bluuebunny/crossref_metadata_2025 huggingface dataset:
Gastroenteritis in Suckling Mice Caused by Human Rotavirus Can Be Prevented with Egg Yolk Immunoglobulin (IgY) and Treated with a Protein‐Bound Polysaccharide Preparation (PSK)
Authors: Ebina Takusaburo , Tsukada Keiko , Umezu Kouji , Nose Makoto , Tsuda Ken , Hatta Hajime , Kim Mujo , Yamamoto Takehiko
Published: November 2013

Oral inoculation of human rotavirus MO strain (serotype 3) into 5‐day‐old BALB/c mice cause gastroenteritis characterized by diarrhea. Clinical symptoms, histopathological changes in the small intestine, and the detection of rotavirus antigen in enterocytes were all characteristic of rotavirus‐induced gastroenteritis. Using this small animal model, passive protection of suckling mice against human rotavirus infection was achieved with the use of immunoglobulin (IgY) from the yolks of eggs of rotavirus‐immunized hens. When IgY against a rotavirus strain homotypic to the challenge virus (MO strain) was administered in the mice, complete protection against rotavirus infection was achieved. On the other hand, with oral administration of IgY against a heterotypic strain (serotype 1, Wa strain), a lower protective effect was nevertheless obtained. The four different strains of human rotavirus (Wa, KUN, MO, and ST3) were inactivated in vitro by treatment with PSK, a protein‐bound polysaccharide preparation, in a dose‐dependent manner. Oral administration of 2.5 mg of PSK caused a therapeutic effect on experimentally MO‐infected suckling mice. The antiviral effect of PSK was indicated by the reduction of the duration of diarrhea.
Localization of a Determinant Mediating Partial Macrolide Resistance in Staphylococcus aureus
Authors: Matsuoka Mayumi , Endou Kikutarou , Nakajima Yoshinori
Published: November 2013

Four out of more than 8,200 Staphylococcus aureus strains isolated in Japan between 1961 and 1980 were constitutively resistant to a variety of macrolide antibiotics except tylosin and rokitamycin, but susceptible to lincosamide and streptogramin type B antibiotics (PM). The data obtained by agarose gel electrophoresis, CsCl‐ethidium bromide density gradient analysis, diagnosis with ATP‐dependent deoxyribonuclease, and a test transducing into a rec− mutant with phage 80L2 propagated on PM‐resistant S. aureus all suggested that the determinant for the PM‐resistance is located in chromosome.
Protective Effect of Recombinant Human Granulocyte Colony‐Stimulating Factor (rG‐CSF) against Various Microbial Infections in Neutropenic Mice
Authors: Matsumoto Masahiko , Matsubara Shuzo , Matsuno Takashi , Ono Masayoshi , Yokota Takeshi
Published: November 2013

Protective effect of recombinant human granulocyte colony‐stimulating factor (rG‐CSF) on microbial infections was studied in cyclophosphamide (CPA)‐induced neutropenic mice. The neutropenic mice showed severely decreased resistance against systemic infections of Pseudomonas aeruginosa, Escherichia coli, Serratia marcescens, Staphylococcus aureus, and Candida albicans. When such mice were injected subcutaneously with rG‐CSF on four consecutive days beginning the day after CPA injection, the decreased anti‐microbial resistance of the mice was restored to the level of that in normal mice. The anti‐infective effect of rG‐CSF was dose‐dependent and the 50% effective doses (ED50) in various microbial infections tested were 1 10 μg/kg/day. The results suggest that rG‐CSF is useful for protection of neutropenic patients from microbial infections.
Hemagglutination Activities of Purified Pertussis Toxin and Filamentous Hemagglutinin against Erythrocytes from Various Animals
Authors: Gupta Rajesh K. , Saxena S.N. , Sharma Suniti B. , Ahuja Subhash
Published: November 2013

The hemagglutinating (HA) activities of purified pertussis toxin (PT) and filamentous hemagglutinin (FHA) were evaluated against unfixed and glutaraldehyde‐fixed erythrocytes from ox, goose, horse, monkey, sheep, chicken, and rabbit. Both PT and FHA showed HA activities against fixed and unfixed erythrocytes from all the animals studied. The HA titers of FHA were higher than those of PT. The HA activities of FHA and PT were not destroyed completely even after heating these preparations at 56 C for 30 min. A simple test for the assay of PT in culture supernatants of Bordetella pertussis on the basis of HA activity has been described.
Isolation of a Serologically Different Compact‐Colony‐Forming Active Substance from Strains of Staphylococcus aureus
Authors: Yoshida Kosaku , Ohtomo Toshichika , Suganuma Masaru
Published: November 2013

To observe the possible serological heterogeneity of compact‐colony‐forming active substance (CCFAS), heat‐killed vaccines were prepared by two strains of Staphylococcus aureus, strains SMU 1–46 and SMU 7931, cultured in 0.03 M tris‐hydrochloride‐buffered brain heart infusion, pH 8.4. After immunization with the vaccine in rabbits, antibody responses were observed during a period of six weeks after the immunization either by homologous and heterologous organisms using alkaline serum‐soft agar technique. The results showed that remarkable antibody production was shown only against homologous strain, but not against heterologous strain. The antibodies were absorbed out only with highly purified preparation of CCFAS extracted from homologous strain and not with heterologous CCFAS. Differences of the major chemical composition of the substances showed that highly purified CCFAS extracted from strain SMU 7931 contained 2.84 and 2.04 times higher amounts of galactose and 2‐amino‐2‐deoxy‐D‐galacturonic acid than those of CCFAS obtained from strain SMU 1–46.
Reassessment of Antigenic Determinant of Saccharomyces cerevisiae Serotype Ia
Authors: Nishikawa Akemi , Serine Takeshi , Ikeda Reiko , Shinoda Takako , Fukazawa Yoshimura
Published: November 2013

