bluuebunny/crossref_metadata_2025_split huggingface dataset:
Electrophoretic Karyotypes of Candida Yeasts Recurrently Isolated from Single Patients
Authors: Doi Matsuko , Mizuguchi Ikuyo , Homma Michio , Tanaka Kenji 
Published: November 2013

AbstractCandida strains were isolated repeatedly from single patients during recurrent episodes of Candida infection in a hospital, and their electrophoretic karyotypes were analyzed by pulsed‐field gel electrophoresis using CHEF system. When only C. albicans (in 6 patients) or C. glabrata (in 1 patient) were recurrently isolated, their karyotypes from each patient were almost identical to one another, suggesting that they carried single type of the yeast. When multiple species were recovered from single patients (6 cases), the karyotypes of the most frequently recovered yeast species were almost identical with respect to each patient. The electrophoretic karyotype analysis has been proved to be useful for epidemiological studies because the method can tell not only the species identification but also the differences among the strains of the same species.
Helicobacter pylori: A Fickle Germ
Authors: Cellini Luigina , Allocati Nerino , Di Campli Emanuela , Dainelli Benedetto 
Published: November 2013

AbstractThe morphologic changes from bacillary to coccoid forms of Helicobacter pylori were studied. These form changes were analyzed by bacterial growth in Brucella broth plus 2% fetal calf serum. The coccoid forms were observed at five days of incubation and a rapid decrease of CFU/ml was recorded. At two weeks of microaerophilic incubation, all coccoid forms observed were not culturable in vitro. The coccoid morphology was observed earlier when the culture of H. pylori was incubated in aerobic conditions and with subinhibitory concentrations of omeprazole and roxithromycin. To evaluate the possibility of resistance of coccal forms, before plating, the cultures were heated to 80 C for 10 min and sonicated. In the absence of these treatments the cultures did not show growth in vitro. The proteic patterns of the same strains of two different morphologies were studied revealing significant differences.
The Transposon‐Like Structure of IS26‐Tetracycllne, and Kanamycin Resistance Determinant Derived from Transferable R Plasmid of Fish Pathogen, Pasteurella piscicida
Authors: Kim Eun‐heui , Aoki Takashi 
Published: November 2013

AbstractTetracycline (pp‐tet), and kanamycin (pp‐kan) resistance genes were cloned from a transferable R plasmid of fish pathogen Pasteurella piscicida, and complete nucleotide sequences were determined. The pp‐tet was a class D Tet determinant constructed with the tetA resistance gene of 1,182 bp encoding a protein with a deduced molecular mass of 41 kDa and the tetR repressor gene of 654 bp encoding a product of 24 kDa. The pp‐tet was highly homologous to the tet(D) of plasmid RA1 isolated from Aeromonas hydrophila with two nucleotide differences in the tetR, and of plasmid pIP173 from Salmonella ordonez with two nucleotide differences in the tetA. The pp‐kan contained 813 bp encoding a 31 kDa protein of 271 amino acids, and was classified into type aph‐Ic. It was identical to the aphA7 in the IAB operon of pBWH77, in which was originally found an isolate of Klebsiella pneumoniae, in its nucleotide sequences and hybrid promoter construction. The genes were connected by an insertion sequence IS 26 of 820 bp, and were flanked by repeated copies in direct orientation at the 3′ flanking region of the pp‐tetA and in inverted orientation at the 3′ flanking region of the pp‐kan. The genetic elements are organized like a complex transposon by close linkage of the IS 26 and the pp‐tet and ‐kan.
Evaluation of the Sensitivity and Specificity of Polymerase Chain Reaction Test Kit, AMPLICOR Chlamydia trachomatis
Authors: Miyashita Naoyuki , Iijima Yoshio , Matsumoto Akira 
Published: November 2013

