bluuebunny/crossref_metadata_2025_split huggingface dataset:
Detection of Methicillin‐Resistant Staphylococcus aureus (MRSA) with Antibodies against Synthetic Peptides Derived from Penicillin‐Binding Protein 2′
Authors: Sekiguchi Kiyoshi , Saito Michihiro , Yajima Ryuichi 
Published: November 2013

AbstractTen kinds of peptides (21 to 32 amino acids in length) were synthesized based on the reported amino acid sequences of the penicillin‐binding protein 2′ (PBP2′) of methicillin‐resistant Staphylococcus aureus (MRSA). Antibodies against these synthetic peptides (SPs) were generated by immunizing rabbits. The antibodies raised against all the peptides except for one reacted to PBP2′ of MRSA and to SPs used for immunization but not to any other protein of MRSA or methicillin‐susceptible S. aureus (MSSA) tested by ELISA and Western blotting. A sandwich immunoradiometric assay (IRMA) for the detection of PBP2′ was developed using these antibodies. The method could detect PBP2′ extracted from as few as 3 × 104 cells of a clinical MRSA isolate, and a good correlation between cell number and signal radio‐count was observed. IRMA was positive for all 51 methicillin‐resistant staphylococci isolated from patients, and was negative for all the 28 methicillin‐susceptible ones and 19 strains of other bacterial species. IRMA could be a simple and reliable method for MRSA detection in the clinical bacterial laboratory.
Segmented Filamentous Bacteria Are Indigenous Intestinal Bacteria That Activate Intraepithelial Lymphocytes and Induce MHC Class II Molecules and Fucosyl Asialo GM1 Glycolipids on the Small Intestinal Epithelial Cells in the Ex‐Germ‐Free Mouse
Authors: Umesaki Yoshinori , Okada Yasushi , Matsumoto Satoshi , Imaoka Akemi , Setoyama Hiromi 
Published: November 2013

AbstractIn ex‐germ‐free mice conventionalized by association with fecal microorganisms, the induction of major histocompatibility complex class II molecules and fucosylation of asialo GM1 glycolipid occur in the small intestinal epithelial cells (IEC). The intestinal intraepithelial lymphocytes (IEL), especially αβ T‐cell receptor‐bearing ones, also remarkably expand and show cytolytic activity. In this study, we investigated the immunological and physiological characteristics of the small intestine induced by a kind of indigenous bacteria of the small intestine, segmented filamentous bacteria (SFB), among chloroform‐resistant intestinal bacteria. Monoassociation of SFB with germ‐free mice was confirmed by the determination of the base sequences of polymerase chain reaction products of 16S rRNA genes of the fecal bacteria of these mice and in situ hybridization using fluorescein‐labeled probes based on them. SFB increased the number of αβTCR‐bearing IEL and induced Thy‐1 expression and cytolytic activity of IEL. The induction of MHC class II molecules and fucosyl asialo GM1 glycolipids and the increases in the mitotic activity and the ratio of the number of columnar cells to those of goblet cells also occurred in the small intestinal epithelial cells on monoassociation of these bacteria. SFB are important indigenous bacteria for the development of the mucosal architecture and immune system in the small intestine, at least in mice.
Inactivation of Lactobacillus Bacteriophage PL‐1 by Microwave Irradiation
Authors: Kakita Yukari , Kashige Nobuhiro , Murata Kunihiko , Kuroiwa Ataru , Funatsu Michiko , Watanabe Kenji 
Published: November 2013

AbstractThe effect of microwave irradiation on the survival of bacteriophage PL‐1, which is specific for Lactobacillus casei, was studied using a commercial 2,450 MHz microwave oven. The phages were inactivated by microwave irradiation according to almost first‐order reaction kinetics. The rate of phage inactivation was not affected by the difference in the continuous or intermittent irradiation, nor by the concentrations of phages used, but was affected by the volume of phage suspensions, which prevented the loss of generated heat. Microwave irradiation of phage suspensions produced a number of ghost phages with empty heads, but fragmentation of the tail was hardly noticed. The breakage of phage genome DNA was primarily caused by the heat generated by microwave irradiation, whereas the phage DNA was not affected by the same temperature achieved by heat from outside. Thus we concluded that the phage‐inactivating effect of microwave irradiation was mainly attributed to a thermal microwave effect, which was much stronger than a simple thermal exposure.
Humoral Immunity to Commensal Oral Bacteria: Quantitation, Specificity and Avidity of Serum IgG and IgM Antibodies Reactive with Actinobacillus actinomycetemcomitans in Children
Authors: Cole Michael F. , Fitzsimmons Sean P. , Sheridan Michael J. , Xu Yong 
Published: November 2013