We examined the immunochemical structure of the antigenic determinant of S. cerevisiae serotype Ia. The specific factor serum for S. cerevisiae serotype Ia was obtained either from factor 18 serum by adsorption with heat‐killed cells of Candida glabrata, or from anti‐S. cerevisiae Ia (M 6001) serum by adsorption with heat‐killed cells of S. cerevisiae Ib (IFO 0751). We designated this adsorbed serum as factor 18a. Acetolysis of S. cerevisiae cell wall D‐mannan gave five oligosaccharides. Signals of 1H‐nuclear magnetic resonance spectra of mannooligosaccharides derived from S. cerevisiae mannan were assigned for their linkages by the aid of those of α‐1,3′‐linked mannooligosaccharides derived from glucuronoxylomannan of capsule of Cryptococcus neoformans serotype A‐D. Agglutination‐inhibition experiments revealed that the mannopentaose from S. cerevisiae mannan was the most effective inhibitor. Moreover, inhibitory activities of α‐1,3′‐linked mannotriose, mannotetraose, and mannopentaose which were derived from glucuronoxylomannan of C. neoformans were shown to be higher than those of mannotetraose with one terminal α‐(l‐3) linkage from homologous S. cerevisiae mannan. These results indicate that mannopentaose with terminal two α‐(l‐3) linkages is responsible for the specificity of S. cerevisiae Ia.
Enzyme‐Linked Immunosorbent Assay Employing Monoclonal Antibodies for Direct Identification of Enteric Adenoviruses (Ad40,41) in Feces
Authors: Nishio Osamu , Ooseto Mituaki , Takagi Kenzi , Yamasita Yasutaka , Ishihara Yuichi , Isomura Shin
Published: November 2013

For detection and identification of enteric adenovirus (Ad) types 40 and 41 in stool specimens, enzyme‐linked immunosorbent assay (ELISA) was developed with the use of three monoclonal antibodies: Ad group‐specific, Ad40 type‐specific, and Ad41 type‐specific antibodies. Of 860 fecal samples from patients with acute gastroenteritis, 44 strains of Ad were isolated using Graham 293 cell cultures. Of these isolates, 20 were typed as Ad40, 18 were Ad41, and 6 were other Ads by neutralization tests with cell cultures. Results of the ELISA tests on these 860 fecal samples resulted in good agreement to those with the cell culture method. The ELISA tests using Ad type‐specific monoclonal antibodies proved to be a specific and rapid technique for laboratory diagnosis of acute gastroenteritis caused by enteric Ads.
Adherence and Pathogenesis of Salmonella enteritidis in Mice
Authors: Aslanzadeh Jaber , Paulissen Leo J.
Published: November 2013

Adherence of many pathogenic organisms to the host cells has been associated with the presence of fimbriae. The exact role of these organelles in the adherence and pathogenesis of Salmonella enteritidis is not well established. Utilizing hemagglutination tests, S. enteritidis was shown to possess type 1 and type 3 fimbriae. Polyacrylamide gel electrophoresis of the isolated fimbriae showed that type 1 and 3 fimbriae of S. enteritidis had subunit M.r of 17 and 22 kDa, respectively. In vitro adherence assays suggested that S. enteritidis utilized type 1 fimbriae to adhere to human buccal and mouse small intestine epithelial cells. In addition, antibody produced against type 1 and type 3 fimbriae protected the mice from infection with a lethal dose of S. enteritidis. These results suggest that type 1 and possibly type 3 fimbriae are involved in the adherence and pathogenesis of S. enteritidis. The data further suggest that they may have a role in the adherence and pathogenesis of the other enteric organisms.
The Growth of Mycobacteria in Liquid Medium Is Influenced by the Dimension of Glass Wall in Contact with the Medium
Authors: Tsukamura Michio
Published: November 2013

The growth of the type strain of Mycobacterium aurum was compared between flask and tube, both of which contained the same concentration of bacteria in synthetic media and had the same air‐contact dimension but had different dimensions of the glass wall of vessels contacting the medium. Much better growth occurred in the flask, which had a smaller dimension of the glass wall contacting the medium, than in the tube, which had a larger dimension. The finding shows that the growth of bacteria is influenced by the dimension of the glass wall of vessels. It was considered that a high chance of the attack shock for bacteria given by the Brownian movement lowers the multiplication of bacteria.
Protective Immunities Induced by Porins from Mutant Strains of Salmonella typhimurium
Authors: Matsui Katsuhiko , Arai Toshihiko
Published: November 2013

The protective immunity against Salmonella typhimurium‐infection in mice immunized with porins from mutant strains of S. typhimurium was studied. A high level of protection against S. typhimurium infection was achieved in mice immunized with native porins from S. typhimurium LT2 (wild‐type strain) but not from S. typhimurium SH6017, SH6260, or SH5551 (mutant strains), which produce 34K, 35K, or 36K porin, respectively. Moreover, when mice were immunized with mixtures of 34K, 35K, and 36K porins (34K + 35K, 35K + 36K, 34K + 36K, or 34K + 35K + 36K porin) or LT2 porin heated at 100 C for 2 min in 2% SDS (heat‐denatured LT2 porin), the degree of protective immunities in the mice was very much lower than that in the mice immunized with the native LT2 porin. However, antisera raised against these porins showed no significant differences of the antibody titer against LT2 porin or LT2 whole cells. On the other hand, mice immunized with the native LT2 porin—but not 34K, 35K, 36K, 34K + 35K + 36K, and the heat‐denatured LT2 porins—exhibited significant levels of delayed‐type hypersensitivity reaction and interleukin‐2 production when they were elicited with whole cells of S. typhimurium LT2. These observations suggested that the high level of protection induced by the native LT2 porin immunization was dependent on the induction of cell‐mediated immunity.
Characterization of an Escherichia coli Mutant Pleiotropically Altered in Membrane‐Bound Oxidoreductase Activities
Authors: Cox John C. , Goldschmidt Eugene P. , Jurtshuk Peter
Published: November 2013

