bluuebunny/crossref_metadata_2025_split huggingface dataset:
Liver‐Associated Natural Killer Activity in Cirrhotic Rats
Authors: Shirachi Miki , Sata Michio , Miyajima Ichiro , Tanaka Masatoshi , Tanikawa Kyuichi 
Published: November 2013

AbstractAn impaired host defense mechanism is well known in patients with liver cirrhosis (LC). Using a sinusoidal lavage method, lymphocytes were obtained from LC rats that were administered thioacetamide, and natural killer (NK) activity was measured by 5lCr‐release assay. The NK cell count was measured by flow cytometric analysis using monoclonal antibody (Mab) 3.2.3 and/or CD 3‐8+ as markers for NK cells, and by immunohistochemical staining using Mab 3.2.3. Furthermore, interferon (IFN) α was administered to LC rats and the subsequent changes in hepatic NK activity and NK cell count were observed. In the large granular lymphocyte (LGL)‐rich fraction (Fr.1, LGLs: 60‐90%), the NK activity was significantly lower in the LC rats (40.0±3.8%) compared to that in the control rats (48.4±4.3%) (P<0.005). In addition, the number of NK cells in the liver tissues of the LC rats was significantly lower compared to that in the liver tissues of the control rats by morphometric analysis (P<0.05). For LC rats, NK activity of the Fr.1 24 hr after IFNα administration (5×104 IU / 100 g body weight) increased significantly (P<0.005). Hepatic NK activity and NK cell count were reduced in the LC rats, and recovered following IFNα administration. The results obtained in this study may give clues to better understanding the impaired host defense mechanism in LC patients.
Construction of the Chimeric Reverse Transcriptase of Simian Immunodeficiency Virus Sensitive to Nonnucleoside Reverse Transcriptase Inhibitor
Authors: Isaka Yoshitaka , Sato Akihiko , Kawauchi Shinobu , Suyama Akemi , Miki Shigeru , Hayami Masanori , Fujiwara Tamio 
Published: November 2013

AbstractA number of structurally diverse compounds have been shown to be potent inhibitors of the DNA polymerase activity of human immunodeficiency virus type 1 (HIV‐1) reverse transcriptase (RT). The compounds can be grouped into two broad classes; nucleoside analogs and nonnucleoside RT inhibitors. The nonnucleoside RT inhibitors are quite specific for HIV‐1 RT but not human immunodeficiency virus type 2 (HIV‐2) and simian immunodeficiency virus (SIV) RT. We have investigated the property of SIV/HIV‐1 chimeric viruses in which portions of SIVMAC RT were exchanged with the corresponding domain of HIV‐1 RT; amino acids 176‐190, 176‐383 and 176‐495 of HIV‐1 RT. The chimeric virus, which was substituted amino acids 176‐190 of RT, had detectable RT activity, and this chimeric RT was sensitive to three nonnucleoside RT inhibitors [nevirapine, HEPT derivative (E‐EBU‐dM) and TIBO derivative (R82913)]. To further study this chimeric virus, we purified the chimeric RT enzyme expressed in Escherichia coli and determined its kinetic properties; the Km and Vmax values, and the Ki value of HEPT derivative calculated for the DNA polymerase activity. This study reveals that amino acids 176‐190 of SIVMAC RT were important for the enzymatic activity and the SIV/HIV‐1 chimeric RT, which had amino acids 176‐190 of HIV‐1, was sensitive to the nonnucleoside RT inhibitor.
Filamentous Phage fs1 of Vibrio cholerae O139
Authors: Nakasone Noboru , Honma Yasuko , Toma Claudia , Yamashiro Tetsu , Iwanaga Masaaki 
Published: November 2013

AbstractFilamentous phage, fs1, was obtained from Vibrio cholerae O139. The lysogenized strains produced a large amount of fs1 phage in the culture supernatant. This phage was previously reported as novel fimbriae of that organism. The genome of the phage was a 6.5 kb single‐stranded DNA. The capsid of fs1 consists of a small molecule peptide (about 2.5 kDa).
Human Immunodeficiency Virus Type 1 (HIV‐1) Infection and Transcytosis Activity of a HIV‐1 Susceptible Clone from HeLa Cell
Authors: Morizono Kouki , Harada Shinji 
Published: November 2013

