bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of Bacterium Isolated from the Sediment at Coastal Area of Omura Bay in Japan and Several Biological Activities of Pigment Produced by This Isolate
Authors: Nakashima Takuji , Kurachi Maki , Kato Yoko , Yamaguchi Kenichi , Oda Tatsuya 
Published: November 2013

AbstractRecently we discovered a bacterial strain (MS‐02–063) that produces large amounts of red pigment from coastal area of Nagasaki Prefecture, Japan. Comparative 16S rDNA gene sequencing analysis revealed that strain MS‐02–063 was phylogenetically closely related to γ‐proteobacterium Hahella sp. MBIC 3957 that produces prodigiosin. However, some physiological and biochemical differences between strain MS‐02–063 and Hahella sp. MBIC 3957 were observed. The red pigment (RP‐063) produced by this isolate was highly purified from the culture supernatant. It was speculated that RP‐063 might be prodigiosin‐like pigment in physical properties and biological activities such as antibacterial and cytotoxic activity. Antibacterial activity of RP‐063 was examined by an agar dilution method. The results indicated that RP‐063 showed antibacterial activity for specific for pathogenic gram‐positive bacteria such as Staphylococcus aureus. The potency of antibacterial activity against S. aureus was nearly equal to those of tetracycline. Moreover, RP‐063 showed inhibition of the superoxide generation by 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA)‐stimulated mouse macrophage RAW 264.7 cell line. Prodigiosin members have a wide variety of biological properties, including anticancer and antimalarial, etc. Especially, potent immunosuppressive properties have been reported for prodigiosin members with the mechanism of action different from that of the other well known immunosuppressors in atopic dermatitis therapy such as cyclosporin A, FK506 and rapamycin. It is suggested that RP‐063 may be able to arrest the inflammation caused by superantigens secreted from S. aureus, which colonized skin on atopic dermatitis as well as suppression of activated lymphocyte proliferation and superoxide generation from leucocytes.
Human IgM Monoclonal Antibodies Reactive with HIV‐1‐Infected Cells Generated Using a Trans‐Chromosome Mouse
Authors: Okada Noriko , Yin Shuping , Asai Suzuka , Kimbara Noriaki , Dohi Natsuki , Hosokawa Masato , Wu Xiaoshan , Okada Hidechika 
Published: November 2013

AbstractThe trans‐chromosome (TC) mouse that we used harbors human chromosomes 2, 14 and/or 22, and has undergone knock‐out of its endogeneous genes coding for μ‐ and κ‐chains of immunoglobulin. One of these TC mice was immunized with HIV‐1‐infected U937 cells, and spleen cells from the immunized animal were fused with the mouse myeloma cell line to generate hybridoma cells. We selected hybridomas that produce human IgM antibodies (Abs) reactive with HIV‐1‐infected MOLT4 cells but not with uninfected MOLT4 cells. Two hybridoma cell lines were established termed 9F11 and 2G9. Although 0.4 μg/ml of 9F11 was able to induce complement‐mediated cytolysis of the infected cells in the presence of fresh human serum, 2G9 could not. There was no difference between the two monoclonal Abs in the base sequences of cDNAs coding for the constant regions of μ‐ and κ‐chains. Therefore, we speculate that the ability to activate complement on homologous cell membranes might reflect the structural presentation of antigenic molecules, which could facilitate the binding of an IgM Ab to multiple binding sites resulting in escape from restriction by species‐specific inhibitors of complement such as DAF (CD55) and CD59. On the other hand, 2G9 induced apoptosis of HIV‐1‐infected cells, including latently infected OM10.1 cells, although the Ag for 2G9 remains to be identified. Since both of the Abs had reduced reactivity toward HIV‐1‐infected MOLT4 cells following cultivation in the presence of tunicamycin, the responsible antigens would involve a sugar moiety.
Simultaneous Multicolor Detection System of the Single‐Molecular Microbial Antigen with Total Internal Reflection Fluorescence Microscopy
Authors: Hoshino Akiyoshi , Fujioka Kouki , Manabe Noriyoshi , Yamaya Shun‐ichi , Goto Yoji , Yasuhara Masato , Yamamoto Kenji 
Published: November 2013

