bluuebunny/crossref_metadata_2025_split huggingface dataset:
Advantage of Dimeric Peptide Antigens in Serodiagnosis of HIV‐1 Infection
Authors: Gokulan Kuppan , Tripathi Srikant Prasad , Rao D. Nageshwara 
Published: November 2013

AbstractThe reactivity of antibodies with dimeric and monomelic peptide antigens was compared by ELISA. A panel of highly purified synthetic peptides of HIV‐1 representing defined regions, 598–609 and 524 533 (fusion domain) of gp41 and 306–320 of gpl20, were used as antigens in the ELISA. These peptides were selected and synthesized taking into account the level of sequence conservation of various strains and hydrophilicity. The analysis included sera from 52 HIV‐1 infected individuals and 53 HIV‐1 negative controls. Both peptides from gp41 were found to be particularly immunoreactive with sera from HIV‐1 infected individuals. The frequency of reactivity to the selected peptide from gp120 (V3 loop) in infected individuals was 82%. An interesting observation was that the dimeric peptide antigens had a detection rate more than 4‐fold higher than the monomeric antigens. We found that lower levels of antibodies could be detected with dimeric antigens. The peptides reacted with few sera other than HIV‐1 positive sera. These results implicate the potential dimeric peptide antigens to be utilized in the serodiagnosis of HIV‐1 infection.
Identification of a Phosphatase‐Sensitive Epitope of Rabies Virus Nucleoprotein Which Is Recognized by a Monoclonal Antibody 5‐2‐26
Authors: Anzai Jun , Takamatsu Fumihiko , Takeuchi Kenji , Kohno Takashi , Morimoto Kinjiro , Goto Hideo , Minamoto Nobuyuki , Kawai Akihiko 
Published: November 2013

AbstractWe have investigated a phosphatase‐sensitive sequential epitope of the nucleoprotein (N), one of the phosphoproteins of rabies virus, which is recognized by the monoclonal antibody (MAb) #5‐2‐26. The epitope was shared in common by all of the rabies virus strains we tested, including the HEP, ERA, CVS and Japanese strains (Nishigahara and Komatsukawa). Thin layer chromatography of the acid hydrolyzates of 32P‐labeled N protein showed that the protein contained phosphoserine and phospho‐threonine at a molar ratio of about 4 to 1, while no phosphotyrosine was detected. Immunoprecipitation studies with several deletion mutants of the N protein showed that the epitope is located in a region spanning from amino acid 344 to 415. If the phosphatase‐sensitive epitope is located at or near the phosphoamino acid, the location of the latter could be narrowed further to a region from amino acid 354 to 389 by comparing the amino‐acid sequences among the viral strains. To examine this assumption, point mutation was introduced by amino‐acid substitution with alanine at either of five potential phosphorylation sites (i.e., positions 354, 375, 377, 386 and 389) in the 354–389 region. Among those, only one substitution, at position 389, greatly affected the antigenicity. Substitution of serine‐389 by threonine also reduced the antigenicity. These results strongly suggest that serine‐389 is a phosphorylation site and essential for constructing or stabilizing the antigenic structure for MAb 5‐2‐26.
Double‐Negative BV8S3 T Cells Expanded in MRL lpr/lpr Lymph Nodes Conserve TCRBJ Gene Usage of Single‐Positive BV8S3 T Cells
Authors: Fan Jiong , Nanki Toshihiro , Miyasaka Nobuyuki , Kohsaka Hitoshi 
Published: November 2013