AbstractIn order to clarify the transmission process of human immunodeficiency virus type 1 (HIV‐1) through the epithelial cell barrier, HeLa cells susceptible and non‐susceptible to HIV‐1 were cloned and designated as P6 HeLa and N7 HeLa cells, respectively. P6 HeLa cells could be infected with the LAI strain of HIV‐1 and mediated HIV‐1 transcytosis. In contrast, N7 HeLa cells exhibited neither HIV‐1 infection nor transcytosis. CD4 and galactosylceramide as the receptors for HIV‐1 were not detected on P6 HeLa cells, although an anti‐CD4 monoclonal antibody (mAb) blocked HIV‐1 infection. Since HIV‐1‐infected P6 HeLa cells exhibited no fusion and survived, we speculated that the P6 HeLa cells expressed molecules other than CD4 which facilitated HIV‐1 infection. Two mAbs (A‐14 ITK and C57 a9‐9) which inhibited the HIV‐1 infection of P6 HeLa cells were generated. Each mAb recognized distinct molecule(s) as shown by Western blotting. Transcytosis by the P6 HeLa cells was inhibited by C57 a9‐9 but not by A‐14 ITK or anti‐CD4 mAb. Both infection and transcytosis may be responsible for HIV‐1 transmission through epithelial cells in a complex manner. Although infection and transcytosis occurred via different mechanisms, the molecule(s) recognized by C57 a9‐9 mAb may be associated with both processes.
Evidence for a Dual Strategy in the Expression of Cauliflower Mosaic Virus Open Reading Frames I and IV
Authors: Kobayashi Kappei , Tsuge Seiji , Nakayashiki Hitoshi , Mise Kazuyuki , Furusawa Iwao 
Published: November 2013

AbstractStudies have indicated that cauliflower mosaic virus (CaMV) gene expression is mediated by the translation of polycistronic 35S pregenomic RNA, but the involvement of some minor subgenomic RNA species is also suspected. We examined the involvement of the 35S promoter in the expression of CaMV open reading frames (ORFs) I and IV using both 35S RNA‐driven and promoter‐less ORF I‐ and ORF IV‐β‐glucuronidase (GUS) fusion constructs. In addition to the 35S promoter‐dependent expression of both ORF I‐ and IV‐GUS fusions, we detected the 35S promoter‐independent expression of both fusion genes via subgenomic mRNAs, which were detected by Northern blotting in the protoplasts transfected with the 35S promoter‐driven constructs as well as in those transfected with the promoter‐less constructs. These results suggest the involvement of subgenomic RNAs in the expression of CaMV ORFs I and IV, and the operation of a dual strategy in the expression of two viral genes.
Epitope Specificity of a Monoclonal Antibody Generated against the Dissociated CFA/I Fimbriae of Enterotoxigenic Escherichia coli
Authors: De Luna Maria Das Graças , Rudin Anna , Vinhas Solange Alves , de Almeida Darcy Fontoura , de Souza Ferreira Luis Carlos 
Published: November 2013

AbstractA monoclonal antibody (MAb 84) raised against the dissociated CFA/I fimbriae of enterotoxigenic Escherichia coli was characterized with regard to antigen binding and epitope specificity. Enzyme‐linked immunosorbent assay (ELISA) showed that MAb 84 had higher affinity to CFA/I subunits than to intact CFA/I fimbriae and recognized a Salmonella flagellin carrying an insert corresponding to amino acids 32 to 45 of the CFA/I subunit. Fine epitope mapping based on the Pepscan technique showed that the peptide 39TFESY43, derived from the sequence of the mature CFA/I subunit, was specifically recognized by MAb 84. The 39TFESY43 sequence is probably not accessible on the surface of the native CFA/I fimbriae since MAb 84 did not bind to intact fimbriae as evaluated in inhibition ELISA tests. Moreover, MAb 84 did not agglutinate fimbriated ETEC cells nor inhibit CFA/I‐mediated hemagglutination or the adhesion to Caco‐2 cells.
Measurement of Arginine Carboxypeptidase‐Generating Activity of Adult Plasma
Authors: Watanabe Motoi , Ishikawa Yasushige , Campbell William , Okada Hidechika 
Published: November 2013