An Escherichia coli mutant pleiotropically altered in membrane‐bound oxidoreductase activities was isolated following nitrosoguanidine treatment. Mutant R23 was able to grow on glucose, but was unable to grow on succinate or other oxidizable substrates as a sole energy source. Isolated membranes prepared from R23 failed to oxidize succinate and formate; while NADH was oxidized at a reduced rate by membranes. The mutant also exhibited markedly reduced cytochrome content, but normal DL‐lactate PMS reductase and H+‐translocating ATPase activities relative to the parent strain. Bacteriophage P1kc was used to transduce R23 to growth on glycerol, DL‐lactate or succinate; regardless of the selection procedure, each of the 179 transductants had gained the ability to grow on all three substrates. The suc− mutation in R23 appeared to be responsible for the loss of growth on oxidizable substrates, altered membrane‐bound oxidoreductase activities, resistance to neomycin, and reduced levels of cytochrome components. The suc− mutation was localized in the 6 to 6.5 min region of the E. coli chromosome map utilizing episomal transfers.
Induction of Interferons (IFNs) and Tumor Necrosis Factor (TNF) in Mice by a Novel Glycolipid Trehalose 2,3,6′‐Trimycolate from Rhodococcus aurantiacus (Gordona aurantiaca)
Authors: Fujita Takako , Sugimoto Nobuko , Yokoi Fusa , Ohtsubo Yoshikazu , Ikutoh Masatoshi , Kato Yoshiko , Natsuhara Yayoi , Oka Shiro , Yano Ikuya
Published: November 2013

The immunomodifying activity of a novel mycoloyl glycolipid, trehalose 2,3,6′‐trimycolate (GaGM), from a unique psychrophilic acid‐fast bacterium, Rhodococcus aurantiacus, was examined. ICR mice were primed intravenously (i.v.) or intraperitoneally (i.p.) with liposomes containing GaGM (300 μg/mouse), and were administered LPS dissolved in saline (25 μg/mouse, i.v.) 2 weeks later. Two hours after injection of LPS, interferons (IFNs) and tumor necrosis factor (TNF) were induced significantly in mice sera. The increase in activities of IFNs and TNF was approximately paralleled with granuloma formation in spleen of mice primed with GaGM. However, IFNs and TNF were not induced either in mice primed with GaGM but not elicited with LPS, or in those primed with GaGM and elicited by GaGM. Both activities induced were lower in mice primed with trehalose mono‐ or dimycolate from R. aurantiacus (GaTMM, GaTDM) or TDM from Nocardia rubra than in GaGM‐primed mice. Time course study showed that the maximum activity of each interferon (α, β, or γ) was observed at different stages after LPS administration; IFN‐α, IFN‐β, and IFN‐γ appeared 3, 2, and 6 hours most abundantly after LPS administration, respectively.
Demonstration and Characterization of the Cell Wall Carbohydrate and Protein Antigens from Clostridium Botulinum Type E Saroma
Authors: Takumi Kemji , Nagao Shigeki , Takeoka Aya
Published: November 2013

Two different cell wall antigens, carbohydrate (CHO) and protein (P), from Clostridium botulinum type E Saroma were extracted with sodium dodecyl sulfate (SDS) and purified by chromatography on DEAE‐Sepharose CL‐6B and Sephadex G‐75 or G‐100. The CHO antigen was composed of glucose, galactose, glucosamine, galactosamine, alanine and phosphorus with a molar ratio of 1.5: 1.5:0.25:0.25: 1:1. The P antigen was an acidic protein with a molecular weight of 60 kDa, in which the major amino acids were aspartate, glutamate and serine, while the minor ones were cysteine and methionine. Thin sections of the intact or SDS‐extracted cells of the organism demonstrated that the cell wall was composed of a two‐layered structure, an inner layer about 20 nm thick and an outer layer about 10 nm, and by the extraction with SDS, the outer layer disappeared from the cell surface, leaving the inner layer. Immunogel diffusion tests demonstrated that either CHO antigen or P antigen was common among the nonproteolytic strains of C. botulinum.
Inactivation of Kanamycin A by Phosphorylation in Pathogenic Nocardia
Authors: Yazawa Katsukiyo , Mikami Yuzuru , Maeda Akio , Kudo Takuji , Suzuki Ken‐ichiro , Saito Naoki , Kubo Akinori
Published: November 2013

Among the five species of pathogenic Nocardia, i.e., N. asteroides, N. brasiliensis, N. farcinica, N. nova and N. otitidiscaviarum, all strains of N. brasiliensis and N. farcinica showed resistance to an aminoglycoside antibiotic, kanamycin A, showing the MIC (minimum inhibitory concentration) values of more than 100 μg/ml. This species‐specific difference in sensitivity was found to be explained by the production of an inactivation enzyme, aminoglycoside 3′‐phosphotransferase APH(3′). Structural studies by mass and NMR spectroscopy on the inactivated substance produced by a cell‐free extract of the Nocardia confirmed the conversion of kanamycin A to an inactive substance, kanamycin A 3′‐phosphate. The MIC values of N. otitidiscaviarum and N. nova for kanamycin A, on the other hand, ranged from 0.78 μg/ml to 100 μg/ml, and both species were non‐producers of APH(3′). Sensitivity to the antibiotic and APH(3′) productivity of N. asteroides varied depending on the strain.
Serological Studies on Leptospira Strain Ictero No. I Using Monoclonal Antibodies
Authors: Kobayashi Yuzuru , Sada Eiji , Tamai Tomonori
Published: November 2013

In order to elucidate the full serological characteristics of strain Ictero No. I, the first strain of Leptospira isolated by Inada and Ido in 1914, 17 monoclonal antibodies against the strain were produced by cell fusion technology. The antibody‐producing hybridomas were designated IMAs 1 to 17. The reactivities of the monoclonal antibodies produced by the hybridomas were determined by the microscopic agglutination test. One of the 17 monoclonal antibodies, IMA 1, reacted with strains Ictero No. I, LT 1131 and Naam, but not with other strains of the serogroup Icterohaemorrhagiae including strain RGA used in the present study. Moreover, the reactivity of the antigenic determinant of IMA 1 was inactivated by treatment at 56 C for 30 min. The 16 other monoclonal antibodies, IMAs 2 to 17, showed different reaction patterns against Leptospira strains of the serogroup Icterohaemorrhagiae. All of the 16 antibodies reacted with both Ictero No. I and RGA strains. The antigens against antibodies IMAs 2 to 17 were thermostable. The present study thus clarified the presence of a thermolabile antigen in strain Ictero No. I, and allowed correct distinction between the serotype of strain Ictero No. I and that of strain RGA using IMA 1. Therefore, we propose that strain Ictero No. I represents serovar icterohaemorrhagiae, as originally designated by Inada and Ido. Moreover, strain Ictero No. I should be designated as the type strain of Leptospira interrogans.
The Presence of Cis‐9,10‐Methylene Hexadecanoic Acid in Yersinia enterocolitica
Authors: Kanemasa Yasuhiro , Nagamachi Eiko , Shibuya Sei‐ichiro
Published: November 2013

