,chemical and pharmacologic phenomena ,complex mixtures ,Article ,lcsh:QD241-441 ,03 medical and health sciences ,lcsh:Organic chemistry ,In vivo ,medicine ,Animals ,Physical and Theoretical Chemistry ,030304 developmental biology ,Cell Proliferation ,030306 microbiology ,Organic Chemistry ,Molecular biology ,Pepsin A ,staphylococcal enterotoxin A ,chemistry ,Gene Expression Regulation ,Pancreatin ,sense organs - Abstract
Staphylococcal enterotoxin A (SEA) functions both as superantigens that stimulate non-specific T cell proliferation as well as potent gastrointestinal toxins. We previously reported that (&minus, )-epigallocatechin gallate (EGCG) binds to SEA. Therefore, the ability of EGCG to inhibit SEA toxin activity was examined. As a result, EGCG significantly decreased SEA-induced expression and production of interferon gamma (IFN-&gamma, ). In addition, EGCG inhibited SEA-induced spleen cell proliferation. To investigate the role of the galloyl group in EGCG on SEA cytotoxicity in more detail, the effect of the binding of a hydroxyl group at position 3 of the galloyl group in EGCG to SEA on SEA cytotoxicity was examined using two methylated EGCG. SEA cytotoxicity was significantly controlled in both (&minus, )-3&prime, &prime, Me-EGCG and (&minus, )-4&prime, Me-EGCG. These results suggest that EGCG inhibits toxic activity via direct interaction with SEA or without any interaction with SEA. The binding affinity between SEA and EGCG under in vivo conditions was examined using a model solution. Although after treatment under acidic and alkaline conditions, the presence of protein and the digestive tract model solution, EGCG still interacted with SEA. Our studies are the first to demonstrate the effect of the binding of EGCG to SEA on toxin activity.
- Published
- 2020
26. Tomato saponin supplementation ameliorates the development of experimental arthritis by regulating inflammatory responses
- Author
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Yuri Yamamoto, Norio Ohashi, Shinjiro Imai, Noriyuki Miyoshi, Yuko Yoshikawa, Manatsu Kamiya, Yuki Katayanagi, and Ryuta Fukutomi
- Subjects
0301 basic medicine ,Experimental arthritis ,Observation period ,Saponin ,Medicine (miscellaneous) ,Arthritis ,Pharmacology ,Significant negative correlation ,complex mixtures ,Esculeogenin A ,03 medical and health sciences ,0302 clinical medicine ,parasitic diseases ,Medicine ,TX341-641 ,chemistry.chemical_classification ,Nutrition and Dietetics ,Tomato saponin ,Nutrition. Foods and food supply ,business.industry ,food and beverages ,Serum concentration ,Inflammatory cytokines ,medicine.disease ,carbohydrates (lipids) ,030104 developmental biology ,chemistry ,030220 oncology & carcinogenesis ,Collagen-induced arthritis ,business ,Food Science - Abstract
We evaluated the effect of tomato saponin, containing esculeoside A, a major saponin found in Japanese pink-colored tomato, on collagen-induced arthritis DBA/1J mice. Arthritis scores in collagen-injected mice were markedly lower by tomato saponin supplementation than that of the control throughout the 15-days observation period. Notably, there was a significant negative correlation between the arthritis scores and serum concentrations of esculeogenin A, an aglycon of esculeoside A (R2 = 0.84, p
- Published
- 2018
27. Case Report: Clinical Features of a Case of Suspected Borrelia miyamotoi Disease in Hokkaido, Japan
- Author
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Kozue Sato, Kaori Kiyanagi, Mutsubu Sugawara, Norio Ohashi, Takuya Ito, Hiroki Kawabata, Aya Zamoto-Niikura, Kimiaki Yamano, Takashige Saito, and Hirotaka Yamazaki
- Subjects
Adult ,Male ,0301 basic medicine ,Pathology ,medicine.medical_specialty ,relapsing fever ,030231 tropical medicine ,030106 microbiology ,Glycerophosphoryl diester phosphodiesterase ,Borrelia miyamotoi ,Disease ,Tick ,03 medical and health sciences ,0302 clinical medicine ,Japan ,Virology ,medicine ,Animals ,Humans ,Seroconversion ,biology ,business.industry ,Borrelia ,Relapsing Fever ,Articles ,medicine.disease ,biology.organism_classification ,Dermatology ,High fever ,Anti-Bacterial Agents ,Treatment Outcome ,Infectious Diseases ,Tick-Borne Diseases ,Borrelia species ,Parasitology ,business - Abstract
We herein report a case of suspected Borrelia miyamotoi disease in Hokkaido, Japan. The patient complained of lassitude, arthralgia, and high fever after a tick bite. Furthermore, at the time of consultation, the patient exhibited momentary loss of consciousness and low blood pressure. Laboratory tests revealed elevation of liver enzymes, thrombocytopenia, and increased C-reactive protein. Seroconversion to B. miyamotoi glycerophosphoryl diester phosphodiesterase antigen suggested the patient was infected with a relapsing fever group Borrelia species.