AbstractArginine carboxypeptidase (CPR) is a novel carboxypeptidase which was first described by Campbell and Okada. CPR is generated from a stable precursor of CPR (proCPR) during coagulation or under other circumstances and is promptly inactivated at 37 C. Therefore, it is not easy to determine CPR in blood samples. Since proCPR can be separated from the other basic carboxypeptidase (carboxypeptidase N; CPN) by passing plasma through DEAE gel, we have established a method to determine the amount of proCPR after converting it to active CPR by trypsin treatment. We first separated the proCPR from CPN using a filter cup tube (FC tube) packed with DEAE Sephadex, and measured activity after conversion of the enzyme to its active form using trypsin. With this method, no significant decrease in proCPR was noted in the plasma of patients including those with rheumatoid arthritis (RA), although CPR activity in fresh sera has been reported to be decreased. This discrepancy suggests that proCPR is not depleted in most patient sera, but that the level of activity of the enzyme which converts proCPR into active CPR may be compromised in RA patients.
Regulation of Promoter and Intron Enhancer Activity in Immunoglobulin Heavy‐Chain Genes during B‐Cell Differentiation
Authors: Naito Akihiro , Suzuki Yasuhiko , Azuma Takachika 
Published: November 2013

AbstractChloramphenicol acetyltransferase (CAT) transgenic mice, in which the transgene is regulated by the VH promoter and heavy‐chain intron enhancer (Eμ), were examined to investigate the variation of activity of these cis‐acting elements during the late stage of B‐cell development. CAT enzyme activity decreased when resting B cells were stimulated through B‐cell receptors (BCR) with goat anti‐mouse IgM antibodies in vitro. On the other hand, when these B cells were stimulated by lipopolysaccharide (LPS) in vitro, they showed enhanced CAT activity, accompanied by an increase in the number of CD43+ B220+ cells (pro‐plasma and plasma cells). In addition, the CAT activities in CD43+ B220+ and PNAhi B220+ cells from immunized mice were higher than those in CD43− B220+ and PNAlo B220+ cells, respectively. These results suggest that the activity of Eμ in the context of VH promoter was transiently down‐regulated by stimulation through the BCR but enhanced at the pro‐plasma and plasma stages.
Occurrence of Coagulase Serotype among Staphylococcus aureus Strains Isolated from Healthy Individuals —Special Reference to Correlation with Size of Protein‐A Gene—
Authors: Sekr Keiko , Sakurada Junji , Seong Hee Kyong , Murai Miyo , Tachi Hidemi , Ishii Hiroko , Masuda Shogo 
Published: November 2013

AbstractOne‐hundred‐and‐nineteen strains of Staphylococcus aureus isolated from healthy individuals for 3 years between 1991 and 1993 were subjected to an investigation on the producibility of proteins including protein A, coagulase, enterotoxins and toxic‐shock syndrome toxin‐1. Especially, protein A was the center of our interest. Among these strains, 69, 43, 3 and 1 strains were found to have the protein‐A gene containing 5, 4, 3 and 2 IgG‐binding domains, respectively. On the other hand, only one strain was devoid of the protein‐A gene. There were some differences in the profile of the coagulase serotype between the group with 4 IgG‐binding domains and that with 5 IgG‐binding domains. Differences in the profile of toxin production were also observed between the two groups.
Differentiation of Chlamydia Species by Combined Use of Polymerase Chain Reaction and Restriction Endonuclease Analysis
Authors: Yoshida Hiroshi , Kishi Yuichiro , Shiga Sadashi , Hagiwara Toshikatsu 
Published: November 2013

