bluuebunny/crossref_metadata_2025 huggingface dataset:
Anaerospora hongkongensis Gen. Nov. Sp. Nov., a Novel Genus and Species with Ribosomal DNA Operon Heterogeneity Isolated from an Intravenous Drug Abuser with Pseudobacteremia
Authors: Woo Patrick C.Y. , Teng Jade L.L. , Leung Kit‐wah , Lau Susanna K.P. , Woo Gibson K.S. , Wong Amy C.Y. , Wong Michelle K.M. , Yuen Kwok‐yung
Published: November 2013

A bacterium was isolated from the blood culture of an intravenous drug abuser with pseudobacteremia. The cells were strictly anaerobic, straight or slightly curved, sporulating, Gram‐negative rods. It grew on sheep blood agar as non‐hemolytic, pinpoint colonies after 48 hr of incubation at 37 C in an anaerobic environment. It was motile but did not produce catalase or cytochrome oxidase. 16S ribosomal DNA (rDNA) sequencing revealed three different copies of 16S rDNA sequences. More than 90% of the differences among them were due to differences in the lengths of the sequences. Phylogenetically, the bacterium is clustered with Dendrosporobacter, Sporomusa, and Propionispora, the other three genera of anaerobic, sporulating, Gram‐negative rods. There were 8.6–11.1% differences between the 16S rDNA sequences of the bacterium and that of D. quercicolus, 4.7–15.1% differences between the 16S rDNA sequences of it and those of S. acidovorans, S. aerivorans, S. malonica, S. ovata, S. paucivorans, S. silvacetica, S. spaeroides, and S. termitida, and 7.6–13.1% differences between the 16S rDNA sequences of it and those of P. hippei and P. vibrioides. The G+C content of the bacterium (mean±SD) was 46.8±3.2%. For these reasons, a new genus and species, Anaerospora hongkongensis gen. nov. sp. nov., is proposed, for which HKU15T is the type strain.
Moraxella catarrhalis Does Not Grow on Nutrient Agar without Sodium Chloride Supplementation
Authors: Chida Toshio , Sawabe Etsuko , Ono Emi , Nishiyama Hiroyuki , Ebina Kengo , Sato Kenya , Watanabe Yuko , Kuroki Toshiro , Okamura Noboru
Published: November 2013

None of the 58 Moraxella catarrhalis strains grew on nutrient agar without sodium chloride supplementation, whereas 49 of 51 commensal Neisseria spp. strains tested did. Growth on nutrient agar without sodium chloride supplementation could be used for screening between M. catarrhalis and commensal Neisseria spp.
Intrathymic Effect of Acute Pathogenic SHIV Infection on T‐Lineage Cells in Newborn Macaques
Authors: Suzuki Hajime , Motohara Makiko , Miyake Ariko , Ibuki Kentaro , Fukazawa Yoshinori , Inaba Katsuhisa , Masuda Kyoko , Minato Nagahiro , Kawamoto Hiroshi , Hayami Masanori , Miura Tomoyuki
Published: November 2013

We intrarectally infected newborn macaques with a pathogenic simian/human immunodeficiency virus (SHIV) that induced rapid and profound CD4+ T cell depletion, and examined the early effects of this SHIV on the thymus. After intrarectal infection, viral loads were much higher in the thymus than in other lymphoid tissues in newborns. In contrast, no clear difference was seen in the viral loads of different tissues in adults. Histological and immunohistochemical observations showed severe thymic involution. Depletion of CD4+ thymocytes began in the medulla at 2 weeks post infection and spread over the whole thymus. After in vivo infection, the CD2+ subpopulation, which represents a relatively later stage of T cell progenitors, was selectively reduced and development of thymocytes from CD3–CD4–CD8– cells to CD4+CD8+ cells was impaired. These results suggest that profound and irreversible loss of CD4+ cells that are observed in the peripheral blood of SHIV‐infected monkeys are due to destruction of the thymus and impaired thymopoiesis as a result of SHIV infection in the thymus.
Protective Effect of Vaccination with Toxoplasma Lysate Antigen and CpG as an Adjuvant against Toxoplasma gondii in Susceptible C57BL/6 Mice
Authors: El‐Malky Mohamed , Shaohong Lu , Kumagai Takashi , Yabu Yoshisada , Noureldin Mohamed S. , Saudy Niveen , Maruyama Haruhiko , Ohta Nobuo
Published: November 2013

Infection with the intracellular protozoan parasite Toxoplasma gondii causes serious public health problems to both humans and livestock and of great economic impact worldwide. Oligodeoxynucleotides (ODN) which contain immunostimulatory CG motifs (CpG ODN) can promote Th1 responses, an adjuvant activity that is desirable for vaccination against intracellular pathogens. We investigated the feasibility of using CpG as an adjuvant combined with Toxoplasma lysate antigen (TLA) as a vaccine against toxoplasmosis. Genetically susceptible C57BL/6 mice were vaccinated with TLA with or without CpG ODN as an adjuvant and then challenged with 85 cysts of the moderately virulent RRA (Beverley) strain of T gondii. Prior to challenge infection, immunization with TLA plus CpG ODN directed cellular and humoral immunity toward a Th1 pattern, characterized by enhanced INF7 production by splenic cells in response to TLA, and enhanced production of toxoplasma‐specific IgG and IgG2a antibodies. Consequently, CpG/TLA‐treated mice showed prolonged survival and 64% reduction in brain parasite burden compared to non‐CpG/TLA treated group. Our results suggest that CpG ODN would provide a stable and effective adjuvant for use in vaccination against toxoplasmosis.
Chimeric Structure of omp2 of Brucella from Pacific Common Minke Whales (Balaenoptera acutorostrata)
Authors: Ohishi Kazue , Takishita Kiyotaka , Kawato Masaru , Zenitani Ryoko , Bando Takeharu , Fujise Yoshihiro , Goto Yoshitaka , Yamamoto Saburo , Maruyama Tadashi
Published: November 2013

