bluuebunny/crossref_metadata_2025 huggingface dataset:
Intrinsic Penicillin Resistance in Penicillinase‐Producing Neisseria gonorrhoeae Strains
Authors: Ikeda Fumiaki
Published: November 2013

The minimal inhibitory concentrations (MICs) of ampicillin for fifty strains of beta‐lactamase‐producing Neisseria gonorrhoeae (PPNG) isolated in Japan ranged from 1.56 to 200 μg/ml, and all the strains harbored a 4.5 megadalton plasmid. These strains were classified into two groups: dicloxacillin‐susceptible (28°00) and ‐resistant group (72°00). A linear correlation was found in the dicloxacillin‐susceptible strains between their beta‐lactamase activity and the susceptibility to ampicillin. but not in the dicloxacillin‐resistant strains. This suggests that the high ampicillin resistance in PPNG is due not only to acquiring the beta‐lactamase producing plasmid. but also to some intrinsic resistance of the strains. To investigate a cause of the high ampicillin resistance, the beta‐lactamase‐producing plasmid, pTMS1, was transferred by conjugation to a penicillin‐susceptible gonococcal strain as well as to its isogenic multiply antibiotic‐resistant transformants, and the susceptibility of the transconjugants to ampicillin was determined. Acquisition of pTMS1 by a penicillin‐susceptible strain resulted in a 32‐fold increase in resistance to ampicillin, whereas the increase was 128‐fold for its isogenic strains which contain some chromosomal mutations. These results suggest that reduced permeability of the outer membrane to ampicillin underlies the high ampicillin resistance of PPNG.
Proposals of Sphingomonas paucimobilis gen. nov. and comb. nov., Sphingomonas parapaucimobilis sp. nov., Sphingomonas yanoikuyae sp. nov., Sphingomonas adhaesiva sp. nov., Sphingomonas capsulata comb, nov., and Two Genospecies of the Genus Sphingomonas
Authors: Yabuuchi Eiko , Yano Ikuya , Oyaizu Hiroshi , Hashimoto Yasuhiro , Ezaki Takayuki , Yamamoto Hiroyuki
Published: November 2013

Based on the partial nucleotide sequence analysis of 16S ribosomal ribonucleic acid (rRNA), presence of unique sphingoglycolipids in cellular lipid, and the major type of ubiquinone (Q10), we propose Sphingomonas gen. nov. with the type species Sphingomonas paucimobilis (Holmes et al, 1977) comb. nov. From the homology values of deoxyribonucleic acid‐deoxyribonucleic acid hybridization and the phenotypic characteristics, three new species, Sphingomonas parapaucimobilis, Sphingomonas yanoikuyae, Sphingomonas adhaesiva, and one new combination, Sphingomonas capsulata, are described. S. par apaucimobilis JCM 7510 (=GIFU 11387), S. yanoikuyae JCM 7371 (=GIFU 9882), and S. adhaesiva JCM 7370 (=GIFU 11458) are designated as the type strains of the three new species. Emended description of the type strain of S. capsulata is presented.
Chemical Characterization of Lipopolysaccharides from Proteus Strains Used in Weil‐Felix Test
Authors: Mizushiri Sakae , Amano Ken‐Ichi , Fujii Shumsaku , Fukushi Kazue , Watanabe Mitsuru
Published: November 2013

The lipopolysaccharides (LPS) extracted from Proteus strains OX2, OX19, and OXK used as antigens in the Weil‐Felix test, were characterized by chemical analysis and SDS‐polyacrylamide gel electrophoresis (SDS‐PAGE). To separate the O‐polysaccharide, core‐oligosaccharide, and lipid A moieties, each LPS was treated with 2% acetic acid, centrifuged, and applied to Sephadex G‐50 column. The core‐oligosaccharides contained L‐glycero‐D‐mannoheptose, D‐glycero‐D‐man‐noheptose, glucose (Glc), galactose, 3‐deoxy‐D‐mannooctulosonic acid, uronic acid, phosphate, glucosamine (GlcN), and galactosamine (GalN). The lipid A preparations contained GlcN, GlcN‐phosphate, and three fatty acids (myristic, palmitic, and β‐hydroxymyristic acids). However, the O‐polysaccharides of OX2‐ and OXK‐LPS had different chemical compositions which consisted of Glc, GlcN, and quinovosamine, and Glc, uronic acid, and GalN, respectively, while OX19‐LPS seemed to lack O‐polysaccharide.
Immunological Characterization of Lipopolysaccharides from Proteus Strains Used in Weil‐Felix Test and Reactivity with Patient Sera of Tsutsugamushi Diseases
Authors: Amano Ken‐Ichi , Mizushiri Sakae , Fujii Shunsaku , Fukushi Kazue , Suto Tsunehisa
Published: November 2013

Immunological analyses of lipopolysaccharides (LPS) isolated from Proteus strains OX2, OX19, and OXK used as antigens of Weil‐Felix (WF) test, were performed by quantitative agglutination, enzyme‐linked immunosorbent assay (ELISA), and immunoblotting. Antisera against LPS and whole cells (WC) of the three Proteus strains reacted with homologous LPS but not with heterologous LPS. and the reaction was inhibited by the O‐polysaccharide fraction isolated from the homologous LPS except OX19‐LPS. which lacked O‐polysaccharide moiety. The immunological data support the findings that the O‐polysaccharide moieties of LPS from OX2 and OXK strains possess different chemical composition (Mizushiri, Amano, Fujii, Fukushi, and Watanabe, Microbiol. Immunol. 34: 121 133, 1990). Antisera against Proteus strains reacted weakly with WC of Rickettsia prowazekii, Rickettsia typhi, and Rickettsia tsutsugamushi. Antisera from patients with tsutsugamushi disease reacted with OXK‐WC by WF test when the sera were obtained 13 days after onset of fever. The immunoperoxidase (IP) test titers of these antisera began to rise 6 days after the onset of fever. By ELISA tests these antisera reacted with OXK‐WC and OXK‐LPS independently of the titers of WF or IP tests.
Cytotoxic Component(s) of Klebsiella oxytoca on HEp‐2 Cells
Authors: Higaki Megumu , Chida Toshio , Takano Hideko , Nakaya Rintaro
Published: November 2013