The fatty acids of Yersinia enterocolitica were investigated by Abbas and Card. In their report, they stated that the major fatty acids were C16:0, C16:1, C17:0 and C18:1 and branched or cyclopropane side‐chain fatty acids could not be detected. We, however, found a moderate amount of cyclopropane side‐chain fatty acid. We could determine that this fatty acid was cis‐9, 10‐methylene hexadecanoic acid by gas‐liquid chromatography, gas chromatography‐mass spectrometry, silver‐nitrate‐treated TLC and PtO2 catalyzing hydrogenation method.
Susceptibility of Newly Established Mouse Strain MPS to Mycoplasma pneumoniae Infection
Authors: Sasaki Yuko , Ogura Atuo , Nakayama Kazue , Noguchi Yoko , Matsuno Kumiko , Saito Manabu
Published: November 2013

A newly established mouse strain, MPS, which is more sensitive to Mycoplasma pulmonis than ICR, ddY and other mouse strains was examined for its susceptibility to Mycoplasma pneumoniae. In experimental infections with M. pneumoniae, it was observed that M. pneumoniae attached to tracheas of MPS mice, and M. pneumoniae cells were isolated from tracheas and lungs of MPS mice even after four weeks of infection, while no mycoplasmas were isolated from ICR and ddY mice after one week of infection. Specific antibodies against M. pneumoniae were also observed by the Western blotting in the sera of MPS mice infected with M. pneumoniae. Although any lung lesion could not be observed in this work, this newly established mouse strain MPS may be useful for experiments of M. pneumoniae infection, especially for the analysis of strain differences in susceptibility to M. pneumoniae infection.
Analysis of the tdh Gene Cloned from a tdh Gene‐ and trh Gene‐Positive Strain of Vibrio parahaemolyticus
Authors: Baba Kiyoshi , Shirai Hiromasa , Terai Akito , Takeda Yoshifumi , Nishibuchi Mitsuaki
Published: November 2013

A variant of the gene (tdh) encoding thermostable direct hemolysin (TDH) was cloned from the chromosome of Vibrio parahaemolyticus AQ3860, which gave positive results in the hybridization tests with the tdh gene probe and the trh (tdh‐related hemolysin) gene probe and showed a low level of reaction in an enzyme‐linked immunosorbent assay for TDH. Nucleotide sequence analysis of the cloned gene (tdh5) provided no evidence that tdh5 is evolutionally closer to the trh gene than the other tdh genes. The tdh5 gene was flanked by 40 base‐pair sequences constituting perfect inverted repeats, which may suggest association of the tdh5 gene with insertion sequence‐like structure. These results suggest that the tdh5 gene and the trh gene were not originally produced by gene duplication in AQ3860 but rather that one of the two genes moved into AQ3860 from an external source.
Rapid Method for Isolation and Staining of Bacterial Lipopolysaccharide
Authors: Al‐Hendy Ayman , Toivanen Paavo , Skurnik Mikael
Published: November 2013

Classically bacterial lipopolysaccharide (LPS) purification and silver staining take several days. We designed a simple and fast method for LPS isolation which when combined with silver staining using Pharmacia PhastSystem™ both can be completed in few hours. The purity of LPS isolated by this simple method may not be comparable to that by the phenol‐water method hence we recommend this rapid isolation and staining procedures for simple and fast study of LPS patterns in gels.
Purification and Partial Characterization of a Soluble Hemagglutinin from Yersinia pseudotuberculosis
Authors: Takumi Kenji , Koga Tetsuro , Oka Tatsuzo , Takeoka Aya , Tsubokura Misao , Sheng Yao Jian
Published: November 2013

A soluble hemagglutinin (HA) produced by Yersinia pseudotuberculosis strain Inoue, serotype 5b, was purified by ammonium sulfate precipitation, gel filtration on Sepharose CL‐6B and high performance liquid chromatography on a DEAE‐5PW anion‐exchange column. The purified HA was a 14.5 kDa protein with an isoelectric point of 4.5. Amino acid analysis indicated that the HA consisted of 133 residues, corresponding to the molecular weight of 14,100. The amino acid sequence of N‐terminal 38 amino acid residues showed no homology with that of several fimbrial proteins from Escherichia coli.
Preventive Effects of a Chinese Herb Medicine (Sho‐saiko‐to) against Lethality after Recombinant Human Tumor Necrosis Factor Administration in Mice
Authors: Sakaguchi Shuhei , Tutumi Eiji , Yokota Katushi , Furusawa Shinobu , Sasaki Ken‐ichi , Takayanagi Yoshio
Published: November 2013

We observed the effects of a Chinese herb medicine Sho‐saiko‐to on the lethal and antitumor activities of recombinant human tumor necrosis factor (rhTNF) administered in mice. Sho‐saiko‐to was noted to protect the rhTNF‐induced lethality in galactosamine‐hypersensitized mice, and also Sho‐saiko‐to pretreated mice was protected against the decrease of rectal temperature after rhTNF administration. On the other hand, there was a remarkable enhancement of antitumor activity of rhTNF by Sho‐saiko‐to pretreatment. These results suggest that Sho‐saiko‐to drug may protect mice from severe shock syndrome induced by rhTNF.
Formation of Vesiculated Large Bodies of Staphylococcus aureus L‐Form in a Liquid Medium
Authors: Yabu Kunihiko
Published: November 2013

