bluuebunny/crossref_metadata_2025_split huggingface dataset:
Gene Cloning and Characterization of VcrM, a Na+‐Coupled Multidrug Efflux Pump, from Vibrio cholerae Non‐O1
Authors: Huda Md. Nazmul , Chen Jing , Morita Yuji , Kuroda Teruo , Mizushima Tohru , Tsuchiya Tomofusa 
Published: November 2013

AbstractWe cloned a DNA fragment responsible for drug resistance from chromosome of Vibrio cholerae non‐O1. Nucleotide sequence analysis of this fragment revealed the presence of a single open reading frame encoding a protein consisting of 445 amino acid residues. We designated the gene as vcrM. Hydropathy analysis of the deduced amino acid sequence of VcrM suggests the presence of 12 trans‐membrane segments. A dendrogram showed that VcrM is a member of the DinF‐subfamily within the MATE family of multidrug efflux pumps. Expression of the cloned vcrM gene in drug‐hypersensitive Escherichia coli KAM32 cells made them resistant to acriflavine, 4′, 6‐diamidino‐2‐phenylindole, Hoechst 33342, rhodamine 6G, tetraphenylphosphonium chloride (TPPCl) and ethidium bromide. Efflux of acriflavine due to VcrM was dependent on Na+ or Li+. Moreover, Na+ efflux was observed with VcrM when TPPCl was added to Na+‐loaded cells. Therefore, we conclude that VcrM is a Na+/drug antiporter‐type multidrug efflux pump.
A Novel Murine Model of Oral Candidiasis with Local Symptoms Characteristic of Oral Thrush
Authors: Takakura Natsuko , Sato Yuichi , Ishibashi Hiroko , Oshima Haruyuki , Uchida Katsuhisa , Yamaguchi Hideyo , Abe Shigeru 
Published: November 2013

AbstractA conventional and easy method to establish a murine oral candidiasis model, which has not only a stable yeast population in the oral cavity but also symptoms characteristic of oral thrush, was developed by using a sedative agent. Mice were immunosuppressed with prednisolone and were given tetracycline hydrochloride. They were orally infected with 106 viable cells of Candida albicans by means of a cotton swab and enough chlorpromazine chloride had been injected to keep them in a sedative state about for 3 hr after inoculation. From day 3 to day 7 post inoculation, 105–106 colony forming units of Candida were recovered from the oral cavity of each mouse and whitish, curd‐like patches were observed on most parts of tongue. Microscopically, germ tubes had appeared on the tongue surface. This model would be a useful experimental oral candidiasis for investigating the pathogenesis of C. albicans oral infection and the efficacy of various antifungal agents microbiologically and symptomatically.
Genotype and Subtype Analyses of Hepatitis B Virus (HBV) and Possible Co‐Infection of HBV and Hepatitis C Virus (HCV) or Hepatitis D Virus (HDV) in Blood Donors, Patients with Chronic Liver Disease and Patients on Hemodialysis in Surabaya, Indonesia
Authors: Lusida Maria I. , Surayah  , Sakugawa Hiroshi , Nagano‐Fujii Motoko , Soetjipto  , Mulyanto  , Handajani Retno , Boediwarsono  , Setiawan Poernomo B. , Nidom Chairul A. , Ohgimoto Shinji , Hotta Hak 
Published: November 2013

AbstractFour subtypes (adw, adr, ayw, and ayr) and eight genotypes (A to H) of the hepatitis B virus (HBV) have been identified. They appear to be associated with particular geographic distribution, ethnicity, and possibly clinical outcomes. In this study, hepatitis B surface antigen (HBsAg) subtyping and HBV genotyping were carried out on sera obtained from HBsAg‐positive HBV carriers, including healthy blood donors; patients with acute hepatitis, chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma; and patients on hemodialysis all located in Surabaya, Indonesia. We report here that all HBV isolates tested in Surabaya belonged to genotype B, with more than 90% of them being classified into subtype adw. Our results also revealed that prevalence of hepatitis C virus (HCV) co‐infection among HBV carriers in Surabaya was approximately 10% for healthy blood donors and patients with chronic liver disease, and approximately 60% for patients on maintenance hemodialysis. Interestingly, HBsAg titers were lower in HBV carriers with HCV co‐infection than in those without HCV co‐infection. We also found that prevalence of hepatitis D virus (HDV) co‐infection was <0.5% among HBV carriers in Surabaya.
Molecular Cloning and Characterization of the HmrM Multidrug Efflux Pump from Haemophilus influenzae Rd
Authors: Xu Xing‐Jue , Su Xian‐Zhong , Morita Yuji , Kuroda Teruo , Mizushima Tohru , Tsuchiya Tomofusa 
Published: November 2013