AbstractThe sensitivity and specificity of the polymerase chain reaction (PCR) test kit, AMPLICOR Chlamydia trachomatis, were examined by the use of purified elementary bodies (EBs), cells having inclusions containing reticulate bodies alone and 20 clinical isolates. The numbers of EB and inclusion of C. trachomatis at the detection limit were determined to be approximately 2 to 4 EBs and one inclusion per assay, respectively. No reaction occurred for C. psittaci and C. pneumoniae. All clinical isolates were positively reacted in the PCR assay.
Epidemiological and Ecological Studies of Japanese Encephalitis in Okinawa, Subtropical Area in Japan. II. Prevalence of Japanese Encephalitis Antibody in Residents in Okinawa, Miyako and Ishigaki Islands
Authors: Tadano Masayuki , Kanemura Kazumi , Arakaki Sakae , Makino Yoshihiro , Fukunaga Toshihiko , Hokama Tetsuo , Kyan Takeyoshi , Makiya Hiroshi , Taira Youichi , Ohama Nagateru , Iju Choyu , Sakurai Kaori 
Published: November 2013

AbstractDuring 1989 to 1990, human sera were collected by age groups in Okinawa (the northern, central and southern areas), Miyako and Ishigaki islands and examined for the neutralization (N) antibodies to two strains, Nakayama (vaccine strain) and C307 (Okinawan strain), of Japanese encephalitis (JE) virus. In Okinawa island, the N antibody positive rate to C307 was higher than that to Nakayama, while in Miyako and Ishigaki islands, the positive rate to Nakayama was higher than that to C307, suggesting that JE virus transmission rate was higher in Okinawa than in Miyako and Ishigaki islands. In Okinawa Prefecture, JE vaccine had not been administered to most of residents over 31 years of age at the time of serum collection. In residents over 31 years old, the positive rate to C307 was highest in the north of Okinawa (83.3%) and was lowest in Miyako (26.3%), with the second lowest in Ishigaki (33.3%). The distribution of N antibody titers to C307 gave hyperbolic patterns in the 0–5 age groups in Miyako and Ishigaki, and also in the 31–40, 41–50 age groups in Miyako and the 41–50 age group in Ishigaki, suggesting low rates of natural infection in these 4–5 decades in both islands. In residents of ages subjected to JE vaccine, a characteristic pattern was obtained, in which the curves to Nakayama shifted to higher titers than those to C307, suggesting that the first antigenic stimulation was caused by vaccine, not by natural infection of JE virus.
A Function of 40 kDa Outer Membrane Protein in Serratia marcescens
Authors: Tada Yoshifumi , Yamaguchi Junji 
Published: November 2013

AbstractThe function of one of the outer membrane proteins of Serratia marcescens was investigated. S. marcescens with an abundant 40 kDa outer membrane protein was induced to form spheroplast at a high rate in an isotonic medium in the presence of calcium, although the spheroplasts were generally fragile in the isotonic environment. The degree of spheroplast induction was correlated to the amount of the 40 kDa protein present in the membrane. In the 40 kDa proteinless mutant strains, the spheroplast induction rate was remarkably decreased. Autoradiography of the outer membrane revealed the presence of a calcium‐binding protein as a radioactive band whose position coincided with the 40 kDa protein. These results suggest that the 40 kDa protein has an important role in maintaining the structural integrity of the cell wall against osmotic shock.
Effective Induction of Human NK Cells with OK‐432 and Further Augmentation of Their Cytolytic Function by rIL‐2
Authors: Hu Zhao‐liang , Kubota Eiro , Imamura Hideo , Katano Mitsuo , Katsuki Takeshi 
Published: November 2013

