bluuebunny/crossref_metadata_2025_split huggingface dataset:
IgA Protease Produced by Streptococcus sanguis and Antibody Production against IgA Protease in Patients with Behçet's Disease
Authors: Yokota Kenji , Oguma Keiji 
Published: November 2013

AbstractThe production of IgA protease in twelve strains of Streptococcus sanguis isolated from patients with Behçet's disease (BD) was examined. Protease activity was detected in 10 out of 12 strains. The protease was purified from one representative strain, S. sanguis 113–20, by employing Rotofor and DEAE‐Sephacel chromatography. The molecular mass of the purified protease was approximately 100 kDa, and it cleaved the proline‐threonine site of the IgA. Both IgG and IgA titers against the cells (113–20) and the purified IgA protease in the sera of BD patients and healthy controls, 36 each, were assayed. The IgG titers against the cells and protease were not significant in the BD patients or controls, but the IgA titers against the cells and protease in the BD patients were significantly higher than those of the controls. These data indicate that the BD patients are infected with IgA protease‐producing S. sanguis strains, which cause an increase of IgA titer against these organisms and IgA protease antigen. Since the organisms can proliferate in BD patients for a long period of time (years), it seems that IgA antibodies cannot effectively eliminate the organisms.
Mechanisms of Allografted Tumor Rejection: The Roles of T Cells in Allograft Rejection Mediated by a Type of Bone Marrow‐Derived Macrophage
Authors: Ushio‐Umeda Yumiko , Yoshida Ryotaro 
Published: November 2013

AbstractAllografted tumor rejection does not occur in the absence of T cells, but the main effector cells responsible for the rejection are allograft‐induced macrophages (AIM). We examined the roles of T cells in the AIM‐mediated rejection of Meth A (H‐2) tumor cells from C57BL/6 (H‐2b) mice. Irradiation of C57BL/6 mice abrogated both the induction of AIM and the allograft rejection. Reconstitution of the irradiated mice with F1 (C57BL/6 X C3H/He: H‐2b/k) bone marrow cells led to the appearance of H‐2b/k haplo‐type of AIM exclusively in the rejection site and to allograft rejection, indicating that radiosensitive cells prerequisite for both the induction of AIM and allograft rejection were bone marrow‐derived cells, and that the progenitors of AIM existed in the bone marrow cells to be activated into AIM in the rejection site. To understand the role of T cells in the induction of AIM, we used adult‐thymectomized, X‐irradiated C57BL/6 mice reconstituted with F1 bone marrow (ATXBM). The ATXBM mice could neither induce AIM nor reject allogeneic Meth A cells, whereas adoptive transfer of F1 lymph node T cells to the ATXBM mice restored not only the induction of AIM but also rejection of the allograft. Among the lymph node T cells, CD4+, but not CD8+, cells were found to be essential for the activation of AIM progenitors to AIM; and CD8+ T cells were further required for rejection, at least in part, to enhance the number of AIM in the rejection site.
The Inhibitory Effect of Staphylococcus epidermidis Slime on the Phagocytosis of Murine Peritoneal Macrophages Is Interferon‐Independent
Authors: Shiau Ai‐li , Wu Chao‐liang 
Published: November 2013

