bluuebunny/crossref_metadata_2025 huggingface dataset:
Prophylaxis of Local Vascular Graft Infection with Levofloxacin Incorporated into Albumin‐Sealed Dacron Graft (LVFX‐ALB Graft)
Authors: Osada Takashi , Yamamura Keiko , Fujimoto Katsuhiro , Mizuno Keisuke , Sakurai Tsunehisa , Ohta Michio , Nabeshima Toshitaka
Published: November 2013

An animal model was used to assess the efficacy of levofloxacin (LVFX) incorporated into albumin (ALB)‐sealed Dacron (LVFX‐ALB) graft for the prevention of vascular graft infections caused by Staphylococcus aureus. Under general anesthetic, an interposition graft was placed into dog carotid artery. On completion of the operation, 0.1 ml of normal saline containing 107 colony‐forming units (CFU) of a slime‐producing S. aureus was inoculated directly onto the graft. After 1 day, the samples were sterilely harvested. The antibacterial activity of LVFX into the LVFX‐ALB graft was evaluated by colony counting in bacterial cultures and by the fluorescent antibody method staining bacteria adhesion to the grafts. LVFX‐ALB grafts had a lower infection rate than the control grafts (1/4, 102 CFU vs 4/4, 1.50×105±1.38×105 CFU (mean±SE)). In an immunostaining study, LVFX‐ALB grafts had small fluorescent areas showing S. aureus adhesion, while fluorescence was observed over the entire surface of the control grafts. Therefore, LVFX‐ALB presumably had a bactericidal action and adhesive prevention against inoculated S. aureus. LVFX‐ALB may be useful in preventing graft infections during and immediately after vascular reconstruction.
Detection of Treponema socranskii Associated with Human Periodontitis by PCR
Authors: Sakamoto Mitsuo , Takeuchi Yasuo , Umeda Makoto , Ishikawa Isao , Benno Yoshimi , Nakase Takashi
Published: November 2013

A PCR technique was used to detect and identify Treponema socranskii associated with periodontitis. A species‐specific forward primer was designed for a variable region within its 16S rRNA gene and was used in conjunction with a conserved reverse primer. This primer pair was tested for specificity against 44 oral bacterial strains. Sensitivity was determined using a serial dilution of T. socranskii cells. Amplification products were obtained from all T. socranskii strains tested, but not from other oral bacteria associated with periodontal disease. The detection limit of PCR was 5 T. socranskii cells per PCR. T. socranskii was detected by PCR in subgingival plaque and saliva samples from patients with periodontitis.
Insufficient Resistance of Trehalose‐6,6′‐Dimycolate‐Treated T‐Cell Receptor δ Gene Mutant (TCR δ–/–) Mice against Influenza Virus Infection
Authors: Hoq M. Mozammal , Suzutani Tatsuo , Nakaya Kazuhiro , Yoshida Itsuro , Ogasawara Masahiro , Takeda Yoshinori , Shibaki Taiichiro , Itohara Shigeyoshi , Yamamoto Hiroshi , Azuma Masanobu
Published: November 2013

Normal mice inoculated intravenously with 50 μg trehalose‐6,6′‐dimycolate, a glycolipid component of the cell wall of Mycobacterium, in an oil‐in‐water emulsion (TDM emulsion) acquired a high resistance to intranasal lethal infection of an influenza virus. In contrast, TDM emulsion‐treated T‐cell receptor δ gene mutant (TCR δ /) mice acquired insufficent resistance against the lethal influenza virus infection. The patterns of insufficient resistance were identical to the results obtained previously with mice which were depleted of T‐lymphocytes bearing γδ T‐cell receptors (γδ T‐cells) by in vivo administration of anti‐γδ T‐cell receptor monoclonal antibody (Hoq et al, J. Gen. Virol. 78: 1597‐1603, 1997). These results strongly suggest that the γδ T‐cells play an important non‐specific role in resistance against influenza virus infection.
Molecular Epidemiology of Rabies in Thailand
Authors: Ito Naoto , Sugiyama Makoto , Oraveerakul Kanisak , Piyaviriyakul Prapruddee , Lumlertdacha Boonlert , Arai Yoko T. , Tamura Yutaka , Mori Yoshio , Minamoto Nobuyuki
Published: November 2013

For the purpose of making clear the dynamics of rabies viruses that are prevalent among dogs in Asia, especially Thailand, nucleoprotein (N) genes of isolates derived from Thailand were partially sequenced, and a phylogenetic analysis was performed on the basis of the sequencing data. Firstly, all 27 isolates from Thailand belonged to one group that was distantly related to an isolate from China and was separated into at least six lineages. On the other hand, the isolate from Japan was related to viruses from the Arctic. Secondly, in order to analyze the diversity of the N gene more conveniently, restriction fragment length polymorphism (RFLP) analysis was performed on the N gene of 27 isolates from Thailand. The RFLP analysis could distinguish the lineages of each isolate, and the lineages of additional 34 isolates were deduced by this method. On examination of the geographical distribution of the six lineages, based on the results of phylogenetic and RFLP analyses, it was clear that infection cycles of the rabies virus in Thailand have tended to be maintained endemically.
The Majority of Lymphocytes in the Bone Marrow, Thymus and Extrathymic T Cells in the Liver Are Generated In Situ from Their Own Preexisting Precursors
Authors: Shimizu Takao , Sugahara Satoshi , Oya Hiroshi , Maruyama Satoshi , Minagawa Masahiro , Bannai Makoto , Hatakeyama Katsuyoshi , Abo Toru
Published: November 2013

