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  2. Yoshichika Arakawa

Yoshichika Arakawa

  (荒川 宜親)

Profile Information

Affiliation
School of Medicine, Fujita Health University
College of Pharmacy, Kinjo Gakuin University
Graduate School of Medicine, Nagoya University
National Institute of Infectious Diseases
名誉教授, 名古屋大学, 東海国立大学機構
Degree
医学博士(Mar, 1989, 名古屋大学)
Researcher number
10212622
J-GLOBAL ID
201101032201306103
Researcher ID
P-5997-2015
researchmap Member ID
6000030043

In the 1980s, I found that the chromosomal β‐lactamase of Klebsiella pneumoniae

LEN‐1 showed a very high similarity to the R‐plasmid‐mediated penicillinase

TEM‐1 on the amino acid sequence level, and this strongly suggested the origination

of TEM‐1 from the chromosomal penicillinases of K. pneumoniae or related

bacteria. Moreover, the chromosomal K1 β‐lactamase (KOXY) of Klebsiella oxytoca

was found to belong to the class A β‐lactamases that include LEN‐1 and TEM‐1,

although KOXY can hydrolyze cefoperazone (CPZ) like the chromosomal AmpC type

cephalosporinases of various Enterobacteriaceae that can hydrolyze several

cephalosporins including CPZ. Furthermore, my collaborators and I found plural

novel serine‐type β‐lactamases, such as MOX‐1, SHV‐24, TEM‐91, CTX‐M‐64,

CMY‐9, CMY‐19, GES‐3, GES‐4, and TLA‐3, mediated by plasmids. Besides these

serine‐type β‐lactamases, we also first identified exogenously acquired metallo‐

β‐lactamases (MBLs), IMP‐1 and SMB‐1, in imipenem‐resistant Serratia marcescens,

and the IMP‐1‐producing S. marcescens TN9106 became the index case for

carbapenemase‐producing Enterobacteriaceae. I developed the sodium mercaptoacetic

acid (SMA)‐disk test for the simple identification of MBL‐producing

bacteria. We were also the first to identify a variety of plasmid‐mediated 16S

ribosomal RNA methyltransferases, RmtA, RmtB, RmtC, and NpmA, from various

Gram‐negative bacteria that showed very high levels of resistance to a wide

range of aminoglycosides. Furthermore, we first found plasmid‐mediated quinolone

efflux pump (QepA) and fosfomycin‐inactivating enzymes (FosA3 and FosK).

We also first characterized penicillin reduced susceptible Streptococcus agalactiae (PRGBS),

macrolide‐resistant Mycoplasma pneumoniae, as well as Campylobacter jejuni, and

Helicobacter pylori, together with carbapenem‐resistant Haemophilus influenzae.

