bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of Microfilaments and Microtubules in the Invasion of INT‐407 Cells by Campylobacter jejuni
Authors: Biswas Debabrata , Itoh Kikuji , Sasakawa Chihiro 
Published: November 2013

AbstractThe internalization mechanisms triggered by Campylobacter jejuni were studied by invasion assays conducted with different inhibitors that act on the cytoskeleton structure of eukaryotic cells. The depolymerization of microfilaments by cytochalasin‐D and that of microtubules by colchicines and nocodazole inhibited the uptake of C. jejuni into INT‐407 cells in a dose‐dependent manner. The inhibitory effect of microfilament depolymerization on C. jejuni internalization was more pronounced than that of microtubule depolymerization. By immunofluorescence microscopic observations, it was demonstrated that both microfilaments and microtubules were localized in INT‐407 cells after C. jejuni infection. These data suggest that the internalization mechanism triggered by C. jejuni is associated with the combined effect of microfilaments and microtubules of host cells.
DNA Base Alignment and Taxonomic Study of Genus Malassezia Based upon Partial Sequences of Mitochondrial Large Subunit Ribosomal RNA Gene
Authors: Yamada Yohko , Makimura Koichi , Ueda Kumiko , Nishiyama Yayoi , Uchida Katsuhisa , Yamaguchi Hideyo , Osumi Masako 
Published: November 2013

AbstractThe sequences of the large subunit of mitochondrial ribosomal RNA (LsmtrRNA) gene of Malassezia species were analysed. The sequences of the seven species of Malassezia are well separated in each species. Therefore the LsmtrRNA gene is thought to be one of the gene targets for species identification in the genus Malassezia. The dendrogram obtained from this gene supports the previous study of Malassezia species based upon the chromosomal genes. This is the first report of taxonomic analysis of Malassezia species based upon the mitochondrial gene.
Possible Relationship of PFGE Patterns of Moraxella catarrhalis between Hospital‐ and Community‐Acquired Respiratory Infections in a Community Hospital
Authors: Masaki Hironori , Asoh Norichika , Kawazoe Kaneo , Watanabe Kiwao , Onizuka Tomoko , Shimogama Seiji , Yamaryo Takeshi , Watanabe Hiroshi , Oishi Kazunori , Nagatake Tsuyoshi 
Published: November 2013

AbstractWe describe a prospective study of molecular analysis of Moraxella catarrhalis isolated from a community hospital. Our study was designed to investigate the possible relationship of pulsed‐field gel electrophoresis (PFGE) patterns of M. catarrhalis between hospital‐ and community‐acquired respiratory infections. A nosocomial outbreak of M. catarrhalis was observed between September 2000 and September 2001. During the study period, 40 strains of M. catarrhalis were isolated from a total of 32 patients with respiratory infections (26 strains from 18 inpatients, and 14 strains from 14 outpatients). We compared the PFGE patterns in 40 strains of M. catarrhalis isolated from the respiratory tract of the study patients. The genomic types of M. catarrhalis were classified into three PFGE patterns (A, B, and C). Interestingly, the nosocomial outbreak of M. catarrhalis included two patterns (A and B). Of the three patterns, two patterns (A and B) were found in both inpatients and outpatients. More interestingly, two subtypes of pattern B (B1 and B4) were simultaneously found in both inpatients and outpatients. Our results indicated that PFGE with SmaI chromosomal digestion is a suitable technique to establish the inter‐strain genetic relatedness of M. catarrhalis, and suggested that the outbreak of M. catarrhalis occasionally included miscellaneous PFGE patterns. The results also showed that PFGE patterns of M. catarrhalis isolates were similar between hospital‐ and community‐acquired respiratory infections. Analysis of the subtypes suggested that there might be some association between hospital‐ and community‐acquired respiratory infections caused by M. catarrhalis.
Hepatitis C Virus Core Protein Selectively Inhibits Synthesis and Accumulation of p21/Waf1 and Certain Nuclear Proteins
Authors: Oka Kiyomasa , Nagano‐Fujii Motoko , Yoshida Isao , Hidajat Rachmat , Deng Lin , Akutsu Masato , Hotta Hak 
Published: November 2013

