bluuebunny/crossref_metadata_2025_split huggingface dataset:
Levels of Porphyromonas gingivalis Fimbriae and Inflammatory Cytokines in Gingival Crevicular Fluid from Adult Human Subjects
Authors: Hirose Kimiharu , Isogai Emiko , Miura Hiroko , Ueda Itsuo 
Published: November 2013

AbstractThe Porphyromonas gingivalis fimbriae level was examined in the gingival crevicular fluid (GCF) from adult human subjects using an immunoblot assay with a monoclonal antibody. The cytokines, interleukin‐1α (IL‐1α), IL‐1β, IL‐6 and tumor necrosis factor‐a (TNF‐α) levels in the GCF were quantified by enzyme‐linked immunosorbent assay (ELISA). The reactivity of the GCF samples with the monoclonal antibody against P. gingivalis fimbriae was related to the IL‐1β, IL‐6 and TNF‐α levels. Moreover, the fimbriae content was associated with the gingival index (GI). In contrast, no significant correlation was seen between the fimbriae content and IL‐1α level. These results suggest that there are possible associations between P. gingivalis fimbriae and IL‐1β, IL‐6 and TNF‐α in the gingival crevicular fluid.
Apoptosis of Intestinal Intraepithelial Lymphocytes Induced by Exogenous and Endogenous Glucocorticoids
Authors: Murosaki Shinji , Inagaki‐Ohara Kyoko , Kusaka Hiroaki , Ikeda Hitoshi , Yoshikai Yasunobu 
Published: November 2013

AbstractTo investigate the effect of glucocorticoids on apoptosis in intestinal intraepithelial lymphocytes (i‐IEL), we examined the changes of i‐IEL followed by in vivo treatment with dexamethasone. The fragmented DNA of i‐IEL were significantly increased at 15 hr after dexamethasone treatment and, subsequently, the number of total i‐IEL were decreased by day 4 after treatment. Although all subsets of i‐IEL including CD8α/α+, CD8α/β+, CD4+ and CD4+ CD8+ i‐IEL were decreased after dexamethasone treatment, CD8α/α+ i‐IEL appeared to be relatively resistant to dexamethasone‐induced apoptosis. Consistent with the in vivo findings, CD8α/α+ i‐IEL exhibited less susceptibility to dexamethasone‐induced cell death in vitro than other subsets. To investigate whether this process occurs under physiological conditions, we examined the kinetics of i‐IEL after treatment with 15‐hr water immersion stress. In mice subjected to water immersion stress, plasma glucocorticoids were remarkably elevated soon after the 15‐hr stress. The increase in the fragmented DNA of i‐IEL and subsequent decrease in the number of i‐IEL were observed in the stressed mice in the same kinetics as seen in the dexamethasone‐treated mice. Similar to dexamethasone‐induced cell death, CD8α/α+ i‐IEL appeared to be relatively resistant to stress‐induced apoptosis compared with other i‐IEL subsets. The expression level of Bcl‐2 was significantly higher in CD8α/α+ i‐IEL than in CD8α/β+ i‐IEL. Our results indicate that i‐IEL are subjected to cell death via apoptosis by exogenous and endogenous glucocorticoids and that different sensitivity to steroid‐induced apoptosis may exist among i‐IEL subsets in relation to their Bcl‐2 expression.
In Vitro Selection of Variants of Herpes Simplex Virus Type 1 Which Differ in Cytopathic Changes
Authors: Padilla Jorge , Yamada Masao , Takahashi Yasushi , Tsukazaki Takashi , Nakamura Jun , Yoshida Mariko , Uno Fumio , Arao Yujiro , Nii Shiro 
Published: November 2013