AbstractThe extracellular slime produced by Staphylococcus epidermidis has been shown to interfere with several human neutrophil functions in vitro, such as chemotaxis, degranulation and phagocytosis. Slime production has been suggested as a useful marker for clinically significant infections with coagulase‐negative Staphylococcus. Since the main role of macrophages in defense mechanisms is phagocytosis, the effect of slime on the phagocytic activity of macrophages was investigated. The phagocytic activity of murine peritoneal macrophages treated with slime in vitro decreased in a dose‐dependent fashion. A similar decrease was also observed in macrophages isolated from mice that had previously received intraperitoneal injection of slime. To investigate whether interferon also plays a role in this process, mice were treated with interferon or an interferon inducer, polyinosinic‐polycytidylic acid (poly I:C), together with slime before macrophage isolation. The slime‐suppressed phagocytic activity of macrophages was partially relieved by both agents, and the recovery effect of poly I:C in slime‐suppressed phagocytosis of macrophages in vivo might be attributed to the increased interferon level in peritoneal fluid and sera. However, when slime was given to poly I:C‐pretreated mice, the phagocytic activity remained suppressed. Thus, it appears that slime is able to suppress the phagocytic activity of macrophages regardless of the state of macrophage activation by poly I:C. The results suggest that the inhibition of phagocytosis by S. epidermidis slime may be independent from the activation of interferon.
Development of Reference Procedures for Broth Microdilution Antifungal Susceptibility Testing of Yeasts with Standardized Endpoint Determination
Authors: Makimura Koichi , Sudo Takako , Kudo Michinari , Uchida Katsuhisa , Yamaguchi Hideyo 
Published: November 2013

AbstractStandard guidelines for the broth microdilution antifungal susceptibility testing of amphotericin B, flucytosine, fluconazole, miconazole and itraconazole are reported. These are a modification of the method developed by the National Committee for Clinical Laboratory Standards (NCCLS) on the following two points: standardization of the means of endpoint determination and the inclusion of miconazole and itraconazole in the testing. MIC was determined to be when the positive control had a turbidity of 0.2 at the 630 nm wavelength. The endpoint was 80% inhibition for azoles and 100% inhibition for other drugs. The method provided good reproducibility, and a wide range of MIC distribution was observed in all antifungal agents except amphotericin B.
Elevated Serum Levels of the Soluble Form of gp130, the IL‐6 Signal Transducer, in HTLV‐1 Infection and No Involvement of Alternative Splicing for Its Generation
Authors: Zhou Yongwei , Horiuchi Sankichi , Yamamoto Mikio , Yamamoto Naoki 
Published: November 2013

AbstractBy using an ELISA, increased levels of the soluble form (sgp130) of gp130, the IL‐6 signal transducer, were detected in the sera of various HTLV‐1‐associated conditions (HC, ATL, HAM) as compared to normal healthy individuals. Sgp130 levels seemed to be correlated with disease severity. The 94 KD of sgp130 was specifically precipitated in the sera of HTLV‐1‐infected patients as revealed by Western blot analysis. A reverse transcriptase (RT)‐polymerase chain reaction (PCR) method was used to detect the message for transmembrane (TM) lacking gp130 in mRNA isolated from peripheral blood mononuclear cells (PBMCs) of patients infected with or without HTLV‐1 and those of various hematopoietic cell lines. Two PCR products, 648 and 507 bp were observed in the PBMCs from HTLV‐1‐infected patients. But the 507 bp PCR product was not detected in the PBMCs from normal healthy individuals and HTLV‐1‐positive cell lines although the 648 bp product was equally expressed. A nucleotide sequence analysis of the 507 bp fragment showed deletion of the 141 bp at the region spanning from nucleotide 1702 (G) to 1842 (T) of the 648 bp product that matched completely with a conventional gp130 molecule. This deleted region was located upstream of the transmembrane (TM) domain, but not within the TM region itself. However, no frame shift was observed. These results indicate that the generation of sgp130 may not be due to an alternative splicing mechanism.
Infectivity and Arthritis Induction of Borrelia japonica on SCID Mice and Immune Competent Mice: Possible Role of Galactosylceramide Binding Activity on Initiation of Infection
Authors: Kaneda Kazuhide , Masuzawa Toshiyuki , Simon Markus M. , Isogai Emiko , Isogai Hiroshi , Yasugami Kitsuzo , Suzuki Takashi , Suzuki Yasuo , Yanagihara Yasutake 
Published: November 2013