Research Interests

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Research Areas

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Major Research History

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Education

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Committee Memberships

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Awards

 3

Papers

 296
  • Kirikae T, Mizuguchi Y, Arakawa Y
    ournal of Antimicrobial Chemotherap, 61(3) 612-615-5, 2008  
    OBJECTIVES: To perform a large-scale investigation of Pseudomonas aeruginosa strains with and without drug resistance in Japan. METHODS: We distributed questionnaires to assess isolation rates of P. aeruginosa with and without drug resistance at medical facilities and clinical laboratories throughout Japan during the period January 2003 through June 2006. Completed questionnaires were obtained from 339 medical facilities and 4 clinical laboratories. RESULTS: The total number of P. aeruginosa strains isolated at the medical facilities was 549,746 and that at clinical laboratories was 640,232. Strains resistant to carbapenems, fluoroquinolones (ciprofloxacin or levofloxacin) and amikacin were defined as multidrug-resistant (MDR) strains, and strains resistant to two of these drugs were defined as two-drug-resistant (TDR) strains. The percentage of MDR at medical facilities and clinical laboratories was 2.4% and 1.1%, respectively, and that of TDR isolates was 6.4% and 4.2%, respectively. MDR and TDR isolates were found nationwide. No MDR isolates were found at approximately one-third of the medical facilities each year. The percentages of MDR and TDR isolates increased significantly from 2003 to 2005. P. aeruginosa strains were obtained mainly from the respiratory and urinary tracts, and the percentages of MDR and TDR isolates were particularly increased in the urinary tract during these years. CONCLUSIONS: MDR P. aeruginosa was prevalent nationwide in Japan. The incidence was low, except in a limited number of facilities, but it increased significantly.
  • Nagano N, Nagano Y, Kimura K, Tamai K, Yanagisawa H, Arakawa Y
    Antimicrob Agents Chemother, 52(12) 4258-4267-4267, 2008  
  • Takahashi H, Miya S, Kimura B, Yamane K, Arakawa Y, Fujii T
    Microb Pathog, 45(2) 150-8, 2008  
  • Hironaga M, Yamane K, Inaba M, Haga Y, Arakawa Y
    Jpn J Infect Dis, 61(3) 212-213, 2008  
  • Liu JH, Deng YT, Zeng ZL, Gao JH, Chen L, Arakawa Y, Chen ZL
    Antimicrob Agents Chemothe, 52(8) 2992-2993-2993, 2008  
  • Kimura K, Suzuki S, Wachino J, Kurokawa H, Yamane K, Shibata N, Nagano N, Kato H, Shibayama K, Arakawa Y
    Antimicrob Agents Chemother, 52(8) 2890-2897-2897, 2008  
  • Doi Y, Wachino J, Arakawa Y
    Antimicrob Agents Chemother, 52(6) 2287-2288, 2008  
  • Han HJ, Kamachi K, Okada K, Toyoizumi-Ajisaka H, Sasaki Y, Arakawa Y
    Vaccine, 26(12) 1530-1534-4, 2008  
    Recently, the incidence of reported pertussis cases of adults has dramatically increased in Japan. In the present study, we analyzed seven Bordetella pertussis isolates recovered from adults in Japan using pulsed-field gel electrophoresis (PFGE) and sequencing of their antigenic and virulence-associated proteins, compared with those from children. PFGE analysis demonstrated that the adult strains were closely related to the child strains (78-100% genetic similarity). On the other hand, the genotyping revealed that 71% (5/7) of the adult strains and 47% (25/53) of the child strains had the same combination of antigenic/virulence-associated allelic variants (ptxS1B/prn1/fim2-1/fim3A/fhaB1/tcfA2) as the Japanese vaccine strain Tohama, respectively. In comparison to the child strains, there was no apparent antigenic and genetic shift in the adult strains. Our result suggests that (i) there is no B. pertussis circulating strain specific to adults and (ii) the antigenic/virulence-associated proteins are unrelated to the rise in adult pertussis incidence in Japan.
  • Yamane K, Wachino J, Suzuki S, Arakawa Y
    Antimicrob Agents Chemother, 52(4) 1564-1566-1566, 2008  
  • Nagano N, Isomine S, Kato H, Sasaki Y, Takahashi M, Sakaida K, Nagano Y, Arakawa Y
    J Clin Microbiol, 46(4) 1545-1547, 2008  
  • Kawamura-Sato K, Wachino J, Kondo T, Ito H, Arakawa Y
    J Antimicrob Chemother, 61(3) 568-576, 2008  
  • Yoshihiro Yamaguchi, Wanchun Jin, Kazuyo Matsunaga, Shinnji Ikemizu, Yuriko Yamagata, Jun-Ichi Wachino, Naohiro Shibata, Yoshichika Arakawa, Hiromasa Kurosaki