A cytotoxic component(s) was detected in culture filtrates of Klebsiella oxytoca isolated from patients with antibiotic‐associated hemorrhagic colitis. Eleven of 12 isolates exhibited cytotoxicity on HEp‐2 cells. The cytotoxic activity of K. oxytoca strain MH43‐1 was stable for the treatment of 60 C for 30 min, but inactivated by the treatment of 100 C for 15 min. This cytotoxicity was not destroyed by the treatment with trypsin or pronase, and the component was filtrable through a membrane filter which cut off molecular weight 5,000.
Study of Seroconversion of Antibody to Human T‐Cell Lymphotropic Virus Type‐I in Children of Okinawa, Japan
Authors: Kajiyama Wataru , Kashiwagi Seizaburo , Hayashi Jum , Nomura Hideyuki , Ikematsu Hideyuki , Noguchi Akinori , Nakashima Kouya , Okochi Kazuo
Published: November 2013

From 1983 to 1986, 1,813 children in nursery schools in Ishigaki Island and 1,228 children under 15 years old in the rural area in the Yaeyama District of Okinawa, Japan. were tested for anti‐HTLV‐I; 18 children (1.0%) in Ishigaki Island and 39 children (3.2%) in the rural area were positive. In order to survey when anti‐HTLV‐I developed in these children, their older serum samples were investigated retrospectively for 1 to 5 years. Two cases of seroconversion from anti‐HTLV‐I negative to positive were found between the age of 2 and 4, and there were no cases of seroconversion over 4 years old. Among the children who suffered maternal transmission of HTLV‐I and developed anti‐HTLV‐I before the age of 15, about 80% of the children were considered to have anti‐HTLV‐I before the age of 2, and the remainder developed anti‐HTLV‐I before the age of 4.
Forssman Antigen Expressed on Lymph Node Cells of MRL/MpJ‐lpr/lpr Mice Is of a Glycoprotein Nature
Authors: Kijimoto‐Ochiai Shigeko , Tashiro Akiko , Katagiri Yohko U. , Hatae Taiko , Kobayashi Seiichi , Okuyama Harue
Published: November 2013

This paper reports the nature of abnormally expressed Forssman (F) antigen in the lymph node cells of MRL/MpJ‐lpr/lpr, autoimmune mice, and also reports its autoantibody in sera. By acetylation study of the F antigen with [14C] acetic anhydride, we concluded that the F antigen was not a glycolipid but a glycoprotein. Several bands of F‐active glycoproteins were identified on a nitrocellulose sheet after purification by an anti‐F antibody affinity column. Hemolysis of SRBC by some sera from MRL/MpJ/lpr/lpr was inhibited by purified F glycoprotein and also by F glycolipid. The antibody in the serum, however, seemed to be more specific for F glycoproteins than F glycolipid, but the opposite was the case for rabbit anti‐F glycolipid antibody. No significant difference of the SRBC hemolysis levels was observed between the sera from MRL/MpJ‐lpr/lpr and its congenic MRL/MpJ‐+/ + mice.
Enhancement of DTH Response by Cholera Toxin B Subunit Inoculated Intranasally Together with Influenza HA Vaccine
Authors: Kikuta Kiyoshi , Kurata Hideki , Nagamine Takashi , Aizawa Chikara , Ueno Yoshio , Kurata Takeshi , Tamura Shin‐Ichi
Published: November 2013

The effects of B subunit of cholera toxin (CTB) on delayed‐type hypersensitivity (DTH) response to influenza vaccine derived from influenza virus A/PR/ 8/34 (PR‐8, H1N1) virus were investigated in B10 mice that were immunized intranasally with both influenza vaccine and CTB. The result showed [1] that intranasal inoculation of this combination augmented DTH response to influenza vaccine, which reached its peak 6 days after inoculation, and also induced accelerated DTH response upon a second inoculation of influenza vaccine alone 4 weeks later, [2] that the cross‐reactive DTH response to PR‐8 vaccine was elicited by the injection of the different influenza A‐type virus vaccine into the footpad of the vaccinated mice, but was not by influenza B‐type virus vaccine, [3] that the DTH‐mediating T cells were detected selectively in the lungs of mice that received the nasal inoculation of the vaccine and CTB together, but that subcutaneous inoculation of this combination failed to induce DTH‐mediating T cells in the lungs. These results, together with the previous papers (Tamura et al, Vaccine 7: 257–262; 314–320, 1989), suggest that CTB could augment both humoral and DTH responses against influenza vaccine in the respiratory mucosal tract.
The Polymerase Chain Reaction for Mycoplasma pulmonis
Authors: Harasawa Ryô , Koshimizu Kaoru , Uemori Takashi , Takeda Osamu , Asada Kiyozo , Kato Ikunoshin
Published: November 2013

in vitro DNA amplification by polymerase chain reaction was examined to detect Mycoplasma pulmonis. A pair of synthetic oligonucleotide primers was constructed, and used to amplify a unique sequence of M. pulmonis DNA. Amplified products were detected by agarose gel electrophoresis and verified by blot hybridization with a synthetic oligonucleotide probe. This system detected cellular DNA of M. pulmonis but not M. arthritidis or M. neurolyticum, and thus appears to be useful for M. pulmonis diagnosis.
New Invasive Serotype of Escherichia coli
Authors: Mendes Edilberto Nogueira , de Magalhães Queiroz Dulciene Maria , Toledo Maria Regina Fernandes , Trabulsi Luiz Rachid
Published: November 2013

A new invasive Escherichia coli strain which presented delayed or variable reactions in lactose, mucate, and acetate tests is described. It did not agglutinate with antisera for all known E. coli and Shigella groups A, B, C, D, and provisional Shigella serovar. We propose the designation of E. coli BH until the situation of its 0 antigen is settled.
Production of Monoclonal Antibody to Bovine Leukemia Virus Glycoprotein gp51 by in vitro Immunization
Authors: Sasaki Osamu , Tanaka Masayuki , Okabe Tatsuji , Imanishi Jiro
Published: November 2013