AbstractTo differentiate Chlamydia spp., a primer pair designed to generate a genus‐specific region of the major outer membrane protein (MOMP) gene was used in a PCR to amplify a single DNA fragment of 245‐259 bp. In the PCR, the expected single DNA fragment was amplified from strains of Chlamydia trachomatis, C. psittaci, C. pneumoniae and C. pecorum, respectively. By restriction endonuclease analysis with AluI and PvuII, the amplified products exhibited four distinct patterns, corresponding to the four species. It is, therefore, concluded that one‐step PCR followed by restriction endonuclease analysis as described in this study could be a valuable method for the detection and differentiation of Chlamydia species.
Induction of Cross‐Reactivity in an Endogenous Viral Peptide Non‐Reactive to FBL‐3 Tumor‐Specific Helper T‐Cell Clones
Authors: Uenishi Hirohide , Iwanami Norimasa , Yamagishi Hideo , Nakatani Takako , Kawasaki Toshisuke , Tamamura Hirokazu , Fujii Nobutaka , Kuribayashi Kagemasa 
Published: November 2013

AbstractWe previously reported a helper T‐cell (Th) epitope (peptide i) which corresponded to the sequence ranging from positions 462 to 479 from the N‐terminus of the Friend‐murine leukemia virus (F‐MuLV) envelope protein (env462‐479). Homologous sequences exist in both Moloney‐murine leukemia (M‐MuLV env452‐469) and endogenous AKV (AKV env453‐470) viruses, which differ from F‐MuLV env462‐479 in 5 and 7 amino acids, respectively. However, peptide i‐specific Th clones did not respond to either of the corresponding exogenous or endogenous peptides. One amino acid substitution in M‐MuLV env452‐469 (Asn to Tyr at position 465: N465Y) and three amino acids in AKV env453‐470 (H460S, A466Y and Y468H) endowed both peptides with the reactivity to one of the Th clones, F5‐5, almost to the same degree as peptide i. However, the other Th clones responded differently to each of the modified endogenous peptides substituted by one to three amino acids. The cells responsive to the cross‐reactive peptides occupied only a minor portion, if any, of the bulk cultured lymph node cells from peptide i‐immune mice, and in particular, no significant response to the modified endogenous peptides was observed in repeated experiments. The exchange of at least 3 residues was necessary for the endogenous peptide to acquire sufficient cross‐reactivity to two of the three Th clones. However, it was noticeable that a single substitution of alanine by tyrosine at the dominant T‐cell receptor (TCR) contact position of the peptide ie generated a weak but significant cross‐reactivity to one of the three Th clones in this study. Thus, peptides of endogenous retroviral origin that would be modified by mutational events might become ‘non‐self’ and prime Th cells leading to auto‐antibody production and resulting in autoimmune disease.
Utilization of Proteinase K‐Treated Cells as Lipopolysaccharide Antigens for the Serodiagnosis of Helicobacter pylori Infections
Authors: Amano Ken‐ichi , Yokota Shin‐ichi , Ishioka Tomonori , Hayashi Shunji , Kubota Toru , Fujii Nobuhiro 
Published: November 2013

AbstractWe have evaluated the use of proteinase K (PK)‐treated cells isolated from Helicobacter pylori as lipopolysaccharide (LPS) antigens in an immunoblot assay and an enzyme‐linked immunosorbent assay (ELISA) for the serodiagnosis of H. pylori infection. The sera from patients with chronic gastritis, gastric ulcer, duodenal ulcer or gastric cancer, and from healthy adults with or without H. pylori infection were assayed with three commercial serodiagnostic kits (HM‐CAP, Helico‐G, and G.A.P. II) and novel methods relying on the use of PK‐treated cells. The PK‐treated cells used in these assays were selected on the basis of their possibility to possess a common epitope in the O‐polysaccharides of H. pylori, which is known to be highly immunogenic in humans. Of the sera from these patients, 71‐94% were positive with the commercial kits, 97% with immunoblot assay, and 90% with ELISA. On the other hand, of the healthy adults infected with H. pylori, 72‐97% were positive with the commercial kits, 86% with immunoblot assay, and 72% with ELISA. PK‐treated cells that did not contain the common epitope were unsuitable as an antigen for immunoblot assay or ELISA. Furthermore, the reactivity of these sera reacted specifically with H. pylori PK‐treated cells but not with LPSs from other gram‐negative bacteria, such as Campylobacter, Proteus, Bordetella, and Salmonella. These results demonstrate that the serological assays relying on the use of H. pylori PK‐treated cells possessing a highly antigenic epitope are potentially useful as a serodiagnostic test for H. pylori infection.
Induction of Apoptosis and Tyrosine Phosphorylation of Cellular Proteins in T Cells and Non‐T Cells by Stimulation with Concanavalin A
Authors: Nagase Fumihiko , Abo Tomoko , Hiramatsu Kumiko , Suzuki Shin‐ichi , Du Jun , Nakashima Izumi 
Published: November 2013