    JOURNAL OF MEDICINAL CHEMISTRY, 50(26) 6647-6653, Dec, 2007  
    The VIM-2 metallo-beta-lactamase enzyme from Pseudomonas aeruginosa catalyzes the hydrolysis of most P-lactam antibiotics including carbapenems, and there are currently no potent inhibitors of such enzymes. We found rac-2-omega-phenylpropyl-3-mercaptopropionic acid, phenylC3SH, to be a potent inhibitor of VIM-2. The structure of the VIM-2-phenylC3SH complex was determined by X-ray crystallography to 2.3 angstrom. The structure revealed that the thiol group of phenylC3SH bridged to the two zinc(II) ions and the phenyl group interacted with Tyr67(47) on loop I near the active site, by pi-pi stacking interactions. The methylene group interacted with Phe61(42) located at the bottom of loop1 through CH-pi interactions. Dynamic movements were observed in Arg228(185) and Asn233(190) on loop2, compared with the native structure (PDB. code: 1KO3). These results suggest that the above-mentioned four residues play important roles in the binding and recognition of inhibitors or substrates and in stabilizing a loop in the VIM-2 enzyme.
  • Jun-ichi Wachino, Keigo Shibayama, Hiroshi Kurokawa, Kouji Kimura, Kunikazu Yamane, Satowa Suzuki, Naohiro Shibata, Yasuyoshi Ike, Yoshichika Arakawa
    ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 51(12) 4401-4409, Dec, 2007  
    We have isolated a multiple-aminoglycoside-resistant Escherichia coli strain, strain ARS3, and have been the first to identify a novel plasmid-mediated 16S rRNA methyltransferase, NpmA. This new enzyme shared a relatively low level of identity (30%) to the chromosomally encoded 16S rRNA methyltransferase (KamA) of Streptomyces tenjimariensis, an actinomycete aminoglycoside producer. The introduction of a recombinant plasmid carrying npmA could confer on E. coli consistent resistance to both 4,6-disubstituted 2-deoxystreptamines, such as amikacin and gentamicin, and 4,5-disubstituted 2-deoxystreptamines, including neomycin and ribostamycin. The his tidine-tagged NpmA elucidated methyltransferase activity against 30S ribosomal subunits but not against 50S subunits and the naked 16S rRNA molecule in vitro. We further confirmed that NpmA is an adenine N-1 methyltransferase specific for the A1408 position at the A site of 16S rRNA. Drug footprinting data indicated that binding of aminoglycosides to the target site was apparently interrupted by methylation at the A1408 position. These observations demonstrate that NpmA is a novel plasmid-mediated 16S rRNA methyltransferase that provides a panaminoglycoside-resistant nature through interference with the binding of aminoglycosides toward the A site of 16S rRNA through N-1 methylation at position A1408.
  • E. Sawabe, H. Kato, K. Osawa, T. Chida, N. Tojo, Y. Arakawa, N. Okamura
    EUROPEAN JOURNAL OF CLINICAL MICROBIOLOGY & INFECTIOUS DISEASES, 26(10) 695-703, Oct, 2007  
    Clostridium difficile isolates recovered from patients admitted to a teaching hospital in Japan over a 5-year period were analyzed. Two molecular typing systems, PCR ribotyping and pulsed-field gel electrophoresis (PFGE) analysis, were used. Twenty-six PCR ribotypes were found among the 148 isolates. The predominant type at our hospital appeared to shift during the study period, from PCR ribotype a in 2000 (15/33, 45%) to PCR ribotype f in 2004 (18/28, 64%). By using PFGE with thiourea added to both the gel and running buffer, all 148 Clostridium difficile isolates were successfully classified into 37 types and 61 subtypes. The PCR ribotype f isolates were further classified into four types and 11 subtypes by PFGE. The PFGE patterns of the 11 subtypes differed from each other by only 1 to 4 bands, suggesting that these differences might reflect genetic changes during patient-to-patient transmission over the 5-year period analyzed, and that PCR ribotype f isolates might be outbreak-related. In addition, the PCR ribotype f was identical to the PCR ribotype designated smz, which is reported to have caused multiple outbreaks in Japan. These results confirmed that PCR ribotype f (type smz) has specific virulence or survival factors that make it more likely to cause nosocomial outbreaks at Japanese hospitals. PCR ribotype 027, which has been reported to have caused recent outbreaks in North America and Europe, was recovered from one patient in this study; however, this strain was community-acquired. Our findings emphasize the importance of monitoring specific strains to control and prevent nosocomial infection.
