bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ultrastructural Study on the Adherence of Branhamella catarrhalis to Oropharyngeal Epithelial Cell
Authors: Ahmed Kamruddin , Rikitomi Naoto , Nagatake Tsuyoshi , Matsumoto Keizo 
Published: November 2013

AbstractIn the present study, it was observed that Branhamella catarrhalis adhere to the microplicae of the oropharyngeal epithelial cells. Both long and short microplicae patterns are present on the surface of oropharyngeal epithelial cells and the adherence ability of fimbriated Branhamella catarrhalis also varies according to the microplicae pattern. It was found that Branhamella catarrhalis attached more to one surface of the epithelial cell than to the other, suggesting that the presence of receptors are more on one surface than on the other. Branhamella catarrhalis did not attach to the mucus layer but directly to the epithelial cell surface. Ruthenium red staining specimen showed that Branhamella catarrhalis attached to a granular ruthenium red positive layer on the microplicae and also to a ruthenium red positive component, external to the unit membrane of the epithelial cell membrane.
Immunoglobulin G Fc Fragment‐Binding Proteins in Mycoplasma salivarium Cells
Authors: Sawa Yoshihiko , Watanabe Tsuguo , Shibata Ken‐ichiro 
Published: November 2013

AbstractCell proteins of Mycoplasma salivarium were separated by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis, transferred to membranes, then examined for reactivity with human IgG molecules, the Fc fragment of human IgG, and concanavalin A (ConA). Multiple protein bands bound IgG, and most of them also bound ConA. One (corresponding to a molecular mass of 90 kDa) of the IgG‐ and ConA‐binding bands intensely interacted with the Fc fragment of IgG. The reactivity of proteins eluted from the band with the Fc fragment, tested by dot‐blotting and ELISA, was inhibited (90%) by pre‐incubation with IgG and to a lesser extent (50%), with IgM. Thus, M. salivarium contained a cellular protein with a molecular mass of 90 kDa, that bound the Fc fragment of human IgG.
Focus Formation by the Human Immunodeficiency Virus (HIV) in the Immobilized MT‐4 Cell Culture and Its Application to the Evaluation of Anti‐HIV Agents
Authors: Nakane Masayuki , Kawai Akihiko 
Published: November 2013

AbstractImmunofluorescence studies were performed on the infection of monolayer cultures of immobilized MT‐4 cells with human immunodeficiency virus type 1 (HIV‐1). By using the anti‐viral p24 monoclonal antibody, we could observe formation of foci of p24 antigen‐positive cells within 3 to 4 days when the infection was initiated with a relatively small amount of the virus. Frequency of the focus formation was in proportion to the dose of input virus (ranging from 0.001 to 0.1 PFU/cell), which allowed us to apply this phenomenon to the assay of anti‐HIV agents as well as to the estimation of relative infectivity of the virus stocks. When antiviral agents were added to the infected cultures, number of foci as well as the size of each focus was reduced in a concentration‐dependent manner. The dose required for reducing the number of foci by 50% was calculated to be 6 ng/ml and 8 ng/ml for tunicamycin (TM) and azidothymidine (AZT), respectively. These values are comparable to those obtained by other current assay methods. In addition, focus reduction assay is also useful in searching for such antiviral agents that would inhibit or block the early step of viral replication cycle.
An Enzyme‐Linked Immunosorbent Assay Using a Chaotropic Agent (Sodium Thiocyanate) for Serotype Specific Reaction between Crude Dengue Viral Antigen and Anti‐Dengue Mouse Antibody
Authors: Nawa Masaru 
Published: November 2013