AbstractA high concentration (30 μg/ml or more) of Con A caused the death of not only thymocytes but also splenic cells of BALB/c mice, whereas a moderate concentration (3 μg/ml) of Con A induced proliferation of these cells. A high concentration of Con A also induced the death of splenic cells of athymic BALB/c‐nu/nu mice and the bone marrow cells of BALB/c mice which mainly consist of non‐T cells. However, any concentration (1‐30 μg/ml) of Con A failed to induce the proliferation of these cells. Specific binding of tetrameric Con A to mannose‐containing receptors was required for the induction of cell death. DNA fragmentation was observed by both laser flow cytometry and electrophoresis in Con A‐stimulated T cells and non‐T cells. This indicated that the mechanism of induction of apoptosis with Con A is not necessarily TCR‐dependent. Con A induced tyrosine phosphorylation of a number of proteins in various types of cells. Interestingly, phosphorylation of the 40 kDa protein developed only in the thymocytes and spleen cells that contain T cells, whereas phosphorylation of the 80 and 120 kDa proteins appeared in both T cells and non‐T cells. These results suggested that the Con A‐induced apoptosis of T cells and non‐T cells involves different but possibly mutually related protein tyrosine phosphorylation‐linked signals.
Rapid Analysis of Allelic Variants of the Sheep PrP Gene by Oligonucleotide Probes
Authors: Ishiguro Naotaka , Shinagawa Morikazu , Onoe Sadao , Yamanouchi Kazunori , Saito Toshiro 
Published: November 2013

AbstractA rapid method to determine the allelic variants of the sheep PrP gene was developed. DNA samples from 128 Suffolk sheep (39 rams and 89 ewes) were screened by using polymerase chain reactions and dot‐blot hybridization with 32P‐labeled nine allele‐specific oligonucleotide probes corresponding to the polymorphic PrP codons 112, 136, 154 and 171. Three allelic variants of the PrP gene, PrPMARQ, PrPTARQ and PrPMARR, were found in the flocks. Among those variants, nearly half of the ewes had alleles of the 171‐Arg variant that is closely associated with resistance to natural scrapie. Assessments of allelic mutations of the PrP gene may help to select the scrapie‐resistant progenitors in the flocks.
Regulation of GM‐CSF‐Induced Dendritic Cell Development by TGF‐β1 and Co‐Developing Macrophages
Authors: Yamaguchi Yasunori 
Published: November 2013

AbstractUsing a culture system of bone marrow progenitor cells with GM‐CSF and TGF‐β1, a study was performed to analyze the effect of TGF‐β1 on the development of dendritic cells (DC) and to elucidate the regulatory role of macrophages co‐developing with dendritic cells. The results demonstrate that DC generated in the presence of TGF‐β1 were immature with respect to the expression of CD86, nonspecific esterase activity and cell shape. Such inhibitory effects of TGF‐β1 were dependent on FcR+ macrophages, which were depleted by panning. TGF‐β1 did not appear to inhibit the commitment of progenitor cells to the DC lineage. In addition, TGF‐β1 also acted directly on the intermediate stage of DC to prevent their over‐maturation, which results in a preferential decrease in MHC class II, but not in CD86, in the presence of TNF‐α. FcR+ suppressive macrophages were also shown to facilitate DC maturation when stimulated via FcR‐mediated signals even in the presence of TGF‐β1. These results indicate that TGF‐β1 indirectly and directly regulate the development of DC and that co‐developing macrophages have a regulatory role in DC maturation.
Developmental Regulation by Cytokines of Bone Marrow‐Derived Dendritic Cells and Epidermal Langerhans Cells
Authors: Yamaguchi Yasunori 
Published: November 2013