- Published
- 2017
28. Effects of Short-Term Intake of Wheat Bran with Different Particle Sizes on the Murine Intestinal Environment
- Author
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Katsuki Iwai, Ryuta Fukutomi, Kenichi Asada, Yuko Yoshikawa, Norio Ohashi, and Noriyuki Miyoshi
- Subjects
0301 basic medicine ,Marketing ,Bran ,Chemistry ,General Chemical Engineering ,Short-chain fatty acid ,Industrial and Manufacturing Engineering ,Term (time) ,03 medical and health sciences ,030104 developmental biology ,Particle ,Dietary fiber ,Food science ,Food Science ,Biotechnology - Published
- 2017
29. Case of Human Infection with Anaplasma phagocytophilum in Inner Mongolia, China
- Author
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Shuichi Masuda, Norio Ohashi, Jianchang Luo, Naoya Takamoto, Yuhua Lu, Shengchun Guo, Masahiko Shimada, Yuk Shimamura, Hongru Su, Chunlian Ding, Xuhong Yin, Minzhi Cao, Shuji Ando, Gaowa, Hiroki Kawabata, and Wulantuya
- Subjects
Microbiology (medical) ,Orientia tsutsugamushi ,Human granulocytic anaplasmosis ,Ehrlichia ,030231 tropical medicine ,General Medicine ,Biology ,Ixodes persulcatus ,Tick ,bacterial infections and mycoses ,biology.organism_classification ,medicine.disease ,Virology ,Anaplasma phagocytophilum ,Spotted fever ,03 medical and health sciences ,0302 clinical medicine ,Infectious Diseases ,Rickettsia ,parasitic diseases ,medicine ,030212 general & internal medicine - Abstract
Anaplasma phagocytophilum is an obligate intracellular bacterium that causes febrile illness in humans and livestock. A 49-year-old woman was suffering from feverish symptoms, fatigue, arthralgia, general body pain, and anorexia for 2 weeks. Later, she visited the Bayannur Centers for Disease Control and Prevention Hospital in Inner Mongolia, China. Molecular-based diagnostic analysis of the patient's blood revealed that A. phagocytophilum p44 DNA was positive, but Brucella omp31, spotted fever group Rickettsia gltA, Orientia tsutsugamushi 16S rDNA, and Ehrlichia p28 were negative. The amino acid sequences of 9 A. phagocytophilum p44 clones obtained from the patient shared 44-100% similarity among them and were closely related to those of previously identified p44 clones from Canis familiaris (accession no. KJV64194) and from Ixodes persulcatus tick (no. BAN28309). Serological tests using the patient's serum showed that immunoglobulin M (IgM) and IgG titers to A. phagocytophilum antigens were 160 and 20, respectively, determined using indirect immunofluorescence assay, and the reaction to recombinant P44 proteins (rP44-1, rP44-18ES, and/or rP44-47) was confirmed using Western blot analysis. Thus, the results obtained in this study strongly suggest that the patient was infected with A. phagocytophilum. To our knowledge, this is the first case of human anaplasmosis infection in the Inner Mongolia Autonomous Region.
- Published
- 2018
30. Intracellular proliferation of Anaplasma phagocytophilum is promoted via modulation of endoplasmic reticulum stress signaling in host cells
- Author
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Yuko Yoshikawa, Kei Sugimoto, Norio Ohashi, and Yoshitsugu Ochiai
- Subjects
X-Box Binding Protein 1 ,animal diseases ,Immunology ,Apoptosis ,Biology ,Protein Serine-Threonine Kinases ,Microbiology ,Cell Line ,03 medical and health sciences ,eIF-2 Kinase ,Virology ,parasitic diseases ,Endoribonucleases ,Humans ,Protein kinase A ,030304 developmental biology ,0303 health sciences ,Gene knockdown ,Host Microbial Interactions ,030306 microbiology ,ATF6 ,Kinase ,Endoplasmic reticulum ,fungi ,Ehrlichiosis ,bacterial infections and mycoses ,biology.organism_classification ,Endoplasmic Reticulum Stress ,Anaplasma phagocytophilum ,Cell biology ,Activating Transcription Factor 6 ,Unfolded protein response ,bacteria ,Intracellular - Abstract
Anaplasma phagocytophilum, an obligate intracellular bacterium that propagates within host granulocytes, is considered to modify the host intracellular environment for pathogenesis. However, the mechanism(s) underlying such host modifications remain unclear. Here, we aimed to investigate the relation between A. phagocytophilum and endoplasmic reticulum (ER) stress in THP-1 cells. A. phagocytophilum activated the three ER stress sensors: inositol-requiring enzyme-1 (IRE1), protein kinase RNA-like endoplasmic reticulum kinase (PERK), and activating transcription factor-6 (ATF6). IRE1 activation occurred immediately after host cell invasion by A. phagocytophilum; however, the activated IRE1-induced splicing of X-box-binding protein 1 was not promoted during A. phagocytophilum infection. This suppression was sustained even after the doxycycline-mediated elimination of intracellular A. phagocytophilum. IRE1 knockdown accelerated A. phagocytophilum-induced apoptosis and decreased intracellular A. phagocytophilum. These data suggest that A. phagocytophilum utilizes IRE1 activation to promote its own intracellular proliferation. Moreover, PERK and ATF6 partially mediated A. phagocytophilum-induced apoptosis by promoting the expression of CCAAT/enhancer-binding protein homologous protein, which induces the transcription of several proapoptotic genes. Thus, A. phagocytophilum possibly manipulates the host ER stress signals to facilitate intracellular proliferation and infection of surrounding cells before/after host cell apoptosis.
- Published
- 2019
31. Influence of Muscle Fiber Direction on Migration of Salmonella Enteritidis, Staphylococcus aureus, and Escherichia coli into Raw Chicken Breast
- Author
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Yuko Shimamura, Yusuke Tsuchiya, Momoka Shinke, Shuichi Masuda, Norio Ohashi, Mizuki Egawa, and Miki Hiraishi
- Subjects
Staphylococcus aureus ,Meat ,Salmonella enteritidis ,Muscle Fibers, Skeletal ,Colony Count, Microbial ,Breast Neoplasms ,Food Contamination ,medicine.disease_cause ,Microbiology ,03 medical and health sciences ,chemistry.chemical_compound ,medicine ,Escherichia coli ,Animals ,Humans ,Fiber ,030304 developmental biology ,0303 health sciences ,Growth medium ,biology ,030306 microbiology ,Chemistry ,Inoculation ,Pathogenic bacteria ,biology.organism_classification ,Food Microbiology ,Chickens ,Bacteria ,Food Science - Abstract
The influence of muscle fiber direction (parallel or perpendicular) in relation to the inoculation surface on migration of Salmonella Enteritidis, Staphylococcus aureus, and Escherichia coli into raw chicken breasts was examined. Chicken breast samples with two types of surface fibers (running parallel or perpendicular to the surface) were inoculated with cultures of each bacterium. Inoculated samples were stored for 5 min, 1 h, or 24 h at 4°C. After storage, the samples were divided into segments, and bacterial counts were determined in different regions (inoculation surface, inoculation surface to 1 cm, 1 to 2 cm, 2 to 4 cm, and 4 to 6 cm). The migration of bacteria did not change at 5 min or 1 h regardless of fiber direction. However, after 24 h each bacterium was detected at 4 to 6 cm in the pieces of sample with a perpendicular muscle fiber surface cut. Although these bacteria were detected at 4 to 6 cm in samples with muscle fibers perpendicular to the inoculated surface, these results do not clearly indicate that bacteria migrated into the chicken breast. To monitor actual migration of bacteria into the chicken breast, the tops of the perpendicular muscle fibers of the breast sample were inoculated with bioluminescent E. coli Xen-14. Various regions of the breast sample (inoculation surface and cut surfaces at 1, 2, 4, and 6 cm) were stamped directly on growth medium. Culture revealed that the bacteria migrated directly under the contaminated site and dispersed along the surface of the chicken breast segments. More bacteria distributed laterally than migrated directly below the contamination site. These results suggest that the direction of the muscle fibers is a major factor influencing migration of pathogenic bacteria into chicken breast. HIGHLIGHTS