A method is described in which cells of Staphylococcus aureus can be converted to vesiculated large bodies of L‐form. When coccal cells were incubated in a liquid growth medium containing D‐cycloserine, N‐acetylmuramidase and subtilisin, a large number of vesiculated large bodies were formed. Electron microscopy revealed that development of internal vesicles arose after 6 hr of incubation. When growth inhibitory concentrations of rifampicin, novobiocin, or chloramphenicol were added to the culture at 6 hr of incubation, small‐sized nonvesiculated bodies were produced instead of vesiculated forms. The viability of cultures was reduced by rifampicin and novobiocin but not by chloramphenicol.
Expression of Clumping and Fibrinogen‐Binding Activities of Staphylococcus aureus at Various Growth Stages
Authors: Yonemasu Kunio , Sasaki Takako , Ohmae Riichi , Kashiba Syuzo
Published: November 2013

Clumping and fibrinogen‐binding activities of 4 Staphylococcus aureus strains (Cowan I, Newman D2C, Wood 46 and NCTC 5655) were assayed with a semiquantitative clumping test and an enzyme‐linked immunosorbent assay (ELISA), respectively. Distinct positive clumping was detected with whole cells of the 3 strains except Wood 46. Amounts of fibrinogen required for a definite clumping depended greatly on strains as well as on their growth phases. On the other hand, fibrinogen‐binding activities were detected both in culture supernatants and in cell lysates of all the 4 strains, and the levels were rather comparable with one another and relatively steady through their growth cycles. No significant correlation was thus found among expression behavior of clumping and fibrinogen‐binding activities.
Characterization of Antibody Response to Pseudomonas aeruginosa in Patients with Wound Infections
Authors: Trafny Elzbieta Anna , Grzybowski Jacek , Patzer Jan , Popiel Danuta , Wrembel‐Wargocka Jolanta , Dzierzanowska Danuta , Zawistowska‐Marciniak Irena , Kobus Kazimierz
Published: November 2013

ELISA was used to measure the amount and avidity of IgG antibodies to exotoxin A (ExA) and 7 Fisher's immunotypes of lipopolysaccharide (LPS) in the sera of 13 patients with mild or moderate Pseudomonas aeruginosa infections. Changes in the specificity of tested sera during the course of infection were demonstrated. A statistically significant increase was seen in the amount and avidity of the antibodies to ExA in a majority of the sera, and an increase was seen in amount of antibodies to LPS immunotype 4 in the sera of patients with moderate infections.
Factors Influencing the in vitro Growth of Mycobacterium leprae: Effect of Oxygen
Authors: Dhople Arvind Madhav , Lamoureux Laurie Christine
Published: November 2013

In our efforts to evaluate factors responsible for in vitro growth of Mycobacterium leprae in DH medium and also to improve the system, effects of oxygen tension on in vitro growth of M. leprae were determined. This was achieved by varying the ratio between DH medium and free air space above the medium in the culture tubes. Growth‐competent M. phlei (ATCC 11758) could tolerate all the oxygen it can get in the medium. On the other hand, M. leprae seemed to be of microaerophilic nature. In vivo‐grown M. leprae cells were more sensitive than their counterparts that were adapted to in vitro environment. In vivo‐grown cells grew better when 70oo of the space in culture tube was occupied by DH medium. These in vitro‐adapted cells gave optimum growth in subcultures when the air spaces in the culture tubes were 40–50oo. The role of oxygen tensions in the development of lesions in leprosy patients and armadillos has been discussed.
Intratypic Variations in Neutralizable Epitopes among Herpes Simplex Virus Type 2 Isolates
Authors: Bhattarakosol Parvapan , Yoosook Chalobon , Matangkasombut Pornchai
Published: November 2013

Intratypic variation among 94 isolates of herpes simplex virus type 2 (HSV‐2) was investigated using 4 different monoclonal antibodies (MAbs). By neutralization test, these MAbs appeared to be directed to at least 2 distinct epitopes on the viral glycoprotein D (gD), i.e., 6G6.G9 and 6E8.F11 which did not require complement (C MAb) and gD‐105 and gD‐110 whose neutralizing activities could be enhanced by complement (C+MAb). The C−MAb pairs each separately could detect significant intratypic variations among the isolates. Whether these variations also existed in the gD epitope(s) recognized by C+MAbs remains to be elucidated. The results suggested that intratypic variation occurred on at least one of the neutralizable (thus related to protective immunity) epitopes on gD of HSV‐2.
Cloning and Expression of cDNAs from Enterically‐Transmitted Non‐A, Non‐B Hepatitis Virus
Authors: Ichikawa Munetaka , Araki Masatake , Rikihisa Tetsuji , Uchida Toshikazu , Shikata Toshio , Mizuno Kyosuke
Published: November 2013

The fragment gene of enterically‐transmitted non‐A, non‐B hepatitis virus (ET‐NANBHV) was cloned as a cDNA and inserted into an expression vector pUEX2. The recombinant protein was expressed in Escherichia coli HB101 as a fusion protein with β‐galactosidase (β‐Gal). The fusion protein reacted with the sera of infected cynomolgus monkeys and of patients from Myanmar. This reaction was highly related with ET‐NANBHV infection, and obviously demonstrates in that the recombinant protein can be used for the detection of ET‐NANBHV infection.
Stimulation of Phagocytic Processes and Antibody‐Dependent Cellular Cytotoxicity of Human Neutrophils by Cefmetazole
Authors: Rodriguez Ana Beatriz , Barriga Carmen , de la Fuente Monica
Published: November 2013

Interactions between antimicrobial agents and phagocytic cells, especially neutrophils, have a potential role in the treatment of infections. The in vitro effects of cefmetazole, a novel β‐lactarn antibiotic, at a therapeutic concentration reached in plasma (50 μg/ml) on phagocytic and cytotoxic functions of human neutrophils have been studied. In human neutrophils, adherence capacity to nylon fiber and to substrate, chemotaxis, attachment to and ingestion of Candida albicans (with serum, with decomplemented serum and without serum), ingestion of inert particles (latex beads), candidicidal activity and superoxide anion production were all stimulated by cefmetazole. Cefmetazole at this dose was a chemotactic agent for neutrophils. Antibody‐dependent cellular cytotoxicity (ADCC) was also increased by this antimicrobial agent.
Lysis of Murine Macrophages by Lymphokine‐Activated Killer‐Like Cells Induced by BCG in vitro
Authors: Chen Ming‐feng , Suzuki Hidehiko , Yano Saburo
Published: November 2013