AbstractBy using a vaccinia virus‐T7 expression system, possible effects of hepatitis C virus (HCV) core protein on synthesis and accumulation of host cellular proteins transiently expressed in cultured cells were analyzed. Immunoblot and immunofluorescence analyses revealed that synthesis and accumulation of certain nuclear proteins, such as p21/Waf1, p53, proliferating cell nuclear antigen and c‐Fos, were strongly inhibited by HCV core protein. On the other hand, synthesis and accumulation of cytoplasmic proteins, such as 2′‐5′‐oligoadenylate synthetase (2′‐5′‐OAS), RNase L and MEK1, were barely affected by HCV core protein. Northern blot analysis showed that the degrees of mRNA expression for those proteins did not differ between HCV core protein‐expressing cells and the control, suggesting that the inhibition occurred at the post‐transcription level. Pulse‐labeling analysis suggested that HCV core protein strongly inhibited synthesis of p21/Waf1 at the translation level. Once being accumulated in the nucleus, p21/Waf1 stability was not significantly affected by HCV core protein. Mutants of HCV core protein C‐terminally deleted by 18 or 41 amino acids (aa), which were localized almost exclusively in the nucleus, lost their ability to inhibit synthesis/accumulation of p21/Waf1 whereas another mutant C‐terminally deleted by 8 aa still maintained the same properties (subcellular localization and the inhibitory effect) as the full‐length HCV core protein of 191 aa. Taken together, our present results suggest that expression of HCV core protein in the cytoplasm selectively inhibits synthesis of p21/Waf1 and some other nuclear proteins at the translation level.
Discrimination of West Nile Virus and Japanese Encephalitis Virus Strains Using RT‐PCR RFLP Analysis
Authors: Shirato Kazuya , Mizutani Tetsuya , Kariwa Hiroaki , Takashima Ikuo 
Published: November 2013

AbstractWest Nile (WN) virus is a mosquito‐borne flavivirus that induces lethal encephalitis in humans and horses. Since an outbreak of WN encephalitis in humans and horses occurred in New York City in late August 1999, the possibility exists that WN virus will invade regions that have close links with the United States, such as Japan. We developed a genetic diagnostic method that discriminates between strains of WN virus and Japanese encephalitis (JE) virus. The method involves RT‐PCR restriction fragment length polymorphism (RFLP) analysis with a RT‐PCR primer set, a nested PCR primer set, and a restriction enzyme. We detected WN and JE viruses in experimentally infected animal brain, spleen, and serum samples. Our method is useful in distinguishing WN viruses from the endemic background of JE viruses, and in discriminating the highly virulent WN strain, which was isolated in New York in 1999, from other WN virus strains.
The First Isolation of Ustilaginomycetous Anamorphic Yeasts, Pseudozyma Species, from Patients' Blood and a Description of Two New Species: P. parantarctica and P. thailandica
Authors: Sugita Takashi , Takashima Masako , Poonwan Natteewan , Mekha Nanthawan , Malaithao Kaewjai , Thungmuthasawat Benjaporn , Prasarn Soem , Luangsook Pakeenee , Kudo Toshiaki 
Published: November 2013

AbstractThe genus Pseudozyma is ustilaginomycetous anamorphic yeasts, and are mainly isolated from plants. We isolated three Pseudozyma strains from the blood of patients in Thailand. While one isolate was identified as P. antarctica by rDNA sequence analysis, the other two were considered to be new species and were named P. parantarctica and P. thailandica. The three isolates proved to be resistant to 5‐flucytosine, and P. thailandica was also resistant to fluconazole and itraconazole. As far as we know, this is the first isolation of Pseudozyma strains from humans. The two new species are described.
Coenzyme Q1‐Catalyzed Luminol Chemiluminescent Assay for Rapid Antimicrobial Susceptibility Testing of Mycobacterium bovis
Authors: Yamashoji Shiro 
Published: November 2013

AbstractCoenzyme Q1 is herein proposed as the best catalyst among coenzymes Q and vitamins K for quinone‐catalyzed luminol chemiluminescent assays applied to rapid determination of viability or rapid antimicrobial susceptibility tests of Mycobacterium bovis. Luminol chemiluminescence intensity (LCI) was determined 10 min after the incubation of M. bovis with coenzyme Q1, and was proportional to CFU (colony‐forming unit)/ml in the range of 9,000 to 2,250,000. LCI depended on the the production of the superoxide anion (O2−) rather than H2O2 during a 10‐min incubation of M. bovis with coenzyme Q1, as superoxide dismutase reduced LCI more effectively than catalase. The minimal inhibitory concentrations (MICs) of 10 kinds of antituberculous agents estimated on the basis of decrease in LCI after one or two days' cultivation were in good agreement with MICs determined by turbidity analysis, which requires upwards of 1 week to complete.
The Role of Glucan‐Binding Proteins in the Cariogenicity of Streptococcus mutans
Authors: Matsumura Mieko , Izumi Tomohiro , Matsumoto Michiyo , Tsuji Masato , Fujiwara Taku , Ooshima Takashi 
Published: November 2013

