bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cytokine Secretion by Stimulated Monocytes Depends on the Growth Phase and Heat Treatment of Bacteria
Authors: Haller Dirk , Bode Christiane , Hammes Walter P. 
Published: November 2013

AbstractThe consumption of food containing lactic acid bacteria (LAB) has been shown to exert immunomodulatory effects in humans. The specific cellular interaction of these bacteria with immuno‐competent cells has not yet been fully understood. Since the TNF‐α secretion of stimulated monocytes is an important initial response to a bacterial challenge, we investigated the potential of LAB originating from the human intestine or fermented food in comparison to the effect of invasive pathogens. The challenge of monocytes with three LAB strains, Listeria monocytogenes or enterohaemorrhagic Escherichia coli (EHEC) elicited a strain specific, dose‐dependent biphasic TNF‐α secretion. The concentration (EDmax) of bacteria or bacterial cell wall components necessary to induce maximal TNF‐α secretion (TNFmax) by monocytes was mathematically approximated. It was shown for exponentially growing LAB strains that the maximal TNF‐α secretion (TNFmax) was stronger (57 to 78%) upon stimulation with living bacteria than with heat killed cells. In contrast to log‐phase bacteria, the maximal TNF‐α secretion of monocytes (TNFmax) was higher (15 to 55%) after the stimulation with heat killed, stationary‐phase bacteria when compared to that of live LAB. Thus, monocyte stimulation was clearly affected by the growth phase of bacteria. Purified cell walls of LAB straines revealed only a limited potential for monocyte stimulation. LPS exhibited a higher capacity to stimulate monocytes than purified Gram positive cell walls or muramyldipeptide. In comparison to pathogenic bacteria, the maximal secretory TNF‐α response (TNFmax) was up to 2 fold higher with LAB strains. In general, the amount of bacteria (EDmax) necessary to induce maximal TNF‐α secretion (TNFmax) was approximatly 1 to 3 log higher for heat killed bacteria when compared to live bacterial cells illustrating the significant lower potential of heat killed bacteria to activate monocytes.
Role of the Cytomegalovirus (CMV)‐Antigenemia Assay as a Predictive and Follow‐Up Detection Tool for CMV Disease in AIDS Patients
Authors: Hoshino Yoshihiko , Nagata Yoichi , Taguchi Hitomi , Masunaga Atsuko , Fujino Yujiro , Mochizuki Manabu , Nakamura Tetsuya , Iwamoto Aikichi 
Published: November 2013

AbstractForty‐two patients were evaluated to determine the value of the CMV antigenemia (CMV‐Ag) test as a follow‐up marker as well as a prediction marker of CMV disease. Twenty patients were positive for at least one positive CMV‐Ag assay and 9 of them developed CMV retinitis. With the threshold value (10 positive cells), sensitivity was 56% and specificity was 94%. The CMV‐Ag assay, with the threshold value, produced high specificity, positive predictive value and negative predictive value but relatively poor sensitivity. Eight patients experienced CMV disease relapse a total of 16 times. At relapse, 8 of the 16 times showed negative for CMV‐Ag assay; 7 underwent systemic maintenance while 1 underwent local maintenance. It is inferred that the CMV‐Ag test is a poor follow‐up marker to detect the relapse of CMV disease, particularly in patients undergoing systemic maintenance.
Characterization of the Cyclophilin of Trichophyton mentagrophytes
Authors: Kano Rui , Nakamura Yuka , Watanabe Shinichi , Tsujimoto Hajime , Hasegawa Atsuhiko 
Published: November 2013

