bluuebunny/crossref_metadata_2025_split huggingface dataset:
Specific Detection of a Verotoxin 2 Variant, VT2vp1, by a Bead‐Enzyme‐Linked Immunosorbent Assay
Authors: Cao Chunyu , Yamasaki Shinji , Lin Zaw , Kurazono Hisao , Takeda Yoshifumi 
Published: November 2013

AbstractA bead‐enzyme‐linked immunosorbent assay to specifically detect a Verotoxin 2 variant, VT2vpl, was developed. The sensitivity of the bead‐ELISA was 200 pg/ml of the purified VT2vpl and it did not react with 20 ng/ml of the purified VT2. The specificity of the bead‐ELISA was examined with 107 strains of Verocytotoxin‐producing Escherichia coli that include VT1‐, VT2‐, VT2vha‐, VT2vhb‐ and VT2vp1‐producing E. coli, and only VT2vp1‐producing E. coli that were confirmed by VT2vp1‐specific polymerase chain reaction gave positive results. It was noted that all 58 VT2vp1‐producing E. coli strains were from pigs, but not from cows and humans.
Existence of a Novel Hemagglutinin Having No Protease Activity in Vibrio mimicus
Authors: Alam Munirul , Miyoshi Shin‐ichi , Maruo Ikuyo , Ogawa Chiemi , Shinoda Sumio 
Published: November 2013

AbstractThe protease elaborated by Vibrio mimicus is known to possess hemagglutinating ability to chicken erythrocytes, the well‐known HA/protease. A non‐protease hemagglutinin (HA) with strong agglutinating ability towards rabbit erythrocytes was obtained from 32 hr culture supernatant of a pathogenic environmental strain of V. mimicus. This HA (V. mimicus HA: VMHA) appeared stable at relatively higher temperature and agglutinated the erythrocytes from rabbit, guinea pig and mouse but not the erythrocytes from chicken, bovine, horse and sheep. Simple sugars, metal ions and chelating agents failed to inhibit the activity of VMHA. The activity of VMHA was found to be sensitive to digestion by proteolytic enzymes including HA/protease. These results provide evidence for the existence of novel HA other than HA/protease in V. mimicus.
Detection and Characterization of Antibodies to Bacterial Heat‐Shock Protein 60 in Sera of Patients with Primary Biliary Cirrhosis
Authors: Yamaguchi Hiroyuki , Miura Hitoshi , Ohsumi Kazuoki , Ishimi Norikuni , Taguchi Haruhiko , Ishiyama Narihiro , Shiraishi Yasumori , Yamamoto Tomoko , Ogata Sachio 
Published: November 2013

AbstractThe enzyme‐linked immunosorbent assay (ELISA) with bacterial heat‐shock protein 60 (HSP60) purified from Yersinia enterocolitica (Ye) revealed that the antibodies directed against YeHSP60 existed in sera of patients with primary biliary cirrhosis (PBC). To characterize the epitope specificity of the antibodies in patients, the epitope mapping of HSP60 by means of the antibodies was performed. The results have suggested that the epitope recognized with anti‐HSP60 antibodies in PBC relates to the amino acid sequence of YeHSP60 molecule as follows: DLGQA‐KRVVINKDTTIIIDGVGDEAAIQGRLAQIRQQIEEATSDYDKEK.
Genotyping of Staphylococcus epidermidis by Small‐Fragment Restriction Endonuclease Analysis and Pulsed‐Field Gel Electrophoresis of Genomic Restriction Fragments
Authors: Haertl Rolf , Bandlow Gert 
Published: November 2013