- Published
- 2019
32. Insight of diagnostic performance using B-cell epitope antigens derived from triple P44-related proteins of Anaplasma phagocytophilum
- Author
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Norio Ohashi, Hiroshi Morita, Shinya Kamiyama, Shuji Ando, Kunihiko Umekita, Takashi Kanda, Yasuaki Kawarasaki, Fuyuki Abe, Akihiko Tokaji, Saori Oishi, Mutsuyo Gokuden, Takeshi Kawaguchi, Fumiko Nakadouzono, Kenichi Ikeda, Yuuki Rikitake, Hiroki Kawabata, Hirohisa Nose, Hongru Su, and Keisuke Ito
- Subjects
0301 basic medicine ,Microbiology (medical) ,Anaplasmosis ,Human granulocytic anaplasmosis ,animal diseases ,030106 microbiology ,Blotting, Western ,Sensitivity and Specificity ,Epitope ,law.invention ,03 medical and health sciences ,0302 clinical medicine ,Western blot ,Antigen ,law ,parasitic diseases ,medicine ,Humans ,Serologic Tests ,030212 general & internal medicine ,Amino Acid Sequence ,Immunoassay ,Antigens, Bacterial ,biology ,medicine.diagnostic_test ,General Medicine ,bacterial infections and mycoses ,medicine.disease ,biology.organism_classification ,Virology ,Anaplasma phagocytophilum ,Antibodies, Bacterial ,Infectious Diseases ,biology.protein ,Recombinant DNA ,Epitopes, B-Lymphocyte ,Antibody ,Bacterial Outer Membrane Proteins - Abstract
Human granulocytic anaplasmosis (HGA) is caused by Anaplasma phagocytophilum. Indirect immunofluorescence assay (IFA) is generally used for HGA serodiagnosis. A. phagocytophilum immunodominant P44 major outer membrane proteins are encoded by p44/msp2 multigene family, responsible for IFA reactivity. However, because multiple P44-related proteins may involve immunoreactivity in IFA, the available diagnostic antigens remain obscure. In this study, we identified 12 B-cell epitopes on triple P44-related proteins using peptide array that reacted with 4 HGA patients' sera. Then, peptide spot immunoassay using 14 synthetic peptides derived from those 12 epitopes as antigens was applied for the detection of antibody to A. phagocytophilum from patients with fever of unknown origin. The sensitivities and diagnostic efficiencies of this immunoassay were higher than those of Western blot analysis using 3 recombinant proteins previously developed. Thus, the immunoassay using our epitope-derived antigens, which has higher diagnostic performances, may have significant benefit for HGA serodiagnosis.
- Published
- 2019
33. Molecular Detection and Characterization of p44/msp2 Multigene Family of Anaplasma phagocytophilum from Haemaphysalis longicornis in Mie Prefecture, Japan
- Author
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Hongru, Su, Ayaka, Sato, Eri, Onoda, Hiromi, Fujita, Shigetoshi, Sakabe, Shigehiro, Akachi, Saori, Oishi, Fuyuki, Abe, Takashi, Kanda, Yuko, Shimamura, Shuichi, Masuda, and Norio, Ohashi
- Subjects
Ticks ,Natural Cytotoxicity Triggering Receptor 2 ,Multigene Family ,Animals ,Genetic Variation ,Amino Acid Sequence ,Polymerase Chain Reaction ,Anaplasma phagocytophilum ,Bacterial Outer Membrane Proteins - Abstract
Anaplasma phagocytophilum is an obligate intracellular bacterium that causes human granulocytic anaplasmosis (HGA), an emerging tick-borne infectious disease. This bacterium expresses various 44-kDa major outer membrane proteins encoded by the p44/msp2 multigene family to avoid the host immune system. We previously detected A. phagocytophilum p44/msp2 from the tick Haemaphysalis longicornis in Mie Prefecture, Japan in 2008. In this study, we further investigated a total of 483 H. longicornis ticks (220 adults and 263 nymphs) collected from the Mie Prefecture by PCR targeting p44/msp2 to characterize the p44/msp2 multigene family of A. phagocytophilum. Six of the 483 ticks tested were PCR-positive for A. phagocytophilum p44/msp2, and these positive individuals were at the nymph stage of the tick life cycle. Cloning, sequencing, and phylogenetic analyses of the amplicons revealed that the 11 p44/msp2 clones obtained from the positive ticks shared a 54.9%-99.3% amino acid sequence similarity with the 27 previously identified clones from HGA patients in Japan. In particular, 6 p44/msp2 clones displayed the highest similarities (97.2%-99.3%) with 3 previously identified clones (FJ417343, FJ417345, FJ417357). Thus, the data from this study provide important public health information regarding A. phagocytophilum infection transmitted by H. longicornis ticks, especially at the nymph stage.
- Published
- 2019
34. Predominant Shift of Different P44-Expressing Anaplasma phagocytophilum in Infected HL-60, THP-1, NB4, and RF/6A Cell Lines
- Author
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Masahiko, Shimada, Naoya, Takamoto, Hongru, Su, Haruka, Sasahara, Yuko, Shimamura, Shuji, Ando, and Norio, Ohashi
- Subjects
Sequence Analysis, RNA ,Gene Expression Profiling ,Animals ,Humans ,Serial Passage ,Macaca mulatta ,Anaplasma phagocytophilum ,Bacterial Outer Membrane Proteins ,Cell Line - Abstract
Anaplasma phagocytophilum, an agent of human granulocytic anaplasmosis, is an obligatory intracellular bacterium that dominantly produces P44 outer membrane proteins encoded by the p44/msp2 multigene family, which are major antigens for serodiagnosis. However, A. phagocytophilum antigens from cultures with different cell lines seem to have varying reactivities with sera. In this study, we performed RNA-seq to investigate the P44 expression of A. phagocytophilum propagated in 4 cell lines. In infected HL-60 cells, the P44-2b transcript was predominant in the first RNA-seq analysis (HL-60.1). However, the P44-23 transcript was predominant in the second RNA-seq analysis at 1 month after additional passages (HL-60.2). We further analyzed the P44 expression of A. phagocytophilum cultured in THP-1, NB4, and RF/6A cells through consecutive passages in the same cell lines for 1 year after transferring A. phagocytophilum from infected HL-60 cells to the respective cell lines. In the long-term cultures, P44-18, P44-78, and P44-51 were predominantly transcribed in infected THP-1, NB4, and RF/6A cells, respectively. Therefore, the predominant shifts of different P44-expressing transcripts of A. phagocytophilum might occur during cell culture even in the same cell line at different time points of sample harvest (HL-60.1 and HL-60.2), which may be attributed to host cell adaptation/selection/interaction.