Parabiotic pairs of B6.Ly5.1 and B6.Ly5.2 mice were used to investigate how lymphocytes in various organs and various lymphocyte subsets mixed with partner cells. The origin of partner cells was determined by using anti‐Ly5.1 mAb in conjunction with immunofluorescence tests. Parabiosis was also produced after the irradiation of B6.Ly5.2 mice at various doses to prepare an immunosuppressive partner. Irrespective of irradiation, lymphocytes and other hematopoietic cells in the bone marrow and lymphocytes in the thymus showed a low mixture of partner cells in comparison with those of all other organs tested. On the other hand, lymphocytes in the blood, spleen, and lymph nodes became a half‐and‐half mixture of their own cells and partner cells by 14 days after parabiosis. Among lymphocyte subsets, intermediate CD3 cells (i.e., CD3int cells) and NKT cells (i.e., NK1.1+ subset of CD3int cells) in the liver also showed a low mixture of partner cells. The present results raise the possibility that lymphocytes in the bone marrow and thymus, and extrathymic T cells in the liver might be in situ generated from their own preexisting precursor cells. Another observation was that, after irradiation, partner cells showed accelerated mixture even if they showed a low mixture under non‐irradiated conditions. However, only lymphocyte subsets with the same phenotype as those of preexisting cells entered the corresponding sites.
Role of Interferon Alpha (IFN‐α) and Interferon Gamma (IFN‐γ) in the Control of the Infection of Monocyte‐Like Cells with Human Cytomegalovirus (HCMV)
Authors: Delannoy Anne‐Sophie , Hober Didier , Bouzidi Ahmed , Wattre Pierre
Published: November 2013

The role of cytokines in the control of HCMV infection has been studied in THP‐1 cells, a macrophage‐like cell model and in MRC‐5 cells. HCMV replication was studied by immune detection of viral immediate‐early antigens (IEA) and virus yield was evaluated in MRC‐5 cells by immunoperoxidase staining. Pretreatment of MRC‐5 and phorbol 12‐myristate 13‐acetate (PMA)‐treated THP‐1 cells with IFN‐α or IFN‐γ for 24 hr prior to the infection reduced the number of infected cells and virus yield. A synergistic anti‐CMV activity in synthesis of early proteins was obtained with these cytokines in combination with TNF‐α in differentiated THP‐1 cells only. Treatment of HCMV‐infected differentiated THP‐1 cells or MRC‐5 cells with IFN‐α or IFN‐γ alone had no inhibitory effect on virus replication, however the virus yield was reduced with ganciclovir. A synergistic anti‐CMV activity in virus yield was obtained only when infected differentiated THP‐1 cells were treated with ganciclovir in combination with IFN‐γ. The current study shows that IFN‐α and IFN‐γ can play a role in the reduction of HCMV replication in macrophage‐like cells and in the efficiency of therapies with ganciclovir in this cell type and that the anti‐CMV effect of cytokines may be different in fibroblasts and in macrophage‐like cells.
Highly Selective Antibacterial Activity of Novel Alkyl Quinolone Alkaloids from a Chinese Herbal Medicine, Gosyuyu (Wu‐Chu‐Yu), against Helicobacter pylori In Vitro
Authors: Hamasaki Norio , Ishii Eiji , Tominaga Kazunari , Tezuka Yasuhiro , Nagaoka Takema , Kadota Shigetoshi , Kuroki Tetsuo , Yano Ikuya
Published: November 2013

To elucidate the antibacterial activity of Gosyuyu, the crude extract from the fruit of Evodia rutaecarpa, a Chinese herbal medicine, has been fractionated chromatographically, and each fraction was assayed for antibacterial activity against Helicobacter pylori (H. pylori) in vitro. As the result, a single spot having marked antibacterial activity against H. pylori was obtained and the chemical structure was analyzed. The isolated compound was revealed to be a novel alkyl quinolone alkaloid based on the solubility, IR spectra, NMR analysis and mass spectrometric data after purification by TLC of silica. We compared the antimicrobial activity of this compound with that of other antimicrobial agents and examined susceptibility of various intestinal pathogens. As the result, the new quinolone compounds obtained from Gosyuyu extracts were found to be a mixture of two quinolone alkaloids, 1‐methyl‐2‐[(Z)‐8‐tridecenyl]‐4‐(1H)‐quinolone and 1‐methyl‐2‐[(Z)‐7‐tridecenyl]‐4‐(1H)‐quinolone (MW: 339), reported previously. The minimum inhibitory concentration (MIC) of these compounds against reference strains and clinically isolated H. pylori strains were less than 0.05 μg/ml, which was similar to the MIC of amoxicillin and clarithromycin that are used worldwide for the eradication of H. pylori, clinically. Furthermore, it was noted that the antimicrobial activity of these compounds was highly selective against H. pylori and almost non‐active against other intestinal pathogens. The above results showed that these alkyl methyl quinolone (AM quinolones) alkaloids were useful for the eradication of H. pylori without affecting other intestinal flora.
Coprobacillus catenaformis Gen. Nov., Sp. Nov., a New Genus and Species Isolated from Human Feces
Authors: Kageyama Akiko , Benno Yoshimi
Published: November 2013

Three strains of Eubacterium‐like isolates from human feces were characterized by biochemical tests and 16S rDNA analysis. The phenotypic characteristics of the three strains resembled those of the genus Collinsella transferred from the genus Eubacterium recently. However, Eubacterium‐like strains were phylogenetically members of the Clostridium subphylum of Gram‐positive bacteria, and these showed a specific phylogenetic association with Clostridium ramosum and C. spiroforme. C. ramosum and C. spiroforme are Gram‐positive, anaerobic, spore‐forming bacteria that belong to the genus Clostridium, and the G + C contents are 26.0 and 27.4 mol%, respectively. However, the three Eubacterium‐like strains had G + C contents of 32.1 to 33.1 mol% and were non‐spore‐forming rods. Based on phenotypic characteristics, we can differentiate these species, and furthermore, a 16S rDNA sequence divergence of greater than 9% with a new related genus, Coprobacillus, is proposed for the three strains, with one species, Coprobacillus catenaformis. The type strain of C. catenaformis is JCM 10604T.
Therapeutic Oral Vaccination Induces Mucosal Immune Response Sufficient to Eliminate Long‐Term Helicobacter pylori Infection
Authors: Ikewaki Junko , Nishizono Akira , Goto Takayuki , Fujioka Toshio , Mifune Kumato
Published: November 2013