The lymphokine‐activated killer (LAK)‐like activity was found to be induced in mouse splenocytes cultured together with Bacillus Calmette‐Guérin (BCG). The killer cells induced by BCG were capable of killing both NK‐sensitive (YAC‐1, P388D1) and NK‐resistant (P815) tumor cells. As an important finding, they also lysed syngeneic macrophages (Mφ). The anti‐Mφ killer activity appeared on day 2, and reached a peak on day 5 of culture. Phenotype analysis of the killer cells by depletion techniques using monoclonal antibody (mAb) and complement indicated that the majority of these anti‐Mφ killer cells were Thy‐1+ and asiolo GM1 +. This Mφ cytolysis could be inhibited by the addition of cold Mφ, YAC‐1 tumor cells, and P815 tumor cells, suggesting that the same population of the effector cells recognize Mφ and tumor cells. The addition of anti‐MHC class I, anti‐MHC class II, anti‐L3T4, or anti‐Ly‐2 mAb directly to assay cultures did not affect anti‐Mφ cytolysis, suggesting that the MHC molecules are not involved in the cytolysis of Mφ by the BCG‐induced killer cells. The addition of anti‐LFA‐1 mAb partially inhibited the cytotoxicity, suggesting importance of the contact between targets and effectors in the cytolysis. Our present data suggest that activation of murine lymphocytes with BCG induces LAK‐like cells capable of killing a wide variety of tumor cells as well as Mφ and this anti‐Mφ cytolysis is mediated by nonspecific killer cells.
An Indirect Microhemagglutination Test for Lyme Disease
Authors: Fujita Hiromi , Sato Tadashi , Ohara Yoshiro , Homma Morio
Published: November 2013

We have developed an indirect microhemagglutination test (IMHA) for detecting antibodies to Borrelia burgdorferi using formalin‐fixed chicken erythrocytes sensitized with a sonicated extract of the bacteria. The specificity of IMHA for Lyme disease was compared with that of a conventional indirect immunoperoxidase technique (IP) using sera from the patients with syphilis, randomly‐chosen outpatients unrelated to Lyme disease, and from normal individuals. The IMHA was more specific than IP, giving little cross‐reaction with syphilis. The IMHA also gave little background reactions with the sera of individuals unrelated to Lyme disease or syphilis. Therefore, the IMHA could be a useful test for the serodiagnosis of Lyme disease.
Nucleosomal Structure of Marek's Disease Virus Genome in Transformed Lymphoblastoid Cell Lines, MDCC‐MSB1 and MDCC‐RP1
Authors: Hayashi Masanobu , Nakamura Kazuyo , Isogai Emiko , Namioka Shigeo
Published: November 2013

The latent MDV (Marek's disease virus) genomes are folded into nucleosomal structures in both virus‐productive and ‐nonproductive lymphoblastoid cell lines, MDCC‐MSB1 (MSB‐1) and ‐RP1 (RP‐1), respectively. There was no difference between transcriptionally active and inactive regions of MDV genome with regard to nucleosomal patterns. In order to investigate whether nucleosomal structure is correlated with the repression of the transcription of MDV genome in lymphoblastoid cells, we examined the DNasel sensitivity of nucleosomal MDV DNA in lymphoblastoid cell lines, MSB‐1 and RP‐1. No difference in the DNasel sensitivity between transcriptionally active and inactive MDV DNA regions in lymphoblastoid cells was observed.
Relative Ability of Distinct Isotypes of Human Major Histocompatibility Complex Class II Molecules in Binding Staphylococcal Enterotoxin A
Authors: Saito Shinji , Imanishi Ken'ichi , Araake Minako , Yan Xiao Jie , Igarashi Hideo , Uchiyama Takehiko
Published: November 2013

Relative ability of distinct isotypes of human major histocompatibility complex class II molecules to bind staphylococcal enterotoxin A (SEA) was investigated. SEA‐binding was observed in L cells transfected with DR2 and DQw1 genes. By contrast, it was not detected in L cells transfected with DPw4 and DP (Cp63) genes. All the transfectants supported SEA‐induced IL‐2 production by human T cells. Levels of the accessory activity were low in the DPw4 and DP (Cp63) transfectants compared with the DR2 and DQw1 transfectants. In view of the observation that all the transfectants express well the transfected gene products on their surface, the results indicate that DR and DQ molecules bind SEA with high affinity, while DP molecules bind it with fairly low affinity.
Susceptibility of a Herpes Simplex Virus Ribonucleotide Reductase Null Mutant to Deoxyribonucleosides and Antiviral Nucleoside Analogs
Authors: Yamada Yoshinari , Yamamoto Naohiko , Daikoku Tohru , Nishiyama Yukihiro
Published: November 2013

A herpes simplex virus ribonucleotide reductase (RR) null mutant, ICP6 Δ, exhibited hypersensitivity to hydroxyurea, and to the precursors of allosteric inhibitors of cellular RR. The mutant was also much more sensitive than the parental KOS to all of antiviral nucleoside analogs tested, including acyclovir (ACV), ganciclovir (DHPG) and BVaralJ. Our data indicate that cellular RR is essential for the growth of ICP6 Δ, and suggest that inhibitors of viral RR could act as potentiators of all of anti‐herpetic nucleoside analogs whose targets are viral DNA polymerase.
Serologic Survey of Spotted Fever Group Rickettsiosis on Hainan Island of China
Authors: Hui‐Min Feng , Tian‐Sheng Chen , Bi‐Hu Lin , Yin‐Zi Lin , Peng‐Fei Wang , Qun‐Hao Su , Hai‐Bo Xia , Kumano Keiko , Uchida Takahiro
Published: November 2013