- Published
- 2018
35. Prevalence of Borrelia miyamotoi in Ixodes persulcatus in Irkutsk City and its neighboring territories, Russia
- Author
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Danchinova Ga, Toshiyuki Masuzawa, Norio Ohashi, Ai Takano, Hiroki Kawabata, and Khasnatinov Ma
- Subjects
0301 basic medicine ,Veterinary medicine ,030231 tropical medicine ,030106 microbiology ,Borrelia miyamotoi ,Tick ,Ixodes persulcatus ,Polymerase Chain Reaction ,Microbiology ,Russia ,03 medical and health sciences ,Human health ,0302 clinical medicine ,RNA, Ribosomal, 16S ,Prevalence ,Animals ,Humans ,Phylogeny ,Lyme Disease ,Ixodes ,biology ,Borrelia ,Sequence Analysis, DNA ,biology.organism_classification ,Infection rate ,Infectious Diseases ,Insect Science ,Arachnid Vectors ,Parasitology ,Nested polymerase chain reaction ,Flagellin ,Lyme Disease Borrelia - Abstract
Adult Ixodes persulcatus were collected in highly populated districts in Irkutsk city, Russia, and in popular recreational and professional areas in its neighboring territories. Borrelia miyamotoi infection in I. persulcatus was examined using multiplex Taqman-PCR targeting 16S rDNA, and nested PCR and sequencing analyses targeting flaB and 16S rDNA. B. miyamotoi and Lyme disease Borrelia species were detected in 13 (infection rate, 2.9%) and 77 (17.3%) out of 445 I. persulcatus ticks, respectively, collected from 4 sites around the Baikal Lake. The 16S rDNA and flaB sequences of these amplicons were closely related to those of B. miyamotoi detected and/or isolated from I. persulcatus in Japan and Far Eastern Russia, and clustered separately from those of Europe and North America. These results indicate that additional surveillance for B. miyamotoi infection is needed in order to determine how it affects human health in Irkutsk City and its neighboring territories.
- Published
- 2016
36. Detection and Characterization of the Emerging Relapsing Fever Pathogen, Borrelia miyamotoi, from the Ixodes ricinus Tick in the Rural Trakya (Thrace) Region of Northwestern Turkey
- Author
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Keiko Sakakibara, Toshiyuki Masuzawa, Ece Şen, Norio Ohashi, Kozue Sato, and Hiroki Kawabata
- Subjects
DNA, Bacterial ,0301 basic medicine ,Ixodes ricinus ,Turkey ,relapsing fever ,030231 tropical medicine ,Borrelia miyamotoi ,Tick ,Communicable Diseases, Emerging ,Microbiology ,03 medical and health sciences ,0302 clinical medicine ,RNA, Ribosomal, 16S ,Virology ,Borrelia ,parasitic diseases ,medicine ,Animals ,Humans ,Phylogeny ,Ixodes ,biology ,Ricinus ,Relapsing Fever ,biology.organism_classification ,medicine.disease ,RNA, Bacterial ,030104 developmental biology ,Infectious Diseases ,Nested polymerase chain reaction ,Flagellin - Abstract
The hard tick-borne relapsing fever agent, Borrelia miyamotoi infection in Ixodes ricinus ticks sampled from Istanbul and the countryside of Kirklareli in northwestern Turkey, was examined by TaqMan-PCR targeting 16S rDNA, nested PCR targeting 16S rDNA, the flagellin gene (flaB), and the 16S and 23S rDNA intergenic spacer (IGS), and sequencing analyses of these amplicons. B. miyamotoi was detected in 1 out of 248 I. ricinus ticks (infection rate 0.4%). The tick infected with B. miyamotoi was collected in Longos, Kirklareli province on the European side of Turkey near the Bulgarian border. The 16S rDNA, flaB, and IGS sequences from the infected tick showed high similarities to those of B. miyamotoi detected in I. ricinus in Europe.
- Published
- 2016
37. Evaluation of Diagnostic Assay for Rickettsioses Using Duplex Real-Time PCR in Multiple Laboratories in Japan
- Author
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Hiroko Sato, Seigo Yamamoto, Naota Monma, Shuichi Masuda, Hiromi Fujita, Yukiko Tamaki, Shuji Ando, Hiroshi Morita, Asaka Ikegaya, Yuko Shimamura, Naoya Takamoto, Y. Shimazu, Hongru Su, Masahiko Shimada, Fumihiko Kawamori, and Norio Ohashi
- Subjects
0301 basic medicine ,Microbiology (medical) ,Adult ,Male ,Orientia tsutsugamushi ,Adolescent ,030106 microbiology ,Eschar ,Real-Time Polymerase Chain Reaction ,Sensitivity and Specificity ,03 medical and health sciences ,Young Adult ,Japan ,Medicine ,Humans ,Rickettsia ,Child ,Aged ,Aged, 80 and over ,Rickettsia japonica ,biology ,business.industry ,Infant ,Rickettsia Infections ,General Medicine ,Tsutsugamushi disease ,Middle Aged ,bacterial infections and mycoses ,biology.organism_classification ,medicine.disease ,Virology ,Spotted fever ,030104 developmental biology ,Infectious Diseases ,Rickettsiosis ,Real-time polymerase chain reaction ,Molecular Diagnostic Techniques ,Child, Preschool ,Japanese spotted fever ,Female ,medicine.symptom ,business ,Multiplex Polymerase Chain Reaction - Abstract
Tsutsugamushi disease and Japanese spotted fever are representative rickettsioses in Japan, and are caused by infection with Orientia tsutsugamushi and Rickettsia japonica, respectively. For molecular-based diagnosis, conventional PCR assays, which independently amplify respective rickettsial DNA, are usually used; however, this approach is time-consuming. Here, we describe a new duplex real-time PCR assay for the simultaneous detection of O. tsutsugamushi and spotted fever group rickettsiae, and its evaluation using several PCR conditions in 6 public health laboratories. The detection limit of the assay was estimated to be 102 copies and the sensitivity was almost identical to that of 3 conventional PCR methods. A total of 317 febrile patients were selected as clinically suspected or confirmed cases of rickettsioses. The detection efficiency of this assay for O. tsutsugamushi from blood or skin (eschar) specimens appeared to be almost the same as that of the conventional PCR method, even when performed in different laboratories, whereas the efficiency for spotted fever group rickettsiae tended to be higher than that of the 2 traditional double PCR assays. Our duplex real-time PCR is thus a powerful tool for the rapid diagnosis of rickettsioses, especially at the acute stage of infection.