AbstractLow concentrations of exogenously added recombinant interleukin 2 (rIL‐2) were able to augment OK‐432‐induced natural killer (NK) cell activity. This kind of augmenting effect depended on the dose of rIL‐2 and manifested itself only in PBMC stimulated with OK‐432 (OK‐MC) followed by rIL‐2; augmentation did not happen in the reverse order. The existence of CD16+/CD25+ (IL‐2 receptor positive; IL‐2R+) and CD57+/CD25+ double positive cells which possess NK cell surface markers in OK‐MC markedly increased in a long‐term culture (12 days). A strong positive correlation was observed between the IL‐2‐dependent augmentation of NK activity and the quantitative changes in cell populations that possessed NK cell phenotypes. Treatment of the day‐12‐OK‐MC with monoclonal anti‐CD56 antibody plus complement could almost completely abrogate the augmented NK cytotoxicity. Furthermore, this augmenting effect was detectable within 4 hr after addition of rIL‐2 at single cell level, suggesting that the effect did not require NK cell's DNA synthesis. Thus it was suggested that OK‐432 could promote and upregulate the expression of IL‐2 receptor (CD25) on CD56+ NK cell populations. Moreover, it was considered that the interaction of low concentration rIL‐2 with IL‐2 receptors on OK‐432‐activated NK cells could augment their lytic function.
Pili of a Vibrio cholerae O139
Authors: Nakasone Noboru , Yamashiro Tetsu , Albert M. John , Iwanaga Masaaki 
Published: November 2013

AbstractThe pili of a strain of Vibrio cholerae O139 were purified and characterized. They were morphologically, electrophoretically and immunologically indistinguishable from the pili with 16 kDa subunit protein of V. cholerae O1. All 22 strains of V. cholerae O139 examined possessed the pili. The pili were different in hemagglutination inhibition pattern from V. cholerae O1 16K pili.
Purification and Characterization of Membrane Protein (90 kDa) from Mycoplasma salivarium, Which Binds Immunoglobulin (Ig) G Fc Fragment
Authors: Sawa Yoshihiko , Shibata Ken‐ichiro , Noda Mamoru , Watanabe Tsuguo 
Published: November 2013

AbstractA 90 kDa protein of Mycoplasma salivarium was released from cell membranes of the organism with Triton X‐100 and purified by ion‐exchange chromatography and chromatofocusing. The protein was eluted at pH 5.5 by chromatofocusing. The protein was shown to react with the Fc fragments of IgG from human and nine different animal species and did not distinguish between IgG from different species, while protein A, tested for comparative purposes, displayed a strong specificity for human and swine IgG. Furthermore, the protein reacted with antigen specific goat IgG (specific for gamma chains of human IgG), sheep red blood cells (SRBC) sensitized with rabbit antiserum to SRBC, that is, the Fc part of rabbit IgG, and concanavalin A as well. These findings may suggest that the protein is a lectin which binds the carbohydrate moiety of the Fc part of IgG.
Experimental Induction of Lyme Arthritis in Outbred Mice
Authors: Masuzawa Toshiyuki , Kurita Toru , Kawabata Hiroki , Suzuki Harumi , Yanagihara Yasutake 
Published: November 2013

AbstractOutbred ddY mice inoculated with live cells ofBorrelia burgdorferistrain 297 into hind footpad displayed swelling of the footpad at days 7 to 11 after inoculation. Marked neutrophilic infiltration was observed in the subcutaneous tissue and the part of bone tissue which was partially destroyed, and synovial layer of articular capsule was thickened and protruded into the joint space in the histopathological examination of footpad inoculated with liveBorreliacells. The inflammation peaked at day 7 andB. burgdorferiwas cultured from bladder and heart of the mice at day 14 after inoculation. The mice inoculated with heat‐inactivated cells at 56 C for 30 min did not show any significant histopathological change. In this mice model, nontreated littermates were not infected in contact with infected littermates for 14 days of experimental period. The outbred ddY mice model is useful for evaluating the effectiveness of vaccination against Lyme disease.
“Silent” Hepatitis B Virus Mutants Are Responsible for Non‐A, Non‐B, Non‐C, Non‐D, Non‐E Hepatitis
Authors: Uchida Toshikazu , Shimojima Masakazu , Gotoh Kenichirou , Shikata Toshio , Tanaka Eiji , Kiyosawa Kendo 
Published: November 2013