The monoclonal antibody against glycoprotein gp51 of bovine leukemia virus (BLV) envelope antigen was produced by in vitro immunization. This monoclonal antibody reacted with viral antigen was observed at the 69 kilodalton (kDa) glycoprotein. However, this monoclonal antibody was not involved in neutralizing. It was shown that in comparison with in vivo immunization, in vitro immunization has some advantages, namely a short immunization period and a small antigen quantity.
Effect of Cytokines on Japanese Encephalitis Virus Production by Human Monocytes
Authors: Hasegawa Hitoshi , Satake Yoichirou , Kobayashi Yuzuru
Published: November 2013

Japanese encephalitis (JE) virus was shown to grow in in vitro cultures of human monocytes. Interferon (IFN)‐α and IFN‐γ inhibited JE virus production by the infected monocytes in the absence of anti‐JE virus antibody, but interleukin (IL)‐1α, IL‐2, IL‐3, granulocyte‐macrophage colony‐stimulating factor (GM‐CSF), granulocyte‐CSF (G‐CSF), and tumor necrosis factor (TNF)‐α did not show a significant inhibition. Antibody against JE virus increased the JE virus production by the infected monocytes probably by enhanced uptake of virus‐antibody complexes via Fc receptors. IFN‐γ and GM‐CSF increased JE virus production by monocytes in the presence of anti‐JE virus antibody, whereas IFN‐α inhibited JE virus production even in the presence of the antibody. The other 5 cytokines (IL‐1α, IL‐2, IL‐3, G‐CSF, and TNF‐α) did not show a significant effect on JE virus production by monocytes in the presence or absence of the antibody.
Function of the Hydrophobic Transmembrane Portion of Thy‐1 Antigen
Authors: Tate Genshu
Published: November 2013

Thy‐1 antigen is anchored in the cell membrane by glycophosphatidyl inositol linkages instead of hydrophobic protein domains. The hydrophobic portion of Thy‐1 antigen is cleaved by putative “transamidase.” Mutated genes were constructed by using site‐directed mutagenesis. One mutant gene codes Thy‐1 antigen lacking carboxy terminal amino acids from 112Cys to 143Leu including cell membrane binding amino acid 112Cys. The other mutant gene codes Thy‐1 antigen lacking from 124Trp to 143Leu that includes leucine core portion. DNA transfection analysis and Northern blot analysis revealed that hydrophobic portion of Thy‐1 antigen is essential to express Thy‐1 molecule onto the cell surface.
Characterization of Gentamicin‐Resistant Respiratory‐Deficient (Res−) Variant Strains of Staphylococcus aureus
Authors: Lewis Leslie A. , Li Koibong , Bharosay Mohanie , Cannella Maria , Jorgenson Vincent , Thomas Robert , Pena Danny , Velez Madeline , Pereira Beryl , Sassine Anne
Published: November 2013

Exposure of sensitive cells of Staphylococcus aureus to concentrations of gentamicin higher than the minimal inhibitory concentration, results in the recovery of low level resistant strains with a greatly altered phenotype (variants). Because the phenotypic alteration in these strains is so great the expected diagnostic characterization of these variants as S. aureus is obscured. Starting with a genetically‐marked parent strain, a comprehensive cytological, physiological, morphological, genetic and biochemical analysis of the variants isolated from it was carried out. The genetic lineage of the variants to the parent was also established by DNA/DNA hybridization. Variants result from mutations in the hemin biosynthesis locus, the effect of which is to disrupt the synthesis of components of the electron transport system, lipid synthesis and selected nucleotide synthesis. Thus the strains are defective in aerobic and anaerobic respiration, (res−), in active transport of aminoglycosides (which confers low level resistance), export of characteristic exoenzymes, and in cell wall composition and structure.
Comparison of Two Kinds of Cell Packets of Micrococcus luteus by Scanning Electron Microscopy: Outermost Layer Maintaining the Packet Structure
Authors: Monodane Toshihiko , Tokunaga Michiko , Koike Hirotami , Kotani Shozo , Matsuhashi Michio
Published: November 2013

Two kinds of cell packets of Micrococcus luteus, one having teichuronic acids (TUA) in the cell wall and the other lacking TUA. have been independently reported by two groups of workers. A comparison by scanning electron microscopy of these packets provided a possibly consistent interpretation for the seemingly conflicting opinions whether TUA were involved in packet induction. It was strongly suggested that the packets having TUA in the wall were rigidly maintained by a bridging structure of the outermost layer of the peripheral wall, while the packets lacking TUA showed low contribution of the outermost layer to the bridging structure probably due to the absence of TUA.
Enhancement of Natural Resistance of Mice to Rhodococcus equi Infection by Dextran Sulfate 500
Authors: Takai Shinji , Yokoyama Yasushi , Tsubaki Shiro
Published: November 2013

The effect of intraperitoneal injection of Dextran Sulfate 500 (DS‐500, 500,000 Mr) on the intravenous clearance and host resistance to Rhodococcus equi in mice was studied. Contrary to the results seen with Listeria monocytogenes, DS‐500 enhanced murine resistance to R. equi and altered organ distribution and clearance.
Genetic Transformation of Vibrio parahaemolyticus, Vibrio alginolyticus, and Vibrio cholerae Non O‐1 with Plasmid DNA by Electroporation
Authors: Hamashima Hajime , Nakano Toshikazu , Tamura Shinobu , Arai Taketoshi
Published: November 2013