In the Pacific common minke whale (Balaenoptera acutorostrata), a new variant of Brucella has been detected using the polymerase chain reaction. Detailed analysis of the porin protein genes, omp2a and omp2b from the whale Brucella showed that these two genes have some motifs in common with Atlantic marine strains in the 5′‐terminal one‐third region. On the other hand, the nucleotide sequences in the 3′‐terminal two‐thirds region of the two genes were almost identical to the respective genes of terrestrial strains. Thus, Pacific whale Brucella omp2 genes are chimeras between marine and terrestrial strains.
Identification of Platelet Receptors for the Streptococcus gordonii DL1 Sialic Acid‐Binding Adhesin
Authors: Yajima Ayako , Takahashi Yukihiro , Konishi Kiyoshi
Published: November 2013

Aggregation of human platelets by Streptococcus gordonii DL1, an interaction implicated in the pathogenesis of infective endocarditis, requires the expression of hsa, the gene encoding the sialic acid‐binding adhesin (Hsa) of this organism. To identify the sialoglycoproteins on the platelet surface as the receptors for Hsa, intrinsic membrane proteins were assessed by bacterial overlay assay. S. gordonii DL1 adhered to 130‐140‐kDa proteins, a reaction that was abolished by neuraminidase treatment of immobilized platelet surface proteins. These sialoglycoproteins were identified as platelet glycoprotein Ibα (GPIbα) and glycoprotein IIb (GPIIb) by immunoprecipitation with specific monoclonal antibody against each glycoprotein.
Serological Evidence of the Persistence of Infection with Leptospira interrogans Serovar Hardjo in Cattle in Mongolia
Authors: Odontsetseg Namsraijav , Sakoda Yoshihiro , Kida Hiroshi
Published: November 2013

Serum samples were collected randomly from Mongol breed cattle in three geographically distinct provinces of Mongolia. Antibodies specific to Leptospira interrogans serovar Hardjo were detected by microscopic agglutination test (MAT) at titres of 100 or more in 80.4% (86/107) of the samples from Dornod Province, but in only 28.9% (13/45) in Arkhangai and 23.5% (12/51) in Khuvsgul Provinces, respectively. Treatment of 9 serum samples from Dornod, positive to serovar Hardjo in MAT and negative in the homologous immunoglobulin G (IgG) enzyme‐linked immunosorbent assay (ELISA), with 2‐mercaptoethanol (2‐ME) indicated that those animals were recently infected.
Genetic Comparison of H5N1 Influenza A Viruses Isolated from Chickens in Japan and Korea
Authors: Mase Masaji , Kim Jae‐Hong , Lee Youn‐Jeong , Tsukamoto Kenji , Imada Tadao , Imai Kunitoshi , Yamaguchi Shigeo
Published: November 2013

Outbreaks of highly pathogenic avian influenza (HPAI) caused by H5N1 virus occurred during 2003 to 2004 in Korea and Japan. The H5N1 viruses isolated in both countries were genetically similar at >99% identity in the nucleotide sequences of all eight RNA segments, indicating that they belong to genotype V and are distinct from HPAI viruses prevalent in southeast Asia that belong to genotype Z. These findings indicate that the H5N1 viruses that caused the HPAI outbreaks in both Korea and Japan were derived from a common ancestor.
Role of Lipooligosaccharide in the Attachment of Moraxella catarrhalis to Human Pharyngeal Epithelial Cells
Authors: Akgul Gulcan , Erturk Ali , Turkoz Mustafa , Turan Tolga , Ichinose Akitoyo , Nagatake Tsuyoshi , Ahmed Kamruddin
Published: November 2013

The goal of this study was to determine the role of lipooligosaccharide in the attachment of Moraxella catarrhalis to human pharyngeal epithelial cells. Strain 2951 and its Pk mutant strain 2951 galE were used in this study. This study suggests that the Pk epitope of LOS is not an adhesin for M. catarrhalis, but plays a crucial role by its surface charge in the initial stage of attachment.
Gene Cloning and Characterization of Four MATE Family Multidrug Efflux Pumps from Vibrio cholerae Non‐O1
Authors: Begum Anowara , Rahman M. Mushfequr , Ogawa Wakano , Mizushima Tohru , Kuroda Teruo , Tsuchiya Tomofusa
Published: November 2013

There are six putative genes for multidrug and toxic compound extrusion (MATE) family multidrug efflux pumps in the chromosome of Vibrio cholerae. We have so far analyzed two MATE family pumps in V. cholerae non‐O1 NCTC4716. Here we cloned four remaining genes for putative MATE family efflux pumps by the PCR method from this microorganism and designated them as vcmB, vcmD, vcmH and vcmN. Each one of the four genes was introduced and expressed in the drug hypersusceptible host Escherichia coli KAM32 cells. We observed elevated MICs of multiple antimicrobial agents, such as fluoroquinolones, aminoglycosides, ethidium bromide and Hoechst 33342 in the transformants. Energy‐dependent efflux of substrate was observed with the transformed cells. We found that efflux activities of VcmB, VcmD and VcmH were Na+‐dependent, but that of VcmN was Na+‐independent. Thus, all six of the MATE family multidrug efflux pumps of V. cholerae non‐O1 have been characterized. We also found that all six genes were expressed in cells of V. cholerae non‐O1.
Prospective Surveillance of Community‐Onset and Healthcare‐Associated Methicillin‐Resistant Staphylococcus aureus Isolated from a University‐Affiliated Hospital in Japan
Authors: Piao Chuncheng , Karasawa Toshiko , Totsuka Kyoichi , Uchiyama Takehiko , Kikuchi Ken
Published: November 2013