AbstractSmall‐fragment restriction endonuclease analysis (SF‐REA) was established as a typing tool for Staphylococcus epidermidis. A total of 60 isolates comprising 48 epidemiologically nonrelated strains and 12 putatively linked isolates from 7 patients in 2 wards were analyzed. Nonrelated isolates were characterized by unique fingerprints when DNA was cleaved with EcoRI or ClaI, electrophoretically separated in a polyacrylamide gel, and silver stained. Three blood culture isolates from one patient in an intensive care unit, 4 isolates obtained from a child over a span of 2 weeks, and 5 isolates from 5 newborns in the same ward were grouped into 3 DNA pattern types, indicating identity of sequential isolates from 2 patients and nosocomial transmission of one Staphylococcus epidermidis strain between 5 babies. Results from pulsed‐field gel electrophoresis of SmaI and SacII DNA digests and conventional marker systems such as antibiogram and plasmid profile were in accordance with these interpretations, whereas slight variation was observed in the biotypes of several strains. From the results of this study, we conclude that SF‐REA is a precise and efficient method for the genotypic characterization of Staphylococcus epidermidis strains that can be used as a rapid and reliable typing tool.
Pig Intestinal Membrane‐Bound Receptor (Guanylyl Cyclase) for Heat‐Stable Enterotoxin: cDNA Cloning, Functional Expression, and Characterization
Authors: Wada Akihiro , Hirayama Toshiya , Kitao Saori , Fujisawa Jun‐ichi , Hidaka Yuji , Shimonishi Yasutsugu 
Published: November 2013

AbstractA cDNA encoding the receptor protein for a heat‐stable enterotoxin (STa) produced by enterotoxigenic Escherichia coli was cloned from intestinal epithelial cells of a 10‐week‐old pig. The cDNA had an open reading frame of 3,219 base pairs and coded for a protein with 1,073 amino acid residues. The mature protein consisted of 1,050 amino acid residues with a molecular mass of ca. 121 kDa and was 87% and 82% identical with the human and rat protein, respectively. The CHO cell line overexpressing the pig recombinant STa receptor specifically bound to a photoaffinity‐labeled analog of STa and showed marked elevation of the cellular content of cGMP in response to STa.
Stimulation of Macrophage Oxygen Free Radical Production and Lymphocyte Blastogenic Response by Immunization with Porins
Authors: Tabaraie Bahman , Sharma Bal Krishan , Sharma Praveen Rishi nee , Sehgal Rakesh , Ganguly Nirmal Kumar 
Published: November 2013

AbstractPorins were prepared from smooth strain of Salmonella typhi 0–901 and chemotype rough mutant of S. typhimurium Ra‐30. Porins could significantly stimulate the immune systems of mice. Immunization of mice with the porins provoked synthesis of anti‐porin antibodies. Macrophages from the immunized mice showed increased capacity to generate oxygen free radicals, and lymphocytes from these mice showed proliferative response to the porins. Thus porins may play a role in providing protection from salmonellosis by stimulating the antibody production and increasing the capacity of macrophages to generate oxygen free radicals along with stimulation of lymphocytes.
Analysis of Major Surface Polypeptides of Rickettsia japonica
Authors: Uchiyama Tsuneo , Uchida Takahiro , Walker David H. 
Published: November 2013

AbstractMajor surface polypeptides of Rickettsia japonica migrated to the position of 120, 135, and 145 kDa on sodium dodecyl sulfate‐polyacrylamide gel electrophoresis, when the organisms were solubilized at room temperature. Two major bands at the position of 135 and 185 kDa were seen, when the organisms were solubilized by heating before electrophoresis. Heat‐denaturation of the 120‐ and 145‐kDa polypeptides in excised gel bands changed their mobility and caused them to migrate to 135‐ and 185‐kDa positions, respectively. Two polypeptides at the 120‐kDa position were demonstrated: one is a major heat‐modifiable polypeptide and the other a minor heat‐stable. Peptide mapping was performed to determine the identity between native and denatured polypeptides.
Molecular Comparison of Dengue Type 1 Mochizuki Strain Virus and Other Selected Viruses Concerning Nucleotide and Amino Acid Sequences of Genomic RNA: A Consideration of Viral Epidemiology and Variation
Authors: Zulkarnaini Edward , Hotta Susumu , Takegami Tsutomu 
Published: November 2013