A serosurvey for antibodies to Rickettsia japonica was conducted on Hainan Island of China. Serum specimens were collected from 1,030 outpatients at hospitals in different parts of the island regardless of their diagnosis. Only two among 538 serum specimens collected in Baoting and Tongshi counties, located in the southern part of the island, were demonstrated to contain antibodies reactive with R. japonica at a high dilution. The specimens also reacted with R. rickettsii at the same titer as with R. japonica. These two specimens reacted with other pathogenic spotted fever group (SFG) rickettsiae to a lesser extent. On the other hand, the specimens were shown to possess antibodies reactive with R. typhi at a significantly lower dilution or were not reactive at all. The findings suggested the occurrence of an SFG rickettsiosis on Hainan Island. More than half of the serum specimens collected from patients with suspected rickettsial infections in the southern area were found to contain IgM and IgG antibodies to R. typhi, indicating a high incidence of murine typhus.
Serological Survey for Lyme Disease in Sika Deer (Cervus nippon yesoensis) by Enzyme‐Linked Immunosorbent Assay (ELISA)
Authors: Isogai Emiko , Isogai Hiroshi , Masuzawa Toshiyuki , Yanagihara Yasutake , Sato Nanao , Hayashi Shunji , Maki Toshiyuki , Mori Mamoru
Published: November 2013

Seventy‐six wild sika deer (Cervus nippon yesoensis) from areas endemic for Borrelia burgdorferi during 1988 to 1989 had the IgG antibody to the bacteria in enzyme‐linked immunosorbent assay. The percentage of seropositive deer was 25.0% to strain HO14 and 22.4% to strain HP3, respectively. Specific IgG antibody titers were high in summer but low in winter. In summer, seropositive deer were 75.0%. Similar results were obtained in serum specimens obtained monthly from 4 farmed deer. It was suggested that the sika deer could be one of the wild reservoirs for B. burgdorferi in Hokkaido, Japan. The transmission risk of B. burgdorferi is considered to be greatest during late spring to early summer.
An Aberrant Hemolysin of Vibrio cholerae Non‐O1
Authors: Iwanaga Masaaki , Ichinose Yoshio
Published: November 2013

An aberrant hemolysin produced by a Vibrio cholerae non‐O1 strain N037 (N037‐hly) was purified and characterized. N037‐Hly was antigenically very similar to E1 Tor hemolysin but differed in molecular weight (48,000 vs. 60,000), interaction with glucose, and hemolytic activity. Of 100 V. cholerae non‐O1 strains other than the N037 strain examined, none produced this aberrant hemolysin. The N‐terminal amino acid sequence of N037‐hly was highly homologous to that of E1 Tor hemolysin.
Sensitive Detection of Viral Antigens with a New Method, “Laser Magnet Immunoassay”
Authors: Mizutani Hiroko , Suzuki Miwako , Mizutani Hiromichi , Fujiwara Koichi , Shibata Shuichi , Arishima Koichi , Hoshino Mitsutoshi , Ushijima Hiroshi , Honma Hitoshi , Kitamura Takashi
Published: November 2013

A new method, “laser magnet immunoassay” (LMIA), has been developed for sensitive detection of viral antigens. Target viruses captured on microbeads were made to react with antibodies labeled with magnetite particles. In a magnetic field, magnetically labeled antigens dispersed in water were attracted to and concentrated at one point on the surface, resulting in the lifting up of a small surface area. A laser beam which was incident on the point reflected, making an interference fringe. The intensity of the fringe indicates the amount of the magnetite conjugated with antigen. A very low concentration of antigens, such as 5 particles of influenza virus and 0.1 pg/ml of human immunodeficiency virus (HIV) p24 antigen in human serum, could be detected by this method. Application of this method to diagnoses of viral diseases in early stages is discussed.
B‐B Cell Interactions in the Spontaneous Activation of B Cells in Autoimmune NZB Mice
Authors: Saito Kazuyoshi , Tanaka Yoshiya , Ota Toshiyuki , Eto Sumiya , Yamashita Uki
Published: November 2013

We analyzed the mechanism of spontaneous B cell activation in lupus mice by using anticlass‐II antibody in vitro. The in vitro culture of B cells from old NZB mice markedly produced Ig without any stimulation, while B cells from NZW mice did not. The addition of anticlass‐II antibody (anti‐Iad antibody) to the culture inhibited Ig production of NZB B cells in a concentration‐dependent manner. On the other hand, the addition of anticlass‐I antibody (anti‐H‐2Dd antibody) and anticlass‐II antibody with different specificity (anti‐Iak) gave no effect on the Ig production of NZB B cells. When mitomycin C‐treated B cells were added to in vitro culture of responder B cells as a stimulator, Ig production of responder B cells was enhanced in a concentration‐dependent manner. However, the enhancing effect of the stimulator B cells was abrogated by the pretreatment with anticlass‐II antibody. The stimulator B‐cell activity to NZB B cells was marked in NZB B cells, moderate in NZB/W F1 B cells, and weak in NZW B cells. Furthermore, the stimulator B‐cell activity with regard to NZB B cells was marked in old female NZB B cells, moderate in old male NZB B cells, and weak in young NZB B cells. The expression of class II antigens on the surface of old female NZB B cells was significantly higher than that of old male NZB and young NZB B cells. These results suggest that in lupus mice the spontaneous B‐cell activation is induced by an abnormal B‐B cell interaction mediated by class II antigens.
Comparison of Immunochemical Specificities of Vibrio parahaemolyticus O10 and O12 Antigens Using Monoclonal Antibodies
Authors: Ogawa Setsuko , Koshikawa Tomihiko , Kawahara Atsumi , Watabe Kazuhito , Torii Mitsuo
Published: November 2013