AbstractWe investigated the relationship between the binding activity to galactosylceramide (GalCer) and the arthritis induction activity of Borrelia japonica. The B. japonica strains maintained the ability to induce arthritis in inbred C3H/HeN and immunodeficient SCID mice, but the ability was lower than that of Borrelia burgdorferi sensu stricto virulent strain 297. Histopathological changes were restricted to the joints, and a marked effusion of polymorphonuclear neutrophils into the joint space was found. The binding activity of B. japonica strains to GalCer was lower than that of the virulent strain 297 but higher than that of the high‐passage strain 297. The lower infectivity and virulence of B. japonica may explain its lower binding ability to GalCer.
Kinetical Analysis of Tumor Cell Death‐Inducing Mechanism by Polymorphonuclear Leukocyte‐Derived Calprotectin: Involvement of Protein Synthesis and Generation of Reactive Oxygen Species in Target Cells
Authors: Mikami Masaaki , Yamazaki Masatoshi , Yui Satoru 
Published: November 2013

AbstractWe have previously shown that calprotectin, the most abundant cytosolic protein existing in polymorphonuclear leukocytes (PMNs), induces apoptotic cell death in various tumor cells, suggesting that calprotectin is an effector molecule against tumor cells in PMNs. To explore the cell death‐inducing mechanism of the factor, we examined the involvement of target protein synthesis and generation of reactive oxygen species (ROS) in the reaction. Calprotectin induced cell death in MM46 mouse mammary carcinoma cells after a 14‐16 hr lag time. When the factor was removed from the medium up to about 12 hr after culturing, the effect was diminished. The induction of cell death by calprotectin was markedly inhibited by the presence of the RNA synthesis inhibitor actinomycin D or the protein synthesis inhibitor cycloheximide. However, the addition of these inhibitors after 12 hr of culturing was unable to inhibit the reaction. Up to 12 hr of culturing, the net protein synthesis of MM46 cells was augmented by the presence of calprotectin, but thereafter was impaired. The induction of cell death was also inhibited by the antioxidative reagents N‐acetyl‐l‐cysteine (NAC) or propyl gallate. The addition of NAC even 15 hr later significantly attenuated the calprotectin effect. Flow cytometry analysis showed that calprotectin began to increase the ROS content in MM46 cells after 8‐12 hr of culturing, and that the increase was abrogated by the antioxidants. Thus, protein synthesis and ROS generation may be essential elements in the early or later phases of the cell death‐inducing reaction of calprotectin, respectively.
Zymographic Characterization of Staphylococcus aureus Cell Wall
Authors: Ohta Kouji , Komatsuzawa Hitoshi , Sugai Motoyuki , Suginaka Hidekazu 
Published: November 2013

AbstractSusceptibilities of several preparations of Staphylococcus aureus cells to various peptidoglycan hydrolases with known bond specificity were analyzed by zymography. The substrates were intact S. aureus cells, cells boiled in the presence of SDS and cells treated with trichloroacetic acid after treatment with boiling SDS solution (TCA‐cells). Twofold dilutions of lysostaphin (LS), lysozyme (LZ), S. aureus 51 kDa glucosaminidase (GL) or S. aureus 62 kDa amidase (AM) were electrophoresed, and the minimal enzyme dose showing a visible bacteriolytic band was defined as MBD (minimal bacteriolytic dose). Under the same experimental conditions, this method gave reproducible results. As the substrate for zymogram, TCA‐cells were the most sensitive to LS, LZ and AM, whereas the three substrate were equally sensitive to GL. A zymographic analysis of methicillin‐resistant S. aureus treated with methicillin together with previous studies suggest that this method can be used for the preliminary characterization of S. aureus cell wall peptidoglycan.
Molecular Characterization of Low‐Molecular‐Weight Component Protein, Flp, in Actinobacillus actinomycetemcomitans Fimbriae
Authors: Inoue Tetsuyoshi , Tanimoto Ichiro , Ohta Hiroyuki , Kato Keijiro , Murayama Yoji , Fukui Kazuhiro 
Published: November 2013