  • Masaki Ochiai, Akihiko Yamamoto, Michiyo Kataoka, Hiromi Toyoizumi, Yoshichika Arakawa, Yoshinobu Horiuchi
    BIOLOGICALS, 35(4) 259-264, Oct, 2007  
    Histamine-sensitization test method based on histamine-sensitizing death is widely used for controlling residual activity of pertussis toxin in acellular pertussis vaccines. The test method evaluates the residual activity according to the death of mice injected with a test vaccine after histamine challenge and the test result, therefore, depends on the sensitivity of mice. A highly sensitive test method based on change in rectal temperature of mice has been used in Japan for many years but has limited feasibility in other countries. We examined the possibility of a test method using dermal temperature measured by infrared thermometer to reduce animal suffering instead of rectal temperature. The dermal temperature method was shown to be as sensitive as the rectal temperature method. Furthermore, the dermal as well as rectal temperature methods can evaluate the activity of a test vaccine in relative to a reference preparation so as to allow direct comparison of the test results among different laboratories. The activity by means of the dermal temperature method was also found to be well consistent with that by the rectal temperature method. (C) 2007 The International Association for Biologicals. Published by Elsevier Ltd. All rights reserved.
  • Kunikazu Yamane, Jun-Ichi Wachino, Satowa Suzuki, Kouji Kimura, Naohiro Shibata, Haru Kato, Keigo Shibayama, Toshifumi Konda, Yoshichika Arakawa
    ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 51(9) 3354-3360, Sep, 2007  
    Plasmid-mediated Qnr and AAC(6')-Ib-cr have been recognized as new molecular mechanisms affecting fluoroquinolone (FQ) resistance. C316, an Escherichia coli strain demonstrating resistance to various FQs, was isolated in Japan. Resistance to FQs was augmented in an E. coli CSR2 transconjugant, but PCR failed to detect qnr genes, suggesting the presence of novel plasmid-mediated FQ resistance mechanisms. Susceptibility tests, DNA manipulation, and analyses of the gene and its product were performed to characterize the genetic determinant. A novel FQ-resistant gene, qepA, was identified in a plasmid, pHPA, of E. coli C316, and both qepA and rmtB genes were mediated by a probable transposable element flanked by two copies of IS26. Levels of resistance to norfloxacin, ciprofloxacin, and enrofloxacin were significantly elevated in E. coli transformants harboring qepA under AcrB-TolC-deficient conditions. QepA showed considerable similarities to transporters belonging to the 14-transmembrane-segment family of environmental actinomycetes. The effect of carbonyl cyanide m-chlorophenylhydrazone (CCCP) on accumulation of norfloxacin was assayed in a qepA-harboring E. coli transformant. The intracellular accumulation of norfloxacin was decreased in a qepA-expressing E. coli transformant, but this phenomenon was canceled by CCCP. The augmented FQ resistance level acquired by the probable intergeneric transfer of a gene encoding a major facilitator superfamily-type efflux pump from some environmental microbes to E. coli was first identified. Surveillance of the qepA-harboring clinical isolates should be encouraged to minimize further dissemination of the kind of plasmid-dependent FQ resistance determinants among pathogenic microbes.
  • K. Shibayama, J. Wachino, S. Suzuki, Y. Arakawa
    HELICOBACTER, 12(4) 412-412, Aug, 2007  
  • Yohei Doi, Yoshichika Arakawa
    CLINICAL INFECTIOUS DISEASES, 45(1) 88-94, Jul, 2007  
    Methylation of 16S ribosomal RNA ( rRNA) has recently emerged as a new mechanism of resistance against aminoglycosides among gram-negative pathogens belonging to the family Enterobacteriaceae and glucose-nonfermentative microbes, including Pseudomonas aeruginosa and Acinetobacter species. This event is mediated by a newly recognized group of 16S rRNA methylases, which share modest similarity to those produced by aminoglycoside-producing actinomycetes. Their presence confers a high level of resistance to all parenterally administered aminoglycosides that are currently in clinical use. The responsible genes are mostly located on transposons within transferable plasmids, which provides them with the potential to spread horizontally and may in part explain the already worldwide distribution of this novel resistance mechanism. Some of these organisms have been found to coproduce extended-spectrum beta-lactamases or metallo-beta-lactamases, contributing to their multidrug-resistant phenotypes. A 2-tiered approach, consisting of disk diffusion tests followed by confirmation with polymerase chain reaction, is recommended for detection of 16S rRNA methylase-mediated resistance.