AbstractThe levels, specificity and avidities of serum IgM and IgG antibodies reactive with Actinobacillus actinomycetemcomitans (Aa) serotypes a, b and c were determined in periodontally healthy (PH) children and compared with subjects with localized juvenile periodontitis (LJP). All PH children exhibited IgM and IgG Aa‐reactive antibodies whether or not Aa was detected subgingivally but the antibodies were not specific for Aa. In contrast, LJP sera contained high concentrations of IgM and IgG antibodies reactive with Aa that were largely specific for this bacterium. IgM and IgG antibodies in both PH and LJP subjects were of low avidity. With one exception, the avidities of IgG anti‐Aa antibodies were significantly greater than those of IgM antibodies in both PH and LJP subjects. However, although the LJP subjects had as much as 115‐fold more Aa‐reactive IgG antibody than did the PH subjects the avidities of their IgG antibodies were no greater than those of the PH group. The induction by the host of low‐avidity antibodies, that are ineffective in immune elimination, may be a reason why commensal bacteria persist at mucosal surfaces and why persons with LJP fail to eliminate Aa from their periodontal pockets.
Alteration of Bacteriolytic Enzyme Profile of Staphylococcus aureus during Growth
Authors: Komatsuzawa Hitoshi , Sugai Motoyuki , Nakashima Seiji , Suginaka Hidekazu 
Published: November 2013

AbstractProfiles of cell‐associated bacteriolytic activities and those in the culture supernatant of Staphylococcus aureus FDA209P at various stages of growth were analyzed using sodium dodecyl sulfate‐polyacrylamide gels containing Micrococcus luteus or S. aureus. In the logarithmic growth phase, the cell‐associated bacteriolytic activities extracted with Triton X‐100 contained a number of bacteriolytic proteins, the profiles of which were similar to those we reported elsewhere (Sugai, M., Akiyama, T., Komatsuzawa, H., Miyake, Y., and Suginaka, H.(1990) J. Bacteriol., 172, 6494‐6498). The proteins include P1, P2, P7, P9, PX, P13, P18 and other minor components. At the stationary growth phase, the bacteriolytic band‐profile of the Triton X‐100 extract changed dramatically. P1, P7 and P9 disappeared, and the other minor bands had markedly decreased band intensities. On the other hand, P2, PX, P13, and P18 retained their band intensities during the stationary growth phase. The band intensities of P7, P13, PX, and P18 increased in the supernatant during the logarithmic growth phase. These results indicated that the bacteriolytic band‐profile changes during growth.
Antiviral Activity of Trichothecene Mycotoxins (Deoxynivalenol, Fusarenon‐X, and Nivalenol) against Herpes Simplex Virus Types 1 and 2
Authors: Tani Naoto , Dohi Yoshiko , Onji Yoshiki , Yonemasu Kunio 
Published: November 2013

AbstractThe effect of trichothecene mycotoxins, deoxynivalenol (DON), fusarenon‐X (FX) and nivalenol (NIV), on plaque formation of herpes simplex virus types 1 and 2 (HSV‐1 and HSV‐2) in HEp‐2 cells was examined. The 50% effective concentrations (EC50) of DON, FX, and NIV for HSV‐1 plaque formation were 160, 56, and 120 ng/ml, respectively. Those for HSV‐2 plaque formation were 94, 26, and 50 ng/ml, respectively. These three mycotoxins showed about 2‐fold higher selectivity to HSV‐2 than to HSV‐1. Plaque formation of HSV‐1 was not inhibited with trichothecenes at concentrations completely inhibiting plaque formation when cells were treated during virus adsorption period or 15 hr before infection. These results indicate that trichothecenes affect replication of HSV‐1 after virus adsorption, but not before or during virus adsorption to the host cells.
Suppression of Cytokine Production in T Helper Type 2 Cells by Nitric Oxide in Comparison with T Helper Type 1 Cells
Authors: Nukaya Ikuei , Takagi Kuniaki , Kawabe Takefumi , Suketa Yasunobu 
Published: November 2013