AbstractEnlarged lymph nodes of mice with lpr mutation consist predominantly of CD4−CD8− (double‐negative: DN) T cells. Among them, TCRBV8S3 (Vβ 8.3) T cells are overrepresented as compared to those in single‐positive (SP) T cells. To address the question of whether the expansion of oligoclonal T cells is responsible for the increase in TCRBV8S3 cells, we examined the TCRBJ gene repertoires of BV8S3 DN and SP T cells from multiple MRL lpr/lpr mice. The BJ repertoires of BV3 (Vβ3), BV8S1 (Vβ8.1) and BV8S2 (Vβ8.2) were studied for comparison with those of BV8S3 T cells. The employed method, which was based on a PCR‐ELISA technique, was newly developed and allowed us to make a precise quantitation of TCRBJ gene usage of the multiple lymphocyte samples. The results showed that there were no biases of the BJ gene usage by BV8S3 DN T cells as well as other BV T cells. Furthermore, the BJ gene usage of CD4 and CD8 BV8S3 T cells was conserved by the DN T cells. It is suggested that the BV8S3 DN T cells were not expanded by specific antigens. The expansion may result from aberrant regulation specific to the BV8S3‐expressing T cells.
Role of Platelet‐Actlvatlng‐Factor (PAF) on Cellular Responses after Stimulation with Leptospire Lipopolysaccharide
Authors: Isogai Emiko , Hirose Kimiharu , Kimura Koichi , Hayashi Shunji , Kubota Toru , Fujii Nobuhiro , Isogai Hiroshi 
Published: November 2013

AbstractLeptospire lipopolysaccharide (LPS) stimulated the adherence of polymorphonuclear neutrophils (PMNs) to human umbilical vein endothelial cells (HUVEC). Enhanced PMN adherence in response to leptospire LPS can be mediated by platelet‐activator‐factor (PAF), because a PAF antagonist reduced adherence. Leptospire LPS also induced the adherence platelets or U937. The second experiment involved leptospire LPS elicited platelet aggregation in a PMN‐platelet mixture, because leptospire LPS stimulated human PMN but not the human platelets. The platelet response was observed only in the mixture system and was inhibited by a PAF antagonist. PAF could be an important pathogenic factor in human leptospirosis.
Polymerase Chain Reaction Test for Differentiation of Five Toxin Types of Clostridium perfringens
Authors: Yamagishi Takayoshi , Sugitani Kayo , Tanishima Kiyoo , Nakamura Shinichi 
Published: November 2013

AbstractIn order to avoid the use of experimental animals, the polymerase chain reaction (PCR) method was applied to differentiate Clostridium perfringens into five toxin types. Twenty‐two out of 23 strains tested produced the toxin(s) corresponding to the toxin gene(s) identified by PCR, and vice versa. Consequently, the gene typing was consistent with conventional typing by animal tests. Twenty‐five strains were identified as types different from original ones by the PCR method as well as a toxin neutralization test. These findings suggest that the PCR method, which is easy and timesaving, is applicable to identify the toxin types of C. perfringens as an alternative to animal tests, and that beta‐, epsilon‐ and iota‐toxin genes might be lost by long‐term preservation. The reasons why the strains lost the genes are discussed.
Unique Expression of HRF20 (CD59) in Human Nervous Tissue
Authors: Akatsu Hiroyasu , Yamada Tatsuo , Okada Noriko , Yamamoto Takayuki , Yamashina Manabu , Okada Hidechika 
Published: November 2013

AbstractDamage to autologous tissue by complement is limited by several widely distributed membrane‐associated glycoproteins which restrict the action of the complement in homologous species. These include decay accelerating factor (DAF), membrane cofactor protein (MCP) and 20 kDa homologous restriction factor (HRF20,CD59). Using immunohistochemical techniques, we examined the localization of these proteins in the centra] nervous system (CNS) and peripheral nervous system (PNS) using non‐neurological human nervous tissue since some complement components have been demonstrated to be synthesized in the CNS. There was no evidence of parenchymal staining by anti‐DAF or anti‐MCP antibodies in either type of tissue except for the staining of the endothelium in capillaries. On the other hand, anti‐HRF20 antibody clearly stained myelinated axons in the CNS as well as Schwann cells in the PNS. In addition, we detected positive staining by anti‐DAF antibody in the PNS of a Paroxysmal nocturnal hemoglobinuria (PNH) patient who is genetically deficient in HRF20.
Overproduction of Campylobacter Ferritin in Escherichia coli and Induction of Paracrystalline Inclusion by Ferrous Compound
Authors: Wai Sun Nyunt , Nakayama Koji , Takade Akemi , Amako Kazunobu 
Published: November 2013