AbstractFimbriae preparation from Actinobacillus actinomycetemcomitans was found to contain an abundant low‐molecular‐weight protein (termed Flp) with an apparent molecular mass of approximately 6.5 kDa, in addition to a small amount of 54‐kDa protein. Immunogold electron microscopy localized the Flp protein at the bacterial fimbriae but not at the cell surface. The DNA fragment including the flp gene was cloned from A. actinomycetemcomitans 304‐a and its nucleotide sequence was determined. An open reading frame of the flp gene was composed of 225 bp encoding a protein of 75 amino acids. Comparison of the translated amino acid sequence with the sequence of native Flp determined by Edman degradation indicated that the N‐terminal part of 26 amino acids is leader peptide. The N‐terminal sequence of mature Flp exhibited some similarity to type‐IV pilin. Furthermore, the processing site of premature Flp is also similar to that of type‐IV prepilin, and a gene encoding a protein homologous to type‐IV prepilin‐like protein leader peptidase was found downstream of the flp gene. These findings indicate that Flp is the major component protein of A. actinomycetemcomitans fimbriae.
Epidemiologic Study of Shigella sonnei from Sequential Outbreaks and Sporadic Cases Using Different Typing Techniques
Authors: Matsumoto Masakado , Suzuki Yasumoto , Saito Makoto , Ishikawa Naohisa , Ohta Michio 
Published: November 2013

AbstractWe noted that eight outbreaks of Shigella sonnei from an unknown source occurred sequentially in Aichi Prefecture, Japan, between October 1992‐June 1993. For comparative purposes we analyzed 53 outbreak‐related isolates of Shigella sonnei using different subtyping methods and studied the epidemiology of the outbreaks. It appeared from our study that DNA‐based techniques such as plasmid typing and pulsed‐field gel electrophoresis (PFGE) were more useful tools for subtyping Shigella sonnei than colicin typing and the antimicrobial susceptibility test. Moreover, according to PFGE analysis, four genetically related isolates of Shigella sonnei were responsible for the eight sequential outbreaks. To further investigate the epidemiology of outbreaks, 58 sporadic isolates of Shigella sonnei from overseas travelers with shigellosis during the same period were also examined. We found that some sporadic isolates from travelers in Asia were genetically related to those of the outbreak‐related isolates, indicating that genetically related isolates prevailed in Asia during this period, probably because of the extensive movement of people or food.
Studies of Escherichia coli Cultured on RainbowTM Agar O157 with Particular Reference to Enterohaemorrhagic Escherichia coli (EHEC)
Authors: Bettelheim Karl A. 
Published: November 2013

AbstractRainbowTM Agar O157 is designed for the rapid isolation and identification of enterohaemorrhagic Escherichia coli (EHEC), particularly O157, characterised by black colonies. Five‐hundred‐eighty‐five E. coli strains, including O157, O111 and O113 serogroups from many sources were examined on RainbowTM Agar O157. EHEC O157 could readily be isolated and recognized uniquely by typical black colonies. Some other EHEC also stand out as blue‐black, whereas O113 and some other EHEC strains were mauve, red or pink and indistinguishable from SLT‐negative strains of E. coli.
Diagnostic Value of the Amplicor PCR Assay for Initial Diagnosis and Assessment of Treatment Response for Pulmonary Tuberculosis
Authors: Iinuma Yoshitsugu , Ichiyama Satoshi , Yamori Sadaaki , Oohama Jinya , Takagi Norio , Hasegawa Yoshinori , Shimokata Kaoru , Nakashima Nobuo 
Published: November 2013