AbstractWe cloned a gene responsible for norfloxacin resistance from the chromosomal DNA of Haemophilus influenzae Rd, and designated the gene as hmrM. HmrM showed sequence similarity with NorM of Vibrio parahaemolyticus and YdhE of Escherichia coli and others that belong to the MATE family multidrug efflux pumps. The recombinant plasmid carrying the hmrM gene conferred elevated resistance not only to norfloxacin but also to acriflavine, 4′,6‐diamidino‐2‐phenylindole, doxorubicin, ethidium bromide, tetraphenylphosphonium chloride, Hoechst 33342, daunomycin, berberine, and sodium deoxycholate in Escherichia coli KAM32, a drug‐hypersensitive strain. We observed an Na+‐dependent efflux of ethidium and an ethidium‐induced efflux of Na+ in E. coli KAM32 cells harboring the plasmid carrying the hmrM gene. These results indicate that HmrM is an Na+/drug antiporter‐type multidrug efflux pump. A difference in substrate preference was observed between HmrM, NorM, and YdhE.
Homologous and Heterologous Antibody Responses to Lipopolysaccharide after Enterohemorrhagic Escherichia coli Infection
Authors: Tsutsumi Reiko , Ichinohe Naoko , Shimooki Osamu , Obata Fumiko , Takahashi Kiomi , Inada Katsuya , Sasaki Mika , Sato Shigehiro , Chida Shoichi 
Published: November 2013

AbstractTo evaluate antibody responses against lipopolysaccharide (LPS: O157, O26, and O111) in enterohemorrhagic Escherichia coli (EHEC) infection, sera of 24 schoolchildren associated with the Morioka outbreak in 1997 and of 74 sporadic patients suspected of having EHEC infection were examined. Using a positive standard serum, quantitative evaluation of LPS antibodies by an enzyme‐linked immunosorbent assay (ELISA) was established. High levels of specific IgM and IgA antibodies against homologous E. coli LPS were present in the acute period and are characteristic of EHEC. This could be used for the serological diagnosis of EHEC infection, except for early infants and the elderly. In addition to the specific homologous response, multiple antibody responses against different serotypes other than those isolated were demonstrated in many cases by qualitative analysis using Western blotting.
CD72 Stimulation Modulates Anti‐IgM Induced Apoptotic Signaling through the Pathway of NF‐κB, c‐Myc and p27Kip1
Authors: Fujiwara Naruyoshi , Fusaki Noemi , Hozumi Nobumichi 
Published: November 2013

AbstractEngagement of mIgM induces G1 arrest and apoptosis in immature B cells. The biochemical mechanism(s) regulating the cell death process are poorly understood. Cross‐linking of CD72 (a B cell co‐receptor) with anti‐CD72 antibody was shown to protect B cells from apoptosis. We investigated the molecular mechanism involved in apoptosis preventing signaling mediated by CD72 ligation using a derivative (WEHIδ) of the WEHI231 cell line which is representative of immature B cells. Apoptotic WEHIδ cells following cross‐linking of mIgM demonstrate a dramatic loss of c‐Myc protein after transient up‐regulation. In contrast, pre‐ligation of CD72 was able to sustain c‐Myc expression after transient up‐regulation. Cross‐linking of mIgM of WEHIδ cells causes accumulation of the Cdk inhibitor, p27Kip1. CD72 pre‐ligation was shown to inhibit the accumulation of p27Kip1 protein. Moreover, NF‐κB activity was not suppressed in WEHIδ cells after mIgM cross‐linking when the cells were pre‐treated with anti‐CD72 antibody. These results strongly suggest that the apoptosis preventing signal evoked by CD72 ligation is delivered through the pathway of NF‐κB, c‐Myc, p27Kip1 and cyclin.
First Isolation of Bartonella henselae Type I from a Cat‐Scratch Disease Patient in Japan and Its Molecular Analysis
Authors: Maruyama Soichi , Izumikawa Koichi , Miyashita Mayumi , Kabeya Hidenori , Mikami Takeshi , Yamanouchi Hirotsugu , Sasaki Eisuke , Yoshida Hiroshi , Izumikawa Kinichi 
Published: November 2013