AbstractStreptococcus mutans produces glucan‐binding proteins (Gbps), which appear to contribute to the virulence of S. mutans. GbpA and GbpC genes were inactivated by the insertion of antibiotic‐resistant genes into each gbp gene of S. mutans MT8148 to generate Gbp‐defective mutants. Sucrose dependent adherences of the GbpA‐ and GbpC‐defective mutants were found to be significantly lower than those of their parent strains MT8148. Caries inducing activity of the mutants in rats was significantly lower than that of strain MT8148R (streptomycin‐resistant strain of MT8148). These results suggest that GbpA and GbpC participate in cellular adherence to tooth surfaces and contribute to the cariogenicity of S. mutans.
Correlation between the Avidity Maturation of Anti‐HSP70 IgG Autoantibody and Recombination Activating Gene Expressions in Peripheral Lymphoid Tissues of Toxoplasma gondii‐Infected Mice
Authors: Chen Mei , Aosai Fumie , Mun Hye‐Seong , Norose Kazumi , Piao Lian Xun , Yano Akihiko 
Published: November 2013

AbstractThe avidity maturation of anti‐TgHSP70 IgG antibody produced by B‐2 cells of BALB/c mice (a resistant strain) and that of anti‐mHSP70 IgG autoantibody produced by B‐1 cells of C57BL/6 mice (B6; a susceptible strain) was observed after Toxoplasma gondii infection. Recombination‐activating genes (RAGs) were predominantly expressed in B‐1 cells from peritoneal exudate cells (PECs) of T. gondii‐infected B6 mice, while RAGs were expressed in B‐2 cells from PECs of BALB/c mice. These results suggest that the involvement of RAG gene activations in the peripheral lymphoid tissues in the avidity maturation of anti‐TgHSP70 IgG antibody and anti‐mHSP70 IgG autoantibody in T. gondii‐infected mice.
Effects of Melanin upon Susceptibility of Cryptococcus to Antifungals
Authors: Ikeda Reiko , Sugita Takashi , Jacobson Eric S. , Shinoda Takako 
Published: November 2013

AbstractMelanin is a recognized virulence factor in Cryptococcus neoformans; several pathogenetic mechanisms have been suggested. We studied melanin as an antifungal resistance factor. The growth of laccase‐active strains of C. neoformans and C. albidus in L‐DOPA resulted in the production of black pigment. The formal minimal inhibitory concentrations (MICs) of amphotericin B and fluconazole were not changed by melanization. However, when we examined those wells which contained inhibited cells, we found live cells only in wells containing melanized C. neoformans. In contrast, melanization did not protect C. albidus from killing by amphotericin B. In an amphotericin B time‐kill study of C. neoformans, significantly more melanized cells than non‐melanized survived for the first few hours. Fluorescence microscopy and flow cytometry analyses showed that fewer melanized cells were stained with the fluorescent dye MitoRed. Incubation of MitoRed (the model) or amphotericin B with melanin extracted from C. neoformans decreased the free concentrations of these substances. Fluconazole, in contrast, was not removed from solution by melanin. This suggests that neoformans cryptococcal melanin deposited amphotericin B in the cell wall binds, reducing its effective concentrations.
Cell‐Mediated Fas‐Based Lysis of Dendritic Cells Which Are Apparently Resistant to Anti‐Fas Antibody
Authors: Yokota Aya , Oikawa Atsuhiko , Matsuda Chieko , Shinohara Nobukata , Eshima Koji 
Published: November 2013

AbstractIn this report, the controversy concerning the sensitivity of dendritic cells (DCs) to Fas‐dependent induction of apoptosis was examined using murine DCs. Although DCs could not be lysed when exposed to an anti‐Fas antibody, Jo2, the observed resistance turned out to reflect their lack of the expression of FcγR necessary for crosslinking the antibody, rather than their intrinsic resistance. Thus, at least a fraction of DCs was sensitive to Jo2 in the presence of FcγR‐expressing by‐standers. Consistently, a significant fraction of DCs was sensitive to Fas‐dependent lysis mediated by T cells including the antigen‐specific killing by CD4+ T cells. Both immature (class II MHClow) and mature (class II MHChigh) DCs were sensitive to the Fas‐based induction of apoptosis.
Probable Laboratory Contamination of Clinical Specimens with Leptospira meyeri
Authors: Koizumi Nobuo , Kawabata Hiroki , Watanabe Haruo 
Published: November 2013