We examined the efficacy of therapeutic oral vaccination using Helicobacter pylori‐whole cell sonicate and cholera toxin (CT) in mice persistently infected with H. pylori. Efficacy was determined by bacterial culture and microscopic examination of gastric tissues for the persistence of bacteria at 6 weeks after the last vaccination. Vaccination of H. pylori‐whole cell sonicate combined with CT eradicated bacteria in 10/16 mice (62.5%). Interestingly, oral vaccination with CT alone also eliminated the bacteria in 8/17 mice (47.1%). However, a therapeutic intraperitoneally administered vaccine failed to eradicate H. pylori from the stomach (1/17 mice, 5.9%). Identification of the type of immunity involved in the eradication process showed that oral vaccination enhanced the antigen‐specific IgA in the feces and saliva. The efficacy of eradication of H. pylori correlated well with increases in IgA secretion in mucosal tissue and a higher labeling index of IgA‐positive lumina of pyloric glands. Moreover, the expression of IL‐4 mRNA in the stomach of mice with eradicated bacteria was higher than in the uneradicated group. Our results suggest that the efficacy of vaccination depends on the mucosal IgA response in the gastrointestinal tract against H. pylori via Th2 cell activation and that therapeutic oral vaccination induces a mucosal immune response sufficient to eradicate long‐term infection with H. pylori.
The DsbA‐DsbB Disulfide Bond Formation System of Burkholderia cepacia Is Involved in the Production of Protease and Alkaline Phosphatase, Motility, Metal Resistance, and Multi‐Drug Resistance
Authors: Hayashi Shoko , Abe Mitsuko , Kimoto Mitsuaki , Furukawa Susumu , Nakazawa Teruko
Published: November 2013

In a previous study, we isolated a dsbB mutant of Burkholderia cepacia KF1 and showed that phenotypes of protease production and motility are dependent on DsbB, a membrane‐bound disulfide bond oxidoreductase. We have now isolated a dsbA mutant by transposon mutagenesis, cloned the dsbA gene encoding a periplasmic disulfide bond oxidoreductase, and characterized the function of the DsbA‐DsbB disulfide bond formation system in B. cepacia. The complementing DNA fragment had an open reading frame for a 212‐amino acid polypeptide with a potential redox‐active site sequence of Cys‐Pro‐His‐Cys that is homologous to Escherichia coli DsbA. The dsbA mutant, as well as the previously isolated dsbB mutant, was defective in the production of extracellular protease and alkaline phosphatase, as well as in motility. In addition, mutation in the DsbA‐DsbB system resulted in an increase in sensitivity to Cd2+ and Zn2+ as well as a variety of antibiotics including β‐lactams, kanamycin, erythromycin, novobiocin, ofloxacin and sodium dodecyl sulfate. These results suggested that the DsbA‐DsbB system might be involved in the formation of a metal efflux system as well as a multi‐drug resistance system.
Characterization and Distribution of a New Enterotoxin‐Related Superantigen Produced by Staphylococcus aureus
Authors: Abe Jun , Ito Yasuhiko , Onimaru Michiko , Kohsaka Takao , Takeda Tae
Published: November 2013

Staphylococcal enterotoxins (SEs) are a family of structurally related pyrogenic exotoxins consisting of the five prototypic SEs (types A to E) and three newly characterized SEs (types G to I) produced by Staphylococcus aureus (S. aureus). They also work as superantigens and cause food poisoning and shock symptoms in humans. In this study, we cloned a new variant gene of the seg and characterized its superantigenic properties and distribution among the clinical isolates of S. aureus. The gene encodes a 233 amino acid protein which is highly homologous to SEG (97.7%). The variant SEG (SEGv) expressed by the cloned gene exerted mitogenic activity on human peripheral blood mononuclear cells at the concentration of 100 pg/ml. T cells bearing Vβ3, 12, 13.1, 13.2, 14 and 15 were preferentially expanded after stimulation with the recombinant protein. The mRNA of the variant seg gene was detected in the total RNA of the organisms bearing this gene. By PCR, 27 out of 48 clinical isolates of S. aureus (56%) possessed either the seg or variant seg gene. These findings suggest that SEG, or SEGv, is one of the most frequently produced superantigen exotoxins by S. aureus and may participate in the inflammatory process of the host by activating a distinct set of Vβ families of T cells.
Staphylococcus aureus Produces Autolysin‐Susceptible Cell Walls during Growth in a High‐NaCl and Low‐Ca2+ Concentration Medium
Authors: Ochiai Toshiro
Published: November 2013

The growth of Staphylococcus aureus 209P becomes unusually sensitive to a high‐NaCl concentration by decreasing the Ca2+ concentration in growth media, and cells either autolyze or transform into protoplast‐like forms when grown standing in high‐NaCl and low‐Ca2+ concentration media below 37 C (Ochiai, T., Microbiol. Immunol. 43 (7): 705–709, 1999). To assess the role of Ca2+ in the salt tolerance of this organism, cells grown in the presence of different concentrations of Ca2+ were treated with boiling SDS, and their susceptibilities to crude autolysin (3 M LiCl extract of S. aureus 209P cells) were evaluated by turbidimetric assay and zymographic analysis. Susceptibilities of SDS‐treated cells (SDS‐cells) to crude autolysin were significantly influenced by Ca2+ concentration in the culture, and SDS‐cells prepared from cultures grown in high‐NaCl and high‐Ca2+ concentration media exhibited marked resistance to crude autolysin when the assay system contained a high concentration of NaCl. On the contrary, SDS‐cells prepared from cultures grown in high‐NaCl and low‐Ca2+ concentration media were rather susceptible to crude autolysin under the same assay conditions. A zymographic analysis revealed that the constitution of cell‐associated autolysins was not influenced by the concentration of exogenous Ca2+. These results suggested that at least part of the mechanism of salt‐induced autolysis in S. aureus 209P might be related to the synthesis of an autolysin susceptible cell wall.
The Complete DNA Sequence of the O Antigen Gene Region of Plesiomonas shigelloides Serotype O17 Which Is Identical to Shigella sonnei Form I Antigen
Authors: Chida Toshio , Okamura Noboru , Ohtani Kiyoshi , Yoshida Yoko , Arakawa Eiji , Watanabe Haruo
Published: November 2013