AbstractDendritic cells (DC) are specialized antigen‐presenting cells involved in T cell‐mediated immune responses. Differentiation and functional maturation of the DC are now known to be regulated by various cytokines, including TGF‐β1. The experiments of this study examined the effect of other cytokines, such as IL‐4, IL‐10 and IL‐6, on the differentiation and maturation of bone marrow (BM)‐derived DC (BM‐DC) and epidermal Langerhans cells (LC). When IL‐6 or IL‐10 was added to cultures of BM cells in the presence of GM‐CSF, both cytokines, as in the case of TGF‐β1, suppressed the maturation of DC in terms of the expression of adhesion and costimulatory molecules and T cell‐stimulating activity. In contrast, IL‐4 was not suppressive but rather supportive for the differentiation of DC. However, these suppressive cytokines hardly counteracted the maturation‐inducing activity of TNF‐α when added to cultures of immature DC. In addition, they appeared to block the overmaturation of DC, which is characterized by a loss of MHC class II molecules. Regarding LC maturation in epidermal cell cultures, IL‐6 and IL‐10 were inhibitory for the expression of CD86 and CD80 in a dose‐dependent fashion. Unlike BM‐DC, LC maturation was slightly enhanced by TGF‐β1. The protein antigen‐presentation by LC to Th1 clone was not affected by IL‐6, but slightly reduced by IL‐10. These results suggest that each cytokine contributes to regulate the differentiation and maturation of DC at a different developmental stage.
Rapid Serodiagnosis of Mycobacterium avium‐intracellulare Complex Infection by ELISA with Cord Factor (Trehalose 6, 6′‐Dimycolate), and Serotyping Using the Glycopeptidolipid Antigen
Authors: Enomoto Kyoko , Oka Shiro , Fujiwara Nagatoshi , Okamoto Takashi , Okuda Yoshinari , Maekura Ryoji , Kuroki Tetsuo , Yano Ikuya 
Published: November 2013

AbstractMycobacterium avium‐intracellulare complex (MAC) is one of the most important opportunistic pathogens, particularly in patients with acquired immunodeficiency syndrome (AIDS). The aim of this study was to determine whether an enzyme‐linked immunosorbent assay (ELISA) using trehalose 6,6′‐dimycolate (TDM) as an antigen can be used for the rapid serodiagnosis of MAC infection. We also identified MAC serotypes by ELISA using serotype‐specific glycopeptidolipid (GPL) antigen. To confirm our findings, the thin‐layer chromatographic (TLC) behavior of serotype‐specific GPL of the strains isolated from MAC‐infected patients was also tested. Forty patients infected with MAC and 30 healthy controls were tested. Thirty‐two of the 40 MAC‐infected patients had higher titers of serum antibodies against MAC TDM than against MTB TDM, while all 30 healthy control sera were unreactive to MAC TDM and MTB TDM. Results of the GPL ELISA indicated that 20 of the 40 MAC‐infected patients' sera were reactive against serotype 4 GPL, 3 against serotype 8 GPL, and 1 against serotype 16 GPL. A TLC analysis of the GPL of the 40 MAC isolates showed that 16 strains were of serotype 4, 5 of serotype 8, and 2 of serotype 16. Results of the GPL ELISA were in good accord with those of the TLC analysis for most patients. Our findings suggest that ELISA using TDM is useful for rapid serodiagnosis of MAC infection, and that complementary ELISA testing using serotype‐specific GPL gives additional detailed information concerning MAC serotypes.