AbstractA genetic approach to cyclophilins in a dermatophyte, Trichophyton mentagrophytes, was carried out. The nucleotide and deduced amino acid sequences of the cyclophilin of T. mentagrophytes shared about 70% sequence similarity with those of Schizosaccharomyces pombe, Saccharomyces cerevisiae and Candida albicans. However, the first 21 amino acid and the C‐terminal amino acid regions of 188 to 226 of the T. mentagrophytes cyclophilin were distinct from those of the other fungal cyclophilins. The recombinant glutathione 5‐transferase (GST)‐T. mentagrophytes cyclophilin fusion protein produced by Escherichia coli was purified. The protease digest of the fusion protein had a molecular weight of about 13 kDa and peptidyl‐prolyl cis‐trans isomerase (PPI) activity. This digest protein from T. mentagrophytes was confirmed to be cyclophilin by proving PPI activity.
Exacerbation of Vascular Endothelial Injury in the Generalized Shwartzman Reaction by the Administration of Anti‐E‐Selectin Antibody
Authors: Koide Naoki , Kato Yutaka , Sugiyama Tsuyoshi , Chakravortty Dipshika , Mu Mya Mya , Yoshida Tomoaki , Yokochi Takashi 
Published: November 2013

AbstractPreviously, we reported that the consecutive administration of lipopolysaccharide (LPS) into LPS‐sensitized mice for the generalized Shwartzman reaction (GSR) induced systemic injury of vascular endothelial cells. The aim of this study was to investigate the participation of vascular adhesion molecules in the vascular endothelial injury of GSR. The administration of anti‐E‐selectin antibody in GSR‐induced mice resulted in massive apoptosis of vascular endothelial cells and congestion in blood vessels. Further, marked hemorrhage was found in the pulmonary alveoli of those mice. GSR, especially lung injury, was definitely exacerbated by the administration of anti‐E‐selectin antibody. On the other hand, the administration of anti‐VCAM‐1 antibody did not induce such injury of vascular endothelial cells. The possible role of E‐selectin in the exacerbation of vascular endothelial injury in GSR is discussed.
Intranasal Sensitization of Japanese Cedar Pollen by the Co‐Administration of Low Doses of Cholera Toxin but Not Its Recombinant B Subunit to Mice
Authors: Hirai Terumasa , Hashiguchi Shuhei , Torigoe Naohiko , Toda Yoshihiro , Ito Yuji , Sugimura Kazuhisa 
Published: November 2013

AbstractWe evaluated the effects of cholera toxin (CT) and the B subunit of cholera toxin (CTB) on the intranasal sensitization of Japanese cedar pollen (JCP) in mice. JCP suspended in phosphate‐buffered saline was administered into the nostrils of mice in combination with varying doses of CT or recombinant CTB(r‐CTB) once a week for 5 weeks. Antibody responses specific to sugi basic protein (SBP) were monitored by ELISA for seven weeks. The sensitization of JCP alone did not induce IgGl, IgG2b, IgG2a, IgE or IgA. In contrast, sensitization of JCP in combination with CT (JCP/CT) elicited the prominent production of SBP‐specific IgG1 and low levels of IgG2b and IgG2a on Day 49. IgE production was detected only in the serum of mice which were treated with JCP/CT, and not under any other protocol. Using spleen cells from these mice, cytokine production was examined by ELISA in culture supernatants after they had been stimulated in vitro with major cedar pollen allergens, Cry j 1, Cry j 2 or SBP. Notable responses were an increase of IFN‐γ as well as IL‐4 in JCP/CT‐sensitized cells stimulated with Cry j 2, but not in those stimulated with Cry j 1. No significant differences were detected in IL‐5 production among the experimental groups. Histopathological examination, however, showed that eosinophil infiltration was evident in the nasal mucosa of the JCP/CT‐sensitized mice following challenge with JCP/CT, but weak with BSA/CT or CT alone. Thus, the immunological and histological analyses indicated that the co‐administration of a low dose of CT in combination with JCP allows the induction of pollen‐allergic states in mice.
Autocrine Interferon‐β Stimulation Augments Nitric Oxide Production by Mouse Macrophage J774A.1 Cells Infected with Herpes Simplex Virus Type 1
Authors: Fujioka Noboru , Ohashi Kunihiro , Ikeda Masao , Kurimoto Masashi 
Published: November 2013