AbstractDengue‐1 (D1) Mochizuki strain was examined for its nucleotide and amino acid sequences of genomic RNA and the data obtained were compared with those of other selected virus strains reported previously. Genomic regions corresponding to C, preM and M proteins were the major subjects of study. Parts of E protein were additionally examined. Among the D1 viruses investigated, the Mochizuki virus which was isolated in 1943 in Japan was shown to be close to Philippine 836‐1 strain isolated in 1984 and Nauru Island strain isolated in 1974 at the respective places, in contrast with Thai AHF 82‐80 strain isolated in 1980 and Caribbean CV1636/77 strain isolated in 1977. At the same time, a difference was noted between the Mochizuki and Philippine/Nauru strains at the cleavage site of preM/M junction: Mochizuki possessed RRGKR/S sequence whereas the Philippine/Nauru had RRDKR/S. The glycosylation site in preM and hydrophobic regions at the carboxyl termini of M and E were well conserved. Significances of the data are discussed in connection with viral epidemiology and variation.
Detection of DNA of Causative Agent of Spotted Fever Group Rickettsiosis in Japan from the Patient's Blood Sample by Polymerase Chain Reaction
Authors: Tange Yoshiki , Matsumoto Mitsuru , Okada Takanori , Hasegawa Hitoshi , Kobayashi Yuzuru 
Published: November 2013

AbstractThe polymerase chain reaction (PCR) was applied for the etiological diagnosis of spotted fever group (SFG) rickettsiosis in Japan. Nucleotide primers derived from the 17‐kDa antigen gene of Rickettsia rickettsii primed a rickettsia‐specific 246‐base‐pair product for all of the Katayama, Abe, Misaka and Kojima strains, which we had isolated previously. Moreover, we were able to detect the same product by PCR amplification from the peripheral blood of a patient in the acute stage of the illness. The PCR method is considered to be useful for rapid etiological diagnosis of SFG rickettsiosis in Japan.
Comparison of Cytokine Induction by Lipopolysaccharide of Bacteroides fragilis with Salmonella typhimurium in Mice
Authors: Jotwani Ravi , Tanaka Yasunori , Watanabe Kunitomo , Tanaka‐Bandoh Kaori , Kato Naoki , Ueno Kazue 
Published: November 2013

AbstractComparison of cytokine stimulation by lipopolysaccharide (LPS) of Bacteroides fragilis and Salmonella typhimurium was done to study the early events occurring in vivo. Mice injected intraperitoneally with either LPS demonstrated endogenous production of all the cytokines studied (tumor necrosis factor‐alpha, interferon‐gamma and interleukin‐6) within 6 hr in the bloodstream. However induction of all the cytokines by B. fragilis LPS (50 μg/mouse) was much weaker compared with S. typhimurium LPS (50 μg/mouse). Even a dose of S. typhimurium LPS 40 times smaller (1.2 μg/mouse) induced cytokines more strongly compared with B. fragilis LPS. Thus, a weak biological response to B. fragilis LPS as evidenced by chick embryo lethality, limulus lysate gelation, LD50 for mice and rabbit pyrogenicity could be due to weak induction of bioactive mediators by LPS.
Determination of the Agretopic Residues of a Peptide Co‐Restricted to Different Class II Isotypes, I‐Au and I‐Eu, and Its Application for Preparation of a Synthetic Peptide Vaccine against Influenza Virus A/Aichi/2/68
Authors: Takami Kimitaka , Ogasawara Kazumasa , Itoh Yasushi , Kajino Kiichi , Naruse Hirohito , Onoé Kazunori 
Published: November 2013

AbstractWe have defined that residues 46 and 54 on a synthetic peptide composed of residues 43–58 of pigeon cytochrome c (p43–58) work as agretopes (sites bound to an MHC molecule) in I‐Ab mice. Substitution of amino acid residues on these positions altered the peptide to bind with the other MHC molecules. Furthermore, by substituting the agretopic residues with a variety of amino acids, we could determine the class II binding motif for each MHC molecule. In the present study, immunogenicity of a peptide, 46R50V54A, carrying valine (V) at epitopic (site bound to TCR) position 50, arginine (R) and alanine (A) at agretopic positions 46 and 54 of the p43–58, respectively has been analyzed in B10.PL (H‐2u) mice. We found that this peptide bound to two different class II isotypes, I‐Au and I‐Eu. Arginine at position 46 or alanine at position 54 of the 46R50V54A was shown to be critical for binding to I‐Au or I‐Eu, respectively. Further, on the basis of this class II binding motif we could prepare potent peptide vaccines against influenza A/Aichi/2/68 virus in B10. PL mice.
CD8+ T Lymphocytes Are the Major Cell Population Involved in the Early Gamma Interferon Response and Resistance to Acute Primary Toxoplasma gondii Infection in Mice
Authors: Shirahata Toshikazu , Yamashita Takumi , Ohta Chikako , Goto Hitoshi , Nakane Akio 
Published: November 2013