AbstractTo analyze the mechanisms for in vitro emergence of the syncytial variants of herpes simplex virus type 1 (HSV‐1), several cell lines were infected with a mixture of equal amounts of two HSV‐1 variants, one syncytial and the other non‐syncytial, and changes in their relative abundance were monitored during passage. With a combination of two variants of the Miyama strain of HSV‐1, the syncytial variant became dominant during passage in Vero, RK‐13 and FL cells. On the other hand, the ratios of the two variants remained around 1:1 during the passage in HEp‐2, MGC and HEL cells. In another set of variants of the SKO strain of HSV‐1, the outcomes were different from those of the Miyama strain in the FL, MGC and HEp‐2 cells. The ratios of the two variants remained around 1:1 during passage in FL cells, while the non‐syncytial variant became dominant during passage in MGC and HEp‐2 cells. In addition, we examined the effects of a complement and interferon‐β (IFN‐β) on the outcome of the selection. As a result, the complement slowed the selection of a syncytial variant, whereas IFN‐β facilitated it.
Thiadiazole Derivatives: Highly Potent and Selective Inhibitors of Human Immunodeficiency Virus Type 1 (HIV‐1) Replications In Vitro
Authors: Fujiwara Masatoshi , Ijichi Katsushi , Hanasaki Yasuaki , Ide Teruhiko , Katsuura Kimio , Takayama Hiromitsu , Aimi Norio , Shigeta Shiro , Konno Kenji , Yokota Tomoyuki , Baba Masanori 
Published: November 2013

AbstractWe have recently reported that thiadiazole (TDA) derivatives are highly potent inhibitors of human immunodeficiency virus type 1 (HIV‐1) replication. These compounds belong to the family of nonnucleoside reverse transcriptase inhibitors (NNRTIs). In an attempt to develop more effective and pharmacologically favorable compounds, novel TDA derivatives have been synthesized and examined for their anti‐HIV‐1 activity in vitro. Among them, RD4‐2217 was found to be the most potent inhibitor of HIV‐1 replication. It inhibited replication of the HTLV‐IIIB strain in MT‐4 cells at a concentration of 6 nM. RD4‐2217 was also inhibitory to clinical isolates and zidovudine‐resistant mutants of HIV‐1. The combination of RD4‐2217 with zidovudine or the protease inhibitor A‐75925 synergistically inhibited HIV‐1 replication. Studies on the emergence of drug‐resistant mutants revealed that, although much higher concentrations (1‐10 μM) were required, RD4‐2217 completely suppressed the breakthrough of HIV‐1 in the supernatants during long‐term culturing of infected cells. Furthermore, RD4‐2217 at low concentrations (10 or 100 nM), in combination with zidovudine, also completely inhibited viral breakthrough. In addition, RD4‐2217 had lower lipophilicity and improved protein binding as compared to its congener RD4‐2024 and loviride. These results suggest that RD4‐2217, one of the TDA derivatives, is worth pursuing as a candidate drug for the treatment of HIV‐1 infections.
Breast Milk Is Not a Significant Source for Early Epstein‐Barr Virus or Human Herpesvirus 6 Infection in Infants: A Seroepidemiologic Study in 2 Endemic Areas of Human T‐Cell Lymphotropic Virus Type I in Japan
Authors: Kusuhara Koichi , Takabayashi Akira , Ueda Kohji , Hidaka Yasufumi , Minamishima Ikuko , Take Hiromichi , Fujioka Katsuyoshi , Imai Shosuke , Osato Toyoro 
Published: November 2013

AbstractIn order to evaluate the possibility of Epstein‐Barr virus (EBV) and human herpesvirus 6 (HHV‐6) transmission via breast milk, a total of 331 serum specimens collected from bottle‐fed and breast‐fed children and their mothers, in 2 endemic areas of human T‐cell lymphotropic virus type I (HTLV‐I) in Japan, were assayed for antibodies to EBV and HHV‐6. The seroprevalences of EBV and HHV‐6 were over 95% both in the mothers of bottle‐fed children and in those of breast‐fed children. The seroprevalence of EBV at 12–23 months of age was 54.5% (36/66) and 55.8% (24/43) in breast‐fed children and bottle‐fed children, respectively. The seroprevalence of HHV‐6 at 12–23 months of age was 90.9% (60/66) and 93.0% (40/43) in breast‐fed children and bottle‐fed children, respectively. No difference was observed between the seroprevalences of EBV and HHV‐6 in breast‐fed and bottle‐fed children at 12–23 months of age. Our seroepidemiologic data indicate that breast milk is not a significant source of early EBV or HHV‐6 infection in infancy.
Further Clonal Expansion of T Cells upon Rechallenge of Superantigen Staphylococcal Enterotoxin A
Authors: Aoki Yoshiyasu , Yoshikai Yasunobu 
Published: November 2013