AbstractLeptospira meyeri were isolated from the blood or the pleural effusion cultures of four patients at a commercial clinical laboratory. All isolates were identical in their 16S rDNA sequences and NotI or SfiI restriction profiles of the chromosome DNA on pulsed‐field gel electrophoresis. However, intraperitoneal inoculation of the isolate (NIID1) failed to kill hamsters and none of the patients' sera reacted with L. meyeri strains isolated, indicating leptospiral contamination occurred during laboratory investigation.
3‐Methylcholanthrene Activates Human Immunodeficiency Virus Type 1 Replication via Aryl Hydrocarbon Receptor
Authors: Ohata Hirokazu , Tetsuka Toshifumi , Hayashi Hidetoshi , Onozaki Kikuo , Okamoto Takashi 
Published: November 2013

AbstractWe found that 3‐methylcholanthrene (3‐MC) could induce the reactivation of human immunodeficiency virus type 1 (HIV‐1) replication in OM 10.1 cell, promyelocytic cell line latently infected with HIV‐1. Transient luciferase expression experiments have revealed no particular transcription factors that are responsible for the effect of 3‐MC in inducing HIV‐1 gene expression as HIV‐1 LTR mutants lacking various upstream transcriptional activators similarly responded to 3‐MC. In addition, there was no effect of 3‐MC on the DNA binding activity of nuclear factor‐kappa B (NF‐κB) that was previously reported to be crucial for the effect of 2, 3, 7, 8‐tetrachlorodibenzo‐p‐dioxin (TCDD), a chemical homologue of 3‐MC. However, overexpression of wild type aryl hydrocarbon receptor (AhR), a nuclear receptor of polycyclic aromatic hydrocarbons (PAHs) such as 3‐MC, augmented the effect of 3‐MC in the induction of gene expression from HIV‐1 LTR. Moreover, a dominant negative mutant of AhR dramatically reduced the 3‐MC‐mediated activation of HIV‐1 LTR. These findings suggest that 3‐MC stimulates HIV‐1 transcription by interacting with general transcription factors. Our observations indicate that chronic exposure of the HIV‐1 infected individuals to PAHs may be contributable to the clinical development of acquired immunodeficiency syndrome (AIDS) among the individuals infected with HIV.
Mapping of the Low pH‐Sensitive Conformational Epitope of Rabies Virus Glycoprotein Recognized by a Monoclonal Antibody #1‐30‐44
Authors: Kankanamge Pushpa Jenette , Irie Takashi , Mannen Kazuaki , Tochikura Tadafumi S. , Kawai Akihiko 
Published: November 2013

AbstractMonoclonal antibody (mAb) #1‐30‐44 recognized an acid‐sensitive conformational epitope of rabies virus glycoprotein (G). The antigenicity of G protein exposed on the cell surface was lost when the infected cells were exposed to pH 5.8. By comparing the deduced amino acid sequence of G protein between the HEP‐Flury strain and the epitope‐negative CVS strain as well as the mAb‐resistant escape mutants, two distant sites that contained Lys‐202 and Asn‐336 were shown to be involved in the epitope formation. Lys‐202 is located in the so‐called neurotoxin‐like sequence, while Asn‐336 is included in antigenic site III and is very near the amino acid at position 333, which is known to affect greatly the neuropathogenicity of rabies virus when changed. Consistent with this finding, antigenicity of a neurovirulent revertant of the HEP‐Flury strain, in which Gln‐333 of G protein was replaced by Arg, was also affected as shown by its greatly decreased reactivity with mAb #1‐30‐44 compared to that of the original avirulent HEP virus. Based on these results, we hypothesize that the neurotoxin‐like domain and some amino acids in antigenic site III come into contact with each other to form a conformational epitope for mAb #1‐30‐44, and such a configuration would be lost when exposed to acidic conditions to perform a certain low pH‐dependent function of G protein.
Distribution of Human Rotaviruses, Especially G9 Strains, in Japan from 1996 to 2000
Authors: Zhou Yumei , Li Lei , Okitsu Shoko , Maneekarn Niwat , Ushijima Hiroshi 
Published: November 2013