AbstractSince the recent introduction of diagnostic kits for hepatitis C and E, some cases of nonA, non‐B, non‐C, non‐D, non‐E hepatitis (so‐called hepatitis F) have been revealed. We attempted to demonstrate that so‐called hepatitis F is caused by hepatitis B virus (HBV) variants. Polymerase chain reaction (PCR) was used to amplify serum HBV DNAs from 20 patients with acute hepatitis and 20 patients with chronic hepatitis who had been diagnosed as having so‐called hepatitis F on the basis of conventional serological markers. The PCR technique successfully amplified HBV DNAs in 18 (90%) cases of acute hepatitis and 17 (85%) cases of chronic hepatitis. Sequencing of HBV DNAs of six patients (acute 3, chronic 3) revealed equally a T‐to‐C mutation of DR2 and an 8‐nucleotide deletion of the 3′‐terminus of the X gene coding region, giving rise to the generation of a C‐terminally truncated X protein and probable damage to the enhancer II/core promoter elements. These mutations of the X gene coding region may lead to suppression of replication and expression of HBV DNAs. Thus virtually all cases of so‐called hepatitis F appear to be caused by “silent” HBV mutants, at least in Japan.
Detection of Antibody‐Coated Bacteria in Expectorated Sputum for Diagnosis of Lower Respiratory Infections
Authors: Matsumoto Tetsuya , Kaku Mitsuo , Tateda Kazuhiro , Furuya Nobuhiko , Hirakata Yoichi , Yamaguchi Keizo 
Published: November 2013

AbstractWe evaluated antibody‐coated bacteria (ACB) in expectorated sputum to discriminate contaminating or colonizing organisms from true pathogens. We examined 60 expectorated sputum samples from 51 patients with lower respiratory infections (chronic obstructive pulmonary disease 25, pneumonia 20, purulent tracheobronchitis 6). All samples were examined with quantitative culture and immunofluorescent demonstration of ACB. From the results of quantitative culture, we divided specimens into pathogen‐isolated and pathogen‐free samples. Among pathogen‐isolated samples, in which we isolated accepted pathogenic organisms at ≥ 107 colony‐forming units per ml, 16 of 23 samples were ACB‐positive (69.5%). In contrast, among pathogen‐free samples, in which we isolated accepted pathogens at < 107 colony forming units per ml or only upper respiratory flora, only 3 of 37 samples were ACB‐positive (8.1%). The ACB‐positive rate was significantly higher in pathogen‐isolated than in pathogen‐free samples (P < 0.001). Consequently, detecting ACB in expectorated sputum shows good potential as another criterion for distinguishing contaminating or colonizing organisms from true pathogens.
Helicobacter pylori Induces Inflammation in Mouse Urinary Bladder and Pelvis
Authors: Isogai Hiroshi , Isogai Emiko , Kimura Koichi , Fujii Nobuhiro , Yokota Kenji , Oguma Keiji 
Published: November 2013

AbstractHelicobacter pylori was transurethrally inoculated into the mouse urinary tract. The organism established infection and induced inflammation in the urinary bladder and pelvis. During the infection, urinary pH was elevated, probably due to the production of NH3 by bacterial urease. H. pylori was recovered from the urinary bladder, kidney and urine of the infected mice. Histopathologically, severe neutrophil infiltration was observed in the mucosal layer of both organs. H. pylori was detected on the surface of the epithelial cells. These results indicate that low pH and bacterial flora were not essential factors in establishing the mucosal infection with H. pylori. This experimental system is useful to investigate the pathogenicity of H. pylori in mucosal organs.
The Rice Culture Filtrate of Bacillus cereus Isolated from Emetic‐Type Food Poisoning Causes Mitochondrial Swelling in a HEp‐2 Cell
Authors: Sakurai Naomi , Koike Kazuko A. , Irie Yuji , Hayashi Hideo 
Published: November 2013