We conducted a prospective comparative study of community‐onset (CO) and healthcare‐associated (HA) methicillin‐resistant Staphylococcus aureus (MRSA) strains between 2000 and 2001 at Tokyo Women's Medical University Hospital (1,500 beds) in Japan. Of the 172 consecutive MRSA isolates analyzed, 13 (8%) were categorized as CO‐MRSA. The mean age of patients with CO‐MRSA was significantly younger than that of patients with HA‐MRSA. Most CO‐MRSA strains were isolated from skin and more likely to be susceptible to erythromycin, clindamycin, tetracycline, levofloxacin, and spectinomycin compared to HA‐MRSA isolates. Pulsed‐field gel electrophoresis (PFGE) analysis, staphylococcal cassette chromosome mec (SCCmec) typing, and multi‐locus sequence typing (MLST) revealed that CO‐MRSA strains were divided into the following multi‐clones: 3 clone A: II: ST5 (PFGE type: SCCmec type: MLST sequence type); 1 L: II: ST5; 1 H: IV: ST1; 1 I: IV: ST81; 2 D: IV: ST8; 1 B: IV: ST89; 1 B: IV: ST379; and 3 B: IV: ST91. Of the 159 HA‐MRSA strains, 124 (78%) belonged to a single clone (PFGE clone A: SCCmec type II: tst and sec positive: coagulase type II: multi‐drug resistance). Four CO‐MRSA strains belonging to PFGE clone B: SCCmec type IV: MLST clonal complex 509 (ST89, 91, 379) had the exfoliative toxin B (etb) genes, but all CO‐MRSA and HA‐MRSA strains did not possess the Panton‐Valentine leukocidin (pvl) genes. These results demonstrate that multiple lineages of CO‐MRSA have the potential for dissemination in the community in Japan.
Molecular Epidemiology of Adenovirus Infection among Pediatric Population with Diarrhea in Asia
Authors: Li Lei , Phan Tung Gia , Nguyen Tuan Anh , Kim Kyo Sun , Seo Jeong Kee , Shimizu Hideaki , Suzuki Eiko , Okitsu Shoko , Ushijima Hiroshi
Published: November 2013

A total of 3,577 fecal specimens from infants and children with acute gastroenteritis in Japan, Korea and Vietnam during 1998 and 2001 were tested for adenovirus by the ELISA method. Of these, adenovirus was detected in 158 (4.4%). The detection rate of adenovirus was highest in Korea (8.7%, 20/231) followed by 5.0% (100/1,991) in Japan and 2.8% (38/1,355) in Vietnam. All adenoviruses were further serotyped by PCR‐RFLP. The diversity of adenovirus serotypes including Ad2, 3, 5, 8, 31, 40 and 41 was demonstrated. Worth of note was a decrease in the rate of isolation of Ad40 (7.6%, 12/158) and a concomitant increase of Ad41 (63.9%, 101/158) to become the predominant serotype. Another interesting feature of the study was the presence of Ad3 (4.0% and 30%) and Ad8 (8.0% and 20%) in Japan and Korea, respectively, which is mainly associated with keratoconjunctivitis worldwide. Our result underscored the importance of adenovirus in association with acute gastroenteritis in Asian countries.
Comparisons of the Laccase Gene among Serotypes and Melanin‐Deficient Variants of Cryptococcus neoformans
Authors: Tanaka Eiichiro , Ito‐Kuwa Shoko , Nakamura Kenjirou , Aoki Shigeji , Vidotto Valerio , Ito Masaaki
Published: November 2013

Cryptococcus neoformans is a fungus causing life‐threatening infections in immunocompromised hosts. Melanin production is a major virulence factor of this fungus and the initial steps of dihydroxyphenylalanine (DOPA)‐melanin biosynthesis pathways are catalyzed by laccase. To understand phylogenetic relationships among serotypes of three varieties, partial sequences (about 600 bases) of the laccase gene (CNLAC1) were determined in a total of 64 strains, including 10 melanin‐deficient variants. The phylogenetic tree constructed from the nucleotide sequence grouped the 64 strains into the clusters corresponding to the three varieties. The diversity of the fragment sequences was very minor among strains of each of var. grubii and var. neoformans. Strains in var. gattii, however, were subdivided into two groups, although differences between serotypes B and C were not large. The sequences of the melanin‐deficient variants were almost completely homologous to those of the melanin‐producing strains in the same serotype. Results of laccase assay and northern blot analysis suggested that the lower melanin production in the variants was associated with lower transcription of the laccase gene.
Preparation of Monoclonal Antibodies Cross‐Reactive with Orthopoxviruses and Their Application for Direct Immunofluorescence Test
Authors: Kitamoto Noritoshi , Kobayashi Takayuki , Kato Yoji , Wakamiya Nobutaka , Ikuta Kazuyoshi , Tanaka Tomoyuki , Ueda Shigeharu , Miyamoto Hiroyuki , Kato Shiro
Published: November 2013

Variola virus (smallpox virus), vaccinia virus (VV), cowpox virus (CPV) and ectromelia virus (EV) belong to the genus Orthopoxvirus of the family Poxviridae. To establish the possible diagnosis for smallpox infection, monoclonal antibodies (MAbs) against VV and CPV were produced. The cross‐reactivity of seven MAbs with cells infected with various strains of the orthopoxviruses (CPV, VV and EV) was confirmed by an immunofluorescence (IF) test and other immunological analyses. Four and three MAbs reacted with the common antigen of all poxviruses (probably NP antigen) and the antigen involved in neutralization, respectively. We developed the IF test using these MAbs. The direct IF test required only 45 min to perform. Smallpox infection is now eradicated, but it is important to prepare for the diagnosis of smallpox in an emergency. The direct IF assay using MAbs cross‐reactive with orthopoxviruses is rapid, simple, specific, applicable for multiple samples, and will make it possible to screen for and detect orthopoxviruses that include variola virus with tissue impression smears from skin lesions in most laboratories or institutes.
Observation of the Intracellular Behavior of Recombinant Yersinia pseudotuberculosis Invasin Protein
Authors: Koga Hisashi , Izawa Ichiro , Araki Norie , Saya Hideyuki , Mimori Tatsuyuki
Published: November 2013