AbstractWe isolated Bartonella henselae from an inguinal lymph node of a 36‐year‐old male patient with cat‐scratch disease. The patient had many areas of erythema on his body, swelling of the left inguinal lymph nodes with pain and slight fever. The diagnosis was made on the basis of polymerase chain reaction for B. henselae DNA from the lymph node biopsies and blood sample, and isolation of the organism, histology of the lymph node and serology with an indirect immunofluorescent antibody test. We also analyzed the genome profiles for five strains of 90 isolates from the lymph node by pulsed‐field gel electrophoresis after NotI endonuclease digestion. We found two different genomic profiles. These results suggest that the patient had been either co‐infected or re‐infected with two genetically different strains of B. henselae.
Emergence and Serovar Transition of Vibrio parahaemolyticus Pandemic Strains Isolated during a Diarrhea Outbreak in Vietnam between 1997 and 1999
Authors: Chowdhury Ashrafuzzaman , Ishibashi Masanori , Thiem Vu Dinh , Tuyet Dinh Thi Ngoc , Van Tung Tran , Chien Bui Trong , von Seidlein Lorenz , Canh Do Gia , Clemens John , Trach Dang Duc , Nishibuchi Mitsuaki 
Published: November 2013

AbstractWe characterized 523 Vibrio parahaemolyticus strains isolated during a survey of diarrhea patients in Khanh Hoa province, Vietnam between 1997 and 1999. Forty‐nine percent of the strains were judged to belong to the pandemic strains that emerged around 1996 and spread to many countries. These strains were positive in the GS‐PCR assay and carried the tdh gene. The ORF8 of the f237 phage genome, a possible marker of the pandemic clone, was absent in 10% of these strains. Eleven O:K serovars were detected among the pandemic strains and the strains representing all 11 serovars of pandemic strains were shown to be closely related regardless of the ORF8 genotype using arbitrarily primed PCR and pulsed field gel electrophoresis analyses. It was clear that a transition of major serovars occurred among the pandemic strains represented by the emergence of O3:K6 in 1997, O4:K68 in 1998, and O1:K25 in 1998 and 1999.
Characterization of a Shiga‐Toxin 1‐Resistant Stock of Vero Cells
Authors: Sekino Takaomi , Kiyokawa Nobutaka , Taguchi Tomoko , Takenouchi Hisami , Matsui Jun , Tang Wei‐Ran , Suzuki Toyo , Nakajima Hideki , Saito Masahiro , Ohmi Kazuhiro , Katagiri Yohko U. , Okita Hajime , Nakao Hiroshi , Takeda Tae , Fujimoto Junichiro 
Published: November 2013

AbstractShiga toxins (Stxs, also referred to as verotoxins) were first described as a novel cytotoxic activity against Vero cells. In this study, we report the characterization of an Stx1‐resistant (R‐) stock of Vero cells. (1) When the susceptibility of R‐Vero cells to Stx1 cytotoxicity was compared to that of Stx1‐sensitive (S‐) Vero cells by methylthiazolyldiphenyl‐tetrazolium bromide (MTT) assay, cell viability after 48‐hr exposure to 10 pg/ml of Stx1 was greater than 80% and less than 15%, respectively. (2) Although both a binding assay of fluorescence‐labeled Stx1 and lipid analysis indicated considerable expression of Gb3Cer, a functional receptor for Stxs, in both Vero cells, anti‐Gb3Cer monoclonal antibodies capable of binding to S‐Vero cells failed to effectively label R‐Vero cells, suggesting a conformational difference in the Gb3Cer expressed on R‐Vero cells. (3) The lipid analysis also showed that the R‐Vero cells contained significant amounts of Gb4Cer. In addition, introduction of exogenous Gb4Cer into S‐Vero cells slightly inhibited Stx1 cytotoxicity, suggesting some correlation between glycosphingolipid composition and Stx1 resistance. (4) Both butyrate treatment and serum depression eliminated the Stx1 resistance of R‐Vero cells. (5) The results of the analysis by confocal microscopy suggest a difference in intracellular transport of Stx1 between R‐Vero and S‐Vero cells. Further study of R‐Vero cells may provide a model of Stx1 resistance via distinct intracellular transport of Stx1.
Comparison of Real‐Time and Nested PCR Assays for Detection of Herpes Simplex Virus DNA
Authors: Kawada Jun‐ichi , Kimura Hiroshi , Ito Yoshinori , Hoshino Yo , Tanaka‐Kitajima Naoko , Ando Yoshihiro , Futamura Masahide , Morishima Tsuneo 
Published: November 2013