  • Kunikazu Yamane, Jun-ichi Wachino, Satowa Suzuki, Naohiro Shibata, Haru Kato, Keigo Shibayama, Kouji Kimura, Kumiko Kai, Satoshi Ishikawa, Yoshiyuki Ozawa, Toshifumi Konda, Yoshichika Arakawa
    EMERGING INFECTIOUS DISEASES, 13(4) 642-646, Apr, 2007  
    To investigate the exact isolation frequency of 16S rRNA methylase-producing, gram-negative pathogenic bacteria, we tested 87,626 clinical isolates from 169 hospitals. Twenty-six strains from 16 hospitals harbored 16S rRNA methylase genes, which suggests sparse but diffuse spread of pan-aminoglycoside-resistant microbes in Japan.
  • Keigo Shibayama, Jun-ichi Wachino, Yoshichika Arakawa, Massoud Saidijam, Nicholas G. Rutherford, Peter J. F. Henderson
    MOLECULAR MICROBIOLOGY, 64(2) 396-406, Apr, 2007  
    gamma-Glutamyltranspeptidase (GGT) is a periplasmic enzyme of Helicobacter pylori implicated in its pathogenesis towards mammalian cells. We have cloned and expressed the H. pylori strain 26695 recombinant GGT protein in Escherichia coli and purified it to homogeneity. The purified protein exhibited hydrolysis activity with very high affinities for glutamine and glutathione shown by apparent K-m values lower than 1 mu M. H. pylori cells were unable to take up extracellular glutamine and glutathione directly. Instead, these substances were hydrolysed to glutamate by the action of GGT outside the cells. The glutamate produced was then transported by a Na+-dependent reaction into H. pylori cells, where it was mainly incorporated into the TCA cycle and partially utilized as a substrate for glutamine synthesis. These observations show that one of the principle physiological functions of H. pylori GGT is to enable H. pylori cells to utilize extracellular glutamine and glutathione as a source of glutamate. As glutamine and glutathione are important nutrients for maintenance of healthy gastrointestinal tissue, their depletion by the GGT enzyme is hypothesized to account for the damaging of mammalian cells and the pathophysiology of H. pylori.
  • Keigo Shibayama, Keiko Mochida, Tetsuya Yagi, Shigetaro Mori, Yoshichika Arakawa, Saburo Yamamoto
    BIOLOGICALS, 35(2) 139-143, Apr, 2007  
    Japanese bacillus Calmette-Guerin (BCG) vaccine preparation contains two types of variant strains, Type I, which has a 22-base-pair deletion in the RD16 region, and Type II, which has an identical sequence to those of other BCG strains. In this study, we established a method to quantify the percentage of variant strain Type II contained in freeze-dried BCG product with real-time PCR. With this method we examined the master seed lot Tokyo 172, two secondary seed lots, Tokyo 172-1 and Tokyo 172-2, which were produced from Tokyo 172, and four commercial lots produced form Tokyo 172-1. Tokyo 172, Tokyo 172-1, and Tokyo 172-2 contained 55.1%, 19.5%, and 3.6% of Type II variant strain, respectively. Commercial lots contained 1.5%, 4.5%, 7.4%, and 4.3% of Type II variant strain, respectively. These results indicated that the two variant strains contained in the master seed lot continued to coexist in subsequently produced lots with a decrease in population of variant strain Type II. This method would be useful for quality control of commercial Japanese BCG vaccine preparations. (c) 2006 The International Association for Biologicals. Published by Elsevier Ltd. All rights reserved.
  • Moriguchi N, Itahashi Y, Tabata N, Yamazumi T, Furuta I, Shibata N, Arakawa Y, Miyata H
    J Infect Chemother, 13(4) 263-266, 2007  
  • Kato H, Ito Y, van den Berg RJ, Kuijper EJ, Arakawa Y
    Euro Surveill, 12(1) E070111.3., 2007  
  • Masaaki Iwaki, Yoshinobu Honiuchi, Takako Komiya, Tadashi Fukuda, Yoshichika Arakawa, Motohide Takahashi
    JOURNAL OF IMMUNOLOGICAL METHODS, 318(1-2) 138-146, Jan, 2007  