AbstractAn enzyme‐linked immunosorbent assay (ELISA) has been developed to detect serotype specific reaction between crude dengue viral antigen and anti‐dengue mouse hyperimmunized antibody under the stringent condition in the presence of a Chaotropic agent, sodium thiocyanate (NaSCN), in the reaction mixture of antigen and antibody. Rapidly sedimenting hemagglutinin (RHA) derived from type 2 dengue virus‐infected mosquito cell culture fluid reacted to the antibody for both type 2 and type 3 dengue viruses in the ELISA. In contrast, its reactivity was reduced after the addition of NaSCN in the ELISA. Soluble complement‐fixing antigen (SCF) derived from type 2 dengue virus‐infected mosquito cell culture fluid reacted serotype specifically to anti‐dengue type 2 antibody, and was relatively stable for the NaSCN treatment in the ELISA. Anti‐type 2 RHA mouse antibody reacted to both type 1 and type 2 dengue viral antigens and its reactivity was reduced after the addition of NaSCN in the ELISA. Anti‐type 2 SCF antibody reacted serotype specifically to type 2 dengue viral antigen with and without NaSCN in the ELISA.
Antibody Response to Oral Streptococci in Behçet's Disease
Authors: Yokota Kenji , Hayashi Syunji , Fujii Nobuhiro , Yoshikawa Kouji , Kotake Satoshi , Isogai Emiko , Ohno Shigeaki , Araki Yoshio , Oguma Keiji 
Published: November 2013

AbstractThe serum antibody titers against oral streptococci were studied by enzyme‐linked immunosorbent assay (ELISA) both in patients with Behçet's disease (BD) and control groups. The patients with BD showed significantly higher antibody titers to S. sanguis strains 113‐20, 114‐23, and 118‐1 which were isolated from patients with BD, in comparison with control groups. Also, the reactions of hightitered sera to the crude cell wall and soluble (or membrane) fractions of the 113‐20 strain were observed by western blot test. The sera of the patients with BD demonstrated strong bands of approximately 36 kDa, 82 kDa, and 87 kDa in the crude cell wall fractions, and many bands of 80 kDa to 150 kDa in the membrane fractions, indicating that these proteins are the ones leading the high antibody titers to this bacterium in the sera of patients with BD.
A Prospective Study on Correlation between the Decrease in Anti‐P17 Antibody Level and Progression to AIDS in Asymptomatic Carriers of HIV
Authors: Choudhury Ahmed Murtaza , Yamada Osamu , Wakamiya Nobutaka , Kurimura Takashi 
Published: November 2013

AbstractAs the majority of human immunodeficiency virus (HIV) carriers are in asymptomatic stage for a long period of time, it is important to investigate the factors or surrogate markers for conversion from asymptomatic to symptomatic stage. Our study is designed to evaluate the relationship among virus isolation rate, anti‐p17 antibody status and progression to AIDS. We studied anti‐p17 antibody status along with virus isolation in 56 asymptomatic carriers and 46 AIDS cases. Progression to AIDS was markedly associated with high rate of virus isolation and loss of anti‐p17 antibody. In order to know the meaning of loss of anti‐p17 antibody during the clinical course, 15 anti‐p17 antibody positive and 16 anti‐p17 antibody negative cases were followed up prospectively for the development of AIDS. None of the anti‐p17 antibody positive cases developed AIDS while 6 out of 16 anti‐p17 negative cases developed AIDS during observation period (P<0.05). Progression to AIDS was associated with loss of anti‐p17 antibody. Identification of cases losing anti‐p17 antibody in peripheral blood during asymptomatic period may help high‐risk group who are in need of chemoprophylaxis. Moreover, study of anti‐p17 antibody may be helpful in designing vaccine in future if it works as a neutralizing antibody to HIV in vivo.
Effects of Gamma‐Interferon and FK506 on Resting B Cell Proliferation of New Zealand Black/White F1 Mice
Authors: Minoda Masahide , Ohno Motoki , Tomioka Yoshiki , Hamada Kinya , Yamazoe Yasushi , Higashikawa Mitsuhiro , Sugishima Hitoshi , Higashitani Sumihiko , Funauchi Masanori , Horiuchi Atsushi 
Published: November 2013