AbstractGamma interferon (IFN‐γ) is known to be a major mediator influencing host defense against Toxoplasma (T.) gondii. To evaluate lymphocyte populations involved in this cytokine‐mediated early resistance to T. gondii, the effects of in vivo administration of monoclonal antibodies (MAbs) against T‐cell subsets and anti‐asialo GM1 antibody on the course of infection and IFN‐γ response were investigated in mice infected acutely with this parasitic protozoan. A single injection of anti‐CD8 MAb on day −1 or day 4 severely exacerbated the infection, in accordance with a marked suppression of endogenous IFN‐γ production. Moreover, the administration of anti‐IFN‐γ MAb on day 0 but not later than day 4 resulted in a total abrogation of resistance to T. gondii, suggesting that endogenous IFN‐γ produced during the first several days of infection is critical for the generation of antitoxoplasmal resistance in mice. In contrast, no significant increase in mortality was observed when injected with either anti‐CD4 MAb or anti‐asialo GM1 antibody on day − 1, while these antibodies reduced significantly the ability of mice to produce IFN‐γ. Indeed, simultaneous depletion of CD4+ and CD8+ cells had no greater suppressive effect on host defense and endogenous IFN‐γ production than depletion of CD8+ cells alone. Together, these results suggest that CD8+ T cells play a central role for resolution of acute toxoplasmosis by participating in endogenous IFN‐γ production. The possible role of early produced IFN‐γ in the development of protective immune response to T. gondii is also discussed.
Human Immunodeficiency Virus Vpx Is Required for the Early Phase of Replication in Peripheral Blood Mononuclear Cells
Authors: Kawamura Meiko , Sakai Hiroyuki , Adachi Akio 
Published: November 2013

AbstractFunctional importance of Vpx protein of human immunodeficiency virus type 2 was evaluated in various types of cells. In 8 lymphocytic or monocytic cell lines tested, vpx mutant virus grew as well as wild‐type virus. Only in primary peripheral blood mononuclear cell cultures, severely retarded growth of mutant virus was observed. No replication of vpx‐minus virus was detected in primary macrophage cells. A highly sensitive single‐round replication assay system was used to determine the defective replication phase in primary mononuclear cells of vpx mutant virus. In all cell lines examined, vpx mutant displayed no abnormality. In contrast, the vpx mutant was demonstrated to be defective at an early stage of the infection cycle in primary cell cultures. No evidence of a replication‐defect at a late phase in primary cells of the vpx mutant was obtained by a transfection‐coculture method. These results indicate that the virion‐associated Vpx protein is essential for early viral replication process in natural target cells such as primary macrophages.
Inhibition of T‐Cell Proliferation Induced by a Cell‐Free Salmonella typhimurium Extract Does Not Involve a Nitric Oxide‐Mediated Mechanism
Authors: Matsui Katsuhiko , Arai Toshihiko 
Published: November 2013

AbstractIn a previous study, we observed that a cell‐free Salmonella typhimurium extract induced suppression of mitogen‐induced T‐cell proliferation and that this suppression involved non‐responsiveness of T‐cells to interleukin‐2 (IL‐2) and augmentation of IL‐2 receptor (IL‐2R) expression. In this study, we found that inhibition of phytohemagglutinin (PHA)‐stimulated murine spleen cell proliferation induced by a cell‐free S. typhimurium extract was reversed by treatment with an anti‐interferon‐γ monoclonal antibody (anti‐IFN‐γ Ab), but not by interleukin‐4 or NG‐monomethyl‐l‐arginine, which is known to inhibit nitric oxide (NO)‐secretion from spleen cells in culture. However, IL‐2R expression was augmented by treatment with the extract, although this was independent of an NO‐mediated mechanism. Only anti‐IFN‐γ Ab treatment reduced the augmented IL‐2R expression to a normal level. These results suggest that the suppression of T‐cell proliferation induced by the Salmonella cell‐free extract is associated with augmentation of IL‐2R expression in an NO production‐independent manner.
Inducement of Respiratory Mucosal Immunogenicity for Nasal Vaccine Due to Amine‐ and Amide‐Inactivation of Parainfluenza Virus Type 1, Sendai
Authors: Miyamae Takeo 
Published: November 2013