AbstractImmunological diagnostic methods have been widely performed and showed high performance in molecular and cellular biology, molecular imaging, and medical diagnostics. We have developed novel methods for the fluorescent labeling of several antibodies coupled with fluorescent nanocrystal QDs. In this study we demonstrated that two bacterial toxins, diphtheria toxin and tetanus toxin, were detected simultaneously in the same view field of a cover slip by using directly QD‐conjugated antibodies. We have succeeded in detecting bacterial toxins by counting luminescent spots on the evanescent field with using primary antibody conjugated to QDs. In addition, each bacterial toxin in the mixture can be separately detected by single excitation laser with emission band pass filters, and simultaneously in situ pathogen quantification was performed by calculating the luminescent density on the surface of the cover slip. Our results demonstrate that total internal reflection fluorescence microscopy (TIRFM) enables us to distinguish each antigen from mixed samples and can simultaneously quantitate multiple antigens by QD‐conjugated antibodies. Bioconjugated QDs could have great potentialities for in practical biomedical applications to develop various high‐sensitivity detection systems.
Interleukin‐8 Secretion by Epithelial Cells Infected with Diffusely Adherent Escherichia coli Possessing Afa Adhesin‐Coding Genes
Authors: Arikawa Kentaro , Meraz Ismail Mustafa , Nishikawa Yoshikazu , Ogasawara Jun , Hase Atsushi 
Published: November 2013

AbstractEscherichia coli that adhere sparsely to human epithelial (HEp‐2) cells are known as diffusely adherent E. coli (DAEC) and considered potentially diarrheagenic. The role of the afimbrial adhesive sheath (Afa)—identified originally as a uropathogenic factor—in diffuse adhesion is now understood. However, the role of DAEC in diarrheal disease remains controversial. Recently, ability to induce interleukin‐8 (IL‐8) secretion from intestinal epithelial cells has been suggested as one of the properties of enterovirulent bacteria. In this study, we examined whether DAEC strains possessing Afa genes induced IL‐8 in cultures of human carcinoma epithelial cells (e.g., HEp‐2, Caco‐2, and T84). Nineteen afa‐positive DAEC strains were examined for their ability to induce IL‐8 secretion, and only 7 strains (37%; 7/19) induced IL‐8 as much as enteroaggregative E. coli did. No marked differences in adhesion were observed between high and low inducers. Diffusive adhesiveness itself is unlikely to be sufficient to induce IL‐8. All high inducers were motile and others were nonmotile. Additional stimulation by flagella may be required to cause high levels of chemokine induction. Motility or presence of flagella can be an important criterion to predict DAEC diarrheagenicity at clinical laboratories.
Molecular Identification of Ehrlichia Species and ‘Candidatus Neoehrlichia mikurensis’ from Ticks and Wild Rodents in Shizuoka and Nagano Prefectures, Japan
Authors: Naitou Hirotaka , Kawaguchi Daizoh , Nishimura Yuusaku , Inayoshi Megumi , Kawamori Fumihiko , Masuzawa Toshiyuki , Hiroi Midori , Kurashige Hideaki , Kawabata Hiroki , Fujita Hiromi , Ohashi Norio 
Published: November 2013

AbstractA total of 293 ticks and 111 wild rodents that were collected in Shizuoka and Nagano Prefectures, Japan, were examined for infection of Ehrlichia species and ‘Candidatus Neoehrlichia mikurensis.’ The 16S rDNA or the omp‐1 gene of these bacterial DNAs were detected from the spleens of tick‐inoculated mice (5 positive/total 29 mice) or from the spleens of wild rodents (25 positive/total 111 rodents) by PCR amplification. Sequencing of the 16S rDNA revealed Ehrlichia spp. from the 5 tick‐inoculated mice and 8 wild rodents, and ‘Candidatus N. mikurensis’ from 17 wild rodents. The data suggest the presence of additional genetic variants, and potential vectors and/or reservoirs for these bacteria in central Japan.
Proteomic Surveillance of Autoantigens in Relapsing Polychondritis
Authors: Tanaka Yasuhiko , Nakamura Manabu , Matsui Toshihiro , Iizuka Nobuko , Kondo Hirobumi , Tohma Shigeto , Masuko Kayo , Yudoh Kazuo , Nakamura Hiroshi , Nishioka Kusuki , Koizuka Izumi , Kato Tomohiro 
Published: November 2013