AbstractThe pathogenic roles of nitric oxide (NO) in mouse models have been reported for herpes simplex virus type 1 (HSV‐1)‐induced pneumonia as well as endotoxin shock. We compared the mechanism of NO production induced by HSV‐1 with that induced by lipopolysaccharide (LPS) using a mouse macrophage cell line, J774A.1. Both HSV‐1 and LPS induced NO production as well as antiviral activity, which were attenuated by anti‐interferon (IFN)‐β treatment. These results suggest that autocrine IFN‐β plays a role in NO release by J774A.1 cells stimulated with HSV‐1 or LPS.
Reduction of Thymocyte Numbers in Transgenic Mice Expressing Viral FLICE‐Inhibitory Protein in a Fas‐Independent Manner
Authors: Ohyama Takahiro , Tsukumo Shin‐ichi , Yajima Nobuyuki , Sakamaki Kazuhiro , Yonehara Shin 
Published: November 2013

AbstractA viral FLIP (FLICE/caspase‐8‐Inhibitory Protein), equine herpesvirus type 2 E8 protein, has been shown to inhibit Death receptor‐induced apoptosis by suppressing the activation of FLICE/caspase‐8. We generated transgenic mice specifically expressing E8 in thymocytes under the control of lck‐proximal promoter. Although E8‐expressing thymocytes were resistant to Fas‐mediated apoptosis, the total number of thymocytes in 4‐8‐week‐old E8 transgenic mice was more than 3‐fold less than that in control littermates. This reduction was also observed in E8 transgenic mice with a Fas–/– background suggesting the reduction to be independent of Fas. The thymocytes of the transgenic mice, however, could similarly respond to CD3‐mediated stimulation, indicating that the reduction of thymocyte numbers might be independent of T cell receptor complex‐mediated stimulation. Thus, the Death receptor‐mediated signaling pathway is too complex to be regarded as only an executor for apoptosis.
The Role of Osteopontin in the Development of Granulomatous Lesions in Lung
Authors: Chiba Satoru , Rashid Mohammod Mizanur , Okamoto Hiroshi , Shiraiwa Hirotake , Kon Shigeyuki , Maeda Masahiro , Murakami Masaaki , Inobe Manabu , Kitabatake Akira , Chambers Ann F. , Uede Toshimitsu 
Published: November 2013

AbstractOsteopontin (OPN) has been shown to be expressed by cells in granulomas of various origins, but whether it plays a functional role in granuloma formation is not known. Here we used a cardiomyopathic hamster (T02) model, to test the hypothesis that OPN contributes functionally to granuloma development. We immunized cardiomyopathic and normal hamsters by subcutaneous injection of bovine serum albumin in complete Freund's adjuvant, and assessed various tissues for both OPN RNA expression and granuloma formation. Cardiomyopathic hamsters expressed OPN, and formed granulomatous lesions, in heart tissue in both immunized and untreated animals. In addition, immunization induced expression of OPN in lung and lymph nodes of cardiomyopathic (but not normal) hamsters, and also induced granuloma formation in these organs. To test whether OPN expression could play a functional role in inducing granulomas, we produced an adenoviral vector containing the murine OPN gene, and introduced this vector intratracheally into the lungs of normal hamsters. The OPN‐containing vector, but not the control vector, induced pulmonary granuloma formation. These studies provided direct in vivo evidence that OPN can contribute functionally to the formation of granulomatous lesions, and suggest that OPN expression may be a common factor involved in formation of granulomas of various origin.
Detection of Hantaviral Antibodies among Patients with Hepatitis of Unknown Etiology in Japan
Authors: Kariwa Hiroaki , Yoshimatsu Kumiko , Araki Koichi , Chayama Kazuaki , Kumada Hiromitsu , Ogino Michiko , Ebihara Hideki , Murphy Michael E. , Mizutani Tetsuya , Takashima Ikuo , Arikawa Jiro 
Published: November 2013