An electroporation procedure for the plasmid‐mediated transformation of the genus Vibrio was performed, as part of an effort to develop recombinant DNA techniques for genetic manipulation of the genus Vibrio. Vibrio parahaemolyticus, V. alginolyticus, and V. cholerae non O‐1 (9 different strains) were transformed with 3 vector plasmids (pACYC184, pHSG398, and pBR325). The efficiency of transformation was highly dependent on three parameters: the concentration of plasmid DNA; the strength of the electric field; and the combination of plasmid DNA and recipient strain. The drug‐resistance genes on the vector plasmid were expressed in the Vibrio strains.
Characterization of Plasmids that Confer Inducible Resistance to 14‐Membered Macrolides and Streptogramin Type B Antibiotics in Staphylococcus aureus
Authors: Jánosi László , Nakajima Yosinori , Hashimoto Hajime
Published: November 2013

During a period from 1978 to 1989, 413 Staphylococcus aureus strains were isolated at 27 different geographical regions in Hungary; they exhibited an inducible resistance to the 14‐membered macrolides and streptogramin type B antibiotics, but not to the 16‐membered macrolides and lincosamides: this resistance is referred to as PMS resistance phenotype. The isolates were mostly associated with patients suffering from staphylococcal diseases and with hygienic screenings in hospitals and closed communities. They were rarely isolated from food‐poisoning cases, food hygienic screenings, or animal sources. Strains with PMS resistance phenotype were resistant to penicillin (99.0%), tetracycline (78.7%), and chloramphenicol (63.0%); however, they were susceptible to oxacillin. Most of them (94.2%) belonged to the phage type 52‐complex. The determinant for PMS phenotype was located on plasmids, which also encoded beta‐lactamase production and cadmium ion resistance, but not arsenate resistance. Three types of plasmid with molecular size of 50 kilobases (kb), 23.8 kb, and 16.8 kb. were found among the strains with PMS resistance phenotype. and the 50 kb and 23.8 kb plasmids also encoded mercury resistance. The 16.8 kb and 23.8 kb plasmids belonged to incompatibility group 1.
Systemic Group B Streptococcal Disease in the Neonate: Characterization of an Oral Colonization Model Using the Suckling Rat
Authors: Weisman Leonard E. , McKinney LuAnne , Villalobos Raphael
Published: November 2013

Aspiration or ingestion of contaminated amniotic fluid or vaginal secretions has been suggested as a cause of systemic group B streptococcal (GBS) infection in the neonate. Suckling rat studies disagree on whether systemic disease will develop after an oral challenge of GBS. Our goal was to determine if systemic GBS disease would occur following oral colonization in the suckling rat and the effect of bacterial, host and environmental factors. Suckling rat littermates received oral inoculation on one of the first four days of life with varying doses and strains of GBS. Studies confirmed gastric inoculation without aspiration. Mortality and bacteremia decreased with age, increased with dose, varied with strain, and increased with asphyxia. Autopsy confirmed sepsis, intestinal colonization, meningitis, and pneumonia. Bacteremia was associated with an abnormal immature: total neutrophil ratio at 24 hr, thrombocytopenia at 48 hr, and neutropenia at 72 hr after inoculation. GBS can cause systemic infection in the host after oral colonization which appears age‐, dose, strain‐, and environment‐dependent. Evaluation of GBS entry in the susceptible host may facilitate therapies directed toward preventing mucosal invasion.
Surface Properties of Staphylococcus aureus Affecting Chemiluminescence Response of Human Phagocytes
Authors: Yamada Sakuo , Matsumoto Akira
Published: November 2013

To assess the surface properties of Staphylococcus aureus affecting the response of human phagocytes, the effects of the organisms with different surface properties on the chemiluminescence (CL) response of human phagocytes were examined. The magnitude of the phagocytic CL response to hydrophobic strains was significantly greater than that to hydrophilic strains, while no significant difference in the CL response was seen between protein A‐deficient strains and their parent strains. The CL response to the hydrophilic organisms prepared from a hydrophobic strain by trypsin treatment decreased significantly. These results suggest that the phagocytic CL response to staphylococci depends on the hydrophobicity of the surface, but not on the presence of protein A. Two protein A‐deficient strains which were isolated from protein A‐positive strains showed identical hydrophobicity with their parent strains. All of the hydrophilic strains isolated from hydrophobic strains possessed protein A identical to that of their parent strains. Moreover, a hydrophilic strain could be isolated from a protein A‐deficient, hydrophobic strain. These results strongly suggest that protein A is not solely responsible for the surface hydrophobicity of S. aureus.
Enhancement of the Interferon‐Induced Double‐Stranded RNA‐Dependent Protein Kinase Activity by Sindbis Virus Infection and Heat‐Shock Stress
Authors: Saito Sakura
Published: November 2013

In extracts of FL cells that were infected with Sindbis virus or treated with heat‐shock stress, dsRNA‐dependent phosphorylation of 77K protein was markedly increased. The 77K phosphoprotein was indistinguishable from the autophosphorylated and activated form of interferon (IFN)‐induced dsRNA‐dependent protein kinase (PK‐I) by two‐dimensional gel electrophoresis, and was immunologically related to P68 (Galabru, J. and Hovanessian, A., J. Biol. Chem. 262, 15538 (1987)), the HeLa cell counterpart of PK‐I. Immunoblotting experiments using monoclonal antibody against PK‐I revealed that control cell extracts contained a substantial amount of PK‐I protein, although they showed no measurable PK‐I activity even when dsRNA was added. The amount of PK‐I protein did not increase during a transient dsRNA‐dependent enhancement of PK‐I activity caused by Sindbis virus infection and heat‐shock stress. This implies that the conversion of PK‐I protein from a dsRNA‐unresponsive form to a responsive form may be important in the regulation of PK‐I activity. A similar mode of PK‐I regulatory mechanism was operative in the early stages of IFN treatment, although after a prolonged treatment a net synthesis of the PK‐I protein did take place.
Cloning of Genes for a Hemolytic Factor of Leptospira interrogans Serovar Autumnalis Strain Congo 21‐543
Authors: Fukunaga Masahito , Mifuchi Ichiji , Yanagihara Yasutake
Published: November 2013