AbstractRice culture filtrates of Bacillus cereus SA‐50, an emetic‐type strain, produced a toxin which caused cytoplasmic vacuole formation in HEp‐2 and HeLa cells. Electron microscopic observation revealed that the apparent vacuoles in HEp‐2 cells seen under a light microscope were actually swollen mitochondria. The oxygen consumption of HEp‐2 cells was accelerated by the addition of the rice culture filtrate as was measured with a polarographic oxymeter; a respiratory control ratio was 1.0 for control cells, while 1.4 for ones with the filtrates. The culture filtrates showed a similar effect on the isolated mouse liver mitochondria; respiratory control ratios for the mitochondria with and without the filtrates were 3.6 and 1.0, respectively. The affecting manner of the culture filtrates on the oxygen consumption of mitochondria was similar to that of 2,4‐dinitrophenol, suggesting that the culture filtrate contains a toxin acting as an uncoupler of oxidative phosphorylation in mitochondria. It is likely that the culture filtrates containing the emetic toxin of B. cereus causes mitochondrial swelling with a close relationship to the uncoupling of the oxidative phosphorylation of mitochondria.
Regulation of Iron on Bacterial Growth and Production of Thermostable Direct Hemolysin by Vibrio parahaemolyticus in Intraperitoneal Infected Mice
Authors: Wong Hin‐chung , Lee Yeong‐sheng 
Published: November 2013

AbstractPathogenesis of Vibrio parahaemolyticus is not clearly understood. Effects of iron on the bacterial proliferation and production of thermostable direct hemolysin (TDH) in intraperitoneal infected mice were studied. Injection of bacterial culture in the presence of ferric ammonium citrate (100 μg/ml) significantly enhanced the lethality for mice, and simultaneously activated bacterial proliferation in vivo. The iron‐limited cultures showed better proliferation than those iron‐rich cultures in response to the addition of supplementary iron source. Production of TDH by the hemolytic strains ST550 and D62 was higher in the iron‐limited cultures than the iron‐rich cultures. Production of TDH by both the iron‐limited or iron‐rich cultures was inhibited by the addition of iron. In conclusion, the virulence enhancement effect of iron in V. parahaemolyticus was probably by activating bacterial proliferation in vivo and not by stimulating the production of TDH. V. parahaemolyticus precultured in iron‐limited condition may be more adaptable to in vivo environment.
Cross‐Reactivity and Neutralizing Ability of Monoclonal Antibodies against Microcystins
Authors: Saito Shinji , Nakano Yasunobu , Kushida Katsutoshi , Shirai Makoto , Harada Ken‐ichi , Nakano Masayasu 
Published: November 2013

AbstractMonoclonal antibodies (MAbs) against the microcystin‐leucine‐arginine variant (MCYST‐LR), a cyclic peptide toxin of the freshwater cyanobacterium Microcystis aeruginosa, were prepared from cloned hybridoma cell lines. The specificity of the MAbs and their ability to neutralize the toxin were investigated by an indirect enzyme‐linked immunosorbent assay (ELISA) and by a neutralizing test in mice, respectively. All MAbs reacted with MCYST‐LR and also with the microcystin‐arginine‐arginine variant (MCYST‐RR), 3, 7‐didesmethylmicrocystin (MCYST‐3, 7‐dDMLR) and 7‐desmethylmicrocystin (MCYST‐7‐DMLR). Furthermore, the antibodies reacted with cell‐extracts of toxic and non‐toxic M. aeruginosa strains. The MAbs can apparently recognize the common configuration, but not the variant‐specific structure, in the microcystin molecules. The non‐toxic strains apparently contain some substance(s) related antigenically to microcystin. The in vivo toxin‐neutralizing ability of MAbs was minimal.
Construction of Mutant Genes for a Non‐Toxic Verotoxin 2 Variant (VT2vp1) of Escherichia coli and Characterization of Purified Mutant Toxins
Authors: Cao Chunyu , Kurazono Hisao , Yamasaki Shinji , Kashiwagi Keiko , Igarashi Kazuei , Takeda Yoshifumi 
Published: November 2013