AbstractWe performed a real‐time PCR assay to detect herpes simplex virus (HSV) DNA, and compared it prospectively with a nested PCR assay in 164 clinical samples (109 cerebrospinal fluid and 55 sera) from patients suspected of having neonatal HSV infection or HSV encephalitis. In 25 of 164 samples, HSV DNA was detected by the nested PCR assay. All samples positive for HSV DNA in the nested PCR assay were also positive in the real‐time PCR assay, and all but two samples negative for HSV DNA in the nested assay were negative in the real‐time assay. The real‐time PCR assay thus had a sensitivity of 100% and a specificity of 99%, when compared with the nested assay. Sequential assays in a case of disseminated HSV showed that a decrease in HSV DNA paralleled clinical improvement. Quantification of HSV DNA by real‐time PCR was useful for diagnosing and monitoring patients with HSV encephalitis and neonatal HSV infection.
Characterization of Ehrlichia Species from Ixodes ovatus Ticks at the Foot of Mt. Fuji, Japan
Authors: Inayoshi Megumi , Naitou Hirotaka , Kawamori Fumihiko , Masuzawa Toshiyuki , Ohashi Norio 
Published: November 2013

AbstractA total of 390 adult ticks (288 Ixodes ovatus and 102 I. persulcatus) collected at the foot of Mt. Fuji and two near cities in Shizuoka prefecture, Japan, were examined for Ehrlichia infection by isolation with laboratory mice from whole tick tissues. Ehrlichial DNAs were detected from the spleens of mice inoculated with tissues from I. ovatus, but not I. persulcatus. The prevalence of ehrlichiae in the ticks was estimated to be ca. 3%. The 16S rDNA analysis revealed that the sequences of 8 ehrlichial isolates (termed “Shizuoka” isolates) obtained were identical, and they were very similar, but not identical, to those of two Ehrlichia species strain variants recently isolated in Japan, followed by Ehrlichia chaffeensis in the US. Analysis of parts of the omp‐1 multigene family specific for monocytic ehrlichiosis agents showed that the Shizuoka isolates were distinct from other ehrlichial organisms. The Shizuoka isolates caused death in immunocompetent laboratory mice, suggesting that they are highly pathogenic in mice. The data show that the Shizuoka isolates are likely to be a new strain variant of Ehrlichia species in Japan. Further characterization and surveillance will be required in Japan due to the presence of these human ehrlichiosis agent‐like organisms.
The Throat Flora and Its Mitogenic Activity in Patients with Kawasaki Disease
Authors: Horita Norihisa , Yokota Shin‐ichi , Fuse Shigeto , Takamuro Motoki , Tomita Hideshi , Sato Kiyoshi , Fujii Nobuhiro , Tsutsumi Hiroyuki 
Published: November 2013

AbstractThe etiology of Kawasaki disease (KD) remains unknown, although some infectious organism has been suggested as the cause. Recent studies suggest that some bacterial toxins with superantigen activity are involved in its pathogenesis, but no specific bacterial toxin has yet been identified. Throat swabs for bacterial culture were obtained from 21 patients with KD and 20 with other febrile illnesses as controls. Mitogenic activity in culture supernatants obtained from individual bacterial strains was measured by lymphocyte proliferation assay. Sixty‐one bacterial strains were isolated from KD patients, and 62 strains from control patients. There was no apparent difference in bacterial species in the throat flora between KD patients and febrile controls. Moreover, total and individual mitogenic activity of strains from KD patients was no greater than that of strains from febrile controls. The bacterial superantigen activity of throat flora may not play a major role in the pathogenesis of KD.
Roles of Toxoplasma gondii‐Derived Heat Shock Protein 70 in Host Defense against T. gondii Infection
Authors: Ahmed Azza Kamal , Mun Hye‐Seong , Aosai Fumie , Piao Lian‐Xun , Fang Hao , Norose Kazumi , Yano Akihiko 
Published: November 2013

AbstractC57BL/6 mice receiving intraperitoneal injection of Toxoplasma gondii‐derived heat shock protein 70 (T.g. HSP70) on day 3 post T. gondii infection succumbed by day 9 post infection, while vector protein‐injected control mice survived more than 6 months. The deteriorating effect of T.g.HSP70 on host immune responses was dose‐dependent. By T.g. HSP70 injection, T. gondii loads increased in various organs of T. gondii‐infected mice. Th2 cytokines such as IL‐4 and IL‐10 were continuously produced from spleen and peritoneal exudate cells of T. gondii‐infected mice by injection of T.g. HSP70. Furthermore, nitric oxide production from peritoneal macrophages in T. gondii‐infected mice was reduced by T.g. HSP70.
Role of p38 Mitogen‐Activated Protein Kinase (MAPK) for Vacuole Formation in Lipopolysaccharide (LPS)‐Stimulated Macrophages
Authors: Hassan Ferdaus , Islam Shamima , Koide Naoki , Mu Mya Mya , Ito Hiroyasu , Mori Isamu , Yoshida Tomoaki , Yokochi Takashi 
Published: November 2013