AbstractHantaviral antibodies were detected in the sera from patients with hepatic disease of unknown etiology in Japan by several different serological diagnostic methods. A total of 105 sera from diseased patients which were negative to AG hepatitis virus infections in the Tokyo area were tested. Among them, 3 out of 73 sera from patients with chronic hepatic disease were positive to hantaviral antibody by enzyme‐linked immunosorbent assay (ELISA), indirect immunofluorescent antibody assay (IFA) and Western blot analysis (WB). Neutralizing antibody titers of the 3 sera to Seoul virus (SEO) were 4 to 8 times higher than those to Hantaan virus (HTN). However, all of the 32 sera from patients with acute hepatitis were negative for hantaviral antibody. Among the 60 patients with chronic hepatitis in Hokkaido which were serologically negative to B and C hepatitis virus infection, one was positive for hantaviral antibody by ELISA and WB. In contrast, the sera from healthy adults in Japan, 550 from the Honshu and Kyushu regions, and 1,000 from the Hokkaido region, were negative for hantavirus antibody. These results show that hantaviral antibodies are more frequently detected in patients with hepatic disease than in healthy adults. However, the observation that no positive sera were detected from patients with acute hepatitis implies that hantavirus might not be directly related to hepatitis.
Protection Induced in Mice Vaccinated with Recombinant Collagen‐Binding Protein (CnBP) and Alpha‐Toxoid against Intramammary Infection with Staphylococcus aureus
Authors: Mamo Wubshet , Fröman Gunnar , Müller Hans‐peter 
Published: November 2013

AbstractMice vaccinated with a combination of two Staphylococcus aureus antigens consisting of a recombinant collagen‐binding protein (CnBP) and alpha‐toxoid (α‐toxoid) were significantly protected from intramammary challenge infection with S. aureus. The average number of bacteria recovered from the glands of mice vaccinated with the combination of CnBP/α‐toxoid was significantly lower compared to the average number of bacteria recovered from the glands of mice vaccinated with only CnBP or α‐toxoid or controls (P ≤ 0.01). Histopathological examination of mammary glands of mice vaccinated with CnBP together with α‐toxoid showed no pathological changes, whereas glands of mice vaccinated with CnBP or α‐toxoid alone developed severe mastitis and showed both focal and disseminated necrosis.
Sequence of the Gene Encoding an Alkaline Serine Proteinase of Bacillus pumilus TYO‐67
Authors: Aoyama Misako , Toma Claudia , Yasuda Masaaki , Iwanaga Masaaki 
Published: November 2013

AbstractThe complete nucleotide sequence of the gene encoding an alkaline serine proteinase (aprP) of Bacillus pumilus TYO‐67 was determined. The sequence analysis showed an open reading frame of 1,149 bp (383 amino acids) that encoded a signal peptide consisting of 29 residues and a propeptide of 79 residues. The deduced 3 amino acid residues, D32, H64, and S221, were identical with 3 essential amino acids in the catalytic center of subtilases. The sequence around these residues revealed that APRP was a new member of the true subtilisin subgroup of the subtilisin family. The highest homology was found in subtilisin NAT at 64.4% in the DNA sequence. The residue S189 of APRP was different from those of other subtilases.
The IgM Antibody Level against Ganglioside GM2 Correlates to the Disease Status of HIV‐1‐Infected Patients
Authors: Wu Xiaoshan , Okada Noriko , Goto Mieko , Iwamoto Aikichi , Okada Hidechika 
Published: November 2013

AbstractHIV‐1 infection induces the expression of high level of GM2 ganglioside on infected cells and IgM antibody (Ab) against GM2 can cause complement (C)‐mediated cytolysis of HIV‐1‐infected cells. Since GM2 is immunogenic in human, we proposed that an anti‐GM2 IgM Ab may be produced by some HIV‐1‐infected patients and the titer of this Ab might provide some insight into the progress of the disease. On this premise, the amount of IgM Ab against GM2 was determined in 124 HIV‐1‐infected patients and 111 seronegative donors. As expected, the anti‐GM2 IgM Ab titers of the patients was significantly higher than that of the seronegative donors while the total IgM levels remained unchanged. In addition, we determined the CD4+ cell count and the HIV‐RNA load in the HIV‐1‐infected patients. The results showed a positive correlation between the anti‐GM2 IgM Ab titer and CD4+ cell count but a negative correlation between the anti‐GM2 IgM Ab titer and HIV‐RNA load. These suggest that anti‐GM2 IgM Ab induced and/or enhanced by HIV‐1 infection causes C‐mediated cytolysis of HIV‐1‐infected cells in vivo to a certain extent, and may help lower the plateau level of the HIV‐RNA load. Therefore, the amount of IgM Ab against GM2 may be related to the prognosis of HIV‐1 infected patients.
Development of Hyperfimbriated Strains of Vibrio cholerne O1
Authors: Ehara Masahiko , Iwami Mamoru , Ichinose Yoshio , Hirayama Toshiya 
Published: November 2013