AbstractWe evaluated the Amplicor PCR assay as an initial diagnostic tool on the basis of clinical diagnosis, and assessed this assay as a follow‐up test for patients with pulmonary tuberculosis during chemotherapy. Of the 208 specimens from 155 patients who were bacteriologically and/or clinically diagnosed with active tuberculosis before chemotherapy, 144 were Amplicor PCR‐positive (sensitivity, 69.2%), which was equal to the results of culturing. Among 89 specimens which showed positive results by smear and culturing, the Amplicor PCR assay detected 87 (97.8%), whereas among 55 specimens which showed smear‐negative but culture‐positive results, the Amplicor PCR assay detected 46 (83.6%)(P = 0.003). No false positive results were found in the two systems (specificity, 100%, 120/120). The Amplicor PCR assay was also evaluated as a follow‐up test using 926 specimens from 207 patients receiving active tuberculosis chemotherapy. Among 433 specimens which showed Amplicor‐PCR positive, 222 (51.3%) were culture‐negative. On the other hand, among 233 culture‐positive specimens, only 12 (5.2%) were Amplicor PCR‐negative. Therefore, this assay is useful for the rapid diagnosis of tuberculosis. The duration of Amplicor PCR‐positive after culture‐negative conversion was significantly associated with the presence of cavitary lesion, smear‐positive specimens before treatment, and smear‐positive specimens with negative cultures during chemotherapy.
The 21‐kDa Polypeptide (VAP21) in the Rabies Virion Is a CD99‐Related Host Cell Protein
Authors: Sagara Junji , Tochikura Tadafumi S. , Tanaka Hajime , Baba Yoshihiro , Tsukita Shoichiro , Tsukita Sachiko , Kawai Akihiko 
Published: November 2013

AbstractIn our monoclonal antibody (MAb) stocks prepared against the BHK‐21 cell antigens, two (#11875 and 28276) recognized a 21‐kDa polypeptide (referred to as VAP21) which is efficiently incorporated into the rabies virion. By using these MAbs, we isolated the cDNA clones that encoded a polypeptide of 144 amino acids from our BHK‐21 cell cDNA library. Based on the following evidence, the cDNA was assumed to encode a full‐length sequence of VAP21 antigen: i) expression of the cDNA in animal cells resulted in the production of a polypeptide recognized by the two MAbs, and its electrophoretic mobility was the same as that of authentic VAP21 antigen; and ii) immunization with the products from the cDNA‐transformed E. coli cells raised specific antibodies in rabbits that recognized a 21‐kDa polypeptide in the virion. From the deduced amino acid sequence, it is suggested that the VAP21 antigen has a molecular structure of type‐I transmembrane protein containing characteristic proline‐rich and glycine‐rich regions in its ectodomain. Homology searches resulted in finding homologous sequences (totally about 40% homology) in the human MIC2 gene product (CD99; 32‐kDa) of T lymphocytes. These results suggest that the VAP21 antigen in the rabies virion is a cellular CD99‐related transmembrane protein.
Cellular Reaction to Mycobacterium avium Complex (MAC) Clinical Isolates Differing in Hemolytic Activity and Virulence for C57BL/6 Mice
Authors: Brzychcy Marzena , Zwolska Zofia , Andrzejczyk Zofia , Rudnicka Wiesława 
Published: November 2013

AbstractIn this study we showed that Mycobacterium avium complex (MAC) clinical isolates differed by the expression of hemolytic activity. Two hemolytic MAC strains were less susceptible to the mycobactericidal effect of murine macrophages than two unhemolytic MAC isolates. In vivo, hemolytic MAC bacilli survived in the spleens of infected mice for a longer time than unhemolytic MAC strains. This suggested a role of hemolysins in the virulence of MAC strains. There was no difference in the cytotoxicity of T cells from mice immunized with M. bovis BCG towards macrophages infected in vitro with MAC strains expressing or not expressing hemolytic activity.
Morphological Alterations of Saccharomyces cerevisiae Induced by Benanomicin A, an Antifungal Antibiotic with Mannan Affinity
Authors: Watanabe Maki , Nishiyama Yayoi , Inouye Shigeharu , Yamaguchi Hideyo , Kondo Shinichi , Takeuchi Tomio 
Published: November 2013