AbstractThe ferritin gene (cft) of Campylobacter jejuni was overexpressed in cells of Escherichia coli using a T7 RNA polymerase expression system. Many round particles which were the same size as the ferritin particles purified from C. jejuni were observed in the lysate of the cft‐overexpressed E. coli cells. Since most of them were devoid of a central electron dense core consisting of ferric irons, the Campylobacter ferritins overproduced in E. coli seemed to be apoferritin. When large amounts of ferrous iron (supplied as FeSO4) were added to culture medium, the cft‐overexpressed cells formed large inclusion bodies of paracrystalline arrays comprised of ferritin particles with central electron dense cores. The addition of ferric irons did not produce paracrystalline inclusion.
Subcellular Localization of the Major Autolysin, ATL and Its Processed Proteins in Staphylococcus aureus
Authors: Komatsuzawa Hitoshi , Sugai Motoyuki , Nakashima Seiji , Yamada Sakuo , Matsumoto Akira , Oshida Tadahiro , Suginaka Hidekazu 
Published: November 2013

AbstractThe Staphylococcus aureus autolysin gene, atl, encodes a unique 138‐kDa protein (ATL) with amidase and glucosaminidase domains. ATL has been suggested to undergo proteolytic processing to generate two extracellular peptidoglycan hydrolases, 51‐kDa endo‐β‐N‐acetylglucosaminidase (51‐kDa GL) and 62‐kDa N‐acetylmuramyl‐L‐alanine amidase (62‐kDa AM). To investigate cell‐associated bacteriolytic enzymes for atl gene products, proteins were extracted from the cells as follows. The cells were exposed to 3 M LiCl followed by 4% SDS. Thereafter, the cells were disrupted and again extracted with 4% SDS. Whole SDS‐stable cell‐associated bacteriolytic proteins were extracted without disrupting the cells. Exposure to 3 M LiCl released major 138‐, 115‐, 85‐, 62‐ and 51‐kDa bacteriolytic proteins, and subsequent 4% SDS extraction released major 138‐ and 115‐kDa bacteriolytic proteins. These bacteriolytic proteins were missing in extracts of atl mutant RUSAL2 (S. aureus RN450 atl:: Tn551). Immunoblotting studies suggest that these are all atl gene products: the 138‐kDa protein is an ATL with a cleaved signal sequence; the 115‐and 85‐kDa proteins are intermediates; and the 51‐ and 62‐kDa proteins are cell‐associated 51‐kDa GL and 62‐kDa AM, respectively. The trypsin susceptibility of these proteins suggests that they are located outside the cell membrane. Differences in extractability and immunoelectron microscopic studies suggest that atl gene products are associated with cells in two different ways, LiCl extractable and non extractable. We suggest that the 138‐kDa ATL undergoes processing through intermediate proteins (115‐ and 85‐kDa proteins) to mature as the active cell cluster‐dispersing enzymes 51‐kDa GL and 62‐kDa AM on the cell surface.
Pathogenicity of Glycoprotein C‐Deficient Herpes Simplex Virus 1 Strain TN‐1 Which Encodes Truncated Glycoprotein C
Authors: Minagawa Hiroko , Yoshida Ying Liu Tetsuhiko , Hidaka Yasufumi , Toh Yasushi , Mori Ryoichi 
Published: November 2013