We cloned and determined the sequence of a DNA region of approximately 15‐kb containing the cluster of genes required for O17 antigen expression in the Escherichia coli K‐12 strain from the chromosome of Plesiomonas shigelloides serotype O17:H2 strain. The sequencing analysis revealed that the minimum essential region of the P. shigelloides O17 antigen gene cluster had a size of approximately 11.5‐kb and contained 9 contiguous open reading frames (ORFs), which were almost identical to the corresponding ORFs of Shigella sonnei form I antigen gene region, except for IS630 sequence, at the DNA as well as amino acid levels. The putative function of most of the ORFs could be determined on the basis of amino acid sequence similarities and characteristics. In addition, the G + C content of the P. shigelloides O17 antigen genes was lower than that of the chromosomal DNA of P. shigelloides and S. sonnei, suggesting that both P. shigelloides O17 and S. sonnei form I antigen genes had been derived from the same origin with a low G+C content.
Severity‐Related Molecular Differences among Nineteen Strains of Dengue Type 2 Viruses
Authors: Pandey Basu Dev , Igarashi Akira
Published: November 2013

Comparative nucleotide sequencing was carried out on dengue type 2 virus (DEN‐2) strains isolated from patients in Northeast Thailand during the epidemic season in 1993. The patients exhibited different clinical manifestations ranging from dengue fever (DF) to dengue haemorrhagic fever (DHF)/dengue shock syndrome (DSS). The results classified 19 DEN‐2 strains into 3 subtypes according to nonsynonymous amino acid replacements. The strain isolated from a DSS patient eliciting secondary serological response belonged to subtype I, whereas 13 strains isolated from DHF patients with secondary response and 2 strains from DF patients with primary response belonged to subtype II. On the other hand, 3 strains isolated from DF cases evoking either primary or secondary response belonged to subtype III. These results suggest that subtype III virus infection could result in clinically milder manifestation irrespective of the serological response compared with subtype I or II viruses. The RNA secondary structure predicted for the 3′ noncoding region showed 4 different structures (A, B, C, and D). The result also indicates that different subtypes of DEN‐2 serotypes are circulating in a single epidemic in Thailand.
Dietary Bifidobacterium lactis (HN019) Enhances Resistance to Oral Salmonella typhimurium Infection in Mice
Authors: Shu Quan , Lin Hai , Rutherfurd Kay J. , Fenwick Stan G. , Prasad Jaya , Gopal Pramod K. , Gill Harsharnjit S.
Published: November 2013

The ability of a newly identified probiotic lactic acid bacterial strain, Bifidobacterium lactis (HN019), to confer protection against Salmonella typhimurium was investigated in BALB/c mice. Feeding mice with B. lactis conferred a significant degree of protection against single or multiple oral challenge with virulent S. typhimurium, in comparison to control mice that did not receive B. lactis. Protection included a ten‐fold increase in survival rate, significantly higher post‐challenge food intake and weight gain, and reduced pathogen translocation to visceral tissues (spleen and liver). Furthermore, the degree of pathogen translocation showed a significant inverse correlation with splenic lymphocyte proliferative responses to mitogens, blood and peritoneal cell phagocytic activity and intestinal mucosal anti‐S. typhimurium antibody titers in infected mice; all of these immune parameters were enhanced in mice fed B. lactis. Together, these results suggest that dietary B. lactis can provide a significant degree of protection against Salmonella infection by enhancing various parameters of immune function that are relevant to the immunological control of salmonellosis. Thus dietary supplementation with B. lactis provides a unique opportunity for developing immune‐enhancing probiotic dairy food products with proven health benefits.
Phylogenic and Phenotypic Characterization of Some Eubacterium‐Like Isolates from Human Feces: Description of Solobacterium moorei Gen. Nov., Sp. Nov.
Authors: Kageyama Akiko , Benno Yoshimi
Published: November 2013

Three isolated strains from human feces were characterized by biochemical tests and 16S rDNA analysis. Phylogenetic analysis revealed that these isolated strains were members of the Clostridium subphylum of Gram‐positive bacteria. The phenotypic characters resembled those of the genus Eubacterium, but these strains were shown to be phylogenetically distant from the type species of the genus, Eubacterium limosum. The strains showed a specific phylogenetic association with Holdemania filiformis and Erysipelothrix rhusiopathiae. Based on a 16S rDNA sequence divergence of greater than 12% with H. filiformis and E. rhusiopathiae, a new genus, Solobacterium, is proposed for three strains, with one species, Solobacterium moorei. The type strain of Solobacterium moorei is JCM 10645T.
Oxalate‐Degrading Enterococcus faecalis
Authors: Hokama Sanehiro , Honma Yasuko , Toma Claudia , Ogawa Yoshihide
Published: November 2013

An oxalate‐degrading Enterococcus faecalis was isolated from human stools under anaerobic conditions. The bacteria required a poor nutritional environment and repeated subculturing to maintain their oxalate‐degrading ability. The E. faecalis produced 3 proteins (65, 48, and 40 kDa) that were not produced by non‐oxalate‐degrading E. faecalis as examined by SDS‐PAGE. Antibodies against oxalyl‐coenzyme A decarboxylase (65 kDa) and formyl‐coenzyme A transferase (48 kDa) obtained from Oxalobacter formigenes (an oxalate‐degrading anaerobic bacterium in the human intestine) reacted with 2 of the proteins (65 and 48 kDa) from the E. faecalis as examined by Western blottings. This is the first report on the isolation of oxalate‐degrading facultative anaerobic bacteria from humans.
Release of Membrane Vesicles Containing Endotoxic Lipopolysaccharide in Escherichia coli O157:H7 Clinical Isolates
Authors: Meno Yuko , Fujimoto Shuji , Horikawa Kazumi , Yoshida Shin‐ichi
Published: November 2013