- Published
- 2018
38. Binding of Catechins to Staphylococcal Enterotoxin A
- Author
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Sohei Ito, Shuichi Masuda, Takahiro Hosoya, Shogo Nakano, Takeshi Ishii, Mio Utsumi, Yuko Shimamura, Norio Ohashi, Ai Tsuji, Toshiyuki Kan, and Chikako Hirai
- Subjects
0301 basic medicine ,(−)-epigallocatechin gallate ,staphylococcal enterotoxin A ,catechins ,molecular docking ,Pharmaceutical Science ,Calorimetry ,Epigallocatechin gallate ,medicine.disease_cause ,Catechin ,Article ,Analytical Chemistry ,lcsh:QD241-441 ,Enterotoxins ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Western blot ,lcsh:Organic chemistry ,Catalytic Domain ,Spectroscopy, Fourier Transform Infrared ,Drug Discovery ,medicine ,Physical and Theoretical Chemistry ,medicine.diagnostic_test ,Toxin ,Chemistry ,Organic Chemistry ,Isothermal titration calorimetry ,Surface Plasmon Resonance ,Staphylococcal Food Poisoning ,Molecular Docking Simulation ,030104 developmental biology ,Biochemistry ,Chemistry (miscellaneous) ,Docking (molecular) ,Polyphenol ,Thermodynamics ,Molecular Medicine ,030217 neurology & neurosurgery ,Protein Binding - Abstract
Staphylococcal enterotoxin A (SEA) is a toxin protein, and is the most common cause of staphylococcal food poisoning. Polyphenols, such as catechins, are known to interact with proteins. In this study, we investigated the binding of catechins to SEA using SPR (Biacore), Fourier transform infrared spectroscopy (FT-IR), isothermal titration calorimetry (ITC), and protein-ligand docking. We found that (−)-epigallocatechin gallate (EGCG) could strongly bind to SEA. According to thermodynamic parameters, a negative ΔG indicated that the interaction between EGCG and SEA was spontaneous, and the electrostatic force accompanied by hydrophobic binding forces may play a major role in the binding. Data from Western blot analysis and docking simulation suggest that the hydroxyl group at position 3 of the galloyl group in the catechin structure was responsible for binding affinity with the Y91 of the A-6 region of SEA active sites. Our results provide further understanding of the binding interactions between catechins and SEA, and the inhibition of toxin activities by catechins.
- Published
- 2018
39. Case of Human Infection with Anaplasma phagocytophilum in Inner Mongolia, China
- Author
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Gaowa, Wulantuya, Xuhong, Yin, Minzhi, Cao, Shengchun, Guo, Chunlian, Ding, Yuhua, Lu, Jianchang, Luo, Hiroki, Kawabata, Shuji, Ando, Hongru, Su, Masahiko, Shimada, Naoya, Takamoto, Yuk, Shimamura, Shuichi, Masuda, and Norio, Ohashi
- Subjects
China ,Ehrlichiosis ,Humans ,Female ,Middle Aged ,Antibodies, Bacterial ,Anaplasma phagocytophilum - Abstract
Anaplasma phagocytophilum is an obligate intracellular bacterium that causes febrile illness in humans and livestock. A 49-year-old woman was suffering from feverish symptoms, fatigue, arthralgia, general body pain, and anorexia for 2 weeks. Later, she visited the Bayannur Centers for Disease Control and Prevention Hospital in Inner Mongolia, China. Molecular-based diagnostic analysis of the patient's blood revealed that A. phagocytophilum p44 DNA was positive, but Brucella omp31, spotted fever group Rickettsia gltA, Orientia tsutsugamushi 16S rDNA, and Ehrlichia p28 were negative. The amino acid sequences of 9 A. phagocytophilum p44 clones obtained from the patient shared 44-100% similarity among them and were closely related to those of previously identified p44 clones from Canis familiaris (accession no. KJV64194) and from Ixodes persulcatus tick (no. BAN28309). Serological tests using the patient's serum showed that immunoglobulin M (IgM) and IgG titers to A. phagocytophilum antigens were 160 and 20, respectively, determined using indirect immunofluorescence assay, and the reaction to recombinant P44 proteins (rP44-1, rP44-18ES, and/or rP44-47) was confirmed using Western blot analysis. Thus, the results obtained in this study strongly suggest that the patient was infected with A. phagocytophilum. To our knowledge, this is the first case of human anaplasmosis infection in the Inner Mongolia Autonomous Region.