AbstractWe examined the effects of gamma‐interferon (γ‐IFN) and the new immunosuppressant FK506 on resting B cell proliferation of New Zealand black/white F1 hybrid (B/W F1) mice, an animal model of human systemic lupus erythematosus (SLE). γ‐IFN and FK506 inhibited in a dose‐dependent manner both B cell proliferation and autoantibody production of resting B cells respectively. There was a synergistic interaction between γ‐IFN and FK506 in their inhibition and they did not exhibit cell cytotoxicity. This in vitro synergism of γ‐IFN and FK506 may have clinical application in that low doses of γ‐IFN and FK506 combinations may be effective to correct polyclonal B cell activation of patients with SLE.
Involvement of Calcium in Germination of Coat‐Modified Spores of Bacillus cereus T
Authors: Shibata Hirofumi , Miyoshi Shigekatsu , Osato Tomohiko , Tani Isamu , Hashimoto Tadayo 
Published: November 2013

AbstractThe effect of calcium on germination of coat‐modified Bacillus cereus T spores was investigated. Coat‐modified spores produced either by chemical extraction (SDS‐DTT‐treated spores) or by mutagenesis (10LD mutant spores) were unable to germinate in response to inosine. While SDS‐DTT‐treated spores could germinate slowly in the presence of l‐alanine, 10LD mutant spores could not germinate at all. The lost or reduced germinability of coat‐modified spores was restored when exogenous Ca2+ was supplemented to the germination media. The calcium requirement of coat‐modified spores for germination was fairly specific. The simultaneous presence of germinant with Ca2+ was also required for germination of coat‐modified spores. The optimal recovery of germinability was observed in the presence of 1.0 mM of calcium acetate. The calcium requirement itself was remarkably diminished under the condition in which l‐alanine and a certain purine nucleoside analog, adenosine or inosine, coexisted. The lost or diminished germinability observed in SDS‐DTT‐treated spores or 10LD mutant spores may be attributed to the loss of calcium associated with the spore integuments.
IgG Subclass Alterations in Adult Asthma
Authors: Outschoorn Ingrid M. , Natta Clayton L. 
Published: November 2013

AbstractImmunoglobulin levels were measured in serum samples of 12 black adult non‐smoking asthmatic patients, 11 females and 1 male, and compared with 15 age‐, sex‐matched normal controls. Their total IgG, IgA and IgM levels were within the normal range. However, on quantitation of subclasses, IgG1 levels were significantly above normal, while IgG2 and IgG3 levels were significantly lower than those of controls. No significant differences were found between the two groups when IgG4 levels were compared. These studies as well as those of others suggest that immunoglobulin administration, particularly of individual subclasses, might prove to be a beneficial addition in the management of this condition.
DNA from Bacteria, but Not from Vertebrates, Induces Interferons, Activates Natural Killer Cells and Inhibits Tumor Growth
Authors: Yamamoto Saburo , Yamamoto Toshiko , Shimada Shizuo , Kuramoto Etsuro , Yano Osamu , Kataoka Tetsuro , Tokunaga Tohru 
Published: November 2013