AbstractA clinical isolate of herpes simplex virus 1 (TN‐1) from a stromal keratitis patient was found to be defective in the glycoprotein C (gC) gene (UL44), thus resulting in the production of truncated gC upon infection. To study the pathogenetic role of truncated gC, we prepared a recombinant LTN‐8 derived from TN‐1 with deletions of the 1.5 kilobase pairs of the gC gene including the initiation codon. A penetration assay revealed LTN‐8 to be less efficient in its penetration ability than TN‐1, the laboratory strain KOS and RTN‐1‐20‐3, a recombinant derived from TN‐1 with the KOS gC gene. The penetration of LTN‐8 was facilitated by the addition of TN‐1‐infected culture medium. TN‐1 virus preparations had no hemagglutinating activity. However, the animals infected with TN‐1 did develop hemagglutination inhibition (HI) antibodies. The LTN‐8‐infected animals did not develop HI antibodies. The pathogenicity in BALB/c mice following either corneal, intraperitoneal or intracerebral inoculation did not significantly differ among TN‐1, RTN‐1‐20‐3 or LTN‐8. Our results indicate that truncated gC was sufficient for the induction of HI antibodies and was also able to facilitate penetration in vitro. Although truncated gC might be a virulence factor acting as a decoy, both truncated gC and intact gC had little effect on the outcome following intracerebral, intraperitoneal or corneal inoculation.
MHC Class I Presentation of an Exogenous Polypeptide Antigen Encoded by the Murine AIDS Defective Virus
Authors: Yee Sung‐tae , Okada Yoshiaki , Ogasawara Kazumasa , Omura Satoshi , Takatsuki Akira , Kakiuchi Terutaka , Muno Daisaku , Kominami Eiki , Mizuochi Toshiaki 
Published: November 2013

AbstractPeptides derived from endogenous proteins are presented by MHC class I molecules, whereas those derived from exogenous proteins are presented by MHC class II molecules. This strict segregation has been reconsidered in recent reports in which exogenous antigens are shown to be presented by MHC class I molecules in the phagocytic pathway. In this report, the presentation pathway of an exogenously added highly antigenic polypeptide encoded by the murine AIDS (MAIDS) defective virus gag p12 gene is investigated. A 25‐mer polypeptide (P12–25) encoded within the gag p12 region of the MAIDS defective virus was found to be effective in stimulating unprimed B6 (H‐2b) CD8+ T cells in vitro. The presentation of P12–25 is sensitive to cytochalasin B and D, brefeldin A and gelonin, a ribosome‐inactivating protein synthesis inhibitor, but less sensitive or resistant to lactacystin, a highly specific inhibitor of the proteasome. Interestingly, CA‐074, a selective inhibitor of cathepsin B, inhibited presentation of the polypeptide, indicating its involvement in the degradation of the P12–25 polypeptide. In fact, when P12–25 was digested with purified cathepsin B in vitro, a highly antigenic 11‐mer peptide containing the class I (H‐2Db)‐binding motif was obtained. Our results favor the phagosome/macropinosome‐to‐cytosol‐to‐endoplasmic reticulum (ER)‐to‐cell surface pathway for exogenous antigens presented by MHC class I molecules. These findings may be relevant to exploiting peptide vaccines that specifically elicit CD8+ T cell immunity in vivo.
Partial Sequences of Large Subunit Ribosomal DNA of a New Yeast Species, Trichosporon domesticum and Related Species
Authors: Sugita Takashi , Makimura Koichi , Nishikawa Akemi , Uchida Katsuhisa , Yamaguchi Hideyo , Shinoda Takako 
Published: November 2013

AbstractWe determined the partial sequences of large subunit rDNA of a new yeast species, Trichosporon domesticum, which was isolated from the house of a summer‐type hypersensitivity pneumonitis patient. Phylogenetically, T. domesticum was positioned in the taxonomic group containing T. montevideense and T. brassicae, which indicated an identical serotype. A phylogenetic relationship among all species of the genus Trichosporon which belong to the basidiomycetous yeast is clarified.
cDNA Cloning and Characterization of Rat C5a Anaphylatoxin Receptor
Authors: Akatsu Hiroyasu , Miwa Takashi , Sakurada Chikai , Fukuoka Yoshihiro , Ember Julia A. , Yamamoto Takayuki , Hugli Tony E. , Okada Hidechika 
Published: November 2013

AbstractActivation of the complement cascade plays an esssential role in the early stages of inflammation. C5a and its receptor are particularly active in anaphylaxis. To determine the pathological roles played by C5a and C5a receptor (C5aR) in rats, we cloned C5aR cDNA and analyzed distribution of its mRNA in various organs including lung from an LPS‐stimulated rat. Furthermore, we generated a polyclonal antiserum which specifically recognizes rat C5aR, as confirmed by its specific interaction with cells transfected with rat C5aR cDNA.
Cyclophosphamide‐Induced Bacterial Translocation in Escherichia coli C25‐Monoassociated Specific Pathogen‐Free Mice
Authors: Nakayama Masaharu , Itoh Kikuji , Takahashi Eiji 
Published: November 2013