AbstractA 4‐year (1996–2000) survey of rotavirus infection involving 2,218 diarrheal fecal specimens of children collected from five regions of Japan was conducted. A total of 642 (28.9%) specimens were found to be rotavirus positive. A changed prevalence pattern of rotavirus G serotype was found with an increase of G9 and G2 and a decrease of G1, although G1 remained the prevailing serotype. Serotype G9 was unexpectedly determined to be the prevailing serotype in Sapporo (62.5%) and Tokyo (52.9%) in 1998–1999, and in Saga (78.4%) in 1999–2000. G9 strains isolated from 1998–1999 belonged to the P[8]‐NSP4‐Wa‐group with long RNA pattern, while, G9 strains isolated from 1999–2000 belonged to three groups, the P[8]‐NSP4‐Wa‐group with long RNA pattern, the P[4]‐NSP4‐KUN‐group with short RNA pattern and a mixed‐type group (P[4]/P[8]‐NSP4‐KUN/Wa‐group with long RNA pattern). Both sequence and immunological analysis of VP7 revealed that the G9 strains from 1999–2000 were much more closely related to the G9 strains isolated worldwide in the 1990s, including G9 strains found in Thailand in 1997. However, the G9 strains from 1998–1999 were distinct from these and more closely related to the G9 prototype strains F45, AU32 and WI61 discovered in Japan and the US in the 1980s. Thus the G9 strains isolated in 1998–1999 had progenitors common to the G9 prototype strains, while the strains isolated in 1999–2000 did not directly evolve from them but were related to global G9 strains that have emerged in recent years. These data supported our previous report that G9 rotavirus might exist as two or more subtypes with diverse RNA patterns, P‐genotype and NSP4 genogroup combinations (Y.M. Zhou et al., J. Med. Virol. 65: 619–628, 2001) and suggested that G9 rotavirus prevalent in Japan during two successive years belonged to different subtypes. The nucleotide sequences presented in this paper were submitted to DDBJ, EMBL and GenBank nucleotide sequence databases. The accession numbers are: 00‐Ad2863VP7 (AB091746), 00‐OS2986VP7 (AB091747), 00‐SG2509VP7 (AB091748), 00‐SG2518VP7 (AB091749), 00‐SG2541 (AB091750), 00‐SG2864 (AB091751), 00‐SP2737VP7 (AB091752), 99‐SP1542VP7 (AB091753), 99‐SP1904VP7 (AB091754), 99‐TK2082VP7 (AB091755) and 99‐TK2091VP7 (AB091756).
The Size of cagA Based on Repeat Sequence Has the Responsibility of the Location of Helicobacter pylori in the Gastric Mucus and the Degree of Gastric Mucosal Inflammation
Authors: Karita Mikio , Matsumoto Satoru , Kamei Toshiaki 
Published: November 2013

AbstractThe aim of this study was to examine whether there is a relationship between cagA size of Japanese Helicobacter pylori strains and the location of these strains in the mucous layer, the degree of gastric inflammation and acid survival. Upper gastrointestinal endoscopies were done to 144 patients with dyspeptic symptom with informed consent, sera, biopsy specimens and H. pylori strains were obtained, and gastric histology and susceptibility to pH 3 of the strains were evaluated. To determine cagA size of Japanese strains using PCR, cagA of strain CPY3401 was sequenced. 74 H. pylori samples (72 cagA +) were obtained from the body and 56 samples (56 cagA +) obtained from the antrum. cagA size of 72 H. pylori strains from the body was mainly classified into 3 groups (short (48), middle (8), long (9), and others (7)) by PCR and all of that of 56 strains from the antrum except 2 was short. The size of cagA of isolated strains from the body is associated with enhanced gastritis, acid survival, and the location in the mucus. The long size cagA of which strain is acid sensitive, may be a strong selective pressure on strain that colonizes close to the host, which enhanced gastritis.
Genetic Characterization of Rabies Field Isolates from Thailand
Authors: Susetya Heru , Sugiyama Makoto , Inagaki Akio , Ito Naoto , Oraveerakul Kanisak , Traiwanatham Nimit , Minamoto Nobuyuki 
Published: November 2013

AbstractWe sequenced 512 nucleotides in two variable regions of the N gene of 23 rabies isolates from the northeastern part of Thailand by direct sequencing of PCR‐amplified products. The sequencing data revealed two new lineages in these rabies isolates. Based on the results of this study together with the findings of our earlier study, the rabies isolates in Thailand were divided into two genogroups, designated as T1 and T2, which were predominantly localized in the northern and northeastern areas, respectively. Each of these two genogroups consisted of four lineages. There was a correlation between these eight lineages and the geographical origins of the isolates. Two lineages belonging to the T2 genogroup from the northeastern area of Thailand were newly identified in this study. The isolates in both genogroups were also prevalent in the central area of Thailand. Each lineage in the T1 and T2 genogroups was found independently in dogs in the upper and lower southern areas of Thailand, respectively. These genetic data and the historical background suggest that rabies viruses belonging to the T2 genogroups were prevalent many years ago in the central and northeastern areas of Thailand and were later transferred to the lower southern parts of Thailand.