- Published
- 2018
40. Inhibitory effects of food additives derived from polyphenols on staphylococcal enterotoxin A production and biofilm formation by Staphylococcus aureus
- Author
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Yuka Sugiyama, Masatsune Murata, Masaharu Shibata, Yuko Shimamura, Chikako Hirai, Shuichi Masuda, Junya Ozaki, and Norio Ohashi
- Subjects
0301 basic medicine ,Staphylococcus aureus ,food.ingredient ,030106 microbiology ,Enterotoxin ,Biology ,Staphylococcal enterotoxin A ,medicine.disease_cause ,Inhibitory postsynaptic potential ,Applied Microbiology and Biotechnology ,Biochemistry ,Analytical Chemistry ,Microbiology ,03 medical and health sciences ,chemistry.chemical_compound ,Enterotoxins ,food ,Tannic acid ,medicine ,Molecular Biology ,Food additive ,Organic Chemistry ,Biofilm ,Polyphenols ,General Medicine ,Gene Expression Regulation, Bacterial ,030104 developmental biology ,chemistry ,Polyphenol ,Biofilms ,Food Additives ,Biotechnology - Abstract
In this study, we examined the inhibitory effects of 14 food additives derived from polyphenol samples on staphylococcal enterotoxin A (SEA) production and biofilm formation by Staphylococcus aureus. Tannic acid AL (TA), Purephenon 50 W (PP) and Polyphenon 70A (POP) at 0.25 mg/mL and Gravinol®-N (GN), Blackcurrant polyphenol AC10 (BP), and Resveratrol-P5 (RT) at 1.0 mg/mL significantly decreased SEA production by S. aureus C-29 (p
- Published
- 2017
41. Interaction between Various Apple Procyanidin and Staphylococcal Enterotoxin A and Their Inhibitory Effects on Toxin Activity
- Author
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Shuichi Masuda, Norio Ohashi, Mio Utsumi, Yuko Shimamura, Akio Yanagida, Yuka Sugiyama, Masatsune Murata, and Chikako Hirai
- Subjects
0301 basic medicine ,Staphylococcus aureus ,Pentamer ,Health, Toxicology and Mutagenesis ,apple ,lcsh:Medicine ,Degree of polymerization ,Toxicology ,Inhibitory postsynaptic potential ,01 natural sciences ,Article ,Catechin ,Enterotoxins ,Interferon-gamma ,03 medical and health sciences ,Pepsin ,Animals ,Biflavonoids ,Drug Interactions ,Proanthocyanidins ,Cell Proliferation ,biology ,Chemistry ,010401 analytical chemistry ,lcsh:R ,Polyphenols ,Biological activity ,Hydrogen-Ion Concentration ,Pepsin A ,staphylococcal enterotoxin A ,procyanidin ,polymerization ,0104 chemical sciences ,Mice, Inbred C57BL ,030104 developmental biology ,Biochemistry ,Proanthocyanidin ,Polymerization ,Polyphenol ,Malus ,Pancreatin ,biology.protein ,Female ,Spleen - Abstract
In this study, we investigated the interaction between apple polyphenols (AP; mainly consisting of procyanidin (PC) from an apple) and staphylococcal enterotoxin A (SEA), and the inhibitory effects of AP on SEA activity. According to the degree of polymerization, in particularly highly polymerized PC (more than pentamer) strongly interacted with SEA. The binding affinity of AP with SEA molecules was determined using Biacore analysis. AP reacted with SEA immobilized on a Biacore sensor chip. After treatment with pepsin and pancreatin, to examine the changes of binding affinity of AP in intragastric conditions, AP maintained interaction with SEA. We examined whether AP inhibits the proliferation and interferon-γ (IFN-γ) production induced by SEA in mouse spleen cells. AP strongly inactivated the proliferation and IFN-γ production induced by SEA. These results suggest that AP, which has a higher degree of polymerization, inactivates stronger biological activity of SEA through interaction with SEA. Our studies are the first to demonstrate the relationship between the degree of polymerization of AP and the inhibitory effects on SEA activities.
- Published
- 2017
42. Simple and Sensitive Analysis of Histamine and Tyramine in Japanese Soy Sauces and Their Intermediates Using the Stable Isotope Dilution HILIC–MS/MS Method
- Author
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Sachiyo Kitagawa, Yasuhiro Ishii, Tomoyuki Yamanaka, Kenichiro Todoroki, Koichi Inoue, Chiemi Miyauchi, Norio Ohashi, Yuko Yoshikawa, Kuniaki Suzuki, Toshimasa Toyo'oka, and Jun Zhe Min
- Subjects
Spectrometry, Mass, Electrospray Ionization ,Time Factors ,Food Handling ,Industrial Waste ,Tyramine ,Tandem mass spectrometry ,Mass spectrometry ,High-performance liquid chromatography ,chemistry.chemical_compound ,Japan ,Limit of Detection ,Tandem Mass Spectrometry ,Ammonium formate ,Food-Processing Industry ,Detection limit ,Chromatography ,Hydrophilic interaction chromatography ,Selected reaction monitoring ,Reproducibility of Results ,Soy Foods ,General Chemistry ,Food Inspection ,chemistry ,Fermentation ,Condiments ,General Agricultural and Biological Sciences ,Histamine - Abstract
We established a simple, sensitive, and reproducible method to analyze the histamine and tyramine levels in Japanese soy sauce and its mash (called moromi) using hydrophilic interaction liquid chromatography with tandem mass spectrometry (HILIC-MS/MS). Histamine and tyramine quantification was performed using their stable isotopes for electrospray ionization-tandem mass spectrometry in the selected reaction monitoring mode. The sample pretreatment process was a simple, one-step liquid-liquid extraction. HILIC separation was performed with a gradient elution of aqueous ammonium formate and acetonitrile. Because of validation tests, the linearity, the accuracies, and precisions were sufficient. The limit of detection and the limit of quantification were 0.09 and 0.29 ppm for histamine and 0.13 and 0.42 ppm for tyramine, respectively. We successfully applied this method to histamine and tyramine determination in four kinds of commercial Japanese soy sauces and also in moromi samples during soy sauce production.
- Published
- 2014
43. Amelioration of Citrobacter rodentium proliferation in early stage of infection in mice by pretreatment with Lactobacillus brevis KB290 and verification using in vivo bioluminescence imaging
- Author
-
Naoko, Waki, Yuki, Kuwabara, Yuko, Yoshikawa, Hiroyuki, Suganuma, Hiroyuki, Koide, Naoto, Oku, and Norio, Ohashi
- Subjects
Mice ,Colon ,Probiotics ,Antibiosis ,Levilactobacillus brevis ,Luminescent Measurements ,Enterobacteriaceae Infections ,Animals ,Citrobacter rodentium ,Cytokines ,Gene Expression ,Female ,Inflammation Mediators - Abstract
Bioluminescent imaging (BLI) has become a useful tool for monitoring bacterial infections in real time. Citrobacter rodentium and its BLI are widely used as a murine model of enteropathogenic and enterohaemorrhagic Escherichia coli infection. In this study, we evaluated the protective effects of the probiotic Lactobacillus brevis KB290 against C. rodentium infection by the BLI approach. First, we examined several solutions for making the suspension of bioluminescent C. rodentium for an oral inoculation to establish a stable intestinal infection. Three percent NaHCO3 solution was found to be the best. Subsequently, mice were orally administered KB290 once daily for 7 days before inoculation with bioluminescent C. rodentium and for 8 days after infection. The bioluminescence intensity of mice fed with KB290 was significantly lower than that of unfed mice on days 1-3 after infection. The mRNA levels of tumour necrosis factor-α and interferon-γ in the distal colon from KB290-fed mice were shown to be significantly higher than those from unfed mice on day 3 after infection. The results suggested that KB290 intake partially inhibited the proliferation of C. rodentium, especially in the early stages of infection, viathe moderate enhancement of tumour necrosis factor-α and interferon-γ production in the colon.