AbstractThe kinetics of bacterial translocation (BTL) from the intestine to the mesenteric lymph nodes (MLN) and the number of peripheral white blood cells (WBC), macrophages in Peyer's patches (PP) and M‐cells on the surface of cecal PP after cyclophosphamide (CY) injection were examined in penicillin‐G and streptomycin sulfate decontaminated and Escherichia coli C25‐monoassociated specific pathogen‐free mice. WBC were counted to confirm the immunological state of the mice. Until 8 days after CY injection, the number of WBC, bacteria in MLN and macrophages in PP decreased, but then significantly increased on day 14. The levels again decreased to the control levels on day 16. Although the number of M‐cells decreased up to day 8, it did not return to the control level on day 16. These results indicate that BTL is stimulated in an immunopotentiated state after CY injection, and this phenomenon may be closely related to the number of macrophages in the blood and PP.
Genetic Diversity of Borrelia burgdorferi Sensu Lato Isolated in Far Eastern Russia
Authors: Masuzawa Toshiyuki , Iwaki Atsue , Sato Yukita , Miyamoto Kenji , Korenberg Edward I. , Yanagihara Yasutake 
Published: November 2013

AbstractOne‐hundred and fifty‐seven Borrelia isolated from adult ticks, Ixodes persulcatus, and wild rodents, Clethrionomys rufocanus and Apodemus peninsulae, in the far eastern part of Russia were characterized and identified by restriction fragment length polymorphism (RFLP) of the 5S‐23S rRNA intergenic spacer. Some isolates showed unique RFLP patterns and were determined as Borrelia garinii on the basis of a sequence analysis of the intergenic spacer amplicon and reactivity with species‐specific monoclonal antibodies (MAbs). 86.5 and 12.7% of the tick isolates, and 74.2 and 12.9% of the rodent isolates were determined as Borrelia garinii and Borrelia afzelii, respectively, but no Borrelia burgdorferi sensu stricto was detected. This finding is similar to the results obtained from Borrelia surveys of I. persulcatus and wild rodents in Hokkaido, Japan.
Genetic Control of Immune Responses to a Synthetic Fimbrial Antigen of Actinobacillus actinomycetemcomitans
Authors: Honma Kiyonobu , Ishii Norihisa , Kato Tetsuo , Ishihara Kazuyuki , Okuda Kenji , Okuda Katsuji 
Published: November 2013

AbstractThe incidence of infection by Actinobacillus actinomycetemcomitans, one of the important pathogens in human periodontal diseases, has been reported to be associated with racial background and genetic factors. We attempted to determine the genetic regulation of immune responses to A. actinomycetemcomitans fimbriae, an attachment factor, using various inbred strains of mice. For this purpose, we synthesized an oligopeptide antigen using the amino acid sequence of the fimbriae and conjugated this antigen to branched lysine polymer resin beads. After immunization with the synthetic A. actinomycetemcomitans fimbrial antigen, serum antibody levels and the delayed‐type hypersensitivity (DTH) reaction to the antigen were measured by enzyme‐linked immunosorbent assay (ELISA) and footpad swelling responses, respectively. The strains of mice found to be high‐IgG responders to the antigen were B10.HTT, B10.RIII, B10A (5R) and B10.S (9R). These results indicate that mice with Eβs: Eαk, Eβr: Eαr and Eβb: Eαk respond strongly to the synthetic peptide. All of the high‐IgG responders showed a high DTH response. A cell transfer experiment confirmed that CD4 T cells mediated with a DTH response to the synthetic peptide. Thus, the results of this study demonstrate that the immune responses to A. actinomycetemcomitans fimbriae are genetically controlled.
Genomic Diversity in the pfoA Region of Theta‐Toxin‐Deficient Strains of Clostridium perfringens
Authors: Ba‐Thein William , Inui Satoko , Shimizu Tohru , Swe Tint , Banu Sayera , Ohtani Kaori , Oe Misari , Sakurai Naomi , Nakamura Shinichi , Hayashi Hideo 
Published: November 2013