AbstractWe examined the effect of nitric oxide (NO) on cytokine production in T helper (Th) cell subsets, using murine splenic CD4+ T cells and two types of Th clones. Interferon‐gamma‐treated murine peritoneal exudate cells (IFN‐PEC) suppressed DNA synthesis to 60% of the control level in CD4+ T cells stimulated with the anti‐CD3 monoclonal antibody. The production of IL‐2 and IL‐4 in the CD4+ T cells decreased to 63.2% and 9.2%, respectively, of the control value by co‐culture with IFN‐PEC. The addition of NG‐monomethyl‐L‐arginine (L‐NMMA) partially recovered the suppression of DNA synthesis. In the presence of indomethacin, the suppression of DNA synthesis was partially inhibited and the reduction in the cytokine production caused by IFN‐PEC was partially recovered. The simultaneous addition of NG‐monomethyl‐L‐arginine (L‐NMMA) and indomethacin completely inhibited not only the suppression of DNA synthesis but also the reduction in the cytokine production caused by IFN‐PEC. Moreover, DNA synthesis in the Th2 clone was suppressed to a greater extent than that in the Th1 clone by co‐culture with IFN‐PEC. This suppression in the Th1 clone was inhibited by the addition of L‐NMMA, whereas the DNA synthesis in the Th2 clone was not recovered by L‐NMMA. In addition, sodium nitroprusside (SNP) suppressed IL‐4 production in the Th2 clone but had no effect on IL‐2 production in the Th1 clone. In the experiment of the co‐culture with IFN‐PEC, the inhibitory‐effect of NO on T cell activation was not clarified by the influence of prostaglandins. However, in conclusion, cytokine production in Th2 cells may be more susceptible to NO than that in Th1 cells.
Growth Conditions Influence Expression of Cell Surface Hydrophobicity of Staphylococci and Other Wound Infection Pathogens
Authors: Ljungh Åsa , Wadström Torkel 
Published: November 2013

AbstractThe initial adhesion of microbes to tissue and solid surfaces can be mediated by hydrophobic interaction. Expression of microbial cell surface hydrophobicity (CSH) is influenced by growth conditions, and often best expressed after growth under nutrient‐poor conditions, or “starvation.” In the present study, the CSH of 133 strains of Enterobacteriaceae, Staphylococcus aureus, coagulase‐negative staphylococci, Enterococcus faecalis, group A streptococcus, Pseudomonas aeruginosa, Clostridium perfringens, Bacteroides fragilis, Peptococcus magnus, and of 8 Candida albicans strains was measured by the salt aggregation test after growth on hematin agar in a 5% CO2 atmosphere, or under anaerobiosis. Cells of all but 8 strains expressed pronounced or moderate CSH, i.e., they aggregated in 0.01‐2 M ammonium sulfate. When the agar surface was covered by human serum (diluted 1:5) to mimic growth conditions in a wound, 94 strains expressed higher CSH, and 44 strains the same CSH as after growth without serum. The CSH of 12 strains of different species was measured after growth on blood, hematin and PDM agar, with or without serum, and in an aerobic or a 5% CO2 atmosphere. The highest CSH was expressed after growth in 5% CO2 with serum, and the lowest growth after on blood agar in aerobic atmosphere. Identical results were obtained with native and heat‐inactivated (56 C, 20 min) serum. The reduced surface tension obtained in 5% CO2, as well as yet unidentified serum factors, promotes expression of CSH.
Class‐Specific Antibody Response in Acyclovir‐Treated and Adenine Arabinoside‐Treated Patients with Primary Genital Herpes Simplex Virus Infection
Authors: Kawana Takashi , Hashido Madoka , Koizumi Yoshio 
Published: November 2013