The monoclonal antibodies against Vibrio parahaemolyticus O10 and O12 antigens (lipopolysaccharide, LPS) were prepared and specificities of the antibodies were examined. Five of six anti‐O10 antibodies reacted with O10 antigen, but none of them reacted with O12 antigen. On the contrary all of the five anti‐O12 antibodies reacted with O10 antigen as well as homologous O12 antigen. O10 and O12 antigens were subjected to alkali treatment or periodate oxidation, and reactivities of these chemically modified preparations with the monoclonal antibodies were examined. Reactivities of O10 with anti‐O10 and anti‐O12 antibodies were reduced by the above two chemical treatments, but that of O12 with anti‐O12 was not. O‐Deacetylation of O10 LPS by the alkaline treatment was confirmed by NMR spectroscopy. These results suggest contributions to O10‐specificity of O‐acetyl group and periodate sensitive sugar residue. Inhibition experiments of O10 and O12 homologous precipitations were also carried out with various sugars. From the results we concluded that O10 and O12 antigenic determinants were distinct entities, although O10 and O12 antigens have been reported to be similar and cross‐reactive.
Bacterial Translocation from the Gastrointestinal Tract in Various Mouse Models for Human Diabetes
Authors: Ohsugi Takeo , Maejima Kazuyoshi , Urano Toru
Published: November 2013

Bacterial translocation from the gastrointestinal (GI) tract to other internal organs was examined in multiple low‐dose streptozotocin‐injected (M‐STZ), single large‐dose streptozotocin‐injected (S‐STZ), alloxan‐injected (Alloxan), and non‐obese diabetic (NOD) mice. The incidence of bacterial translocation from the GI tract to the tested organs among diabetic mice was in the order of M‐STZ mice>S‐STZ mice>NOD, Alloxan, and control mice. The injections of insulin to M‐STZ mice did not decrease the incidence of translocation. These results suggest that bacterial translocation from the GI tract in diabetic mice is not induced by diabetes.
Novel Cell Surface Antigens Expressed on Mouse Alveolar Macrophages
Authors: Yoshikawa Kazuhiro , Suzuki Yasumoto , Kawai Makoto , Fukada Masako , Yokochi Takashi
Published: November 2013

Two new cell surface antigens expressed on mouse alveolar macrophages were defined by rat monoclonal antibodies. One marker, AVM‐1, was detected on mouse alveolar macrophages, but it was undetectable on resident peritoneal cells, thioglycollate medium‐induced peritoneal cells, and splenic macrophages. Splenic lymphocytes, thymocytes and bone marrow cells were also AVM‐1 negative. Anti‐AVM‐1 monoclonal antibody immunoprecipitated a single polypeptide with a molecular weight of 200,000. Of particular interest was the finding that the anti‐AVM‐1 antibody could inhibit the formation of EA and EAC rosette on macrophage line cells. A second antigen (AVM‐2) was also present on alveolar macrophages. and its molecular weight was 38,000.
New Enzyme Immunoassays for Specific Assay and General Detection of Trypanosoma cruzi, Epimastigotes
Authors: Kitagawa Tsunehiro , Iwamoto Masayuki , Zhao Li‐ping , Kanbara Hiroji
Published: November 2013

A highly specific competitive enzyme‐linked immunosorbent assay for the epimastigote of Tulahuen strain was developed by using the usual 3 immunological reagents, a rabbit antiserum specific for T. cruzi, epimastigote of Tulahuen strain, β‐D‐galactosidase‐labeled goat anti‐rabbit immunoglobulin G and the solid‐phase cell fragments of the epimastigote of Tulahuen strain. A new method, the selected antibody enzyme immunoassay (SAEIA) which generally detected all strains of the epimastigote tested with the same working range, was developed by changing only the solid‐phase antigen to the epimastigote of Y strain among the 3 immunological reagents. Both assays permitted us to measure accurately as little as 1,000 parasites per assay tube. Scope of the SAEIA was limited to the epimastigote. Both life‐cycle forms of T. cruzi which appear in mammals, amastigote and trypomastigote, and other kinetoplastids showed low cross‐reaction values by the assay. The assay principle of the new method and a preliminary study to apply the SAEIA for finding the field T. cruzi‐infected insect vectors were also reported.
Serial Observations of Chronic Rotavirus Infection in an Immunodeficient Child
Authors: Oishi Isao , Kimura Teruo , Murakami Tsukasa , Haruki Kosuke , Yamazaki Kenji , Seto Yoshiyuki , Minekawa Yoshiichi , Funamoto Hitokazu
Published: November 2013

Chronic rotavirus infection of an infant with severe combined immunodeficiency (SCID) was studied by virological examinations in association with long‐term observation of his symptoms and immune status. During eleven months of hospitalization, the patient was suffering from incurable severe diarrhea with persisting excretion of rotaviruses detected by electron microscopy and the reversed‐passive hemagglutination (R‐PHA) test and had transient hepatitis symptom despite multiple administrations of human gammaglobulin and high calorie fluids. The detected viruses were morphologically recognized as rotavirus with double capsid structure. Polyacrylamide gel electrophoretic (PAGE) analysis of their genomic RNAs showed the long electropherotype of group A virus with abnormal migration profiles changing considerably from the early to the late phase of illness: (1) The 11th segment became undetectable; (2) the molecular weight of the 6th segment slightly increased; (3) seven to fourteen extra segments appeared; and (4) PAGE patterns of viral genomic RNAs changed every three or four months. These findings suggest that chronic infection with rotavirus accompanied the generation of extra viral genomic segments and their unusual assortments in an immunodeficient host.
Seroprevalence of Human Herpesvirus 6 Infection in Brazilian and Japanese Populations in the North‐East of Brazil
Authors: Linhares Maria Iêda Siqueira , Eizuru Yoshito , Tateno Seiki , Minamishima Yoichi
Published: November 2013

The seroprevalence of human herpesvirus 6 (HHV‐6) infection was examined for 434 Brazilians and 250 Japanese immigrants living in Recife and its vicinity, in the North‐East of Brazil. A total of 684 sera from the healthy individuals were screened for IgG antibodies to HHV‐6 by anticomplement immunofluorescence (ACIF) test. The seropositivity rate to HHV‐6 showed little difference between the two groups: namely, it was 76.5% for Brazilians and 77.2% for Japanese immigrants. The seropositivity rate was constantly higher in females than in males. The high prevalence of anti‐HHV‐6 antibodies among children indicates that HHV‐6 infection occurs very early in life.