AbstractThe genomic structure of the pfoA‐colA region in six theta‐toxin‐deficient strains of Clostridium perfringens was examined by Southern hybridization using the pfoR, pfoA, pbg, arcABDC and colA genes, encoding regulator for pfoA, theta‐toxin, beta‐galactosidase, arginine metabolism enzymes and kappa‐toxin, respectively, as gene probes. It is suggested that the productivity of theta‐toxin in these strains is diverse because of the multiple genetic backgrounds including single deletion of pfoA, large deletion of the pfoA‐colA region and the putative point mutations.
Interferon‐α and ‐γ Differentially Reduce Rapid Immature T‐Cell Death by Contact with HIV‐1 Carrier Cell Clones In Vitro
Authors: Kinzl Petra , Otani Takeshi , Benz Roland , Minowada Jun 
Published: November 2013

AbstractThe non‐antigen specific rapid cytotoxic (CT) death of immature TdT+CD4+CD8+ T cells due to contact with HIV‐1 carrier T‐cell clones we have found recently is a novel phenomenon. The effects of interferons (IFN) on this CT reaction were studied in vitro. Treatment of the HIV‐1 carrier clones, referred to as “effectors,” with IFN‐α but not IFN‐γ, or of the susceptible immature TdT+CD4+CD8+ T cells, referred to as “targets,” with IFN‐γ but not IFN‐α, for 24 hr prior to CT testing was found to reduce the CT reaction. Simultaneously, a down‐regulated CD8 expression and an up‐regulated antigen expression of both major histocompatibility antigen complex class I (MHC‐I) and HIV‐1 gp120/gp160 in the IFN‐α treated effector (gp120+CD8+ HPB‐ALL/HIV), and/or simultaneously up‐regulated antigen expression of both CD8 and MHC‐I in the IFN‐γ treated target (CD4+CD8+ HPB‐ALL) were found to be associated with reduced CT reaction. However, altered antigen expression in the IFN‐γ treated effectors or IFN‐α treated targets did not affect the ultimate degree of CT reaction. This study thus suggests a possible therapeutic efficacy of IFN by reducing the direct elimination of the T‐cell precursors in HIV‐1 infection.
Negative Finding in Cross‐Protective Activity of JapaneseBorreliaIsolates against Infection with Three Species of Lyme DiseaseBorreliain Outbred Mice
Authors: Masuzawa Toshiyuki , Kurita Toru , Yanagihara Yasutake 
Published: November 2013

AbstractOuter surface protein A (OspA) is the most promising candidate for a component vaccine against Lyme disease caused byBorrelia burgdorferisensu lato. Active cross‐protection using a whole‐cell vaccine prepared from strains belonging various OspA serotypes observed in Japan and worldwide was examined. No cross‐protection was obtained by heterologous OspA‐serotype vaccines. Since OspA is a highly variable protein expressed byBorrelia, this suggests that immunologically different OspA serotypes need to be combined for the development of an effective vaccine in Japan.
Cloning and Characterisation of the aroA and aroD Genes of Shigella dysenteriae Type 1
Authors: Walker John C. , Verma Naresh K. 
Published: November 2013

AbstractThe aroA and aroD genes from Shigella dysenteriae type 1, encoding 5‐enolpyruvylshikimate 3‐phosphate synthase and 3‐dehydroquinase, respectively, were cloned by polymerase chain reaction (PCR). Their nucleotide sequences were determined and predicted to code for 46 kDa and 27.5 kDa proteins, respectively. Protein expressed from these genes using the minicell system, corresponded to the size of the predicted protein products. The cloned genes were shown to be functional by complementation of Escherichia coli aroA and aroD− mutants. The predicted amino acid sequences of the cloned aroA (427 amino acids) and aroD (252 amino acids) genes of S. dysenteriae type 1 were found to be highly homologous to the corresponding genes in other bacterial species, indicating the high conservation of these housekeeping genes. The use of the cloned aroA and aroD genes in the development of a vaccine strain against S. dysenteriae is discussed.