AbstractThe nucleic acid fraction from cells of 6 species of bacterium and 2 kinds of vertebrate, calf and salmon, was extracted and purified by the same procedures as described previously. When the spleen cells from BALB/c mice were incubated with the nucleic acid fraction from either of the bacteria, natural killer (NK) activity of the cells was remarkably elevated and the cells produced factors to activate macrophages and to inhibit viral growth. It was shown that the factor to activate macrophages was interferon (IFN)‐gamma and that to inhibit viral growth was IFN‐alpha/beta. On the other hand, the nucleic acid fraction from either of the vertebrate cells did not show such activities. Pretreatment of the bacterial nucleic acid fraction with DNase, but not with RNase, abrogated completely the biological activities. The activities of the bacterial nucleic acid were not influenced by the presence of polymyxin B, an inhibitor of lipopolysaccharide (LPS), and the spleen cells from not only BALB/c mice but also LPS‐insensitive C3H/HeJ mice were activated, indicating that the activities of the fraction were not ascribed to LPS contaminated possibly into the fraction, but to DNA itself. Intralesional injection with the bacterial DNA fraction caused regression of mouse IMC tumors, but the injection with the vertebrate DNA fraction did not. These findings prompted us to examine the biological activities of DNA samples from a variety of animals and plants, which were provided from other laboratories or purchased from manufacturers. All of the DNA samples from cells of 5 kinds of bacterium, 2 of virus and 4 of invertebrate augmented NK activity and induced IFN, more or less, in mouse spleen calls, while the DNA from 10 kinds of vertebrate, including 3 of fish and 5 of mammal, showed no such activities. The DNA from 2 species of plants, were also inactive. Possible mechanisms to explain the different biological activities of DNA from different cell sources were discussed based on our previous finding that the particular palindromic sequences with a G‐C motif (s) are required for induction of IFNs and activation of NK cells with synthetic 30‐mer oligonucleotides.
Seroepidemiological Survey for Antibody to Borrelia burgdorferi in Cows
Authors: Isogai Hiroshi , Isogai Emiko , Masuzawa Toshiyuki , Yanagihara Yasutake , Matsubara Mitsunori , Shimanuki Masamichi , Seta Toshiyuki , Fukai Kenji , Kurosawa Nobumichi , Enokidani Masafumi , Nakamura Toshio , Tajima Motoshi , Takahashi Kiyoshi , Takahashi Koichi , Fujii Nobuhiro 
Published: November 2013

AbstractAntibody to Borrelia burgdorferi was examined in 380 healthy and 38 clinical cases of cows from Hokkaido and Shizuoka in Japan. In healthy animals, IgG and IgM antibody to B. burgdorferi HO14 strain were found in 44 cows (14.6%) and 24 cows (8.0%) from Hokkaido. In contrast, antibody‐positive case was not observed except for only 1 case which was IgM positive (1/79: 1.3%) in cows from Shizuoka. Mean antibody levels of healthy animals in Hokkaido and Shizuoka were 0.651 and 0.263 (IgG antibody to HO14 strain), 0.642 and 0.169 (IgG to HP3 strain), 0.613 and 0.367 (IgM to HO14 strain) and 0.582 and 0.286 (IgM to HP3 strain). The differences of the antibody levels between cows from Hokkaido and Shizuoka were significant. Seasonal difference was found in seropositive cows from Hokkaido. The rate of seropositive cows was high in summer (23.4% in June and 11.8% in July) but low in winter (0% in January and February). The pattern was discussed to be associated with activation of ticks. One of 4 cows with arthritis showed significantly higher IgG antibody level than that of healthy cows and cows with some disease, although the serum was collected from Shizuoka where antibody‐positive animals for B. burgdorferi were rare among healthy cows. This high IgG antibody may suggest that the arthritis of such cows was caused by infection with B. burgdorferi. Two of 7 cows with unclassified abortion showed positive antibody reaction in Hokkaido. These cases, however, may not be related to the B. burgdorferi infection because the positive rate was similar to those of healthy cows in the same season.
Nucleotide Sequence Analysis of the Class G Tetracycline Resistance Determinant from Vibrio anguillarum
Authors: Zhao Jiang , Aoki Takashi 
Published: November 2013

AbstractThe nucleotide sequence of the class G tetracycline resistance determinant previously isolated from Vibrio anguillarum has been determined. Two open reading frames of divergent polarity were identified. A resistance gene (tet A) encodes a protein of 393 amino acid residues (deduced molecular mass of 40.9 kDa), and a repressor gene (tet R) encodes a protein consisting of 210 amino acids with a calculated molecular mass of 23.6 kDa. Based on the deduced amino acid sequences, the proteins of tet A(G) and tet R(G) are about 60% homologous with those of RP1/Tn1721 (class A) and pSC101/pBR322 (class C), and about 50% homologous with Tn10 (class B). The relationship of the tet (G) sequence to five known tetracycline resistance determinants (class A to E) is discussed.
Impaired NK Response of Cancer Patients to IFN‐α but Not to IL‐2: Correlation with Serum Immunosuppressive Acidic Protein (IAP) and Role of Suppressor Macrophage
Authors: Aso Hisashi , Tamura Keiji , Yoshie Osamu , Nakamura Takehiko , Kikuchi Syuu , Ishida Nakao 
Published: November 2013