In this study, we observed the intracellular behavior of recombinant invasin, a 103‐kDa outer membrane protein of Yersinia pseudotuberculosis. To mimic the in vivo behavior of bacterial invasin, a polyvalent form of invasin was generated by incubation of biotinylated GST‐fused invasin C‐terminal portion protein (GST‐INVS) with avidin. Several experiments confirmed that the recombinant invasin could consistently reproduce the invasin‐mediated entry to mammalian epithelial cells. We analyzed the molecular kinetics of polyvalent INVS by western blotting, 125I‐uptake, and immunofluorescent microscopy. The internalized polyvalent INVS was rapidly translocated to the RIPA‐insoluble (polymerized‐actin enriched) fraction and formed cytoplasmic vesicles, while monovalent invasin did not show such kinetics. From these observations, we concluded that our bacterial‐free system is able to analyze the action of invasin for Yersinia pseudotuberculosis entry.
Hepatitis Induced by an IgM Monoclonal Antibody against Procarboxypeptidase R
Authors: He Lianying , Asai Suzuka , Kawamura Takeshi , Kimbara Noriaki , Tada Toyohiro , Okada Hidechika , Okada Noriko
Published: November 2013

Procarboxypeptidase R (proCPR), also known as thrombin‐activatable fibrinolysis inhibitor (TAFI), is present in plasma and can be activated to carboxypeptidase R (CPR) by trypsin‐like enzymes such as thrombin and plasmin. CPR has the carboxypeptidase B‐like activity that can inactivate the inflammatory peptides such as C5a by removing the C‐terminal arginine and can interfere with fibrinolysis by removing C‐terminal lysine residue of fibrin. In the present study, we conducted to produce monoclonal antibodies (mAbs) by using spleen cells from proCPR‐deficient mice immunized by partially purified mouse proCPR. The mAbs obtained were IgM isotype and reacted with proCPR and interfered with activation of proCPR to CPR by thrombin‐thrombomodulin complex. Some BALB/c mice implanted with the hybridoma died in 7 days, and intravenous injection of the mAb to BALB/c mice induced transient elevation of GOT and GPT in plasma although injection to the deficient mice did not. Furthermore, the histological features showed the focally lesions in liver tissue of BALB/c mice injected with the mAb. Since liver is the major site of proCPR synthesis, IgM mAb to proCPR should have induced local inflammation at the side resulting in induction of hepatitis.
A Rapid and Simple PCR Analysis Indicates There Are Two Subgroups of Vibrio vulnificus Which Correlate with Clinical or Environmental Isolation
Authors: Rosche Thomas M. , Yano Yutaka , Oliver James D.
Published: November 2013

Vibrio vulnificus is an estuarine bacterium which is the causative agent of both food‐borne disease and wound infection. Although V. vulnificus is commonly found in molluscan shellfish at high numbers, the incidence of disease is relatively low, leading to the hypothesis that not all strains of V. vulnificus are equally virulent. Unfortunately, there is currently no easy test to identify virulent strains of this species. We have previously identified a 200 bp randomly amplified polymorphic DNA (RAPD) PCR amplicon associated with clinical isolates. DNA sequence data from this locus in six clinical and four environmental isolates showed that the strains could be divided into two groups, which we termed C‐type (correlates with clinical origin) and E‐type (correlates with environmental origin). We designed PCR primers that could distinguish between the two groups, and typed 55 randomly selected strains. We found that 90% of the C‐type strains were clinical isolates, while 93% of environmental isolates were classified as E‐type. The region directly downstream of this locus contained a heptanucleotide sequence repeated various times depending on the strain. Using a PCR‐based assay to detect the repeat number present in a given strain, we found a statistically significant correlation with the C/E type classification and the number of repeats. The data reported here are consistent with the existence of two genotypes of V. vulnificus, with the C‐type being a strong indicator of potential virulence.
Macrophage Cell Death Due to Salmonella enterica Serovar Typhi and Its Acid Stress Protein Has Features of Apoptosis
Authors: Chander Harish , Majumdar Siddarth , Sapru Sunita , Rishi Praveen
Published: November 2013

Salmonella spp. have been shown to cause apoptosis of various host cell types as a part of their infection process. However, the induction of apoptosis remains to be looked into under the different host environments including acidic stress experienced by the pathogen. In order to simulate the in vivo acidic conditions, we studied the potential of S. typhi and its protein expressed under in vitro acidic conditions to induce apoptosis in macrophages. Murine macrophages were isolated and interacted with serovar Typhi and its acid stress protein for different time periods. The assessment of nucleosomal DNA, and nuclear staining with H‐33342 dye and flow cytometry indicated the occurrence of characteristic features of apoptosis. Analysis of data revealed that S. typhi caused apoptotic cell death in 61% of macrophages whereas stress‐induced protein alone accounted for apoptotic cell death in 45% of macrophages. The present study, for the first time demonstrates the potential of stress‐induced outermembrane component of S. typhi to induce apoptosis. Identification of such factors may offer new insights for understanding the pathophysiology of the disease during the host‐pathogen interactions.
Screening of Pro‐Apoptotic Genes Upregulated in an Experimental Street Rabies Virus‐Infected Neonatal Mouse Brain
Authors: Ubol Sukathida , Kasisith Jitra , Pitidhammabhorn Dhanesh , Tepsumethanol Veera
Published: November 2013

Rabies virus (RABV) is able to induce apoptotic death of target cells. The molecular pathway of RABV‐induced cell death is partially known. In the present study, cDNA array analysis was used as a tool to screen for pro‐apoptotic genes that may be involved in RABV induction. RNA was extracted from the infected CNS and from mock‐infected controls. When the mean gene expression was compared between the infected group and controls, 21 potential apoptotic genes were identified that exhibited more than 2.5‐fold difference in their expression levels. These 21 genes can be grouped into two groups, those genes that participate in the commitment phase and those that play a role as executioners. Examples of genes in commitment phase were death receptors (Fas‐L receptor, TNF‐receptor), lysosomal proteases, calpain, caspase‐1, signaling molecules (ERK, p38MAPK) and bcl‐2 family members. Cytochrome c and caspase‐3 were representatives of executioners. Based on types of genes activated during the commitment phase, two independent apoptotic mechanisms may be activated in response to the RV infection. The first is immune‐mediated death which may operate through the receptor‐ligand pathway activated by caspase‐1 and the pro‐inflammatory cytokine, IL‐1β. The other mechanism is a protease‐mediated process which involves lysosomal proteases and calcium‐dependent neutral proteases. These two stimulating pathways were followed by Bad, Bak, Bid activation and subsequently the upregulation of cytochrome c and caspase‐3. In addition, mobilization of K+ ion and other accessory apoptotic genes such as annexins and clusterin were also upregulated.
XAGE‐1 mRNA Expression in Prostate Cancer and Antibody Response in Patients
Authors: Koizumi Fumihito , Noguchi Yuji , Saika Takashi , Nakagawa Kazuhiko , Sato Shuichiro , Eldib Ali Mohamed Ali , Nasu Yasutomo , Kumon Hiromi , Nakayama Eiichi
Published: November 2013