Membrane vesicles released by E. coli O157:H7 strains were investigated by immuno‐electron microscopy using anti‐O157 antibody. Anti‐O157 antibody enhanced the negative‐staining of vesicles and we found numerous small vesicles clearly formed around bacterial cells. An immunogold‐electron microscopic examination confirmed that lipopolysaccharide (LPS) including the O‐side chain is present on the surface of the vesicles. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis of the purified vesicles showed that the vesicles contained LPS consisting of a lipid‐A and an O polysaccharide. In addition, the endotoxic activity of the vesicle was confirmed by a limulus test. These results suggest that the vesicles may play an important role in the pathogenesis of Escherichia coli O157:H7.
Helicobacter pylori May Have Only a Transient Presence in the Oral Cavity and on the Surface of Oral Cancer
Authors: Okuda Katsuji , Ishihara Kazuyuki , Miura Tadashi , Katakura Akira , Noma Hiroyasu , Ebihara Yoko
Published: November 2013

We used the reverse transcription polymerase chain reaction (RT‐PCR) and culture methods to study the presence of Helicobacter pylori in the gastric and oral samples from a total of 116 gastritis and peptic ulcer patients, including 58 with oral cancer. Detection rates of H. pylori were 46.6% in stomach samples and 12.1 % in oral swab samples. All of the oral cancer surface swab samples were positive for H. pylori, as were their gastric samples suggesting that oral H. pylori derived from the stomach. The culture supernatants of Streptococcus mutans and Prevotella intermedia inhibited the growth of the H. pylori strain and caused the formation of the coccal form. In cases where H. pylori was detected in the oral cavity samples, including the oral cancer surface samples, it was believed that this species had colonized the stomach and were present in the oral cavity only as a transient organism.
Proposal of Sphingomonadaceae Fam. Nov., Consisting of Sphingomonas Yabuuchi et al. 1990, Erythrobacter Shiba and Shimidu 1982, Erythromicrobium Yurkov et al. 1994, Porphyrobacter Fuerst et al. 1993, Zymomonas Kluyver and van Niel 1936, and Sandaracinobacter Yurkov et al. 1997, with the Type Genus Sphingomonas Yabuuchi et al. 1990
Authors: Kosako Yoshimasa , Yabuuchi Eiko , Naka Takashi , Fujiwara Nagatoshi , Kobayashi Kazuo
Published: November 2013

Based on the results of phylogenetic analysis of the 16S rDNA sequences and the presence of N‐2′‐hydroxymyristoyl dihydrosphingosine 1‐glucuronic acid (SGL‐1) and 2‐hydroxymyristic acid (non‐hydroxymyristic acid in Zymomonas) in cellular lipids, a new family, Sphingomonadaceae, for Group 4 of the alpha‐subclass of the class Proteobacteria is herein proposed and a description of the family is given. The family consists of six genera, Sphingomonas, Erythrobacter, Erythromicrobium, Porphyrobacter, Sandaracinobacter and Zymomonas. Thus, all the validly published and currently known genera in Group 4 of the alpha‐subclass of Proteobacteria belong to Sphingomonadaceae fam. nov. Among them, type strains of two species, Porphyrobacter and Erythrobacter, Sandaracinobacter sibiricus and Sphingomonas ursincola, respectively, are facultatively photosynthetic due to bacteriochlorophyll (Bchl)‐a. The type strains of two subspecies of Zymomonas mobilis are facultative anaerobes. The presence of SGL‐1 together with the results of a phylogenetic analysis of the 16S rDNA sequences recommends a new criteria by which to define the new family Sphingomonadaceae. The type genus is Sphingomonas Yabuuchi et al. 1990.
Comparison of the Oral Bacterial Flora in Saliva from a Healthy Subject and Two Periodontitis Patients by Sequence Analysis of 16S rDNA Libraries
Authors: Sakamoto Mitsuo , Umeda Makoto , Ishikawa Isao , Benno Yoshimi
Published: November 2013

The oral bacterial flora in the saliva from two patients with periodontitis and from a periodontally healthy subject were compared using a sequence analysis of 16S rDNA libraries without cultivation. 16S rDNAs were amplified from salivary DNA by PCR and cloned. Randomly selected clones were partially sequenced. On the basis of sequence similarities, the clones were classified into several clusters corresponding to the major phylum of the domain Bacteria. The major phylum in the libraries was the low G+C Gram‐positive bacteria. There was no clonal sequence affiliated with periodontopathic bacteria in the salivary sample from the healthy subject, while a number of periodontal pathogens such as Campylobacter rectus, Prevotella intermedia, Porphyromonas gingivalis and Treponema socranskii were detected in the salivary samples from the patients with periodontitis. In addition, a number of previously uncharacterized and uncultured microorganisms were recognized. These organisms may have some role in periodontal disease. This study reveals some potential for a molecular‐biological technique to analyze the oral microflora associated with periodontal disease, including previously uncharacterized and uncultured microorganisms, without cultivation.
Production of Serine Protease of Aeromonas sobria Is Controlled by the Protein Encoded by the Gene Lying Adjacent to the 3′ End of the Protease Gene
Authors: Okamoto Keinosuke , Nomura Tomohiko , Hamada Masaki , Fukuda Takayuki , Noguchi Yoko , Fujii Yoshio
Published: November 2013