AbstractRelapsing polychondritis (RP) is a systemic inflammatory disease, in which autoimmunity to cartilage‐related components is thought to be involved in its pathogenesis. However, the autoimmune profile in RP has not been studied fully. We therefore investigated autoantibodies/autoantigens in RP comprehensively, by 2‐dimensional electrophoresis (2DE), subsequent western blotting (WB) and mass spectrometry, using cell‐extracted proteins as the antigen source. As a result, we detected 15 autoantigens on 2DE‐WB, and further identified five of them. On average, one RP serum recognized approximately 8 out of the 15 autoantigens. Frequencies of the autoantibodies to the 5 identified antigens of tubulin alpha ubiquitous/6, vimentin, alpha enolase, calreticulin, and colligin‐1/‐2 were 91%, 46%, 36%, 82%, and 36%, respectively. ELISA using recombinant proteins for them revealed that frequencies of the autoantibodies to tubulin alpha ubiquitous, vimentin, alpha enolase, calreticulin, and colligin‐1 were 36%, 64%, 46%, 27%, and 18%, respectively. Our data demonstrated that the autoimmune reaction was not restricted to cartilage‐related components, rather a variety of autoimmune responses occurred in patients with RP, which may be involved in the pathophysiology of RP. In addition, the proteomic approach using cell‐extracted proteins would be a powerful way to investigate autoantigens.
Competitive Study of Monoclonal Antibodies against the HIV‐1 Gp41 Core Structure
Authors: Usami Osamu , Xiao Peng , Ling Hong , Hattori Toshio 
Published: November 2013

AbstractMonoclonal antibodies (MAbs) 50.69, 98.6, and T26 bind specifically to the core structure of the human immunodeficiency virus type 1 (HIV‐1) envelope transmembrane glycoprotein (gp41). To clarify the specificity of the anti‐core structure MAbs, we performed competitive assays using the MAbs to the H9 human T cell line infected with the IIIB strain of HIV‐1 (H9/IIIB). Bound MAb 50.69 inhibited MAb 98.6 binding unidirectionally. The reason for the unidirectional cross competition between MAbs 50.69 and 98.6 is not clear, but these results help to define the antigenic structure of gp41 on the surface of infected cells.
A Novel Mouse Model for MPO‐ANCA‐Associated Glomerulonephritis
Authors: Yumura Wako , Itabashi Mitsuyo , Ishida‐Okawara Akiko , Tomizawa Kazuo , Yamashita Junji , Kaneshiro Yoshiaki , Nihei Hiroshi , Suzuki Kazuo 
Published: November 2013

AbstractWe established a novel model mouse for myeloperoxidase anti‐neutrophil cytoplasmic antibody (MPO‐ANCA)‐associated glomerulonephritis with crescentic formation, which was induced by administering bovine serum albumin (BSA). Neutrophil infiltration into the renal glomeruli began at 8 weeks and crescent formation was observed from 10 weeks after the first BSA injection. Platelet and neutrophil counts significantly increased, and proteinuria was observed from 5 weeks. MPO‐ANCA increased slightly at 4 and markedly at 9 weeks, and the TNF‐α level increased at 11 weeks. Glomerular neutrophil infiltration was correlated with MPO‐ANCA levels. In addition, proteinuria also significantly correlated with MPO‐ANCA levels. Finally, renal crescent formation was associated with an increase of MPO‐ANCA levels and neutrophil infiltration into glomeruli. The glomerular immune deposition of IgG and C3 was observed. These findings indicate that BSA induces neutrophil activation of peripheral blood followed by the elevation of MPO‐ANCA, resulting in the development of crescentic glomerulonephritis in mice.
A Viral Non‐Coding Region Determining Neuropathogenicity of Murine Leukemia Virus A8 Is Responsible for Envelope Protein Expression in the Rat Brain
Authors: Takase‐Yoden Sayaka , Wada Misaho , Watanabe Rihito 
Published: November 2013