To evaluate the feasibility of cancer vaccine targeting XAGE‐1, we investigated the expression of 4 XAGE‐1 transcript variants and the humoral immune response to XAGE‐1 in prostate cancer patients. XAGE‐1a, b, c, d mRNA expression was analyzed in 54 prostate cancer specimens and 8 specimens of benign prostate hypertrophy (BPH) by reverse transcription‐polymerase chain reaction (RT‐PCR). The humoral response to XAGE‐1 was investigated in sera obtained from 278 patients with prostate cancer and 40 healthy volunteers by enzyme‐linked immunosorbent assay (ELISA) using recombinant protein. XAGE‐1b mRNA expression was observed in 14 of 54 (26%) prostate cancer specimens, while XAGE‐1a, c, and d mRNA expressions were observed in 1, 1, and 3, respectively. None of the 4 XAGE‐1 transcript variants was observed in the 8 BPH specimens. Antibody against XAGE‐1 was detected in sera from 2 of 129 stage D2 patients, whereas none of sera from 149 patients with localized prostate cancer or lymph node metastasis had detectable XAGE‐1 antibody. No reactivity to XAGE‐1 was found in sera from the 40 healthy individuals.
Seroprevalence to the Circumsporozoite Protein Peptide Antigen of Plasmodium vivax in Korean Children
Authors: Lim Chae Seung , Yoon Jae Kyun , Chang Eun Ah , Suh In Bum , An Seong Soo A. , Lee Kee‐Hyoung , Chung Ji Tae , Tockgo Young Chang
Published: November 2013

Recently, malaria re‐emerged in the Republic of Korea (ROK), but there have been only a few reports on malaria seroprevalence. For the epidemiological study in children, a total of 1,176 serum samples were obtained from children and adolescent inhabitants from the three different regions, Pajoo, the Guro district in the western part of Seoul (Guro), and Ansan, from June to September 2002, when the transmission rate was high. Anti‐circumsporozoite protein (CSP) antibody levels were assessed in samples using an indirect enzyme‐linked immunosorbent assay (ELISA) method. Among the three regions, the highest IgG seroreactivity against the CSP antigen of Plasmodium vivax was found in the children from Pajoo (8.0%), followed by the children from Guro (3.2%), and those in Ansan (0.02%) had the lowest seroreactivity. In Pajoo, the profile of antibody response showed the highest in age 9, but decreased with age towards 12 years old. We found significant correlation between the seroprevalence and annual incidence in the investigated areas, suggesting that the area‐related patterns reflected the differences of inoculation rates in children.
Confirmation of the Anomeric Structure of Galacturonic Acid in the Galacturonosyl‐Ceramide of Sphingomonas yanoikuyae
Authors: Kawahara Kazuyoshi , Sato Noriko , Tsuge Koichiro , Seto Yasuo
Published: November 2013

The anomeric structure of glycosphingolipids significantly influences their activity to stimulate natural killer T cells. In this study the chemical structure of the galacturonosyl‐ceramide in Sphingomonas yanoikuyae, designated GSL‐1′sy, was re‐examined to prove the anomeric structure of the D‐galacturonic acid (GalA) in the lipid, which was reported as β‐configuration by Naka et al., but was suggested as α‐configuration in our preliminary study. GSL‐1′sy was purified from the bacterial cells with the same procedure as Naka et al. The 1H‐NMR analysis of GSL‐1′sy revealed that the coupling constant of the anomeric proton of GalA was 3.0 Hz, indicating that GalA in GSL‐1′sy is α‐anomer, the configuration active for the stimulation of natural killer T cells.
Isolation and Identification of Methylobacterium Species from the Tap Water in Hospitals in Japan and Their Antibiotic Susceptibility
Authors: Furuhata Katsunori , Kato Yuko , Goto Keiichi , Hara Motonobu , Yoshida Shin‐ichi , Fukuyama Masafumi
Published: November 2013

Contamination of tap water by Methylobacterium species has become a serious concern in hospitals. This study was planned to examine the distribution of Methylobacterium species inhabiting tap water used in Japanese hospitals and antibiotic sensitivity of the isolates in 2004. Species identification of 58 isolates was performed based on the homology of a partial sequence of 16S rDNA. The dominant Methylobacterium species in hospital water were M. aquaticum and M. fujisawaense. To examine the biochemical properties of these isolates, a carbon source utilization was tested using an API50CH kit. The phenotypic character varied widely, and was not necessarily consistent with the results of phylogenic analysis based on the partial 16S rDNA sequence, suggesting that the biochemical properties are not suitable for identification of Methylobacterium species. The isolates were also subjected to antibiotic sensitivity tests. They were resistant to 8 antibiotics, but highly sensitive to imipenem (MIC90=1 μg/ml) and tetracycline (MIC90=8 μg/ml). These findings concerning the isolates revealed the presence of Methylobacterium species with resistance to multiple antibiotics in hospital tap water.
Cytokine GM‐CSF Genetic Adjuvant Facilitates Prophylactic DNA Vaccine against Pseudorabies Virus through Enhanced Immune Responses
Authors: Yoon Hyun A. , Aleyas Abi G. , George Junu A. , Park Seong Ok , Han Young Woo , Lee John Hwa , Cho Jeong‐Gon , Eo Seong Kug
Published: November 2013