AbstractSuperantigens are known to induce clonal anergy and/or deletion in reactive T cells peripherally. This study was undertaken to investigate the T‐cell status early after exposure to staphylococcal enterotoxin A (SEA) in vivo and in vitro. At the peak of clonal expansion following the administration of 5 μg SEA (i.e., 2 days after the injection), C57BL/6 mice were rechallenged with the same dose of SEA in vivo. The secondary stimulation augmented clonal expansion of the T cells bearing Vβ3 and Vβ11 in both CD4+ and CD8+ populations. In vitro restimulation of the spleen cells taken from the SEA‐primed mice also induced further expansion of the Vβ3+ T cells during 2 days of culturing, whereas without restimulation, a marked reduction of Vβ3+ T cells occurred. The spleen cells from the SEA‐primed mice were hyper‐reactive to in vitro restimulation with SEA as measured by 3H‐TdR uptake on day 1 of culturing, but augmented proliferation leveled off thereafter. By day 3, the values of 3H‐TdR uptake were less than 20% of those of the controls in which spleen cells from native mice were stimulated with SEA in vitro. These results suggest that T cells exposed to SEA in vivo are still capable of proliferating upon SEA rechallenge, but subsequently, the proliferation starts to wane.
Analysis of Cytokine Producing Activity of Intestinal Intraepithelial T Cells from TCR β‐Chain and 5‐Chain Mutant Mice
Authors: Kohyama Masako , Hachimura Satoshi , Nanno Masanobu , Ishikawa Hiromichi , Kaminogawa Shuichi 
Published: November 2013

AbstractIntestinal intraepithelial T cells (IELs) expressing either 78 TCR or αβ TCR have been proposed to play an important role in the regulation of intestinal epithelia by producing cytokines that directly influence the adjoining intestinal epithelial cell (IEC) functions. To illuminate this issue, we utilized TCR mutant mice to obtain γδ IELs, αβ IELs and mixed γδ and αβ IELs from corresponding αβ T‐cell‐deficient (β−/‐), γδ T‐cell‐deficient (δ−/‐) and wild‐type (WT) littermate mice. The production of IFN‐γ by these IELs as well as the mRNA for IFN‐γ, TGF‐α, TGF‐β1, TNF‐α and TNF‐β in these IELs, in conjunction with the effect of produced cytokines on the expression of class II MHC molecules by the in vitro cell line IEC‐6, were investigated. IFN‐γ and TGF‐α specific mRNA were detectable in all freshly isolated γδ, αβ and WT IELs. In addition to the IFN‐γ and TGF‐α mRNA, αβ and WT IELs that had been activated in culture plates coated with anti‐CD3 mAb contained mRNA for TGF‐β1 and TNF‐β proteins. In the cultured γδ IELs, however, the signals for IFN‐γ and TGF‐α transcripts were weak, and mRNA for the latter two cytokines was almost undetectable. Supernatants from in vitro culturing of αβ and WT IELs but not γδ IELs induced class II MHC gene expression in IEC‐6, whereas, in the presence of anti‐IFN‐γ mAb, the same culture supernatants failed to do so. In fact, the concentration of IFN‐γ in supernatants from αβ and WT IEL cultures was ten‐ to twentyfold higher than that in the supernatant from the γδ IEL culture. Finally, TGF‐α specific mRNA was not detectable in the γδ and αβ IELs even after in vitro activation. These results indicate that αβ IELs are superior to γδ IELs in the ability to produce IFN‐γ, TGF‐β1, TGF‐α and TNF‐β through TCR crosslinking primary in vitro stimulation.
Experimental Helicobacter pylori Infection in Association with Other Bacteria
Authors: Isogai Hiroshi , Isogai Emiko , Hayashi Shunji , Kimura Koichi , Kubota Tom , Fujii Nobuhiro , Oguma Keiji 
Published: November 2013