    Internationally accepted designations of antigen content for toxoid vaccines are provided by the WHO in Lf (limes flocculationis) units, based on the formation of antigen-antibody complexes. The current assay method for Lf determination involves observation of the complexes by eye, making the development of a more objective system highly desirable. Here we report a novel detection system using a laser light-scattering platelet aggregometer. The system was highly reproducible and more objective than the current method. Only three sets of duplicate data were sufficient for statistically significant determination of toxoid Lf by parabolic regression. (c) 2006 Elsevier B.V. All rights reserved.
  • Kazunari Kamachi, Yoshichika Arakawa
    VACCINE, 25(6) 1000-1006, Jan, 2007  
    A toxic N-terminal 180-amino-acid fragment (C180) of pertussis toxin S1 subunit has the most potent ability to induce protective immunity against pertussis toxin (PT) following DNA-based immunization [Kamachi K, Arakawa Y. Infect Immun 2004;72:4293-6]. For the development of a safer pertussis DNA vaccine, three plasmids encoding mutant C180 (C180-R9K, C180-E129G and C180-R9K/E129G) were constructed and tested for their protective immunogenicity and cytotoxicity. All of the gene gun delivery of the plasmid, performed by inserting the mutant C180 gene into a mammalian expression vector pcDNA3.1, successfully induced anti-PT IgG antibody production without the loss of immunogenicity in mice. The immunizations of mice with the plasmids significantly inhibited leukocytosis-promoting activity by PT. Among stably transfected Chinese hamster ovary (CHO) cells expressing mutant C180, the expression of C180-R9K and C180-R9K/E129G was non-toxic to the transfectants, confirming that these mutant C180s have no cytotoxicity to mammalian cells. These results indicate that C180-R9K and C180-R9K/E129G genes, especially C180-R9K/E129G, are candidates for safe and effective antigen DNAs in the development of pertussis DNA vaccine. (c) 2006 Elsevier Ltd. All rights reserved.
  • Shibayama K, Wachino J, Arakawa Y, Saidijam M, Rutherford N, Henderson P
    Helicobacter, 11(4) 355-356, 2006  
  • Iwaki M, Arakawa Y
    Letters in Applied Microbiology, 43(2) 215-221, 2006  
  • Kamachi K, Sota M, Tamai Y, Nagata N, Konda T, Inoue T, Top EM, Arakawa Y
    Microbiology, 152(12) 3477-3484-3484, 2006  
    The complete 41,268 bp nucleotide sequence of the IncP-1beta plasmid pBP136 from the human pathogen Bordetella pertussis, the primary aetiological agent of whooping cough, was determined and analysed. This plasmid carried a total of 46 ORFs: 44 ORFs corresponding to the genes in the conserved IncP-1beta backbone, and 2 ORFs similar to the XF1596 and XF1597 genes with unknown function of the plant pathogen Xylella fastidiosa. Interestingly, pBP136 had no accessory genes carrying genetic traits such as antibiotic or mercury resistance and/or xenobiotic degradation. Moreover, pBP136 had only two of the kle genes (kleAE) that have been reported to be important for the stability of IncP-1 plasmid in Pseudomonas aeruginosa. Phylogenetic analysis of the Kle proteins revealed that the KleA and KleE of pBP136 were phylogenetically distant from those of the present IncP-1 plasmids. In contrast, IncC1 and KorC, encoded upstream and downstream of the kle genes respectively, and the replication-initiation protein, TrfA, were closely related to those of the IncP-1beta 'R751 group'. These results suggest that (i) pBP136 without any apparent accessory genes diverged early from an ancestor of the present IncP-1beta plasmids, especially those of the R751 group, and (ii) the kle genes might be incorporated independently into the backbone region of the IncP-1 plasmids for their stable maintenance in various host cells.
  • Tanimoto K, Nomura T, Maruyama H, Tomita H, Shibata N, Arakawa Y, Ike Y
    Antimicrob Agents Chemother, 50(11) 3966-3967-3967, 2006  
  • Shibayama K, Nagasawa M, Ando T, Minami M, Wachino J, Suzuki S, Arakawa Y
    J Clin Microbiol, 44(11) 4255-4257-4257, 2006  
  • Wachino J, Yamane K, Kimura K, Shibata N, Suzuki S, Ike Y, Arakawa Y
    Antimicrob Agents Chemother, 50(9) 3212-3215-3215, 2006  
  • Kurosaki H, Yamaguchi Y, Yasuzawa H, Jin W, Yamagata Y, Arakawa Y
    Chem Med Chem, 1(9) 969-972-972, 2006  
  • Park YJ, Lee S, Yu JK, Woo GJ, Lee K, Arakawa Y
    J Antimicrob Chemother, 58(4) 907-908-908, 2006  
  • Lee H, Yong D, Yum JH, Roh KH, Lee K, Yamane K, Arakawa Y, Chong Y