- Published
- 2016
44. Detection and Characterization of p44/msp2 Transcript Variants of Anaplasma phagocytophilum from Naturally Infected Ticks and Wild Deer in Japan
- Author
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null Gaowa, null Wuritu, Dongxing Wu, Yuko Yoshikawa, Norio Ohashi, Fumihiko Kawamori, Kanji Sugiyama, Masayoshi Ohtake, Masataka Ohashi, Seigo Yamamoto, Tomokazu Kitano, Nobuhiro Takada, and Hiroki Kawabata
- Subjects
Microbiology (medical) ,Infectious Diseases ,General Medicine - Published
- 2012
45. Serotype, Shiga Toxin (Stx) Type, and Antimicrobial Resistance of Stx-Producing Escherichia coli Isolated from Humans in Shizuoka Prefecture, Japan (2003^|^ndash;2007)
- Author
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Tetsuya Harada, Naomi Takahashi, Takashi Masuda, Midori Hiroi, Yukiko Hara-Kudo, Natsuko Iida, Kanji Sugiyama, Takashi Kanda, Fumihiko Kawamori, and Norio Ohashi
- Subjects
Microbiology (medical) ,Serotype ,education.field_of_study ,biology ,Nalidixic acid ,Population ,Shiga toxin ,General Medicine ,biochemical phenomena, metabolism, and nutrition ,Fosfomycin ,bacterial infections and mycoses ,Antimicrobial ,Virology ,Microbiology ,Ciprofloxacin ,fluids and secretions ,Infectious Diseases ,Antibiotic resistance ,biology.protein ,medicine ,bacteria ,education ,medicine.drug - Abstract
SUMMARY: The serotype, Shiga toxin (Stx) type, and antimicrobial resistance patterns of 138 Stxproducing Escherichia coli (STEC) strains isolated from humans between 2003 and 2007 in Shizuoka Prefecture, Japan were characterized. The predominant O serogroups of the STEC isolates were O157, O26, and O111. Antimicrobial susceptibility testing of the STEC isolates showed that 31 of the 138 isolates (22.5z) were resistant to antibiotics. Compared to the results reported in the previous studies, a higher rate of STEC O157 isolates were susceptible to all the antimicrobial agents used in this study. However, antimicrobial susceptibility data from this study showed that antimicrobial resistance patterns have increased by 6 compared to the survey performed by Masuda et al. between 1987 and 2002 (Jpn. J. Food Microbiol., 21, 44–51, 2004). This indicates that STEC isolates have evolved to show a variety of antimicrobial resistance patterns. It is important to consider the population of isolates showing decreased susceptibility to clinically relevant drugs such as ciprofloxacin (CPFX) and fosfomycin (FOM). All the 3 STEC isolates resistant to nalidixic acid showed low susceptibility to CPFX (MIC, 0.25–0.5 mg/ml). In addition, a decreased susceptibility to FOM was clearly observed in the E. coli O26 isolates. Our findings also showed that 1 STEC O26 strain could possibly be a chromosomal AmpC blactamase hyperproducer. These results suggest that antimicrobial therapy may be less effective in patients with non-O157 STEC infections than in those with STEC O157 infections.
- Published
- 2012
46. Molecular detection of Anaplasma phagocytophilum and Borrelia burgdorferi in Ixodes ricinus ticks from Istanbul metropolitan area and rural Trakya (Thrace) region of north-western Turkey
- Author
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Ece Şen, Teruki Kadosaka, Norio Ohashi, Yoshiyuki Uchishima, Toshiyuki Masuzawa, Yoshihiro Okamoto, and Takashi Fukui
- Subjects
Ixodes ricinus ,Turkey ,Human granulocytic anaplasmosis ,animal diseases ,Biology ,Microbiology ,Sensu ,RNA, Ribosomal, 16S ,Recreational Parks ,parasitic diseases ,medicine ,Animals ,Borrelia burgdorferi ,Phylogeny ,Ixodes ,Lyme borreliosis ,fungi ,bacterial infections and mycoses ,medicine.disease ,biology.organism_classification ,Metropolitan area ,Virology ,Anaplasma phagocytophilum ,Infectious Diseases ,Insect Science ,bacteria ,Arachnid Vectors ,Parasitology - Abstract
We demonstrated the presence of the agent of human granulocytic anaplasmosis (HGA), Anaplasma phagocytophilum, and the agent of Lyme borreliosis, Borrelia burgdorferi sensu lato, in north-western Turkey. A total of 241 questing Ixodes ricinus adult ticks were sampled by flagging from recreational parks of the Asiatic and European sides of the heavily populated Istanbul metropolitan area and rural forests of Kirklareli located in the Thrace region in 2008. Both tick-borne pathogens were detected and identified by PCR and DNA sequencing analysis. A. phagocytophilum infection rates were 2.7% in Istanbul and 17.5% in the Kirklareli area. B. burgdorferi sensu lato infection rates were 38.7% in Istanbul and 11.4% in the Kirklareli area. Only 3 of 241 ticks were coinfected with A. phagocytophilum and B. burgdorferi sensu lato.