AbstractThe gene encoding a Verotoxin 2 variant, VTvp1, was mutated by oligonucleotide‐directed site‐specific mutagenesis. Among 6 mutant toxins encoded by the mutated genes, E167Q‐R170L (glutamic acid at position 167 and arginine at position 170 from N‐terminus of the A subunit were replaced by glutamine and leucine, respectively) was found to have markedly decreased activities; inhibition of protein synthesis, Vero cell cytotoxicity and mouse lethality of the purified E167Q‐R170L were 1/1,900, 1/125,000 and 1/2,000, respectively, of those of the purified wild‐type VT2vp1. Since the antigenic property of the E167Q‐R170L was demonstrated to be similar to that of the wild‐type VT2vp1 by Ouchterlony double gel diffusion test and by neutralization test of Vero cell cytotoxicity of the VT2vp1, a possibility to use the mutant VT2vp1, E167Q‐R170L, as a toxoid is discussed.
Coaggregation between Porphyromonas gingivalis and Mutans Streptococci
Authors: Kamaguchi Arihide , Baba Hisae , Hoshi Masaaki , Inomata Koshiro 
Published: November 2013

AbstractCoaggregation occurred between Porphyromonas gingivalis and mutans streptococci. The coaggregation was completely inhibited by l‐arginine, Nα‐p‐tosyl‐l‐lysine chloromethyl ketone (TLCK), and a trypsin inhibitor, and weakly inhibited by l‐lysine, N‐ethylmaleimide, lysozyme, and human whole saliva. The results of heat and proteinase K treatment suggested that a heat‐labile proteinaceous substance of P. gingivalis and a heat‐stable substance of mutans streptococci may play a role in the coaggregation. Mutans streptococci also aggregated in the presence of the heat‐labile factor in the supernatant of P. gingivalis. The aggregation was also inhibited by l‐arginine, TLCK, and a trypsin inhibitor.
A Unique Mutation of Glycoprotein Gene of the Attenuated RC‐HL Strain of Rabies Virus, a Seed Virus Used for Production of Animal Vaccine in Japan
Authors: Ito Hiroshi , Minamoto Nobuyuki , Watanabe Tomiyoshi , Goto Hideo , Rong Luo Ting , Sugiyama Makoto , Kinjo Toshio , Mannen Kazuaki , Mifune Kumato , Konobe Takeo , Yoshida Iwao , Takamizawa Akihisa 
Published: November 2013

AbstractAlthough the RC‐HL strain of rabies virus is avirulent in adult mice, the amino acid at position 333 of its G protein is arginine, which is thought to be necessary for virulence in adult mice upon intracerebral inoculation of the virus. This result suggests that besides arginine at position 333, some other positions of G protein might also be involved in determining the virulence of rabies virus.
Detection of Human Immunodeficiency Virus‐1 Nucleic Acid on Inactivated Filter Paper Disks by Polymerase Chain Reaction and Microtiter Plate Assay
Authors: Kunisada Takao , Ando Shuji , Saito Kunihiro , Eshita Yuki , Röder Wolfgang , Kruse Michael , Müller Werner E. G. , Ushijima Hiroshi 
Published: November 2013

AbstractHuman immunodeficiency virus type 1 (HIV‐1) in cultured cells, peripheral blood samples and sera were adsorbed on filter paper disks and inactivated by heat or ethanol. Two procedures, the polymerase chain reaction (PCR) and microtiter plate assay (HMPA) were used to detect the nucleic acid. The sensitivity after different heat treatments with nested PCR for HIV‐1 DNA (or nested reverse transcription‐PCR for HIV‐1 RNA) was identical regardless of whether the samples were examined immediately or one month later. Inactivation by ethanol treatment resulted in a slight loss of sensitivity. The HMPA proved to be as reliable and specific as the conventional PCR technique. We conclude that the heat‐treated filter paper disk assay is suitable for identifying HIV nucleic acid in clinical samples sent to the laboratory from a distance, e.g. in an envelope.