AbstractIn vitro NK responses of cancer patients (N=21) to rIFN‐αA and rIL‐2 were examined. The serum concentration of IAP (immunosuppressive acidic protein) was determined in parallel. Five out of seven patients whose serum IAP contents were within the normal range (270 μg/ml to 470 μg/ml), had their NK activities significantly augmented by rIFN‐αA and rIL‐2. On the other hand, NK cells from ten out of fifteen patients whose serum IAP concentrations were 650 μg/ml or more, were not activated by rIFN‐αA. NK cells of these fifteen patients yet were capable of responding to rIL‐2. NK cells from cancer patients, however, became responsive to rIFN‐αA by either removal of adherent cells or treatment with indomethacin. Therefore, macrophages in PBMC of cancer patients with high serum IAP levels seem to selectively suppress NK response to rIFN‐αA by an indomethacin‐sensitive mechanisms. It was further shown that PGE2 was not the mediator of this suppression.
Sensitivity of Polymerase Chain Reaction Assay for Rickettsia tsutsugamushi in Patients' Blood Samples
Authors: Murai Koichi , Tachibana Nobuyoshi , Okayama Akihiko , Shishime Eiichi , Tsuda Kazunori , Oshikawa Tatsumi 
Published: November 2013

AbstractWe developed a nested polymerase chain reaction (PCR) method to detect Rickettsia tsutsugamushi (R. tsutsugamushi) DNA and determined its sensitivity. Primers were selected from the DNA sequence of the 58‐kDa group‐specific antigen gene of the Karp strain. The target sequence of rickettsial DNA was detectable as the band corresponding to 88 bp in 1.0 μg of the DNA extracted from BS‐C‐1 cells infected with R. tsutsugamushi. Rickettsia‐specific bands were observed not only for the homologous Karp strain, but also for four heterologous strains: two other reference strains (Gilliam and Kato) and two prototype strains prevalent in Miyazaki district (Irie and Hirano). The minimum copy number detectable by this method was estimated to be five rickettsiae. All of nine peripheral blood mononuclear cell samples from patients with tsutsugamushi disease who were seen 2‐11 days after disease onset tested positive for rickettsial DNA. The PCR assay method presented here could be a specific diagnostic tool for tsutsugamushi disease, especially in its early acute stage.
Enteric Adenovirus Type 41 Isolates: Cloning, Physical Maps and Diversity in Restriction Enzyme Cleavage Pattern
Authors: Yamashita Yumi , Hotsubo Tomoyuki , Nakata Shuji , Yamaguchi Taihei , Sanekata Takeshi , Chiba Shuzo , Homma Morio , Fujinaga Kei 
Published: November 2013

AbstractAdenovirus (Ad) type 40 and 41 DNAs were directly extracted from stool specimens of children with gastroenteritis. Two new strains of Ad41, Sanekata and Ehime strain, were cloned and their restriction maps were constructed. The left terminal end of the cloned Ad41 genome, EcoRI‐E fragment of the Sanekata strain and EcoRI‐F fragment of the Ehime strain, had transforming ability in rat 3Y1 cells. Only one of the 35 isolates of Ad40 tested showed a different restriction profile, while three different restriction profiles were found in DNAs from Ad41 isolates.
Polymorphism of Crystals of Salmonella minnesota Re and Ra Lipopolysaccharides
Authors: Kato Nobuo , Ohta Michio , Kido Nobuo , Arakawa Yoshichika , Sugiyama Tsuyoshi , Naito Setsuko , Ito Hideo 
Published: November 2013