AbstractThe protective effects in mice by nasal vaccination of amine‐ and amide‐inactivated Sendai viruses were investigated by a contact exposure experiment, immunofluorescent examination of the entire respiratory tract, and checking the serum HI antibody development. Of 10 monoamines, ethanolamine and 2‐methoxyethylamine vaccines induced complete protection, and methylamine, ethylamine, n‐propylamine, n‐butylamine, 2‐ethoxyethylamine, diethylamine and triethylamine vaccines brought about almost complete protection or lower respiratory infection. The methoxyamine‐treated mouse conferred the least protection. Of 5 diamines, 1, 3‐diaminopropane vaccine inhibited completely the infection, but hydrazine, ethylenediamine, putrescine, and cadaverine vaccines produced regional infection. Two polyamines, spermine and spermidine, did not inactivate the virus. Of 4 amides, only semicarbazide vaccine conferred complete mucosal defense, while acetamide, propionamide, and isonicotinic acid hydrazide vaccines lead to regional infection. Serum HI titers developed by vaccination were low on the whole, following their slight rise, fall or maintenance postexposure. In effect, the 4 vaccines inactivated by a best‐suited interstrand crosslink between phosphate groups in helix of viral RNA brought about the strongest protection, and showed the necessity of a definite length of molecules for inactivants.
Tumor Necrosis Factor Receptor Expression in HIV‐1‐Infected CD4+ T Cells
Authors: Hober Didier , de Groote Donat , Vanpouille Nathalie , Dehart Isabelle , Shen Lu , Wattré Pierre , Maniez‐Montreuil Michèle 
Published: November 2013

AbstractWe investigated whether HIV‐1 can regulate tumor necrosis factor receptor (TNFR) expression in SupT‐1, a CD4 + T‐cell line. The cells were infected with HIV‐1 containing 1,000 cpm RT activity, as early as day 3 after infection and all along the culture the supernatant level of core protein p24 was >250 pg/ml, and on days 6 and 9 after infection, p24 was found in 10 % of the cells as determined by indirect immunofluorescence assay. The cells were growing without loss of viability. The study of TNFR expression was based on a microassay for measurement of binding of 125I‐TNFα to cells, in which free and cell‐bound ligand separation was performed by centrifugation through oil. Scatchard analysis of TNFα binding on days 6 and 9 after infection revealed a 90 % increase in the expression of high‐affinity membrane receptors in HIV + SupT‐1 culture compared with uninfected cells (mean +/‐S.D. = 501 +/‐148.5 vs. 263 +/‐77.8 receptors/cell, n = 9, P< 0.001) with no change in dissociation constants (mean +/− S.D. = 4.36 +/−1.06 vs. 4.00 +/−1.12 × 10−10 m).
Serological Studies of the Antigenic Similarity between Typhus Group Rickettsiae and Weil‐Felix Test Antigens
Authors: Amano Ken‐ichi , Kyohno Kaori , Aoki Sumie , Suto Tsunehisa 
Published: November 2013

AbstractThe sera from two patients with murine typhus reacted with whole cells of Rickettsia prowazekii, R. typhi, and Proteus vulgaris OX19, and with lipopolysaccharides (LPS) from the spotted fever group rickettsia strain TT‐118 and P. vulgaris OX19 in the enzyme‐linked immunosorbent assay. Sera from these patients reacted with ladder‐like bands of LPS from R. prowazekii and R. typhi in the immunoblot, whereas the reactivity of these sera with LPS from P. vulgaris OX19 differed from each other. These results indicate that LPS from the typhus group rickettsiae and P. vulgaris OX19 contain similar epitopes.
Influence of Exogenous Interleukin‐2 Concentration on the Isolation of Human Immunodeficiency Virus
Authors: Zöllner Bernhard , Feucht Heinz‐hubert , Hamann Alf , Laufs Rainer 
Published: November 2013