AbstractFriend murine leukemia virus clone A8 causes spongiform neurodegeneration in the rat brain. A 0.3‐kb fragment containing the R‐U5–5′ leader sequence of A8 is required in addition to the A8‐env gene to induce spongiosis. The A8‐env gene is a primary determinant of neuropathogenicity. Comparative studies of the neuropathogenic virus R7f, which carries the 0.3‐kb fragment of A8 and A8‐env on the background of the non‐neuropathogenic clone 57, and the non‐neuropathogenic virus Rec5, which carries A8‐env on the background of 57, showed that the 0.3‐kb fragment of A8 was responsible for increasing the ratio of Env/Gag expression in the brain, but not in the spleen.
Virulence‐Defective Strains of Salmonella enterica Serovar Typhi as Candidates for Education at Level 2 Facilities
Authors: Huang Xinxiang , Xu Huaxi , Sha Mohammad Monir , Zhao Licheng , Ohkusu Kiyofumi , Kawamura Yoshiaki , Ezaki Takayuki 
Published: November 2013

AbstractThe use of biosafety level 3 pathogens is an essential element of education and training at medical schools. We previously reported on invasion‐defective strains of Salmonella enterica serovar Typhi, GTC 3P408 (ΔinvA, ΔsipB) and GTC 3P409 (ΔinvA, ΔsipB, and ΔviaB), as candidates for use in educational programs. Vi negative strains of S. enterica serovar Typhi became extremely sensitive to complement attack but showed increased invasiveness. Therefore, this study was conducted to construct two virulence‐defective strains, GTC 3P460 (ΔinvA, ΔsipB, and ΔrpoS) and GTC 3P461 (ΔinvA, ΔsipB, ΔviaB, and ΔrpoS), of S. enterica serovar Typhi by deleting rpoS from the GTC 3P409 and GTC 3P408 strains. Stress tests demonstrated that GTC 3P460 and GTC 3P461 are sensitive to conditions of starvation, acid stress and oxidative stress. These results suggest that these virulence‐defective strains have difficulty surviving in the gastric environment and in macrophages, characteristics that make them ideal candidates for education at level 2 facilities. Colony morphology and conventional biochemical features of these strains are identical to the parent strain S. enterica serovar Typhi GIFU 10007.
Determination of Antifungal Activities in Serum Samples from Mice Treated with Different Antifungal Drugs Allows Detection of an Active Metabolite of Itraconazole
Authors: Maki Katsuyuki , Watabe Etsuko , Iguchi Yumi , Nakamura Hideko , Tomishima Masaki , Ohki Hidenori , Yamada Akira , Matsumoto Satoru , Ikeda Fumiaki , Tawara Shuichi , Mutoh Seitaro 
Published: November 2013