AbstractHerpes simplex virus (HSV) class‐specific antibody responses after primary genital herpes infection were evaluated with an enzyme‐linked immunosorbent assay (ELISA) in 16 patients treated with acyclovir (ACV), given orally, and 17 patients treated with adenine arabinoside (Ara‐A), given topically. ACV significantly suppressed the levels of IgM, IgA, and IgG. In the ACV‐treated patients, IgM and IgG were not detected in 4 of the 16 and in 1 of the 16 patients, respectively. We must take into account this suppressive effect of ACV on antibody responses, especially on the IgM response, when serodiagnosis of HSV infection is made.
Cross‐Reactivity of Rickettsia japonica and Rickettsia typhi Demonstrated by Immunofluorescence and Western Immunoblotting
Authors: Uchiyama Tsuneo , Zhao Licheng , Yan Yansheng , Uchida Takahiro 
Published: November 2013

AbstractCross‐reactivity between Rickettsia japonica and R. typhi was observed by immunofluorescence tests using sera from patients with Oriental spotted fever (OSF), from whom the causative agent was isolated and identified as R. japonica. Western immunoblotting with these sera revealed that only the 120‐kilodalton surface polypeptide, i.e., rickettsial outer membrane protein (rOmp) B, has a common antigenicity with the 105‐kilodalton surface polypeptide of R. typhi. In some cases, antibodies specifically reactive with R. typhi were detected in acute‐phase sera followed by a significant rise in titers, possibly because of an anamnestic response to a previous infection with an R. typhi‐like agent; the sera retained reactivity to R. typhi even after absorption by a homologous strain. A lipopolysaccharide (LPS)‐like antigen of R. typhi was found to be reactive with some sera of OSF patients. The ladder bands on Western immunoblot of rickettsial organisms were confirmed to be polysaccharide in nature, which was demonstrated by comparing them with the pattern of silver‐stained gel of proteinase K‐treated rickettsial specimens after sodium dodecyl sulfate‐polyacrylamide gel electrophoresis.
Purification and Characterization of a Fibrinogen‐Degrading Protease in Bacteroides fragilis Strain YCH46
Authors: Chen Yong , Kinouchi Takemi , Kataoka Keiko , Akimoto Shigeru , Ohnishi Yoshinari 
Published: November 2013

AbstractA novel fibrinogenolytic protease was purified from Bacteroides fragilis strain YCH46. The protease was extracted from cells by ultrasonic treatment and was purified 425‐fold with a recovery of 2.1% by sequential procedures using azocasein as a substrate. The purified protease showed a single band on sodium dodecyl sulfate‐polyacrylamide gel electrophoresis with an estimated molecular weight of 100 kDa, which was consistent with the value obtained by gel filtration, indicating a monomeric native structure. Its optimal pH, Km, and Vmax for azocasein were 7.5, 0.2%, and 286 U/min/mg, respectively. The protease activity was completely inhibited by addition of 1 mM Hg2+, Cu2+, Zn2+, diisopropyl fluorophosphate, N‐ethylmaleimide or p‐chloromercuribenzoate but not by the inhibitors of metalloprotease or aspartic protease, suggesting that the enzyme is a serine‐thiol‐like protease. The protease hydrolyzed azocasein, casein, fibrinogen, gelatin, and azocoll, but not bovine serum albumin, ovalbumin, fibrin, fibronectin, immunoglobulins, transferrin, hemoglobin or types I, III, and IV collagen. The enzyme also hydrolyzed the chromogenic substrates alanyl‐alanine p‐nitroanilide, L‐valyl‐alanine p‐nitroanilide, alanyl‐alanyl‐valyl‐alanine p‐nitroanilide, and glycyl‐proline p‐nitroanilide, but was inert toward L‐alanine p‐nitroanilide, alanyl‐alanyl‐alanine p‐nitroanilide, and N‐α‐benzoyl‐DL‐arginine p‐nitroanilide. The protease completely hydrolyzed the α‐chain of fibrinogen at 37 C within 10 hr and at the same time the time required for clotting of protease‐treated fibrinogen by thrombin was prolonged. The fibrinogenolytic activity of a crude extract of B. fragilis was stronger than that of other species of the Bacteroides fragilis group tested: B. ovatus, B. distasonis, B. eggerthii, B. uniformis, and B. thetaiotaomicron. These results suggest that the fibrinogenolytic protease is an important biological factor in Bacteroides infection.
Demonstration and Genotyping of Pestivirus RNA from Mammalian Cell Lines
Authors: Harasawa Ryô , Mizusawa Hiroshi 
Published: November 2013