AbstractA significant increase (P=0.015) in the HIV isolation rate from plasma samples was achieved by use of 10 U/ml exogenous interleukin‐2 compared to 20 U/ml. The sensitivity rose from 0% to 29% on patients negative for p24 core antigen (P=0.031) and from 71% to 86% in patients positive for p24 core antigen in plasma (P>0.05). Titration of infectious HIV revealed that 10 U/ml interleukin‐2 is the optimal concentration to isolate low numbers of infectious particles of HIV.
C3d and Epstein‐Barr Virus (CR2/CD21 Ligands) Stimulate Cells of an HTLV‐I Line, MT‐2
Authors: Kuraya Mikio , Sato Tetsuo , Fujita Teizo 
Published: November 2013

AbstractWe studied the physiological role of complement receptor type II (CR2, C3d/EBV receptor) expressed on T cells using MT‐2 cells. First, we confirmed CR2 expression on MT‐2 cells by flow cytometry and found that the MW of CR2 molecules on these cells and Raji B cells were the same by SDS‐PAGE analysis. When MT‐2 lysates were incubated with anti‐CR2 mAb HB5 and thereafter with 32P‐labeled ATP, 52‐ and 74‐kDa proteins were phosphorylated, suggesting the activation of MT‐2 cells through the complex of CR2 with these proteins. In this respect, we measured lymphotoxin production by MT‐2 cells when incubated with C3d or EBV. The cytotoxicity of the MT‐2 supernatant against L929 cells was elevated in a dose‐ and time‐dependent manner. Next, we confirmed EBNA expression on EBV‐infected MT‐2 cells and attempted to establish an EBV‐positive MT‐2 clone by in vitro EBV infection. However, these clones disappeared during cloning. To clarify this mechanism, we examined the EBV genome in MT‐2 cells. By Southern blot analysis, BamHI digestion of DNA extracts from MT‐2 cells 3 days after EBV treatment gave a 3.0‐kb signal which comigrated with the EBV BamHI‐W probe. The 3.0‐kb signal of genomic EBV‐DNA was detected at 1, 2, 3, 5, and 7 days after EBV treatment, but could not be detected at 14 days. Thus, natural ligands of CR2 stimulate CR2‐positive MT‐2 cells through their functionally active CR2 molecules and in vitro EBV infection of MT‐2 cells might be transient.
Conspicuous Growth of Intravenously Inoculated Staphylococcus aureus in Subcutaneously Established Ehrlich Ascites Tumor Tissue of Mice
Authors: Murai Miyo , Seki Keiko , Sakurada Junji , Usui Akemi , Masuda Shogo 
Published: November 2013

AbstractIntratumoral growth of Staphylococcus aureus was compared with the intrarenal growth, to examine the usefulness of the method as a marker of its pathogenicity. When 5 × 107 CFU/mouse of three derivatives from S. aureus Cowan I with different intrarenal growth were intravenously injected into Ehrlich tumor‐bearing mice, they lodged in the tumor tissue at approximately 103 CFU/0.1 g by 30 min after infection, and grew in the range of 106 CFU/0.1 g to 108 CFU/0.1 g by day 4, regardless of their intrarenal growth capacity. In contrast, S. saprophyticus lodged in both tissues to the same degree as S. aureus, but did not grow at all. The time course of the staphylococcal growth was different between tumor tissue and kidney, suggesting differences in the local responses against S. aureus.
Nutrient Starvation Induces Cross Protection against Heat, Osmotic, or H2O2 Challenge in Vibrio parahaemolyticus
Authors: Koga Tetsuro , Takumi Kenji 
Published: November 2013

AbstractGlucose‐starved cells of Vibrio parahaemolyticus were compared with non‐starved counterparts with respects to heat, osmotic, and oxidative challenges. The starved cells demonstrated greater thermal and oxidative resistance than did the non‐starved cells. The starved cells also showed greater resistance against low osmotic challenge than did the non‐starved cells although both cells showed a comparable resistance against high osmotic challenge.