AbstractWe performed surgical treatment on normal ddY mice before Helicobacter pylori inoculation. The treatment was expected to obstruct bacterial flow out of the stomach and increase the chance of bacterial attachment to the gastric epithelium in mice. The bacterial challenge induced inflammation in the stomach. H. pylori was recovered from the stomach throughout the observation period. Lactobacilli and streptococci tended to relate to the increase in number of H. pylori recovered. Pretreatment with atropine was considered to confuse the gastric flora and affect the number of H. pylori recovered. These results suggested that a certain amount of time is necessary for H. pylori to contact with the gastric epithelium and that the composition of flora is important for the establishment of H. pylori infection.
Moraxella (Branhamella) catarrhalis Adherence to Human Bronchial and Oropharyngeal Cells: The Role of Adherence in Lower Respiratory Tract Infections
Authors: Rikitomi Naoto , Ahmed Kamruddin , Nagatake Tsuyoshi 
Published: November 2013

AbstractTo study the role of Moraxella (subgenus Branhamella) catarrhalis (B. catarrhalis) adherence to airway cells in lower respiratory tract infections, the in vitro attachments of B. catarrhalis to upper airway (oropharyngeal) and lower airway (bronchial) epithelial cells were compared. The adherence of 4 strains (1 nonfimbriated and 3 fimbriated) of B. catarrhalis to respiratory tract epithelial cells collected from 11 patients with chronic pulmonary disease (CPD) and 11 healthy individuals was evaluated. Both the fimbriated and nonfimbriated strains showed increased attachment to oropharyngeal cells in the CPD patients (mean ± SEM; 25.0 ± 3.2/cell; P < 0.01) when compared to the control subjects (12.1 ± 1.1/cell). On the average, the attachment to bronchial cells was 6.1 to 13.6 times greater per surface area (bacteria/μ2) than the attachment to oropharyngeal cells. The fimbriated strains tended to adhere in higher numbers to bronchial cells (19.0 ± 1.8/cell) than the nonfimbriated strain (8.7 ± 1.2/cell), although there was no difference between the CPD and control groups. In conclusion, the attachment of B. catarrhalis to oropharyngeal cells may be an enhancing factor for colonization in the upper respiratory tract in patients with CPD, and elevated adherence of the bacteria to bronchial cells may suggest pathogenic importance when mucociliary function is impaired.
Lambda‐Toxin of Clostridium perfringens Activates the Precursor of Epsilon‐Toxin by Releasing Its N‐ and C‐Terminal Peptides
Authors: Minami Junzaburo , Katayama Seiichi , Matsushita Osamu , Matsushita Chieko , Okabe Akinobu 
Published: November 2013

AbstractThe effect of γ‐toxin, a thermolysin‐like metalloprotease of Clostridium perfringens, on the inactive ε‐prototoxin produced by the same organism was examined. When the purified ε‐prototoxin was incubated with the purified γ‐toxin at 37 C for 2 hr, the 32.5‐kDa ε‐prototoxin was processed into a 30.5‐kDa polypeptide, as determined by SDS‐polyacrylamide gel electrophoresis. A mouse lethality test showed that the treatment activated the prototoxin: the 50% lethal doses (LD50) of the prototoxin with and without γ‐toxin treatment were 110 and 70,000 ng/kg of body weight, respectively. The lethal activity of the prototoxin activated by γ‐toxin was comparable to that with trypsin plus chymotrypsin and higher than that with trypsin alone: LD50 of the prototoxin treated with trypsin and trypsin plus chymotrypsin were 320 and 65 ng/kg of body weight, respectively. The ε‐toxin gene was cloned and sequenced. Determination of the N‐terminal amino acid sequence of each activated ε‐prototoxin revealed that γ‐toxin cleaved between the 10th and 11th amino acid residues from the N‐terminus of the prototoxin, while trypsin and trypsin plus chymotrypsin cleaved between the 13th and 14th amino acid residues. The molecular weight of each activated ε‐prototoxin was also determined by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry. The C‐terminus deduced from the molecular weight is located at the 23rd or 30th amino acid residue from the C‐terminus of the prototoxin, suggesting that removal of not only N‐terminal but also C‐terminal peptide is responsible for activation of the prototoxin.
Endogenous Tumor Necrosis Factor (TNF) α Mediates Neutrophil Accumulation at the Mid‐Phase of a Murine Model of Pseudomonas aeruginosa Pneumonia
Authors: Sonoda Fuminari , Oishi Kazunori , Iwagaki Akitaka , Matsumoto Keizo 
Published: November 2013