We cloned a protease gene of Aeromonas sobria and determined its nucleotide sequence. The protease is composed of 624 amino acid residues and its calculated molecular weight is 66,737.7. The amino acid sequence showed the characteristic features of a bacterial serine protease. We expressed the protease gene in Vibrio parahaemolyticus from which the synthesized protease is secreted into the culture medium as the mature form, and purified the mature protease by successive column chromatographies. The size of the mature protease is 65,000 daltons and the amino acid sequence analysis revealed that a 24‐amino acid peptide at the amino terminal of the precursor is removed from the mature protease. This peptide might function as a signal peptide in translocation across the inner membrane. Subsequently, we found that the protein, designated ORF2 protein, encoded by the gene lying adjacent to the 3′ end of the protease gene plays an important role in production of the protease. Mutation of the ORF2 gene did not affect transcription of the protease gene, but resulted in degradation of the protease in the cell. This shows that ORF2 protein is required for the successful production of the serine protease by cell.
Erythromycin Resistance Genes in Streptococcus pyogenes Isolates in Kanagawa, Japan
Authors: Murase Toshiyuki , Suzuki Rieko , Watanabe Yuko , Yamai Shiro
Published: November 2013

The susceptibility of 224 Streptococcus pyogenes isolates obtained from children in Japan from 1981 to 1997 to treatment with erythromycin was determined by the agar dilution method. A total of 17 isolates belonging to serotype M12T12 were resistant (MICs > 1 μg/ml). Fourteen of the 17 resistant strains obtained from 1982 to 1985 harbored ermB and showed an identical pulsed‐field gel electrophoresis pattern, indicating the spread of a single clone. Two ermTR‐containing isolates were obtained in 1983. mefA gene was found in a strain obtained in 1994 in the present study, although this gene is predominantly associated with recent erythromycin resistance among S. pyogenes strains in many countries.
Lipopolysaccharide (LPS)‐Induced Intra‐Uterine Fetal Death (IUFD) in Mice Is Principally Due to Maternal Cause but Not Fetal Sensitivity to LPS
Authors: Kohmura Yasuhiro , Kirikae Teruo , Kirikae Fumiko , Nakano Masayasu , Sato Ikuo
Published: November 2013

The present study deals with whether lipopolysaccharide (LPS)‐induced intra‐uterine fetal death (IUFD) is related to LPS‐susceptibility of either mother or fetus and how LPS or LPS‐induced TNF causes IUFD. LPS‐susceptible C3H/HeN or ‐hypo‐susceptible C3H/HeJ pregnant mice and the mice mated reciprocally with these mice were used on days 14 to 16 of gestation for experiments. All of fetuses in pregnant C3H/HeN mice mated with either C3H/HeN males [HeN(HeN)] or C3H/HeJ males [HeN(HeJ)] were killed within 24 hr when injected intravenously (i.v.) with 50 or 100 μg of LPS. On the other hand, the majority of fetuses in C3H/HeJ females mated with either C3H/HeJ males [HeJ(HeJ)] or C3H/HeN males [HeJ(HeN)] survived when injected i.v. with even 400 μg of LPS. These findings indicate that LPS‐induced IUFD depends on the maternal LPS‐responsiveness. LPS injected into mothers could pass through placenta to fetuses, since an injection with 125I‐labeled LPS or IgG into pregnant mice resulted in considerable levels of radioactivity in fetuses as well as placenta. Cultured peritoneal macrophages derived from F1 mice of HeJ(HeN) or HeN(HeJ) mice, produced nitric oxide (NO) and tumor necrosis factor (TNF) in response to LPS, although the levels of NO and TNF were lower in comparison with those of C3H/HeN macrophage cultures, suggesting a possibility that the fetus as well as F1 cells might be responsible to LPS. LPS‐induced IUFD was not blocked by treatment with anti‐TNF antibody which inhibited LPS‐induced TNF production in pregnant females, although an injection of recombinant TNFα instead of LPS could induce IUFD, suggesting that the cause of IUFD cannot be attributed to mother‐derived TNF alone. The roles of LPS passed through placenta and LPS‐induced mediators on IUFD were discussed.
Characterization of Vibrio parahaemolyticus Manganese‐Resistant Mutants in Reference to the Function of the Ferric Uptake Regulatory Protein
Authors: Funahashi Tatsuya , Fujiwara Chihiro , Okada Mitsuyo , Miyoshi Shin‐ichi , Shinoda Sumio , Narimatsu Shizuo , Yamamoto Shigeo
Published: November 2013

In many bacteria, the ferric uptake regulatory protein (Fur) has a central role in the negative regulation of genes affected by iron limitation. In this study, Vibrio parahaemolyticus strains carrying mutations in the fur gene encoding Fur were isolated by the manganese selection method to assess the function of Fur in connection with alternations in the coordinate expression of the siderophore vibrioferrin (VF) and iron‐repressible outer membrane proteins (IROMPs). Ten out of 25 manganese‐resistant mutants constitutively produced VF and expressed at least two IROMPs irrespective of the iron concentration in the medium. PCR‐direct DNA sequencing of the fur genes in these mutants identified four different point mutations causing amino acid changes. Moreover, a fur overexpressing plasmid was constructed to prepare antiserum against V. parahaemolyticus Fur. Western blotting with this antiserum revealed that the intracellular abundance of the wild‐type Fur was not significantly affected by the iron concentrations in the growth medium, and that the Fur proteins of the mutant strains occurred at substantially smaller amounts and/or migrated more rapidly in sodium dodecyl sulfate‐polyacrylamide gel electrophoresis than the wild‐type Fur. These data afford an additional insight into the structure‐function relationship of Fur and imply its involvement in the iron acquisition systems of V. parahaemolyticus, although it is yet unknown whether its action on the target genes is direct or indirect.
A VSV‐G Pseudotyped HIV Vector Mediates Efficient Transduction of Human Pulmonary Artery Smooth Muscle Cells
Authors: Li Shou‐lin , Zhang Xiao‐yan , Ling Hong , Ikeda Jun , Shirato Kunio , Hattori Toshio
Published: November 2013