AbstractThe Vibrio cholerae O1 and O139 fimbrillin genes (fimA or mshA) were amplified by polymerase chain reaction and cloned into an Escherichia coli pCR™ vector. These clones were sequenced. The fimA sequences were found to be identical between V. cholerae O1 and O139. One of the plasmids was digested with EcoR I and inserted into the EcoR I site of pGEX‐3X. The plasmid pVPP thus obtained was transferred into strains of wild‐type V. cholerae O1 Bgd17 (classical in biotype) and its fimbriated strain by electroporation. The recombinant plasmid pVPP overexpressed mature fimbriae following induction of the tac promoter with isopropyl‐beta‐d‐thiogalactopyranoside. The cloned gene product was purified to homogeneity by sucrose‐linear gradient centrifugation (7.8 mg of fimbriae/L‐culture). All the properties of the recombinant fimbriae (e.g., subunit structure, hydrophobicity, hemagglutinating activity sensitive to d‐mannose and d‐glucose and immunogenicity) were identical to those of the wild‐type fimbriae. This overexpression system will be extremely useful for rapid, inexpensive preparation of large amounts of fimbriae for vaccine design and development.
Phylogenetic and Taxonomic Heterogeneity of Cryptococcus humicolus by Analysis of the Sequences of the Internal Transcribed Spacer Regions and 18S rDNA, and the Phylogenetic Relationships of C. humicolus, C. curvatus, and the Genus Trichosporon
Authors: Sugita Takashi , Takashima Masako , Ikeda Reiko , Nakase Takashi , Shinoda Takako 
Published: November 2013

AbstractThe phylogenetic and taxonomic heterogeneity of a rare opportunistic yeast pathogen, Cryptococcus humicolus, was revealed by analysis of the sequence of the internal transcribed spacer (ITS) region. Sixteen strains of C. humicolus showed a wide diversity in their ITS sequences. In addition, their 18S rDNA sequences were determined and used to analyze the phylogenetic relationships among C. humicolus and related yeasts. On trees constructed by the Neighbor‐Joining and Maximum Parsimony methods, C. humicolus strains were phylogenetically closely related to each other with the exception of one strain, and they clustered with C. curvatus and Trichosporon species with high bootstrap values. Three C. humicolus strains obtained from humans belonged to the group of Trichosporon serotype I species. The results suggest that C. humicolus is a genetically heterogeneous species which should be reclassified on the basis of DNA sequence data.
Modification of Autolysis by Synthetic Peptides Derived from the Presumptive Binding Domain of Staphylococcus aureus Autolysin
Authors: Takano Mayumi , Oshida Tadahiro , Yasojima Akira , Yamada Masaki , Okagaki Chieko , Sugai Motoyuki , Suginaka Hidekazu , Matsushita Tadahiro 
Published: November 2013