A DNA fragment encoding a hemolytic factor was cloned from the parasitic spirochete Leptospira interrogans serovar autumnalis strain Congo 21–543. Initial clones were isolated by screening a genomic library in pBR322 in Escherichia coli for hemolytic activity. Hemolytic activity was coded by a 4.5 kilobase BamHI‐HindIII fragment. Southern hybridization with DNAs from other strains of Leptospira using this gene as a probe showed that DNAs from non‐parasitic strains failed to hybridize with the probe, whereas those from all parastic strains tested had the sequence which hybridize to the probe.
Microtiter Plate Assay for Selecting “Macrophage Virulent” Strains of Mycobacterium avium intracellulare Mycobacteria in Mouse Pulmonary Alveolar Macrophages
Authors: Straub Leslie , Thomas Dwayne , Gonzalez‐Rothi Ricardo J.
Published: November 2013

Currently used macrophage‐mycobacterial in vitro infection models require substantial numbers of macrophages. We developed a miniaturized version of such a model, using microtiter plates, which is comparable to standardly published methods, is reproducible, and requires fewer macrophages. In addition to its ease of handling and its economy in time, number of animals, and supplies, this method is preferable when limited numbers of macrophages are available. We have used this assay as a means of selecting human derived isolates from patients with M. avium intracellulare pulmonary disease for their ability to infect and multiply in cultured mouse pulmonary alveolar macrophages.
Interaction between Two Distinct Plaque‐Cloned Human Immunodeficiency Viruses (HIVs): The Possible Existence of Heterozygous Virus
Authors: Masuda Takao , Harada Shinji
Published: November 2013

To know the biological significance of the HIV variation, we investigated the interaction between two different HIV clones. Two distinct clones were mixed and propagated by infecting MT‐4 cells. After passaging the mixture viruses 8 times, and cloning by the plaque method, we obtained not only viruses of homogeneous type like each parental clone but also heterogeneous‐type viruses which showed a mixture of two parental viruses with regard to phenotype and genotype. To further segregate a single virus from the mixture, we recloned the heterogeneous viruses using the plaque method. We found that about 40% of recloned viruses from heterogeneous‐type viruses were still heterogeneous.
Evaluation of Live Trivalent Vaccine of Measles AIK‐C Strain, Mumps Hoshino Strain and Rubella Takahashi Strain, by Virus‐Specific Interferon‐γ Production and Antibody Response
Authors: Nakayama Tetsuo , Urano Takashi , Osano Mitsuru , Nakagawa Masaharu , Maehara Nobutoshi , Sasaki Keiko , Yamamura Anna M. , Makino Satoshi
Published: November 2013

A trivalent measles‐mumps‐rubella live virus vaccine, containing measles AIK‐C strain, mumps Hoshino strain, and rubella Takahashi strain, was evaluated in 229 children, aged 1 to 5 years. The vaccine induced a high seroconversion rate: 221 (98.7%) out of 224 subjects initially seronegative for measles virus, 167 (93.3%) out of 179 initially seronegative for mumps virus, and 212 (99.1%) out of 214 initially seronegative for rubella virus. It also induced a sufficient cellular immunity against each of the three viruses in over 90% of the subjects, as judged by virus‐specific interferon‐γ (IFN‐γ) production. Virus‐specific IFN‐γ production was observed 10 days after vaccination by stimulation with measles virus and rubella virus and 14 days after vaccination by stimulation with mumps virus. Mumps‐virus‐specific IFN‐γ production was observed in 7 out of 12 recipients without seroconversion for mumps virus. And measles‐virus‐specific IFN‐γ production was demonstrated in one out of three recipients without seroconversion for measles virus. A significant correlation was observed between the serum antibody and IFN‐γ production six weeks after vaccination for measles virus (r = 0.201, P<0.01) and for mumps virus (r‐0.174, P<0.05) but not for rubella virus (r= ‐0.045, P>0.05). The incidence of febrile reactions of ≧37.5 C was quite low, 14.4%, and that of ≧39 C occurred in only 1.3% of the recipients. These results suggested that the trivalent vaccine induced sufficient humoral and cellular immunity and yet resulted in no more untoward reaction than observed from the measles vaccine alone.
Overproduction of Human Immunodeficiency Virus Type I Reverse Transcriptase in Escherichia coli and Purification of the Enzyme
Authors: Saitoh Atsushi , Iwasaki Hiroshi , Nakata Atsuo , Adachi Akio , Shinagawa Hideo
Published: November 2013

Overexpression of the reverse transcriptase was designed in E. coli. For a high level of expression, HIV protein was expressed as a protein fusion with β‐galactosidase. When the proviral DNA fragment covering the 3′ half of the gag gene and the entire pol gene was ligated to the 3′ end of the lacZ gene to fuse the truncated gag to lacZ in frame, a small quantity of reverse transcriptase was produced, indicating that frameshifting and post‐translational processing have occurred. Much more reverse transcriptase was produced when the entire pol region was directly fused to the lacZ gene. From a one liter culture of bacteria, 1 mg of highly purified reverse transcriptase consisting of approximately equimolar amounts of two species (p64 and p51) was obtained. These proteins had identical N‐termini consistent with the deduced amino acid sequence and therefore, might be correctly processed from the fusion protein in E. coli by the protease encoded by the pol region. The purified reverse transcriptase was enzymatically as active as the enzyme purified from the virus particles, and immunoreactive to the sera of HIV carriers with high sensitivity and specificity.
Comparison of Serological and Chemical Characteristics of Capsular Polysaccharides of Cryptococcus neoformans var. neoformans Serotype A and Cryptococcus albidus var. albidus
Authors: Ikeda Reiko , Matsuyama Hirohisa , Nishikawa Akemi , Shinoda Takako , Fukazawa Yoshimura
Published: November 2013