Granulocyte/macrophage colony‐stimulatory factor (GM‐CSF) is an attractive adjuvant for a DNA vaccine on account of its ability to recruit antigen‐presenting cells (APCs) to the site of antigen synthesis as well as its ability to stimulate the maturation of dendritic cells (DCs). This study evaluated the utility of GM‐CSF cDNA as a DNA vaccine adjuvant for glycoprotein B (gB) of pseudorabies virus (PrV) in a murine model. The co‐injection of GM‐CSF DNA enhanced the levels of serum PrV‐specific IgG with a 1.5‐ to 2‐fold increase. Moreover, GM‐CSF co‐injection inhibited the production of IgG2a isotype. However, it enhanced production of an IgG1 isotype resulting in humoral responses biased to the Th2‐type against PrV antigen. In contrast, the co‐administration of GM‐CSF DNA enhanced the T cell‐mediated immunity biased to the Th1‐type, as judged by the significantly higher level of cytokine IL‐2 and IFN‐γ production but not IL‐4. When challenged with a lethal dose of PrV, the GM‐CSF co‐injection enhanced the resistance against a PrV infection. This suggests that co‐inoculation with a vector expressing GM‐CSF enhanced the protective immunity against a PrV infection. This immunity was caused by the induction of increased humoral and cellular immunity in response to PrV antigen.
Phylogenetic Analysis of Envelope Glycoprotein (E1) Gene of Rubella Viruses Prevalent in Japan in 2004
Authors: Saitoh Mika , Shinkawa Naomi , Shimada Shinichi , Segawa Yukari , Sadamasu Kenji , Hasegawa Michiya , Kato Masahiko , Kozawa Kunihisa , Kuramoto Tsuyoshi , Nishio Osamu , Kimura Hirokazu
Published: November 2013

We performed a molecular epidemiological study on the envelope glycoprotein gene (E1 gene) obtained by PCR amplification from specimens of 17 rubella patients in certain areas (Gunma, Saitama, and Kagoshima prefectures, and Tokyo metropolitan area) in Japan in 2004. In these sequences of partially amplified DNAs (283 bases) within the E1 gene, no nucleotide substitution was observed. They were classified into genotype 1D of clade 1 in the constructed phylogenetic tree. One amino acid substitution was found between the amino acid sequence predicted from these DNAs and those of Japanese strains [To‐336 vaccine strain (To‐336 vac) and its wild progenitor (To‐336 wt)]. The results suggest that the rubella viruses (RV) prevalent in certain areas of Japan in 2004 were highly homologous and were closely related with Japanese vaccine strain.
Characterization of Recombinant Rabies Virus Carrying Double Glycoprotein Genes
Authors: Hosokawa‐Muto Junji , Ito Naoto , Yamada Kentaro , Shimizu Kenta , Sugiyama Makoto , Minamoto Nobuyuki
Published: November 2013

A recombinant rabies virus carrying double glycoprotein (G) genes, R(NPMGGL) strain, was generated by a reverse genetics system utilizing cloned cDNA of the RC‐HL strain, and the biological properties of the virus were compared to those of the recombinant RC‐HL (rRC‐HL) strain. The extents of virus growth in cultured cells and virulence for adult mice of the R(NPMGGL) strain were almost same as those of the rRC‐HL strain, while G protein content of the purified R(NPMGGL) virion and G protein expression level in R(NPMGGL)‐infected cells were 1.5‐fold higher than those of the rRC‐HL strain. As a result of serial passages of the R(NPMGGL) strain in cultured cells, the expression level of G protein in cultured cells infected with the passaged R(NPMGGL) strain was maintained and virus titers rose with adaptation to the cultured cells. Furthermore, analysis of neutralization titers in mice immunized with UV‐inactivated virus suggested that the R(NPMGGL) strain had higher immunogenicity than that of the rRC‐HL strain. The results suggest that the R(NPMGGL) strain carrying double G genes might be a useful candidate for development of a new inactivated rabies vaccine.
Nucleolar Protein B23 Interacts with Japanese Encephalitis Virus Core Protein and Participates in Viral Replication
Authors: Tsuda Yoshimi , Mori Yoshio , Abe Takayuki , Yamashita Tetsuo , Okamoto Toru , Ichimura Tohru , Moriishi Kohji , Matsuura Yoshiharu
Published: November 2013

Japanese encephalitis virus (JEV) core protein is detected not only in the cytoplasm but also in the nucleoli of infected cells. We previously showed that a mutant JEV lacking the nucleolar localization of the core protein impaired viral replication in mammalian cells. In this study, we identified a nucleolar phosphoprotein B23 as a protein binding with the core protein of JEV but not with that of dengue virus. The region binding with JEV core protein was mapped to amino acid residues 38 to 77 of B23. Upon JEV infection, some fraction of B23 was translocated from the nucleoli to the cytoplasm, and cytoplasmic B23 was colocalized with the core protein of wild‐type JEV but not with that of the mutant JEV. Furthermore, overexpression of dominant negatives of B23 reduced JEV replication. These results suggest that B23 plays an important role in the intracellular localization of the core protein and replication of JEV.
Monitoring and Survival of Lactobacillus gasseri SBT2055 in the Human Intestinal Tract
Authors: Takahashi Hidetoshi , Fujita Takashi , Suzuki Yutaka , Benno Yoshimi
Published: November 2013

The monitoring and survival of Lactobacillus gasseri SBT2055 in the human intestinal tract was investigated with seven healthy subjects having a low number of fecal lactobacilli. An increase of fecal lactobacilli (1032–52 CFU/g feces) was recognized after ingestion of yogurt with SBT2055 by the subjects. A high positive rate of L. gasseri in fecal lactobacilli detected from the subjects (over 70% at 2nd weeks of feeding) was also observed during the ingestion period using the species‐specific PCR system. These findings indicate that the SBT2055 strain in yogurt survived in the human intestinal tract and was recovered from human feces.
Identification and Characterization of a Novel Human Collectin CL‐K1
Authors: Keshi Hiroyuki , Sakamoto Takashi , Kawai Takao , Ohtani Katsuki , Katoh Tsuyoshi , Jang Seong‐Jae , Motomura Wataru , Yoshizaki Takayuki , Fukuda Mitsuko , Koyama Satoshi , Fukuzawa Jun , Fukuoh Atsushi , Yoshida Itsuro , Suzuki Yasuhiko , Wakamiya Nobutaka
Published: November 2013