AbstractThe autolytic cell wall hydrolase of Staphylococcus aureus, Atl, contains three highly cationic repeats in the central region of the amino acid sequence, and the repeats are presumed to have the role of binding the enzyme to some components on the cell surface. To explain the possible function of the repeats, we synthesized a number of 10‐ to 30‐mer oligopeptides based on the Atl amino acid sequence (Thr432–Lys610) containing repeat 1, and examined their effects on the autolysis of S. aureus cells. When the peptides were added to a cell suspension of S. aureus under low ionic strength conditions, five peptides, A10, A11, A14, A16 and B9, showed immediate increases in optical density (OD) of the cell suspension accompanied by decreases in viable cell counts. After the immediate increases, the ODs for A10 and A14 changed little in the first 2 hr. In contrast, the ODs for A11 and A16 decreased rapidly. When peptide A10 was added to suspensions of heat‐killed whole cells, crude cell walls and a crude peptidoglycan preparation, their ODs were increased approximately 2‐fold. In contrast, the OD was not increased when the peptide was added to a suspension of pure peptidoglycan from which anionic polymers had been removed. Light microscopic and transmission electron microscopic study showed that A10 and A14 inhibited autolysis and that A11 and A16 induced autolysis earlier than the control. These results suggest strongly that the peptides adsorb to and precipitate on the anionic cell surface polymers such as teichoic acid and lipoteichoic acid via ionic interaction. The effects of peptides on the autolysis may be the results of the modification of S. aureus autolysin activities. These peptides, especially the 10‐mer peptide B9 (PGTKLYTVPW) that represents the C‐terminal half of A10 and N‐terminal half of A11, may be important segments for Atl to bind to the cell surface.
Mumps Virus Can Suppress the Effective Augmentation of HPC‐Induced Apoptosis by IFN‐γ through Disruption of IFN Signaling in U937 Cells
Authors: Hariya Yasushi , Yokosawa Noriko , Yonekura Noriyuki , Kohama Gen‐iku , Fujii Nobuhiro 
Published: November 2013

AbstractCells of the human promonocytic cell line U937 were found to be sensitive to hexadeeylphosphocholine (HPC), which is a potential anticancer drug. Induction of apoptosis was found in U937 cells after treatment with HPC for 24 to 48 hr. The apoptosis in U937 cells exposed to HPC was increased significantly in the presence of interferon‐gamma (IFN‐γ). The augmentation of HPC‐induced apoptosis by IFN‐γ is repressed in cells (U937‐MP) persistently infected with mumps virus. A persistently infected cell line, U937‐MP, showed poor induction of signal transducers and activators of transcription‐1α (STAT‐1α), STAT‐2, p48 and IFN‐regulatory factor‐1 (IRF‐1), which are closely correlated with interferon‐α (IFN‐α) and IFN‐γ signaling pathways. Expression of MHC class‐I or class‐II was augmented by IFN‐α or IFN‐γ in U937 cells, but not in persistently infected cells. Therefore, it is suggested that the IFN‐γ signaling pathway plays an important role in the augmentation of HPC‐induced apoptosis. Mumps virus can suppress the IFN‐γ signaling pathway and subsequent development of apoptosis.
Hepatitis C Virus NS5B RNA Replicase Specifically Binds Ribosomes
Authors: Tanaka Torahiko , Sugiyama Kazuo , Ikeda Masanori , Naganuma Atsushi , Nozaki Akito , Saito Masaki , Shimotohno Kunitada , Kato Nobuyuki 
Published: November 2013

AbstractHepatitis C virus (HCV) non‐structural protein 5B (NS5B) is an RNA replicase. We expressed full‐length NS5B (591 amino acid residues) in Escherichia coli as a fusion protein with maltose binding protein (MBP‐NS5B). MBP‐NS5B was recovered in the soluble fraction after centrifugation at 40,000 × g and affinity‐purified with amylose resin. The purified MBP‐NS5B had a high‐level of poly (A), oligo (U)‐dependent UMP incorporation with a Km of 2 μm for UTP. Surprisingly, the enzymatically active MBP‐NS5B was sedimented by ultracentrifugation at 160,000 × g. The pellet contained 16S and 23S ribosomal RNAs, suggesting that ribosomes were associated with MBP‐NS5B. Ribosomes and MBP‐NS5B were subsequently co‐purified on amylose resin. Deletion study revealed that either the N‐terminal (amino acid residues 1 107) or the C‐terminal (amino acid residues 498–591) region of NS5B were sufficient for this association with ribosomes. We further found that NS5B also bound with human ribosomes. Our results implicate a novel mechanism of coupling between replication and translation of the viral genome in the life cycle of HCV.