The antigenic formula and chemical structure of capsular polysaccharide (CPS) of Cryptococcus albidus var. albidus (C. albidus) were studied in relation to those of C. neoformans var. neoformans serotype A (C. neoformans A). The results of slide agglutination tests with factor sera and reciprocal adsorption experiments showed that antigenic formula of C. albidus was the same as that of C. neoformans A. The soluble CPSs from the two species were obtained from culture supernatants by precipitation with ethanol followed by purification by chromatography on DEAE‐cellulose column. The structural analyses of such CPSs from the two species showed that the antigenic CPS fractions consisted of a backbone of α(l‐3)‐linked D‐mannopyranosyl residues with a single branch of β(1–2)‐xylose or glucuronic acid, and mostly with O‐acetyl groups, in which side chains and O‐acetyl groups were responsible for antigenic specificity. It was found that there was a minor difference between the CPS of C. neoformans A and that of C. albidus; in the former, unsubstituted mannose residues existed in a low frequency, but in the latter none. Moreover, the 1H‐nuclear magnetic resonance spectra of partially hydrolyzed acidic fragments of the two CPSs indicated that two xylose side chains were present between glucuronic acid side chains. Taken together, it was suggested that these two species of C. neoformans A and C. albidus are closely related to each other in their CPSs.
Inhibition of DNA Synthesis in Varicella‐Zoster Virus‐Infected Cells by BV‐araU
Authors: Machida Haruhiko , Watanabe Yoko
Published: November 2013

The inhibitory effect of BV‐araU on DNA synthesis in human embryonic lung cells infected with varicella‐zoster virus (VZV) or herpes simplex virus type 1 (HSV‐1) was compared with that of acyclovir. Cellular uptake of [3H]thymidine and its incorporation into DNA was markedly stimulated by the infection with VZV or HSV‐1, suggesting that the incorporation was mainly due to viral DNA synthesis. DNA synthesis in VZV‐infected cells was dose‐dependently suppressed by BV‐araU and acyclovir, although cellular uptake of [3H]thymidine decreased in cells treated with a high concentration of drugs for an extended time. DNA synthesis in HSV‐1‐infected cells was also markedly inhibited by both drugs in a dose‐dependent manner, without affecting cellular uptake of [3H]thymidine. The concentration of drugs inhibiting DNA synthesis was well correlated to their in vitro anti‐VZV and anti‐HSV‐1 activities. The inhibitory concentration of BV‐araU for DNA synthesis in VZV‐infected cells was one‐thousandth of that of acyclovir. Our results suggest that the antiviral action of BV‐araU against VZV and HSV‐1 is based on the inhibition of DNA synthesis in herpesvirus‐infected cells.
Serovars of Multi‐Antibiotic Resistant Escherichia coli from the Freshwater Environs of Calcutta, India
Authors: Ghosh Asit R. , Nair G. Balakrish , Naik Trailokya N. , Sarkar Swarup K. , Mazumdar Rabin , Pal Sudhir C. , Sen Dipak
Published: November 2013

For a period of one year (March 1987 to February 1988), the incidence of Escherichia coli was determined in water, sediment and plankton collected from two sampling sites in a freshwater lake extensively used by humans and animals. Densities of E. coli associated with plankton was the lowest while sediments, especially at site 2, harbored high densities of the organism. Correlation coefficients revealed that the density of E. coli in water samples was linearly correlated to temperature, pH of water, sediment and humidity. Stepwise multiple regression analysis, however, showed that sediment temperature was the dominant variable which could explain 27% of the observed variation in the numbers of E. coli in the overlying waters (p = <0.001). Of the 150 environmental E. coli strains which were characterized, 31 (20.7%) were found to belong to the classic enteropathogenic E. coli (EPEC) serogroups. Seven of the serogroups among the environmental EPEC strains were also encountered from EPEC strains isolated from human cases during a concurrent clinical study. None of the 150 environmental strains were enterotoxigenic or enteroinvasive but 4 strains possessed HEp‐2 cell adhesive factor. With the exception of one, all the EPEC strains isolated were multi‐drug resistant. From this study, it was evident that the lake is an important source of infection of EPEC and other related diarrheagenic E. coli.
Localization of IpaB Protein in Escherichia coli K‐12 MC1061 Strain Carrying Shigella sonnei Form I Plasmid pSS120
Authors: Ito Ken‐ichiro , Nakajima Takashi , Sasaki Tsuguo , Watanabe Haruo
Published: November 2013

We partially purified antigens which reacted with shigellosis convalescent‐monkey antisera. Hybridomas, which were constructed from mice immunized by the antigens, produced monoclonal antibodies recognizing IpaB protein. Using the monoclonal antibody against IpaB, evidence indicating that IpaB proteins were localized on the cell surface of invasive bacteria (Escherichia coli K‐12 MC1061 harboring pSS120 plasmid) was obtained by immunoelectron microscopy.
Neutralization of Shwartzman‐Inducing Activity by Antibodies Recognizing the Re Core or Lipid A Structures of Lipopolysaccharide from Salmonella minnesota R595 and Pseudomonas vesicularis JCM1477
Authors: Mashimo Jun‐ichi , Mizutani Tomonori , Mita Akira , Kasai Nobuhiko
Published: November 2013

Antibodies recognizing the Re core or lipid A structures of lipopolysaceharide (LPS) derived from Salmonella minnesota R595 and Pseudomonas vesicularis JCM1477 were tested for the ability to neutralize the preparatory activity of endotoxin using the local Shwartzman reaction. Shwartzman‐inducing activity of R595 LPS (Reform) was strongly suppressed when the LPS was incubated with the rabbit anti‐R595 antiserum or the purified IgG antibody which recognizes core region of the LPS. The antiserum also suppressed the preparatory activity of LPS from S. typhimurium SL1102 (Re) and Escherichia coli F515 (Re), but not that of either S. typhimurium LT‐2 (S) LPS or R595 lipid A. Moreover, it was found that the murine monoclonal antibody (MAb), SmRe100G (IgG2a) which recognizes the core region of R595 LPS, significantly suppressed the preparatory activity of R595 LPS. Both conventional antibodies specific to R.595 lipid A, which contains a 1, 4‐bisphosphorylated β‐D‐glucosaminyl‐α‐D‐glucosamine disaccharide structure, and JCM 1477 lipid A, which contains a monophosphorylated 3‐amino‐D‐glucosamine disaccharide structure, neutralized the preparatory activity of homologous and a closely related lipid A, but not that of LPS. In addition, it was observed that MAb Sm5G (IgG2b) specific to enterobacterial lipid A preparations (especially R595 lipid A) neutralized the preparatory activity of R595 lipid A, although the effect was somewhat weak as compared with that of rabbit antiserum. These results suggest that anti‐Re LPS antibody binding to the core of Re LPS is involved in suppressing the endotoxic activity of Re LPS, and that the direct binding of anti‐lipid A antibody to some specific epitopes of lipid A is important in neutralizing the endotoxic activity.
Mechanism of Protective Effect of Recombinant Human Granulocyte Colony‐Stimulating Factor (rG‐CSF) on Pseudomonas Infection
Authors: Matsumoto Masahiko , Tamura Masahiko , Matsubara Shuzo , Matsuno Takashi , Ono Masayoshi , Yokota Takeshi
Published: November 2013