AbstractTo determine the role of endogenous tumor necrosis factor (TNF) α on neutrophil influx into the lungs in acute Pseudomonas aeruginosa pneumonia, we evaluated TNF α activity, inflammatory cell response and neutrophil chemotactic activity in the bronchoalveolar lavage fluids (BALFs) of P. aeruginosa‐infected mice. In the case of fatal pneumonia, the TNF α activity in the BALFs appeared within 3 hr, peaked at 6–12 hr and attenuated within 24 hr after intratracheal challenging, while no TNF α activity was detected in the plasma. The elevation of TNF α activity in the BALFs was closely associated with neutrophil accumulation. Mirroring the TNF α activity response and the influx of neutrophils into the murine airway, the number of neutrophils in the BALFs increased within 3 hr, peaked at 6–12 hr and remained elevated up to 24 hr after challenging. Neutralization of the TNF α activity in the BALFs with anti‐murine TNF antiserum decreased the level of neutrophil migration by BALF 45.0–49.7% at 6 hr and 49.3–54.2% at 12 hr, while the neutralizing antiserum had no effect on the level of neutrophil migration by BALFs at 3 and 24 hr. Furthermore, the intravenous administration of anti‐murine TNF antiserum 2 hr before challenging significantly inhibited neutrophil migration into the lungs of mice with sublethal pneumonia (P < 0.05; compared with mice receiving pre‐immune serum). These data suggest that intra‐alveolar TNF α plays an important role in causing lung neutrophil accumulation at the mid‐phase of murine P. aeruginosa pneumonia.
Intestinal Distribution and Intraluminal Localization of Orally Administered Clostridium butyricum in Rats
Authors: Sato Rumiko , Tanaka Mamoru 
Published: November 2013

AbstractClostridium butyricum has been used as a probiotic in animals and humans for years, however, its fate in the intestine has not been clarified yet. We investigated the intestinal fate of C. butyricum using a selective medium and a monoclonal antibody after orally administering C. butyricum spores to rats. The number of C. butyricum, both viable and dead cells, in the intestinal contents were counted by enzyme‐linked immunosorbent assay (ELISA) at various times after a single oral administration. The total viable number of C. butyricum was counted using a selective medium, and viable resting spores were selectively detected by treating the samples with ethanol. To investigate the intraluminal localization of the C. butyricum cells, frozen intestinal tracts were imprinted onto slides and stained with immunogold‐silver. Total viable spores exceeded the number of viable resting spores by more than 10‐fold from the proximal to middle of the small intestine 30 min after administration. Vegetative cells of C. butyricum were first detected in the distal small intestine after 2 hr, and vegetative growth was observed from the cecum to the colon 5 hr after administration. Dead vegetative cells were detected 9 hr after administration, and C. butyricum cells were not detected in the intestine after 3 days. The C. butyricum cells in the intestinal imprints were stained specifically by immunogold‐silver staining, and proliferative cells were observed in the cecum after 3 hr. These results suggest that the administered C. butyricum germinated in the upper small intestine, grew mainly from the distal small intestine to the colon and were excreted from the rat intestine within 3 days.
A Long‐Term Survey of Methicillin‐Resistant Staphylococcus aureus in the Oral Cavity of Children
Authors: Suzuki Junji , Komatsuzawa Hitoshi , Sugai Motoyuki , Suzuki Takako , Kozai Katsuyuki , Miyake Yoichiro , Suginaka Hidekazu , Nagasaka Nobuo 
Published: November 2013