Attempts were made to infect human vascular smooth muscle cells derived from the pulmonary artery (hPASMC) with two different human immunodeficiency virus (HIV) vector systems. ADA/Luc or HXB2/Luc were generated by cotransfection of luciferase reporter gene vector, pNL4‐3‐Lue‐E–R–, and one of two envelope expressing vectors, pSMADA (R5) or pSMHXB2 (X4). The VSV‐G/Luc or VSV‐G/GFP were produced by a three‐plasmid expression system which consisted of vesicular stomatitis virus G protein (VSV‐G) expressing vector, packaging plasmid, and one of two reporter genes (pHR'‐CMV‐Luc or pHR'‐CMV‐GFP). We used hPASMC, U87.CD4.CCR5 and U87.CD4.CXCR4 for infection. Neither ADA/Luc nor HXB2/Luc could infect hPASMC, though they could infect U87.CD4 with corresponding coreceptors. On the other hand, the transduction of both VSV‐G/Luc and VSV‐G/GFP to hPASMC was remarkable. At day 3, the relative proportion of positive cells of hPASMC infected with VSV‐G/GFP was 15%. The above finding indicates a direct role of HIV‐1 infection in pulmonary hypertension ‘a rare complication of HIV‐1 infection’ and HIV‐based vectors could introduce foreign genes into hPASMC for gene therapy of pulmonary hypertension.
Nitric Oxide Production and iNOS mRNA Expression in Mice Induced by Repeated Stimulation with Live Fusobacterium nucleatum
Authors: Kato Chihomi , Mikami Masato , Saito Kazuko
Published: November 2013

There have been few studies on the detection of direct nitric oxide (NO) production and interferon‐gamma (IFN‐γ) in vivo without using animal cell culture. We questioned whether NO and IFN‐γ could be produced at the site of infection. The peritoneal cavity of mice was used as the local infection model. NO and IFN‐γ in abdominal washings from these mice were measured directly at various times after injection of Fusobacterium nucleatum, a gram‐negative rod periodontal pathogen. The mice were divided into three groups: those treated with live bacteria (LB), those treated with heat‐killed bacteria (HKB) and those untreated: normal (N). These mice were compared on the basis of cell filtration, NO and IFN‐γ production by injection of live bacteria (LFn) or heat‐killed bacteria (HKFn). In the LB group, the total cell number increased corresponding to an increase in neutrophils after injection of both LFn and HKFn. A low level of NO was constantly produced in abdominal washings, but a significant amount of NO was synthesized in the LB group only 12 hr to 24 hr after injection of LFn. At the same time iNOS enzyme activity and iNOS mRNA expression were detected. IFN‐γ, which may contribute to enhance NO production, was also secreted at a high level from peritoneal exudate cells (PEC) at 12 hr and 24 hr in the LB group by stimulation of LFn. At 12 hr and 24 hr, iNOS positive cells in the LB group by infection of LFn were identified and shown to contain mostly macrophages. These findings indicate that live bacteria play important roles in NO production by macrophages. It is suggested that NO may contribute to the inflammatory response during F. nucleatum infection in periodontitis.
A Non‐Surgical Rat Model of Foreign Body‐Associated Urinary Tract Infection with Pseudomonas aeruginosa
Authors: Kurosaka Yuichi , Ishida Yoshihisa , Yamamura Eiko , Takase Hiroyuki , Otani Tsuyoshi , Kumon Hiromi
Published: November 2013

This study established a rat model of foreign body‐associated urinary tract infection. A spiral polyethylene tube (PT) was placed transurethrally into the bladder without surgical manipulation, followed by transurethral inoculation with Pseudomonas aeruginosa. The persistence of P. aeruginosa in the kidneys and bladder was significantly enhanced by placement of the PT, whereas the bacteria were eliminated rapidly from the urinary tract in the animals without the PT. Scanning electron microscopy revealed a thick biofilm on the surface of the PT from the early stage of infection. Histopathologically, acute pyelonephritis was followed by chronic renal inflammation as well as continuous and sporadic polymorphonuclear leukocyte accumulation and hemorrhage in the pelvis and adjacent tissues, suggesting continuous ascending introduction of the bacteria from the biofilm adhering to the PT. We believe our model simulates the pathophysiology of foreign body‐associated urinary tract infection characterized by biofilm formation on the surface of a foreign body.
A Lack of Consistent Amino Acid Substitutions in NSP4 between Rotaviruses Derived from Diarrheal and Asymptomatically‐Infected Kittens
Authors: Oka Toyonori , Nakagomi Toyoko , Nakagomi Osamu
Published: November 2013

Nonstructural glycoprotein NSP4 of group A rotavirus has recently been shown to be a viral enterotoxin, inducing diarrhea in neonatal mice. Literature is conflicting as to whether there is any consistent amino acid substitution between virulent (or symptomatic) and attenuated (or asymptomatic) rotavirus strains. We have sequenced and compared the NSP4 sequences derived from a total of 10 geographically‐ and serologically‐related feline rotavirus strains from both diarrheal and asymptomatically‐infected kittens. These NSP4 sequences were closely related to each other and there were differences at 19 amino acid residues, but none was segregated according to whether the strain was isolated from a diarrheal kitten or not. Thus, this study failed to lend support to the contention that mutations in NSP4 play a significant role in the pathogenesis of rotavirus diarrhea. Involvement of other genes may explain the outcome of infection in cats from which these 10 feline rotaviruses were isolated.
Genomic Map of Clostridium perfringens Strain 13
Authors: Shimizu Tohru , Ohshima Shinobu , Ohtani Kaori , Shimizu Takeshi , Hayashi Hideo
Published: November 2013