AbstractThe role of p38 mitogen‐activated protein kinase (MAPK) on vacuole formation in lipopolysaccharide (LPS)‐stimulated RAW 264.7 cells was examined. LPS definitely induced the formation of vacuoles in RAW 264.7 cells and SB202190 as a p38 specific inhibitor also induced slight vacuole formation. The simultaneous treatment with LPS and SB202190 induced many more vacuoles in RAW 264.7 cells than the treatment with LPS or SB202190 alone, and the vacuoles were extraordinarily large in size. On the other hand, an inactive inhibitor of p38 MAPK did not augment LPS‐induced vacuole formation. Further, the inhibitors of other MAPKs and nuclear factor (NF)‐κB pathways did not affect it. The extraordinarily large vacuoles in RAW 264.7 cells treated with LPS and SB202190 were possibly formed via fusion of small vacuoles. However, SB202190 did not augment vacuole formation in CpG DNA or interferon (IFN)‐γ‐stimulated RAW 264.7 cells. The role of p38 MAPK in the vacuole formation in LPS‐stimulated macrophages is discussed.
The Larval Specific Lymphatic Filarial ALT‐2: Induction of Protection Using Protein or DNA Vaccination
Authors: Ramachandran Sabarinathan , Kumar Mishra Pankaj , Rami Reddy Maryada Venkata , Chinnaiah Harinath Basker , Nutman Thomas , Kaliraj Perumal , McCarthy James 
Published: November 2013

AbstractGenes from the infective stage of lymphatic filarial parasites expressed at the time of host invasion have been identified as potential vaccine candidates. By screening an L3 cDNA library with sera from uninfected longstanding residents of an area endemic for onchocerciasis, so‐called “endemic normals” (EN), we have cloned and characterized one such gene termed the abundant larval transcript two (ALT‐2). The stage specificity of ALT‐2 gene transcription and protein synthesis was confirmed by PCR using gene‐specific primers, and by western blot analysis of protein extracts from various stages of the parasite life cycle using specific antisera. Significant differences in antibody response to the recombinant ALT‐2 were observed in endemic populations with differing clinical manifestations of lymphatic filariasis with an antibody response present in sera from 18 of 25 (72%) EN subjects compared to 9 of 25 (36%) with subclinical microfilaracmia (MF) and 14 of 25 (52%) of those with chronic lymphatic obstruction (CP) (P=0.01 for comparison of EN to CP or to MF). This differential responsiveness suggests that the protective immunity postulated to account for their uninfected status might be associated with a response to this protein. When the utility of ALT‐2 as a vaccine candidate was tested in a murine model using either recombinant protein or a DNA vaccine construct, statistically significant protection was observed when compared to a control filarial gene product expressed across all stages of the parasite lifecycle (SXP‐1; P=0.02 for protein and P=0.01 for the DNA vaccine) or compared to adjuvant alone. This level of protection indicates that this vaccine is a promising candidate for further development.
Release of Shiga Toxin by Membrane Vesicles in Shigella dysenteriae Serotype 1 Strains and In Vitro Effects of Antimicrobials on Toxin Production and Release
Authors: Dutta Shanta , Iida Ken‐ichiro , Takade Akemi , Meno Yuko , Nair Gopinath Balakrish , Yoshida Shin‐ichi 
Published: November 2013

AbstractEffects of various antimicrobials on in vitro Shiga toxin production and release by Shigella dysenteriae serotype 1 was investigated in this study with particular reference to the role of outer membrane vesicles in toxin release by the organism. Five antimicrobials, namely nalidixic acid, ciprofloxacin, norfloxacin, fosfomycin and mitomycin C, were chosen for the study and the toxin titre was measured by the reverse passive latex agglutination (RPLA) method using an available kit. Only mitomycin C was found to induce production of Shiga toxin in the bacteria and its release by outer membrane vesicles. The highest titre of toxin was obtained in vesicle fraction suggesting that the vesicles play an important role in the release of Shiga toxin from periplasmic space by the organism.
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

AbstractA 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

AbstractNone 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.