AbstractTo establish an in vitro method of predicting in vivo efficacy of antifungal drugs against Candida albicans and Aspergillus fumigatus, the antifungal activities of fluconazole, itraconazole, and amphotericin B were determined in mouse serum. The minimum inhibitory concentration (MIC) of each drug was measured using mouse serum as a diluent. For C. albicans, the assay endpoint of azoles was defined as inhibition of mycelial extension (mMIC) and for A. fumigatus, as no growth (MIC). The MICs of amphotericin B for both pathogens were defined as the MIC at which no mycelial growth occurred. Serum MIC or mMIC determinations were then used to estimate the concentration of the drugs in serum of mice treated with antifungal drugs by multiplying the antifungal titer of the serum samples by the serum (m)MIC. The serum drug concentrations were also determined by HPLC. The serum concentrations estimated microbiologically showed good agreement with those determined by HPLC, except for itraconazole. Analysis of the serum samples from itraconazole‐treated mice by a sensitive bioautography revealed the presence of additional spots, not seen in control samples of itraconazole. The bioautography assay demonstrated that the additional material detected in serum from mice treated with itraconazole was an active metabolite of itraconazole. The data showed that the apparent reduction in the itraconazole serum concentration as determined by HPLC was the result of the formation of an active metabolite, and that the use of a microbiological method to measure serum concentrations of drugs can provide a method for prediction of in vivo efficacy of antifungal drugs.
Immunodominant Epitope in the C‐Terminus of a Variable Major Protein in Borrelia duttonii, an Agent of Tick‐Borne Relapsing Fever
Authors: Tabuchi Norihiko , Tomoda Koichiro , Kawaguchi Hiroshi , Iwamoto Hiroyuki , Fukunaga Masahito 
Published: November 2013

AbstractBorrelia duttonii strain Ly was isolated from a child with tick‐borne relapsing fever in Tanzania. B. duttonii produces variable major proteins (Vmps), which undergo antigenic variation. We previously reported transcription of the vmpP gene, which is one of the Vmp genes in strain Ly, detected in vitro cultivation. In the current study, we purified the recombinant non‐lipidated VmpP protein by affinity chromatography and produced VmpP polyclonal antibodies. Antigenicity of VmpP was examined by Western immunoblot analysis and peptide‐based enzyme‐linked immunosorbent assays. Antigenic epitopes were shown to comprise five regions interspersed within the VmpP primary amino acid sequence. Synthetic peptides spanning residues of three of five regions, 232–237 (LASIVD), 280–285 (AGGIAL), and 350–355 (KAADQQ), reacted strongly with the VmpP‐specific antibody and these residues were identified as epitopes. In particular, the C‐terminal domain (KAADQQ) of this protein was immunoreactive. Further research based on our results will promote the development of a recombinant vaccine for B. duttonii infection.
Gingipains Inactivate a Cell Surface Ligand on Porphyromonas gingivalis That Induces TLR2‐ and TLR4‐Independent Signaling
Authors: Kishimoto Mami , Yoshimura Atsutoshi , Naito Mariko , Okamoto Kuniaki , Yamamoto Kenji , Golenbock Douglas T. , Hara Yoshitaka , Nakayama Koji 
Published: November 2013

AbstractArginine‐specific gingipain and lysine‐specific gingipain are two major cysteine proteinases produced by Porphyromonas gingivalis. To clarify the role of gingipains in the interaction between P. gingivalis and the innate immune system, CHO reporter cells expressing TLR2 or TLR4 were stimulated with wild‐type or gingipain‐deficient P. gingivalis cells and activation of nuclear factor‐κB in these cells was examined. While CHO/CD14 cells and 7.19 cells, an MD‐2‐defective mutant derived from CHO/CD14 cells, failed to respond to wild‐type P. gingivalis, they responded to gingipain‐deficient P. gingivalis. On the other hand, CHO/CD14/TLR2 cells responded to both wild‐type and gingipain‐deficient P. gingivalis. These results suggested that gingipains have no effects on TLR2‐dependent signaling from P. gingivalis but have inhibitory effects on TLR2‐ and TLR4‐independent signaling in CHO cells. Indeed, the activity of gingipain‐deficient P. gingivalis to induce the activation of 7.19 cells was diminished after treatment of the bacterial cells with gingipains. We next partially purified bacterial cell components activating 7.19 cells from gingipain‐deficient P. gingivalis. The activity of the partially purified components was diminished by treatment with heat or gingipains. It is also noteworthy that anti‐CD14 mAb inhibited the activation of 7.19 cells induced by the partially purified components. These results indicated that the components of P. gingivalis that were able to induce TLR2‐ and TLR4‐independent signaling were inactivated by gingipains before being recognized by CD14. The inactivation of the components would be helpful for P. gingivalis to escape from the innate immune system.
Characterization of Biological Activities of Brucella melitensis Lipopolysaccharide
Authors: Tumurkhuu Gantsetseg , Koide Naoki , Takahashi Kazuko , Hassan Ferdaus , Islam Shamima , Ito Hiroyasu , Mori Isamu , Yoshida Tomoaki , Yokochi Takashi 
Published: November 2013