AbstractSalmonella minnesota Re and Ra lipopolysaccharides (LPSs) formed three‐dimensional crystals when they were precipitated by the addition of 2 volumes of 95% ethanol containing 375 mM MgCl2 and incubated in 70% ethanol containing 250 mM MgCl2 at 4 C. Besides typical shapes of crystals, hexagonal plates and solid columns, which were already reported (J. Bacteriol. 172: 1516–1528 (1990)), the LPSs thus treated formed crystals possessing various shapes such as square or rectangular plate, lozenge plate, discoid, and truncated hexangular pyramid forms. Electron diffraction patterns from all these crystals except square or rectangular plate crystals obtained by electron irradiation from the direction perpendicular to the basal plane were essentially the same as those from hexagonal plate crystals, indicating that they consist of hexagonal lattices with the lattice constant of 4.62 Å. From these results as well as the results of electron microscopic observations of these crystals, it was concluded that all these crystals except square or rectangular plate crystals are composed of hexagonal plate sheets as the basic structural units. Square or rectangular crystals were assumed to correspond to the {1011} planes of solid hexagonal column crystals.
Enhancement by Recombinant Human Interleukin 2 of Host Resistance to Toxoplasma gondii Infection in Pregnant Mice
Authors: Shirahata Toshikazu , Muroya Naoyoshi , Ohta Chikako , Goto Hitoshi , Nakane Akio 
Published: November 2013

AbstractThe lymphokine production by pregnant mice infected with a lethal dose of Toxoplasma gondii was evaluated in comparison with that by virgin mice infected with a sublethal dose of this protozoan parasite. Splenocytes taken from mice before and on the day after infection produced considerable amounts of IL‐2 in response to concanavalin A (Con A) stimulation, but the titers rapidly declined in both pregnant and virgin mice as infection progressed. A trace amount or undetectable level of IL‐2 was produced by splenocytes from acutely infected mice when stimulated with Toxoplasma lysate antigen (TLA). In contrast to the kinetics of IL‐2 production, the levels of IFN‐γ produced by splenocytes cultured with Con A or TLA increased steadily in the later stage of infection in both pregnant and virgin mice. Thus, the response to Con A or TLA of splenocytes to produce IL‐2 and IFN‐γ differed strikingly in acute toxoplasmosis in mice. The administration of rHuIL‐2 resulted in a significant decrease in the mortality of pregnant mice infected with a lethal dose of Toxoplasma. The combination of rHuIL‐2 and rMuIFN‐γ increased the survival rate slightly but not significantly compared with pregnant mice receiving either rHuIL‐2 or rMuIFN‐γ. Moreover, exogenously administered rHuIL‐2 enhanced the production of both IFN‐α and IFN‐γ in the bloodstreams of pregnant mice, in accordance with the decreased mortality. These results indicate that IL‐2 may play a significant role in modulating the host defense against Toxoplasma infection in pregnant mice.
Neutrophil Response to Nontypable Haemophilus influenzae in Respiratory Infections
Authors: Ahmed Kamruddin , Ichinose Akitoyo , Dai Tran Cong , Takahashi Atsushi , Utsunomiya Yoshiaki , Kawakami Kenji , Nagatake Tsuyoshi , Matsumoto Keizo 
Published: November 2013

AbstractSputa from patients with respiratory infections by nontypable Haemophilus influenzae (H. influenzae) were investigated by electron microscopy. The cell wall of H. influenzae appeared wavy and nonwavy. In the cell wall the peptidoglycan layer was ill‐defined. These patients had adequate IgG response in the serum against H. influenzae. However neither capsule nor fimbriae were found. Different stages of phagocytosis and destruction of the bacteria by polymorphonuclear neutrophils (PMN) were observed. PMNs were also found to phagocytose the debris. Evidences were found that the debris is formed mainly by the destruction of polymorphonuclear neutrophil. Extracellular lysosomes were also observed, which may have a role in destruction of both bacteria and host tissue. It was concluded that nontypable H. influenzae are nonfimbriated and noncapsulated during infection. Debris are the end product of PMN destruction, and phagocytosis of debris by PMNs has a role in the pathogenesis of chronic respiratory diseases.