AbstractWe examined 20 cell lines of various animal origins for the presence of pestivirus contamination by the reverse transcription‐polymerase chain reaction (RT‐PCR), and found 15 (75%) cell lines were positive. The RT‐PCR products of the 5′ untranslated region (UTR) of pestivirus genome were sequenced and subjected to genotyping. Stem‐loop structures at three variable regions in the 5′ UTR render genotyping of the contaminated pestiviruses. Bovine cell lines tested were all contaminated with genotypes I, II, or III of bovine diarrhea virus (BVDV). Cell lines of canine, feline, and primate origin were contaminated with genotype II of BVDV. Cell line Ch1Es of caprine origin was contaminated with border disease virus (BDV).
Extraction and Characterization of the Smooth‐Type Lipopolysaccharide from Fusobacterium nucleatum JCM 8532 and Its Biological Activities
Authors: Onoue Sakura , Niwa Motō , Isshiki Yasunori , Kawahara Kazuyoshi 
Published: November 2013

AbstractThe lipopolysaccharide (LPS) of Fusobacterium nucleatum JCM 8532 was isolated by hot‐phenol water extraction. Most of the LPS was extracted in the phenolic phase and shown to be the smooth‐type, whereas the aqueous phase contained mainly rough‐type LPS. The chemical composition of the LPS was similar to that reported in other studies, but D‐quinovosamine, which may be a major component of O‐antigenic polysaccharide, and 3‐deoxy‐D‐manno‐2‐octulosonic acid (Kdo) were detected for the first time by gas chromatography‐mass spectrometry. The biological activities of smooth‐type LPS, including limulus activity, lethal toxicity, pyrogenicity, and B lymphocyte mitogenicity, were comparable to those of enterobacterial LPS. Smooth‐type LPS inhibited the cell growth and DNA synthesis of adult and fetal human gingival fibroblasts in a dose‐dependent manner, suggesting that LPS may play a role in the occurrence of human gingivitis.
Typing of Verotoxins by DNA Colony Hybridization with Poly‐ and Oligonucleotide Probes, a Bead‐Enzyme‐Linked Immunosorbent Assay, and Polymerase Chain Reaction
Authors: Yamasaki Shinji , Lin Zaw , Shirai Hiromasa , Terai Akito , Oku Yuichi , Ito Hideaki , Ohmura Mari , Karasawa Tadahiro , Tsukamoto Teizo , Kurazono Hisao , Takeda Yoshifumi 
Published: November 2013

AbstractTo identify the type of Verotoxins (VT) produced by Verocytotoxin‐producing Escherichia coli (VTEC), a sensitive bead‐enzyme‐linked immunosorbent assay and polymerase chain reaction with common and specific primers to various VTs (VT1, VT2, VT2vha, VT2vhb, and VT2vp1) were developed. Together with colony hybridization tests with oligo‐ and polynucleotide probes, these methods were applied to VTEC isolates to type the VT produced. The toxin types of 26 of 37 strains were identified, but the reaction profiles in assays of the remaining 11 strains suggested the existence of new VT2 variants. The application of these identification procedures may be useful as a tool for clinical and epidemiological studies of VTEC infection.
Transient Expression of CD20 Antigen (Pan B Cell Marker) in Activated Lymph Node T Cells
Authors: Murayama Yuichi , Mukai Ryozaburo , Sata Tetsutaro , Matsunaga Satoko , Noguchi Atsuo , Yoshikawa Yasuhiro 
Published: November 2013