AbstractBiological activities of lipopolysaccharide (LPS) from Brucella melitensis 16M were characterized in comparison with LPS from Escherichia coli O55. LPS extracted from B. melitensis was smooth type by electrophoretic analysis with silver staining. The endotoxin‐specific Limulus activity of B. melitensis LPS was lower than that of E. coli LPS. There was no significant production of tumor necrosis factor‐α and nitric oxide in RAW 264.7 macrophage cells stimulated with B. melitensis LPS, although E. coli LPS definitely induced their production. On the other hand, B. melitensis LPS exhibited a higher anti‐complement activity than E. coli LPS. B. melitensis LPS as well as E. coli LPS exhibited a strong adjuvant action on antibody response to bovine serum. The characteristic biological activities of B. melitensis are discussed.
Enhancement of Mice Susceptibility to Infection with Listeria monocytogenes by the Treatment of Morphine
Authors: Asakura Hiroshi , Kawamoto Keiko , Igimi Shizunobu , Yamamoto Shigeki , Makino Sou‐ichi 
Published: November 2013

AbstractThe effect of morphine on the susceptibility of BALB/c mice to diarrheagenic Escherichia coli, Shigella flexneri, Listeria monocytogenes, Salmonella Enteritidis, Yersinia enterocolitica, was examined via the intraperitoneal inoculation. Morphine treatment increased the susceptibility to S. Enteritidis and L. monocytogenes, resulting in bacteremia and central nervous system (CNS) invasion (for L. monocytogenes), while the infection with other bacteria did not show the systemic dissemination in the morphine‐treated mice. Notably, L. monocytogenes infection caused 100% mortality with a mean survival time (MST) of 1.3 days in morphine‐treated mice, but untreated mice did not die. The present data suggested that individuals using heroin or treated with morphine derivatives might be at high risk for listeriosis, especially those who are immunocompromised. Recent increasing consumption of morphine may propose the necessity for further epidemiological surveillance on infectious diseases.
Quantitative Analysis of Cutaneous Malassezia in Atopic Dermatitis Patients Using Real‐Time PCR
Authors: Sugita Takashi , Tajima Mami , Tsubuku Hisae , Tsuboi Ryoji , Nishikawa Akemi 
Published: November 2013

AbstractWe quantified the cutaneous Malassezia in patients with atopic dermatitis using a real‐time PCR assay. Seven to 12 times more Malassezia colonized the head and neck compared to the trunk or limbs, and the species M. globosa and M. restricta accounted for approximately 80% of all Malassezia colonization at any body site.
A Slow Spread of Adenovirus Type 7 Infection after Its Re‐Emergence in Yamagata, Japan, in 1995
Authors: Mizuta Katsumi , Abiko Chieko , Aoki Yoko , Murata Toshio , Katsushima Noriko , Sakamoto Michiyo , Itagaki Tsutomu , Hoshina Hitoshi , Ootani Katsumi 
Published: November 2013

AbstractWe have continued the epidemiological study on adenovirus type 7 (Ad7), which re‐emerged in 1995 in Yamagata, Japan. Between 1999 and 2004, we isolated only four strains from 10,778 throat swab specimens among children with acute respiratory infections. A serological survey of 303 specimens revealed the antibody‐positive rate against Ad7 to be 0–7.4% in children under 10 years of age in 2005, although it was 3.3–16.7% in 1997 and 0% in 1993. Our results suggest that a re‐emergence does not always provoke a sudden major outbreak, even if the antibody‐positive rate against Ad7 is low in the local community.