AbstractThe effects of benanomicin A, a mannose‐binding antifungal antibiotic, on yeast cells of Saccharomyces cerevisiae were studied by electron microscopy. Cytological studies using vital stain with methylene blue demonstrated that benanomicin A at 20 and 80 μg/ml killed buds in preference to parent cells. In confirmation, examination by TEM revealed that benanomicin A at 80 μg/ml damaged buds more severely than parent cells. The major effect on the ultrastructure was characterized by severe damage to the cell membrane. In addition, it caused expansion and vacuolation of the endoplasmic reticulum (ER), and partial fragmentation and disappearance of nuclear membranes. The membrane‐disruptive activity of benanomicin A may be closely associated with its membrane affinity.
Serological Differentiation between HCV Subtypes 1a and 1b by a Recombinant Immunoblot Assay
Authors: Schröter Matthias , Feucht Heinz‐hubert , Zöllner Bernhard , Laufs Rainer 
Published: November 2013

AbstractUntil now, no serological assay has been available for the differentiation of HCV subtypes. Since there is evidence that the subtypes differently influence the clinical course of HCV infection and the outcome of interferon therapy, we established a strip immunoblot assay (NS‐4 IBA) with recombinant HCV proteins of the nonstructural 4 (NS‐4) region propagated in Escherichia coli. Using this NS‐4 IBA, we were able to distinguish HCV subtypes la and 1b, which are the most prevalent subtypes in Europe and the U.S.A. The results of the serotyping assay were compared with those obtained by nucleotide sequencing from the NS‐5 region. Concordant results were observed to match 94.9% (n = 100) by the NS‐4 IBA and nucleotide sequencing. Discrepant results were obtained in only 5.1% (n = 6). These data indicate that HCV subtypes can be serologically distinguished, providing the possibility for easier identification of infection with different KCV subtypes.
Application of RT‐PCR Designed from the Sequence of the Local SRSV Strain to the Screening in Viral Gastroenteritis Outbreaks
Authors: Saito Hiroyuki , Saito Shioko , Kamada Kazuko , Harata Seizaburo , Sato Hiroyasu , Morita Morihiro , Miyajima Yoshimichi 
Published: November 2013

AbstractThe Yuri strain of small round structured virus (SRSV) was cloned from a fecal specimen in which virus particles were observed by electron microscopy. The most common RT‐PCR protocol in Japan, however, using 35/36 and NV81/NV82/SM82 nested primer pairs, could not detect the SRSV genome in this specimen. Nucleotide and amino acid sequence analysis revealed that the Yuri strain is genetically close to the genotype II of SRSV. A novel procedure using primer sets designed from the nucleotide sequence of the Yuri strain was applied to the screening of 119 stool samples obtained from subjects with sporadic diarrhea and 46 samples obtained during seven foodborne gastroenteritis outbreaks. Using this novel procedure, PCR bands were detected in 44% and 52% of the samples, respectively. These detection rates were approximately twice those obtained with the 35/36 and NV81/NV82/SM82 nested primers. In particular, more than 40% of positive samples could be detected by using only the Yuri primer sets. Furthermore, three improvements were made in the RNA preparation, cDNA synthesis, and amplification steps to save materials and time. The background, or extra bands, in the amplification reaction resulting from DNA in the fecal specimens was completely removed by DNase I treatment just before cDNA synthesis. Random nonamers were used as universal primers in the reverse transcription. No difference in sensitivity or specificity was noted in the final results when either random nonamers or specific primers were used. The use of a pre‐amplification step under low stringent conditions before standard amplification under highly stringent conditions compensated for any mismatched bases in the primers with respect to the target sequences. Thus our novel procedure using Yuri primer sets may be useful for the screening of SRSV in the recent SRSV outbreaks in Japan.