AbstractIn contrast to the case of peripheral T cells, the surface expression of CD20 antigen and the expression of CD20 mRNA in monkey lymph node (LN) T cells underwent a noticeable increase when they were cultured with mitogen and interleukin‐2 (IL‐2). To confirm in vivo regulation of CD20 expression during the activation of LN T cells, we examined LNs derived from monkeys experimentally inoculated with simian immunodeficiency virus (SIV). Significant expression of CD20 antigen was detected in the T cells of the LNs at the stage of lymphadenopathy. These findings suggest that lymphocyte activation in the LNs induced expression of the CD20 molecule in some T cells.
Role of Intestinal Bacteria in Ileal Ulcer Formation in Rats Treated with a Nonsteroidal Antiinflammatory Drug
Authors: Uejima Motoo , Kinouchi Takemi , Kataoka Keiko , Hiraoka Isao , Ohnishi Yoshinari 
Published: November 2013

AbstractThe role of intestinal bacteria in induction and repression of ulcer formation in the ileum of rats treated with one of the nonsteroidal antiinflammatory drugs (NSAIDs), 5‐bromo‐2‐(4‐fluorophenyl)‐3‐(4‐methylsulfonylphenyl) thiophene (BFMeT), was examined in this study. BFMeT was administered by intragastric gavage once at doses of 500‐1,500 mg/kg of body weight to Wistar rats treated with and without antibiotics (bacitracin, neomycin, streptomycin), germ‐free rats and gnotobiotic rats, and 72 hr later their gastrointestinal tracts were examined for ulcer formation. A single oral administration of BFMeT induced ileal ulcers in specific pathogen‐free rats. However, the rats given antibiotics to reduce the intestinal bacteria had no ulcers. BFMeT‐treated germ‐free rats and gnotobiotic rats mono‐associated with Bifidobacterium adolescentis or Lactobacillus acidophilus also had no intestinal ulcers. However, the drug induced ileal ulcers in gnotobiotic rats mono‐associated with Eubacterium limosum or Escherichia coli. An overnight culture of B. adolescentis or L. acidophilus or yogurt containing Bifidobacterium breve and Streptococcus thermophilus, when given as drinking water, inhibited ulcer formation in the ileum of rats treated with BFMeT. Gram staining of the ileal contents of normal rats revealed that 97.4% of the stained microorganisms were Gram‐positive rods and only 1.2% were Gram‐negative rods. In the group of rats with ulcers induced by BFMeT, the Gram‐positive rods decreased by 56.4% and the Gram‐negative rods including Escherichia coli, Klebsiella, Proteus and Bacteroides increased by 37.3%. However, in the group of rats administered the Bifidobacterium culture, the Lactobacillus culture or yogurt, the percentages of the Gram‐negative rods were decreased. Although Lactobacillus was a major bacterium in the ileum of normal rats, the Gram‐negative facultatively anaerobic rods E. coli, Klebsiella and Proteus were increased in the ulcerated ileum of rats treated with BFMeT, suggesting that these bacteria are associated with ulcer formation in rats treated with NSAIDs, and that Lactobacillus and Bifidobacterium inhibit it by repressing the growth of ulcer‐inducing bacteria.
Expression of Virulence‐Associated Antigens of Rhodococcus equi Is Regulated by Temperature and pH
Authors: Takai Shinji , Fukunaga Nariaki , Kamisawa Kei , Imai Yumiko , Sasaki Yukako , Tsubaki Shiro 
Published: November 2013

AbstractWe recently reported that there are two different virulence‐associated antigens correlated with virulence levels in Rhodococcus equi isolates from AIDS patients: virulent R. equi that kills mice with 106 cells expresses 15‐ to 17‐kDa antigens and intermediately virulent R. equi that kills mice with 107 cells expresses a 20‐kDa antigen. Environmental parameters were evaluated for their effects on the expression of these virulence‐associated antigens in virulent R. equi strains by immunoblotting using monoclonal antibodies in this study. Expression of these two virulence‐associated antigens of R. equi was regulated by pH and temperature; the antigens were produced maximally when the isolates were grown at 38 C and pH 6.5, but were not produced when grown at 38 C and pH 8, nor at temperatures below 30 C. The 20‐kDa antigen was found to be located on the cell surface, as were the 15‐ to 17‐kDa antigens, and showed susceptibility to proteolysis by trypsin. These results indicate that expression of the virulence‐associated antigens of R. equi is dependent on the environmental conditions.