Collectins are a family of C‐type lectins with two characteristic structures, collagen like domains and carbohydrate recognition domains. They recognize carbohydrate antigens on microorganisms and act as host‐defense. Here we report the cloning and characterization of a novel collectin CL‐K1. RT‐PCR analyses showed CL‐K1 mRNA is present in all organs. The deduced amino acid sequence and the data from immunostaining of CL‐K1 cDNA expressing CHO cells revealed that CL‐K1 is expressed as a secreted protein. CL‐K1 is found in blood by immunoblotting and partial amino acid analyses. CL‐K1 showed Ca2+‐dependent sugar binding activity of fucose and weakly mannose but not N‐acetyl‐galactosamine, N‐acetyl‐glucosamine, or maltose, though mannose‐binding lectin (MBL) containing similar amino acid motif. CL‐K1 can recognize specially several bacterial saccharides due to specific sugar‐binding character. Elucidation of the role of two ancestor collectins of CL‐K1 and CL‐L1 could lead to see the biological function of collectin family.
Genotypic Analyses of Escherichia coli Isolated from Chickens with Colibacillosis and Apparently Healthy Chickens in Japan
Authors: Kawano Masanori , Yaguchi Kazuhiko , Osawa Ro
Published: November 2013

A genotypic comparison using pulsed‐fleld gel electrophoresis (PFGE), amplified ribosomal restriction analysis (ARDRA) as well as PCRs targeting virulence associated genes reported elsewhere in avian pathogenic Escherichia coli (APEC) was made between E. coli strains isolated from chickens with colibacillosis and those from the feces of apparently healthy chickens in Japan. The majority (67%) of clinical isolates belonged to a certain phylogenetic ARDRA but not PFGE cluster, with virulence‐related genes carried by ColV plasmid being markedly prevalent. The result suggests that APEC strains originated from the same “ancestor” in the course of E. coli evolution.
Adrenal Participation in Thymocyte Death by Anti‐CD3 Antibodies In Vivo
Authors: Yoshida Mitsuo , Gotoh Ken , Fujii Masato , Shimada Hiroto , Touma Maki , Hosono Masamichi
Published: November 2013

The deletion of CD4‐ and CD8‐double‐positive (DP) cells in the thymus after treatment with anti‐CD3 antibodies has long been considered as a useful model for clonal deletion during T cell development, although it was reported that DP cell death was not observed in neonates where self‐tolerance should be developing. We dealt with the cellular basis of this enigmatic phenomenon in this report Due to the similar susceptibility to the antibody‐treatment in vitro between neonatal and adult thymocytes, critical factors may be outside rather than within the thymus. Indeed, newborn thymus lobes transplanted into recipients of different ages showed an increased susceptibility to the thymo‐toxicity as the age of the recipient increased. The thymo‐toxicity seems to be based on the adrenal function of glucocorticoid (GC) synthesis, because administration of an inhibitor of GC synthesis significantly reduced the DP cell death by the antibody‐treatment Finally, adrenalectomy completely prevented DP cell death by anti‐CD3 antibodies in adult mice. Therefore, the thymocyte death by anti‐CD3 antibodies in vivo may not be due to the T cell‐receptor mediated selection in the thymus.
Shifting Seroepidemiology of Hepatitis A in Japan, 1973–2003
Authors: Kiyohara Tomoko , Sato Tomoko , Totsuka Atsuko , Miyamura Tatsuo , Ito Toshihiro , Yoneyama Tetsuo
Published: November 2013

Background. Hepatitis A infection is caused by hepatitis A virus (HAV) contracted through fecal‐oral transmission. Life‐long immunity is conferred after infection. Improved sanitary conditions have generally resulted in a significant decline in the incidence of hepatitis A. However, a low incidence of infection results in increased HAV susceptibility. The present study investigates the prevalence of anti‐HAV antibody and clarifies the current HAV status and HAV susceptibility in Japan at 2003. Methods. A total of 2,430 serum specimens collected during 2003 from Japanese individuals ranging in age from 0–92 years, were tested for anti‐HAV antibody using an inhibition enzyme linked immunosorbent assay. All specimens were obtained from the WHO and the National Serum Reference Bank/National Institute of Infectious Diseases, Tokyo, Japan. Results. The overall seroprevalence was 12.2%. Anti‐HAV antibodies were rarely detected in individuals between 0–44 years of age. Starting from the age of 45–49 years, seropositivity gradually increased through age 65 years and above. Seroprevalence was not affected by gender, and geographic distribution did not affect age‐specific seroprevalence until the age of 60 years. Conclusions. HAV susceptibility in Japan is increasing annually. Particularly, the prevalence of anti‐HAV antibody in individuals older than 50 years in 2003 was 50.3%, which is significantly lower than that of corresponding studies in 1994 (74.3%), 1984 (96.9%) and 1973 (96.9%). The growing susceptible population of advanced age results in more frequent HAV infection among them. The surveillance of anti‐HAV antibody prevalence is useful for implementing preventive measures and for controlling the spread of HAV.
Characterization of the Inhabitancy of Mouse Intestinal Bacteria (MIB) in Rodents and Humans by Real‐Time PCR with Group‐Specific Primers
Authors: Kibe Ryoko , Sakamoto Mitsuo , Yokota Hiroshi , Benno Yoshimi
Published: November 2013