Decrease in resistance to systemic Pseudomonas infection in cyclophosphamide (CPA)‐induced neutropenic mice was prevented by injections of recombinant human granulocyte colony‐stimulating factor (rG‐CSF). In order to explore mechanism of the prevention of CPA‐induced decrease in the anti‐infectious resistance by rG‐CSF, CPA‐treated and then rG‐CSF‐injected mice were inoculated i.p. with P. aeruginosa, and growth of the infecting bacteria and infiltration of leukocytes in the peritoneal cavity were determined. In the mice who had received 4 daily s.c. injections of rG‐CSF from the day after CPA‐injection, a large number of neutrophils were mobilized into the peritoneal cavity in response to the bacterial inoculation and growth of the infecting Pseudomonas in the cavity was markedly‐inhibited, whereas in CPA‐induced neutropenic mice few neutrophils were mobilized and the infecting bacteria proliferated vigorously in the peritoneal cavity. These results suggest that administration of rG‐CSF prevents CPA‐induced neutropenia and neutrophils circulating at normal level in the number are normally mobilized into the peritoneal cavity in response to Pseudomonas inoculation, and that the mobilized neutrophils inhibit proliferation of the infecting Pseudomonas.
Experimental Infection of Specific‐Pathogen‐Free Mice with Enterohemorrhagic Escherichia coli 0157: H7
Authors: Xin‐He Lai , Xu Jian‐guo , Liu Bing‐yang
Published: November 2013

By subcutaneous inoculation of 108 CFU of enterohemorrhagic E. coli 0157:H7, specific‐pathogen‐free mice revealed most of the symptoms and histological changes observed in patients. The histological changes in intestine were mainly seen in the distal parts of small intestine and the cecum. Vacuolation of villi in the cecum was also observed. The histological changes in the kidneys of the infected mice were featured as the swollen epithelial cells of glomeruli and the marked thickening of glomerular capillaries with barely visible lumens. Unexpected findings in the bronchiole were characterized by sloughing of the epithelial cells of bronchiolar wall, leading to partial or complete obstruction of the lumens. Histological changes in the spleen, liver and lymphnodes were also observed. The bacteria were recovered from the feces, contents of small intestine, and samples taken from kidney, liver, heart, spleen, different parts of small intestine, cecum, and colon. By using peroxidase‐antiperoxidase (PAP) assay with polyclonal antibodies against “O” antigen of E. coli 0157:H7, it was observed that the samples taken from the brain, kidney, ileum, cecum, spleen, and liver gave positive reactions. Feces and contents of small intestine obtained from all of the infected animals were positive by occult blood test. These results show that the experimental infection of E. coli 0157:H7 in this model is systemic in nature.
Electron Microscopic Studies on the in vitro Proliferation of Spotted Fever Group Rickettsia Isolated in Japan
Authors: Amano Ken‐ichi , Hatakeyama Hiroshi , Sasaki Yoshimitsu , Ito Reietsu , Tamura Akira , Suto Tsunehisa
Published: November 2013

Rickettsia was isolated from a patient with Japanese spotted fever, and its proliferation in cultured green monkey kidney cells was observed by electron microscopy. In the course of this study, we observed fusion of infected cells to uninfected cells which may be a way of spreading the rickettsiae from a cell to another. On the other hand, whirlpool‐like, multilayer membranous structures, similar to the mesosomes of gram‐negative bacteria, were sometimes seen in the rickettsial cells. The other profiles common to the other rickettsiae in spotted fever group were observed, such as the electron‐lucent halo zone around the rickettsiae, and external fibrous materials on their surface, but intranuclear multiplication was rarely observed.
Chemical Structure of the 2‐Keto‐3‐Deoxyoctonate (KDO) Region of the Lipopolysaccharide Isolated from O1 Vibrio cholerae NIH 4IR (Ogawa)
Authors: Kondo Seiichi , Haishima Yuji , Hisatsune Kazuhito
Published: November 2013

The chemical structure of the 2‐keto‐3‐deoxyoctonate (KDO) region of the lipopolysaccharide (LPS) isolated from Ol V. cholerae NIH 41R (Ogawa) was elucidated by dephosphorylation, periodate oxidation and methylation analysis. Methylation analysis of KDO in the dephosphorylated LPS revealed the presence of 5‐O‐acetyl‐1,2,4,6,7,8‐hexa‐O‐methyl‐3‐deoxy‐octitol and 2‐keto‐3‐deoxy‐heptulosonic acid was detected in the methanolysate of the periodate‐oxidized and dephosphorylated LPS. These results indicated that the site of binding of KDO to the core oligosaccharide is position C5 as in enteric gram‐negative bacterial LPS, while only one molecule of the KDO residue carrying phosphate on position C4 is present in the inner core region of the LPS in contrast to enteric gram‐negative bacterial LPS in which one molecule of KDO carrying KDO or KDO2→4KDO disaccharide instead of the phosphate group at position C4 is present in its main chain.
Antibody Production Useful for Cytotoxicity against Mouse MM2 Tumor Cells by Peritoneal Macrophages
Authors: Natsuume‐Sakai Shunnosuke , Murayama Tsugiya , Furukawa Toru , Koshimura Saburo
Published: November 2013