- Published
- 2011
47. Novel concentration method for the detection of norovirus and sapovirus from water using minute particles of amorphous calcium phosphate
- Author
-
Kazue Uchida, Sachie Kawahashi, Kyoko Tomioka, Michiyo Shinohara, Yuko Yoshikawa, Toshitaka Minegishi, Norio Ohashi, Noriko Suzuki, and Shin-ichi Shimada
- Subjects
Calcium Phosphates ,Microbiology (medical) ,viruses ,Virus Attachment ,medicine.disease_cause ,Microbiology ,Sapovirus ,chemistry.chemical_compound ,Virology ,medicine ,Humans ,Centrifugation ,Amorphous calcium phosphate ,Feline calicivirus ,Chromatography ,biology ,Chemistry ,Norovirus ,General Medicine ,biology.organism_classification ,Caliciviridae ,Ultrapure water ,Particulate Matter ,Adsorption ,Water Microbiology ,Citric acid - Abstract
A novel concentration method using minute particles of amorphous calcium phosphate (ACP) was developed for the detection of caliciviruses including norovirus and sapovirus, agents of human gastroenteritis, from water. In seeding experiments with feline calicivirus (FCV), ACP particles were able to adsorb efficiently the viruses in water, and the FCV-concentrated solution was obtained by dissolution of the virus-adsorbing ACP particles with citric acid after centrifugation. By quantitative real-time RT-PCR, the recovery efficiencies from 300 ml ultrapure water seeded with 103, 104 and >105 copies of FCV were 48, 68 and >100 %, respectively. A comparative study showed that in the addition of viruses at
- Published
- 2011
48. Detection of the New Ehrlichia Species Closely Related to Ehrlichia ewingii from Haemaphysalis longicornis in Yonaguni Island, Okinawa, Japan
- Author
-
Naoaki Yokoyama, Fumihiko Kawamori, Satoshi Zakimi, Norio Ohashi, Kotaro Matsumoto, Hisashi Inokuma, Gaowa, Mamoru Ooshiro, Yoshito Katagiri, and Toshihiko Takeuchi
- Subjects
Ehrlichia ewingii ,Ixodidae ,Sequence analysis ,Molecular Sequence Data ,Ehrlichia ,Tick ,Polymerase Chain Reaction ,Haemaphysalis longicornis ,law.invention ,Bacterial Proteins ,Japan ,law ,RNA, Ribosomal, 16S ,parasitic diseases ,Animals ,Phylogeny ,Polymerase chain reaction ,Base Sequence ,General Veterinary ,biology ,Sequence Analysis, RNA ,Chaperonin 60 ,biology.organism_classification ,bacterial infections and mycoses ,Virology ,groEL ,Anaplasmataceae ,RNA, Bacterial ,16SrRNA - Abstract
application/pdf, We collected a total of 206 Haemaphysalis longicornis ticks by flagging in pastures in Yonaguni Island, Okinawa, Japan, in April 2008. Four of the 206 tick DNA samples tested were positive in a polymerase chain reaction (PCR) screening for the 16SrRNA gene of Anaplasmataceae. Partial sequences of 4 PCR products were identical to each other. Longer sequences of the 16SrRNA gene were successfully determined in 2 of the 4 tick samples, and the obtained 1,392 bp and 1,300 bp sequences revealed high similarity to the 16SrRNA gene sequences of the validated Ehrlichia species, including Ehrlichia ewingii, E. chaffeensis, and E. canis (98.3–98.6%). We also sequenced 1,304 bp of the groEL gene from the 2 tick samples, and found that these had the highest similarity to sequences of E. ewingii (94.0–94.4%) in the validated ehrlichial species. Based on the 16SrRNA and groEL gene sequences, the ehrlichial agents detected in this study were similar to the Ehrlichia species detected in Asia and may compose a new Ehrlichia species with other Ehrlichia species detected in Asia.
- Published
- 2011
49. Toxicological evaluation of l-proline in a 90-day feeding study with Fischer 344 rats
- Author
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Hiroshi Ando, Norio Yano, H. Takahashi, Akio Ogata, Y. Kubo, Katsuhiro Yuzawa, Norio Ohashi, Akemichi Nagasawa, Dai Nakae, and Yukie Tada
- Subjects
Male ,medicine.medical_specialty ,Proline ,Urinalysis ,Physiology ,Biology ,Kidney ,Toxicology ,chemistry.chemical_compound ,Sex Factors ,Internal medicine ,Toxicity Tests ,medicine ,Animals ,Blood urea nitrogen ,Creatinine ,Hematologic Tests ,Hematology ,medicine.diagnostic_test ,Body Weight ,Organ Size ,General Medicine ,Rats, Inbred F344 ,Rats ,Red blood cell ,medicine.anatomical_structure ,chemistry ,Dietary Supplements ,Toxicity ,Uric acid ,Female ,Spleen - Abstract
L-proline (L-Pro) is a non-essential amino acid, and has become widely used as supplements and health foods, recently. A subchronic oral toxicity study of L-Pro was conducted with groups of 10 male and 10 female Fischer 344 rats fed a powder diet containing 0%, 0.625%, 1.25%, 2.5% and 5.0% of L-Pro for 90 days. No treatment-related clinical signs and mortality were noted. We observed no clear treatment-related effects with regard to body weight, food intake or urinalysis data. The average daily water intakes of the treated female groups were significantly increased compared to the controls. The hematology (red blood cell parameter) and serum biochemistry (glucose, blood urea nitrogen, creatinine or uric acid) of the treated male and/or female groups were lower than those of the control groups. However, these changes were lacked dose-dependence, and no abnormalities were found in corresponding pathological findings. In conclusion, the no-observed-adverse-effect-level (NOAEL) for L-Pro was determined to be a dietary dose of 5.0% (2772.9 mg/kg body weight/day for males and 3009.3mg/kg body weight/day for females) under the present experimental conditions.
- Published
- 2010
50. Proteomic Identification of Serum Proteins Associated with Stress-Induced Gastric Ulcers in Fasted Rats
- Author
-
Tsutomu Nakayama, Hirotaka Naitou, Kato Ayako, Hiroyuki Sakakibara, Masanobu Akimoto, Takeshi Ishii, Kayoko Shimoi, Makoto Namioka, and Norio Ohashi
- Subjects
Male ,medicine.medical_specialty ,Apolipoprotein B ,Enzyme-Linked Immunosorbent Assay ,Apolipoprotein A-IV ,Proteomics ,Applied Microbiology and Biotechnology ,Biochemistry ,Analytical Chemistry ,Rats, Sprague-Dawley ,APOA4 ,Internal medicine ,Immersion ,Animals ,Medicine ,Stomach Ulcer ,Molecular Biology ,Apolipoproteins A ,chemistry.chemical_classification ,biology ,business.industry ,Organic Chemistry ,Proteins ,Blood Proteins ,General Medicine ,Blood proteins ,digestive system diseases ,Rats ,Blot ,Enzyme ,Endocrinology ,chemistry ,Duodenal Ulcer ,biology.protein ,Creatine kinase ,business ,Stress, Psychological ,Biotechnology - Abstract
Several physical and psychological stresses frequently become triggers for gastrointestinal disorders such as ulcer. In this study, we tried to identify serum proteins as potential biomarkers for the evaluation of stress-induced gastric ulcer. By proteomic analysis using rats with gastric ulcer induced by water immersion and restraint (WIR) stress as an animal model, we found quantitative changes in several serum proteins, including creatine kinase muscle M chain (CK-M) and apolipoprotein A-IV (ApoA4) in the stressed rats. On western blotting and enzyme-linked immunosorbent assay (ELISA), we confirmed that serum CK-M was remarkably increased by WIR stress. However, ApoA4 appeared to be decreased by fasting, but not WIR stress, which is usually applied prior to WIR stress. The findings suggest that these two serum proteins might be useful as biomarkers, CK-M for stress-induced gastric ulcer and ApoA4 for starvation.
- Published
- 2010
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