A physical and genetic map of Clostridium perfringens strain 13 was constructed. C. perfringens strain 13 was found to have a 3.1‐Mb chromosome and a large 50‐kb plasmid, indicating that strain 13 has a relatively small genome among C. perfringens strains. A total of 313 genetic markers were mapped on the chromosome of strain 13. Compared with the physical and genetic map of C. perfringens CPN50, strain 13 had a quite similar genome organization, but with a large deletion (~400 kb) in a particular segment of the chromosome. Among several toxin genes, a beta2 toxin gene that is a novel virulence gene in C. perfringens was found to be located on the 50‐kb plasmid.
Vi‐Suppressed Wild Strain Salmonella typhi Cultured in High Osmolarity Is Hyperinvasive toward Epithelial Cells and Destructive of Peyer's Patches
Authors: Zhao Licheng , Ezaki Takayuki , Li Zhi‐Yu , Kawamura Yoshiaki , Hirose Kenji , Watanabe Haruo
Published: November 2013

Salmonella typhi GIFU10007–3 which lost a viaB locus on its chromosome became highly invasive in our previous study. To investigate the phenomenon, we controlled Vi expression in wild strain S. typhi GIFU10007, and studied the invasive phenotype both in vitro and in vivo. When the wild strain of S. typhi was cultured in 300 mm NaCl containing Luria‐Bertani broth (LBH), the expression of Vi antigen was suppressed, but secretion of invasion proteins (SipC, SipB and SipA) was increased. In this condition, wild strain S. typhi became highly invasive toward both epithelial cells and M cells of rat Peyer's patches. When GIFU10007 was cultured under conditions of high osmolarity, the bacteria disrupted Peyer's patches and induced massive bleeding in these structures only 20 min after inoculation into the ileal loop. In contrast, Vi‐encapsulated wild strain GIFU10007 cultured under low osmolarity was not destructive, even after 60 min. To understand the role of the type III secretion system under conditions of high osmolarity, we knocked out the invA and sipC genes of both GIFU10007 and GIFU10007–3. Neither invA nor sipC mutants could invade epithelial cells or M cells in a high osmolarity environment. Our data show that the highly invasive phenotype was only expressed when the wild strain S. typhi was cultured under high osmolarity, which induced a state of Vi suppression, and in the presence of the type III secretion system.
Herpesvirus DNA in Peripheral Blood Mononuclear Cells of Some Patients with Meniere's Disease
Authors: Takahashi Kazuo , Aono Toshiya , Shichinohe Mitsuo , Tamura Manabu , Iwata Yasuhide , Yamanishi Koichi , Shigeta Shiro
Published: November 2013

Herpes simplex virus‐1 (HSV) or varicella zoster virus (VZV) DNA was detected by nested polymerase chain reaction in peripheral blood mononuclear cells of patients with Meniere's disease (one of 28 patients for HSV‐1,2 of 28 patients for VZV) during acute illness (within 5 days after onset). On the other hand, neither HSV‐1 DNA or VZV DNA was detected in PBMCs of 50 age‐ and sex‐matched healthy individuals and 50 pregnant women. These findings may imply that reactivation of HSV‐1 or VZV may be associated with the development of some cases of Meniere's disease.
Coaggregation of Porphyromonas gingivalis and Prevotella intermedia
Authors: Kamaguchi Arihide , Nakayama Koji , Ohyama Tohru , Watanabe Toshihiro , Okamoto Masaaki , Baba Hisae
Published: November 2013

Porphyromonas gingivalis cells coaggregated with Prevotella intermedia cells. The coaggregation was inhibited with L‐arginine, L‐lysine, Nα‐p‐tosyl‐L‐lysine chloromethyl ketone, trypsin inhibitor, and leupeptin. Heat‐ and proteinase K‐treated P. gingivalis cells showed no coaggregation with P. intermedia cells, whereas heat and proteinase K treatments of P. intermedia cells did not affect the coaggregation. The vesicles from P. gingivalis culture supernatant aggregated with P. intermedia cells, and this aggregation was also inhibited by addition of L‐arginine or L‐lysine and by heat treatment of the vesicles. The rgpA rgpB, rgpA kgp, rgpA rgpB kgp, and rgpA kgp hagA mutants of P. gingivalis did not coaggregate with P. intermedia. On the other hand, the fimA mutant lacking the FimA fimbriae showed coaggregation with P. intermedia as well as the wild type parent. These results strongly imply that a heat‐labile and proteinous factor on the cell surface of P. gingivalis, most likely the gingipain‐adhesin complex, is involved in coaggregation of P. gingivalis and P. intermedia.
Inhibitory Role of Neutrophils on Influenza Virus Multiplication in the Lungs of Mice
Authors: Fujisawa Haruo
Published: November 2013

The protective role of neutrophils on intranasal infection of influenza virus was investigated in 3 strains of tumor‐bearing mice with neutrophilic leukocytosis. In vitro multiplication of influenza virus was inhibited by neutrophils from both normal and tumor‐bearing mice, and the inhibitory effect of neutrophils was augmented by an addition of fMLP to the culture. Pulmonary virus infectivities in the early phase after infection decreased in such ICR and BALB/c mice, and virus elimination in the late phase was accelerated in the ICR mice. However, no decrease in pulmonary virus infectivity was observed in tumor‐bearing C57BL/6 mice. Intranasal administration of fMLP into normal and tumor‐bearing C57BL/6 mice after infection significantly inhibited the virus propagation in the lungs. The decrease in neutrophil infiltration into the lung in tumor‐bearing C57BL/6 mice was confirmed from histological observations of the lung and lung lavage after infection and from analysis of the neutrophil chemotactic activity induced by fMLP. This might be responsible for the high level of pulmonary virus titer in tumor‐bearing C57BL/6 mice. Phagocytic activities of alveolar macrophages and productions of neutralizing antibody were suppressed in the 3 strains of tumor‐bearing mice. These observations indicated that neutrophils could be significant effector cells as a host defense mechanism against influenza virus infection in vivo, and infiltration and functional activation of neutrophils could play a significant role in virus elimination from the infected site. Furthermore, the inhibition of virus propagation by neutrophils in vitro was almost completely abrogated by an addition of ZnSO4, suggesting that calprotectin could inhibit influenza virus multiplication.