We applied an antibody‐dependent macrophage‐mediated cytotoxicity (ADMC) test in order to analyze the effector mechanism of the host‐mediated antitumor effects induced by OK‐432. Adherent peritoneal exudate (PE) cells were obtained from each of high (C3H/He) and low (B10) responder mice treated with OK‐432. and 51Cr‐labeled MM2 tumor cells were used as target cells. The cytotoxic activity in vitro coincided well with the results obtained in in vivo antitumor experiments. When adherent PE cells from C3H/He mice reacted with anti‐MM2 serum from B10 mice, the degree of ADMC was significantly lower than that obtained with the anti‐MM2 serum from C3H/He mice. The removal of IgGl from the anti‐MM2 serum induced in B10 mice resulted in the enhancement of ADMC activity. Then mean level of IgG2 in each of anti‐MM2 sera from C3H/He and B10 mice was higher than in normal serum, and the IgG1 level in the antiserum from B10 mice was also higher than that in the serum from normal B10 mice. The present work suggested that the active component(s) in anti‐MM2 serum participating in ADMC was a specific antibody of the IgG2 subclass, and that the inhibiting factor(s) was the IgG1 subclass.
Suppression of Lymphocyte Signal Transduction by Murine Mastocytoma Ascites
Authors: Ding Linna , Isobe Ken‐ichi , Yoshida Tomoaki , Nakashima Izumi
Published: November 2013

The lymphocyte signal transduction, as determined by intracellular free Ca2+ mobilization of concanavalin A‐stimulated T lymphocytes and of antiimmunoglobulin μ chain antibody‐stimulated B lymphocytes, was suppressed in spleen cells from mice injected with murine Pl.HTR mastocytoma‐induced ascites and in spleen cells treated with the ascites in vitro. The suppression was observed both at the peak level and in the reactive pattern of Ca2+ influx. In the suppression, the ascites were replaceable with tumor culture supernatants or tumor homogenates. Correspondingly, primary and secondary cytotoxic T lymphocyte (CTL) responses of DBA/2 mice to allogeneic antigen were also significantly suppressed by injection of the syngeneic Pl.HTR tumor‐derived ascites. This new finding suggested that the mechanism of the tumorous ascites or of the tumor‐derived factor‐mediated immunosuppression involves at least in part the suppression of the early event of the signal transduction for lymphocyte activation.
Platelet Aggregation by Strains of Enterococci
Authors: Usui Yukio , Ichiman Yoshitoshi , Suganuma Masaru , Yoshida Kosaku
Published: November 2013

The platelet aggregation capability of whole cells of Enterococcus faecalis, E. faecium and E. avium was tested. The optimum ratios of bacteria to platelets in E. faecalis (strain SMU‐37), E. faecium (strain SMU‐138) and E. avium (strain SMU‐197) were 1.0, 1.2 and 2.0, respectively. During the platelet aggregation induced by the three strains of enterococci, 65–69% of total serotonin was released. The aggregation was totally inhibited by ethylenediaminetetraacetate (10 mM) and apyrase (1 mg/ml), while no effect was shown by aspirin (10 mM), indomethacin (10 mM) and quinacrine (1 mM). By pretreatment of platelet‐poor plasma with heat (56 C, 30 min) or zymosan, the reactivities with platelets of each strain of species were markedly diminished. These results suggest that enterococci‐induced platelet aggregation was an ion‐dependent, cyclooxygenase‐insensitive event, and plasma component(s) was (were) required for the reaction.
Natural Killer (NK) Cell Activating Factor Produced by a Human T‐Cell Hybridoma
Authors: Kobayashi Yoshiro , Yoshimatsu Kentaro , Osawa Toshiaki
Published: November 2013

A human T‐cell hybridoma (KC8–1.10), whose culture supernatant augments peripheral blood lymphocytes (PBL)‐mediated spontaneous cytotoxicity against K562 cells, was established. This activity [natural killer (NK) cell activating activity] appears to be not due to interferon‐γ (IFN‐γ) and interleukin‐2 (IL‐2) for the following reasons: 1) KC8–1.10 produced negligible or small amounts of IFNs and IL‐2. 2) The NK cell activating activity in the KC8–1.10 culture supernatant was not neutralized by anti‐IFN‐γ antiserum and stable even after pH 2 treatment for 24 hr, which is known to destroy IFN‐γ activity, 3) IL‐2‐dependent cell line absorbed IL‐2 more efficiently than it absorbed the NK cell activating activity, and the latter activity was not retained by Blue Sepharose column in contrast with IL‐2. The NK cell activating factor in the KC8–1.10 culture supernatant appears to be a glycoprotein, because the activity was abolished with pronase treatment or with boiling for 5 min and because the activity was retained by concanavalin A‐ and Pisum sativum agglutinin‐agarose. Finally it was found that the NK cell activating activity requires Leu 11b+ cells to exert its effect.
Inhibitor of Interferon‐Induced Double‐Stranded RNA‐ Dependent Protein Kinase and Its Relevance to Alteration of Cellular Protein Kinase Activity Level in Response to External Stimuli
Authors: Saito Sakura , Kawakita Masao
Published: November 2013

In FL cells, interferon (IFN)‐induced dsRNA‐dependent protein kinase (PK‐I) was found to be present in a form complexed with a potent inhibitor of its dsRNA‐dependent activation. The inhibitor was readily dissociated from PK‐I by DEAE‐cellulose chromatography to yield a dsRNA‐responsive PK‐I. The inhibitor was also dissociated easily from PK‐I by gel filtration through Sephacryl S‐200. The apparent molecular mass of the inhibitor as estimated by gel filtration was more than 160 kilodaltons. Activity of the inhibitor was decreased on IFN treatment for 8.5 hr or on Sindbis virus infection with concomitant increase in the amount of dsRNA‐activatable form of PK‐I. This result implies that the inhibitor may be one of the regulatory factors of cellular PK‐I activity. Longer IFN treatment (24 hr) led to recovery of the inhibitor activity, but it was overridden by an extensive net synthesis of the PK‐I protein.