    Diagn Microbiol Infect Dis, 56(3) 305-312, 2006  
  • Yamazaki T, Narita M, Sasaki N, Kenri T, Arakawa Y, Sasaki T
    Clin Vaccine Immunol, 13(6) 708-710-710, 2006  
  • Kamachi K, Toyoizumi-Ajisaka H, Toda K, Soeung SC, Sarath S, Nareth Y, Horiuchi Y, Kojima K, Takahashi M, Arakawa Y
    J Clin Microbiol, 44(5) 1899-1902-902, 2006  
    We developed a loop-mediated isothermal amplification (LAMP) method to detect Bordetella pertussis infection. This LAMP assay detected B. pertussis with high sensitivity, but not other Bordetella species. Among nasopharyngeal swab samples from subjects with suspected pertussis, LAMP results showed a high level of agreement with results of conventional PCR. This method is a rapid, sensitive, and specific method for diagnosis of B. pertussis infection even in clinical laboratories with no specific equipment.
  • Nagano N, Oana S, Nagano Y, Arakawa Y
    Jpn J Infect Dis, 59(2) 132-134-134, 2006  
  • Phan LT, Ngo TT, Dang DA, Vu TT, Le NM, Tran QC, Matsuoka M, Kamachi K, Yamazaki T, Arakawa Y, Sasaki T
    Jpn J Infect Dis, 59(2) 111-116-116, 2006  
  • Muta T, Tsuruta N, Seki Y, Ota R, Suzuki S, Shibata N, Kato K, Eto T, Gondo H, Arakawa Y
    Jpn J Infect Dis, 59(1) 69-71-71, 2006  
  • Seki N, Sasaki T, Sawabe K, Sasaki T, Matsuoka M, Arakawa Y, Marui E, Kobayashi M
    Jpn J Infect Dis, 59(1) 31-35-35, 2006  
  • Fukuda T, Iwaki M, Hong SH, Oh HJ, Wei Z, Morokuma K, Ohkuma K, Dianliang L, Arakawa Y, Takahashi M
    Jpn J Infect Dis, 59(1) 20-24-24, 2006  
  • Shibata N, Kurokawa H, Doi Y, Yagi T, Yamane K, Wachino J, Suzuki S, Kimura K, Ishikawa S, Kato H, Ozawa Y, Shibayama K, Kai K, Konda T, Arakawa Y
    Antimicrob Agents Chemother, 50(2) 791-795-795, 2006  
  • Suzuki S, Yamazaki T, Narita M, Okazaki N, Suzuki I, Andoh T, Matsuoka M, Kenri T, Arakawa Y, Sasaki T
    Antimicrob Agents Chemother, 50(2) 709-712-712, 2006  
  • Wachino J, Kurokawa H, Suzuki S, Yamane K, Shibata N, Kimura K, Ike Y, Arakawa Y
    Antimicrob Agents Chemother, 50(2) 534-541-541, 2006  
  • Wachino J, Yamane K, Shibayama K, Kurokawa H, Shibata N, Suzuki S, Doi Y, Kimura K, Ike Y, Arakawa Y
    Antimicrob Agents Chemother, 50(1) 178-184-184, 2006  
  • H Kato, T Yokoyama, H Kato, Y Arakawa
    JOURNAL OF CLINICAL MICROBIOLOGY, 43(12) 6108-6112, Dec, 2005  
    We applied the loop-mediated isothermal amplification (LAMP) assay to the detection of the toxin B gene (tcdB) of Clostridium difficile for identification of toxin B (TcdB)-positive C difficile strains and detection of tcdB in stool specimens. tcdB was detected in all toxin A (TcdA)-positive, TcdB-positive (A(+)B(+)) and TcdA-negative, TcdB-positive (A(-)B(+)) C difficile strains but not from TcdA-negative, TcdB-negative strains. Of the 74 stool specimens examined, A(+)B(+) or A(-)B(+) C. difficile was recovered from 39 specimens, of which 38 specimens were LAMP positive and one was negative. Amplification was obtained in 10 specimens that were culture negative, indicating that LAMP is highly sensitive. The LAMP assay was applied to detection of tcdB in DNA extracted by a simple boiling method from 47 of those 74 specimens, which were cultured overnight in cooked-meat medium (CMM). Twenty-two of 24 culture-positive specimens were positive for LAMP on DNA from the culture in CMM. Four specimens were culture negative but positive by LAMP on DNA from CMM cultures. The LAMP assay is a reliable tool for identification of TcdB-positive C difficile as well as for direct detection of tcdB in stool specimens with high sensitivity. Detection of tcdB by LAMP from overnight cultures in CMM could be an alternative method of diagnostic testing at clinical laboratories without special apparatus.
  • K Yamane, JI Wachino, Y Doi, H Kurokawa, Y Arakawa
    EMERGING INFECTIOUS DISEASES, 11(6) 951-953, Jun, 2005  
    Emergence of the newly identified 16S rRNA methylases RmtA, RmtB, and ArmA in pathogenic gram-negative bacilli has been a growing concern. ArmA, which had been identified exclusively in Europe, was also found in several gram-negative pathogenic bacilli isolated in Japan, suggesting global dissemination of hazardous multiple aminoglycoside resistance genes.
  • T Yagi, J Wachino, H Kurokawa, S Suzuki, K Yamane, Y Doi, N Shibata, H Kato, K Shibayama, Y Arakawa
    JOURNAL OF CLINICAL MICROBIOLOGY, 43(6) 2551-2558, Jun, 2005  