Mouse intestinal bacteria (MIB) is a new operational taxonomic unit (OTU) belonging to the Bacteroides subgroup in the Cytophaga‐Flavobacter‐Bacteroides (CFB) phylum recently found in the intestine of mice, rats and humans. However, their characters are still unknown since they have not yet been isolated by culture. To understand their habitat characteristics in intestinal tracts, the quantification assays of MIB were established using MIB group‐specific primers. The MIB population in the intestine was evaluated as a percentage of the number of 16S rRNA gene copy of MIB. A real‐time PCR assay using group‐specific primers showed the fluctuation of MIB inhabitancy and revealed that the MIB population in the small intestine of mice was significantly lower than the large intestinal contents. Moreover, MIB was found in human feces though the number was lower than in murine. This assay using group‐specific primers revealed new information about host‐preference of MIB.
Molecular Survey ofBabesia microti, EhrlichiaSpecies andCandidatusNeoehrlichia mikurensis in Wild Rodents from Shimane Prefecture, Japan
Authors: Tabara Kenji , Arai Satoru , Kawabuchi Takako , Itagaki Asao , Ishihara Chiaki , Satoh Hiroshi , Okabe Nobuhiko , Tsuji Masayoshi
Published: November 2013

A significant number of patients are diagnosed with “fevers of unknown origin” (FUO) in Shimane Prefecture in Japan where tick‐borne diseases are endemic. We conducted molecular surveys forBabesia microti, Ehrlichiaspecies, andCandidatusNeoehrlichia mikurensis in 62 FUO cases and 62 wild rodents from Shimane Prefecture, Japan. PCR using primers specific for theBabesia18S small‐subunit rRNA (rDNA) gene andAnaplasmataceae groESLamplified products from 45% (28/62) and 25.8% (16/62) of captured mice, respectively. Of the 28 18S rDNA PCR positives, 23 and five samples were positive for Hobetsu‐ and Kobe‐typeB. microti, respectively. In contrast, of the 16groESLPCR positives, eight, one and seven samples were positive forEhrlichia muris, Ehrlichiasp. HF565 andCandidatusN. mikurensis, respectively. Inoculation of selected blood samples into Golden Syrian hamsters indicated the presence of Hobetsu‐ and Kobe‐typeB. microtiin four and one sample, respectively. Isolation of the latter strain was considered important as previous studies suggested that the distribution of this type was so far confined to Awaji Island in Hyogo Prefecture, where the first case of transfusion‐associated human babesiosis originated. DNA samples from 62 FUO human cases tested negative forB. microti18S rDNA gene,Anaplasmataceae groESLgene,Rickettsia japonica17K genus‐common antigen gene andOrientia tsutsugamushi56K antigen gene by PCRs. We also conducted seroepidemiological surveys on 62 human sera collected in Shimane Prefecture from the FUO patients who were suspected of carrying tick‐borne diseases. However, indirect immunofluorescent antibody tests usingB. microti‐ andE. muris‐infected cells detected IgG againstE. murisin only a single positive sample. This study demonstrates the presence of several potentially important tick‐borne pathogens in Shimane Prefecture and suggests the need for further study on the causative agents of FUOs.
Detection of Enterotoxin and Toxic Shock Syndrome Toxin 1 Genes in Staphylococcus, with Emphasis on Coagulase‐Negative Staphylococci
Authors: de Lourdes R.S. da Cunha Maria , Calsolari Regina A.O. , Júnior João Pessoa Araújo
Published: November 2013

The detection of staphylococcal enterotoxins is decisive for the confirmation of an outbreak and for the determination of the enterotoxigenicity of strains. Since the recognition of their antigenicity, a large number of serological methods for the detection of enterotoxins in food and culture media have been proposed. Since immunological methods require detectable amounts of toxin, molecular biology techniques represent important tools in the microbiology laboratory. In the present study, polymerase chain reaction (PCR) was used to identify genes responsible for the production of enterotoxins and toxic shock syndrome toxin 1 (TSST‐1) in S. aureus and coagulase‐negative staphylococci (CNS) isolated from patients and the results were compared with those obtained by the reverse passive latex agglutination (RPLA) assay. PCR detection of toxin genes revealed a higher percentage of toxigenic S. aureus strains (46.7%) than the RPLA method (38.3%). Analysis of the toxigenic profile of CNS strains showed that 26.7% of the isolates produced some type of toxin, and one or more toxin‐specific genes were detected in 40% of the isolates. These results suggests the need for further studies in order to better characterize the pathogenic potential of CNS and indicate that attention should be paid to the toxigenic capacity of this group of microorganisms.
Pro‐ and Anti‐Inflammatory Cytokines Produced by Human Monocytes Challenged In Vitro with Paracoccidioides brasiliensis
Authors: Kurokawa Cilmery Suemi , Araujo João P. , Soares Angela M.V.C. , Sugizaki Maria F. , Peraçoli Maria Terezinha S.
Published: November 2013

Monocytes and macrophages play a central role in innate and adaptive immune response against systemic fungal infections. Imbalances in suppressor or stimulatory cytokine secretion caused by these cells may influence disease development, microorganism death, and the nature of the adaptive immune response. This study analyzed the monocyte cytokine profiles of healthy individuals challenged with high and low virulent strains of P. brasiliensis and mRNA cytokine expression kinetics by reverse transcription polymerase chain reaction (RT‐PCR). Peripheral blood monocytes from healthy volunteers were cultured in vitro with and without virulent (Pb18) or low virulence (Pb265) strains from P. brasiliensis viable yeast cells. Interleukin‐1 beta (IL‐1β), IL‐6, IL‐8, IL‐10, tumor necrosis factor‐alpha (TNF‐α), and transforming growth factor‐beta (TGF‐β1) were measured in culture supernatants by enzyme immunoassay (ELISA), and mRNA cytokine expression was determined by RT‐PCR at 0, 4, 8, 12, 18 and 48 hr. Both P. brasiliensis strains induced monocyte production of IL‐1β, IL‐6, IL‐10 and TNF‐α. Pb18 induced higher levels of IL‐1β, IL‐6, and IL‐10 than Pb265. IL‐8 and TGF‐β1 levels were not significantly different from those cultured without stimulus. The mRNA cytokine expression was similar to supernatant cytokines measured by ELISA. In vitro monocyte challenge with virulent P. brasiliensis strain induces earlier and higher levels of pro‐ and anti‐inflammatory cytokines than low virulence strain.