AbstractMethicillin‐resistant Staphylococcus aureus (MRSA), an indigenous bacteria in healthy people, often causes nosocomial infections. If the host human becomes compromised, MRSA can cause a serious infection. The long‐term colonization of MRSA increases this risk. The purpose of this study was to demonstrate the incidence of S. aureus and MRSA colonization in the oral cavities of healthy children, and to examine the stability of identical strains of MRSA over a long‐term period. Fourteen children were examined in two stages (first stage: 1987–88, second stage: 1992–93). Five of the 14 children were negative for S. aureus in both stages, seven children were positive in both stages and two children were positive in only the second stage. The children who were colonized with S. aureus in the first stage always harbored the bacteria in the second stage. Of the seven children that were positive for S. aureus in both stages, three persisted in carrying MRSA. We compared two MRSA strains isolated from the same children in both stages by coagulase typing, antibiogram typing and DNA fingerprinting. In two children, the strains showed the same coagulase types, similar antibiograms and similar DNA fragment profiles. These data strongly suggest that identical strains of MRSA persisted in the oral cavities for more than five years, and that the oral cavity can serve as a reservoir for MRSA with the potential to cause nosocomial infections.
Different Susceptibility of Three Clinically Isolated Strains of Cryptococcus neoformans to the Fungicidal Effects of Reactive Nitrogen and Oxygen Intermediates: Possible Relationships with Virulence
Authors: Xie Qifeng , Kawakami Kazuyoshi , Kudeken Norifumi , Zhang Tiantuo , Qureshi Mahboob Hossain , Saito Atsushi 
Published: November 2013

AbstractWe investigated the susceptibility of three clinically isolated strains of Cryptococcus neoformans with different virulences to reactive nitrogen and oxygen intermediates (RNI and ROI, respectively), representing two important mediators of macrophage microbicidal activity. All mice infected with the highly virulent strain of C. neoformans, YC‐11, died within 3 to 6 weeks because of rapid multiplication of the organism in the lungs and dissemination to the brain. In contrast, a weakly virulent strain, YC‐13, was almost completely eradicated from the lungs and did not disseminate to the brain, leading to survival of all infected animals during the period of observation (15 weeks). The virulence of the third strain, YC‐5, was intermediate between the other two strains. To examine the susceptibility of C. neoformans to the fungicidal effect of nitric oxide (NO) and superoxide anions (O2‐), the organisms were exposed to these oxidants, which were chemically generated in a cell‐free system. Interestingly, the number of live YC‐13 yeast cells was markedly reduced after exposure to NO and O2−. In contrast, YC‐11 was almost completely resistant to the killing effect of these oxidants. YC‐5 showed an intermediate susceptibility. Our results demonstrate that the resistance of C. neoformans to the fungicidal effects of RNI and ROI is related to virulence, and suggest that the resistance to nitrogen‐ and oxygen‐derived oxidants may be one of the factors to determine the outcome of infection with C. neoformans.
Fc Receptor‐Mediated Phagocytosis, Superoxide Production and Calcium Signaling of β2Integrin‐Deficient Bovine Neutrophils
Authors: Nagahata Hajime , Sawada Chikari , Higuchi Hidetoshi , Teraoka Hiroki , Yamaguchi Mamoru 
Published: November 2013

AbstractFc receptor for immunoglobulin G‐mediated phagocytosis, superoxide production and intracellular calcium ([Ca2+]i) signaling of complement receptor type 3 (CR3)‐deficient neutrophils from a heifer with leukocyte adhesion deficiency (BLAD) were compared to those of control heifers. The mean phagocytic activity of IgG‐coated yeasts and aggregated bovine IgG (Agg‐IgG)‐induced superoxide production of CR3‐deficient neutrophils were 10% and 77.9%, respectively, of those of control neutrophils. The [Ca2+]isignals in CR3‐deficient neutrophils stimulated with Agg‐IgG or concanavalin A were different with mean peak [Ca2+]iconcentrations of 78% and 41.9%, respectively, of those of control neutrophils. These findings suggest that Fc receptor‐mediated neutrophil functions are closely dependent on the presence of CR3 (CD11b/CD18) on the neutrophil cell surfaces.