    Detection of the resistance mediated by class C beta-lactamases remains a challenging issue, considering that transferable plasmid-mediated class C beta-lactamases are of worldwide concern. Methods for the identification of strains that produce extended-spectrum beta-lactamases (ESBLs) or metallo-beta-lactamases (MBLs) have been developed and applied for routine use in clinical microbiology laboratories, but no practical methods for identification of plasmid-mediated class C producers have been established to date. We therefore developed three simple methods for clinical microbiology laboratories that allow identification of plasmid-mediated class C beta-lactamase-producing bacteria using a boronic acid derivative, 3-aminophenylboronic acid (APB), one of the specific inhibitors of class C beta-lactamases. Detection by the disk potentiation test was based on the enlargement of the growth-inhibitory zone diameter (by greater than or equal to 5 mm) around a Kirby-Bauer disk containing a ceftazidime (CAZ) or a cefotaxime (CTX) disk in combination with APB. In a double-disk synergy test, the discernible expansion of the growth-inhibitory zone around the CAZ or the CTX disk toward a disk containing APB was indicative of class C beta-lactamase production. A greater than or equal to eightfold decrease in the MIC of CAZ or CTX in the presence of APB was the criterion for detection in the microdilution test. By using these methods, Escherichia coli and Klebsiella pneumoniae isolates producing plasmid-mediated class C beta-lactamases, ACT-1, CMY-2, CMY-9, FOX-5, LAT-1, and MOX-1, were successfully distinguished from those producing other classes of beta-lactamases, such as ESBLs and MBLs. These methods will provide useful information needed for targeted antimicrobial therapy and better infection control.
  • M Ohashi, T Yoshikawa, S Akimoto, A Fujita, S Hayakawa, M Takahashi, Y Arakawa, Y Asano
    PEDIATRIC INFECTIOUS DISEASE JOURNAL, 24(5) 466-467, May, 2005  
    A 4-year-old Japanese girl developed a sore throat and high fever. Her tonsils were enlarged, red and covered with a thick white membrane. There was marked leukocytosis (26,600 leukocytes per mm(3)) and elevated C-reactive protein levels (23.3 mg/dL). Rothia dentocariosa was recovered from the throat swab; many Gram-positive cocci were observed in the smear from the pseudomembrane covering the tonsil.
  • Y Yamaguchi, T Kuroki, H Yasuzawa, T Higashi, WC Jin, A Kawanami, Y Yamagata, Y Arakawa, M Goto, H Kurosaki
    JOURNAL OF BIOLOGICAL CHEMISTRY, 280(21) 20824-20832, May, 2005  
    Metallo-beta-lactamase IMP-1 is a di-Zn(II) metalloenzyme that efficiently hydrolyzes beta-lactam antibiotics. Wild-type (WT) IMP-1 has a conserved Asp-120(81) in the active site, which plays an important role in catalysis. To probe the catalytic role of Asp-120( 81) in IMP-1, the IMP-1 mutants, D120(81)A and D120(81)E, were prepared by site-directed mutagenesis, and various kinetics studies were conducted. The IMP-1 mutants exhibited 10(2)-10(4)-fold drops in k(cat) values compared with WT despite the fact that they contained two Zn(II) ions in the active site. To evaluate the acid-base characteristics of Asp-120( 81), the pH dependence for hydrolysis was examined by stopped-flow studies. No observable pK(a) values between pH 5 and 9 were found for WT and D120( 81) A. The rapid mixing of equimolar amounts of nitrocefin and all enzymes failed to result in the detection of an anion intermediate of nitrocefin at 650 nm. These results suggest that Asp-120( 81) of IMP-1 is not a factor in decreasing the pKa for the water bridging two Zn( II) ions and is not a proton donor to the anionic intermediate. In the case of D120( 81) E, the nitrocefin hydrolysis product, which shows a maximum absorption at 460 nm, was bound to D120( 81) E in the protonated form. The three-dimensional structures of D120( 81) A and D120( 81) E were also determined at 2.0 and 3.0 angstrom resolutions, respectively. In the case of D120(81) E, the Zn-Zn distance was increased by 0.3 angstrom compared with WT, due to the change in the coordination mode of Glu-120(81)OE1 and the positional shift in the conserved His-263(197) at the active site.

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  • 1989 - Present
    医学細菌学、病原細菌学、薬剤耐性菌等  (名古屋大学 [医、保健、工]、群馬大学 [医]、千葉大学 [薬]、東京薬科大学 [薬]、愛知学院大学 [歯・薬]、岐阜薬科大学 [薬]、愛知医科大学[医]、 他)

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