bluuebunny/crossref_metadata_2025_split huggingface dataset:
Japanese Encephalitis Virus Infection in Vero Cells: The Involvement of Intracellular Acidic Vesicles in the Early Phase of Viral Infection Was Observed with the Treatment of a Specific Vacuolar Type H+‐ATPase Inhibitor, Bafilomycin A1
Authors: Nawa Masaru 
Published: November 2013

AbstractThe involvement of intracellular acidic vesicles in the early phase of Japanese encephalitis (JE) virus infection in Vero cells was observed by adding a specific vacuolar type H+ ‐ATPase (V‐ATPase) inhibitor (bafilomycin A1) in the cell culture medium. Studies with the detection of viral envelope (E) protein suggested that bafilomycin A1 inhibited virus infection in the cells. Subcellular distribution of incoming biotinylated virions and 3H‐uridine‐labeled viral RNA were observed in fractions of a Percoll density gradient. At 10 min of the chasing period, virions and viral RNA were found mainly in fractions with a mean density of 1.04 g/ml corresponding to the endosome both in the control and bafilomycin A1‐treated cells. At 60 min of the chasing period, the peak of biotin activity was detected in fractions with a mean density of 1.08 g/ml corresponding to the lysosome, whereas the peak of radioactivity did not run parallel with that of biotin and shifted to fractions with a mean density of 1.05 g/ml and higher than 1.084 g/ml, respectively. At 60 min of the chasing period in bafilomycin A1‐treated cells, the peak of biotin and radioactivity were still found mainly in the fraction with a density of 1.04 g/ml, representing the endosome. Subcellular fractionation by a Percoll density gradient revealed that bafilomycin A1 treatment resulted in the accumulation of virions in the endosome fraction and suggested the prevention of intracellular translocation of the virions which occurs during the early entry process of an infecting virus to the cells.
Biovar Diversity Is Reflected by Variations of Genes Encoding Urease of Ureaplasma urealyticum
Authors: Ruifu Yang , Minli Zhang , Zhaobiao Guo , Xiaohui Wang 
Published: November 2013

AbstractFive oligonucleotide primers derived from the gene encoding urease of Ureaplasma urealyticum were designed to evaluate the relationship between the urease gene and biovar diversity of this organism. Five combinations of these primers were tested by PCR and the result revealed that there were variations in urease genes among different serovars of U. urealyticum. This result, in agreement with other PCRs based on other functionally unrelated (rRNA and MB antigen) genes, may reflect the phylogenetic relationship among organisms taxonomically classified as U. urealyticum.
Phylogenetic Positions and Assignment of Swine and Ovine Corynebacterial Isolates Based on the 16S rDNA Sequence
Authors: Takahashi Tatsufumi , Mori Yukari , Kobayashi Hideaki , Ochi Masaru , Kikuchi Naoya , Hiramune Takashi 
Published: November 2013

AbstractThe nucleotide sequences of the 16S ribosomal RNA gene (rDNA) of swine and ovine corynebacterial strains were determined. The sequences of the strains that identified as Corynebacterium pseudotuberculosis by their biochemical characteristics were homologous with each other. The phylogenetic position of C. pseudotuberculosis strains was closest to C. ulcerans and next closest to C. diphtheriae. The nucleotide sequence of another swine isolate, SC8, was similar to that of a recently proposed species, C. seminale, and a non‐validated species, “C. glucuronolyticum,” with about 0.01 to 0.02 evolutionary distances. Analysis of the predicted secondary structure of the 16S rRNA molecule agreed with the close phylogenetic relationships between C. pseudotuberculosis and C. ulcerans and between C. seminale and strain SC8.
Anti‐Chemotactic Activity of Capsular Polysaccharide of Cryptococcus neoformans In Vitro
Authors: Fujihara Hitomi , Kagaya Keiko , Fukazawa Yoshimura 
Published: November 2013

AbstractIn this study, we demonstrated the anti‐chemotaetic activity of the capsular polysaccharides (CPSs) isolated from each of the heavily (H)‐ and weakly (W)‐encapsulated strains of Cryptococcus neoformans in vitro. The capacity for activation of the alternative complement pathway (ACP) of cells of the two C. neoformans strains in fresh human sera was comparable to that of zymosan (insoluble control), whereas the capacity for generation of the chemotactic factor (CF) of the cells of the two strains in fresh murine sera was markedly lower in the order H‐ < W‐strain than that of zymosan. Conversely, the capacities for ACP activation and CF generation of the CPSs were extremely lower than those of lipopolysaccharide (LPS, soluble control). When zymosan‐activated murine serum was incubated with CPS, both CPSs inhibited CF activity dose dependently. When zymosan‐activated serum was incubated with heat‐killed cells of each strain of C. neoformans, H and W, the CF activity of the treated sera decreased significantly, suggesting that CPS per se did not affect the neutrophils directly, but CPS absorbed CF. On the other hand, both CPSs were shown to possess the O‐acetyl groups in their molecules by 1H‐nuclear magnetic resonance spectroscopy. The de‐O‐acetylation of both CPSs increased the capacity for ACP activation to a level similar to that of LPS, and the de‐O‐acetylated CPS of both strains exhibited a lower ability to inhibit CF than did native CPS. Collectively, these results suggest that the anti‐chemotactic activity of CPS accounts for its ability to absorb the CF which was mostly generated at the sites around the cell wall of whole cells via the ACP, thus suppressing the inflammatory response by preventing dispersal of CF to the extracellular space; and also that the O‐acetyl group is partly, if any, involved in the mechanism for incompetence in ACP activation as well as the inhibition of CF.
Protease‐Induced Multicell Formation in Staphylococcus haemolyticus
Authors: Yabu Kunihiko , Nishiyama Yayoi , Ochiai Toshiro 
Published: November 2013

AbstractMulticellular cells were efficiently induced in Staphylococcus haemolyticus by the addition of protease to exponentially growing cultures at 30 C. Electron microscopy revealed the formation of tetrad‐shaped multicells that were septated but not separated from each other. Incubation of the multicells with extract from the cells grown without protease resulted in a fourfold increase in the number of colony‐forming units as compared with the untreated control. An electrophoretic analysis showed that protease caused a loss of cell wall‐lytic activity of the cell, which possibly led to the formation of multicells through cessation of cross wall separation.
The Host Response to Listeria monocytogenes Mutants Defective in Genes Encoding Phospholipases C (plcA, plcB) and Actin Assembly (actA)
Authors: Rudnicka Wiesława , Kaczmarek Małgorzata , Szeliga Jacek , Germann Tieno , Więckowska Marzena , Różalska Barbara 
Published: November 2013

AbstractSeveral genes involved in the determination of Listeria monocytogenes pathogenesis have been identified. Among them, plcA gene encodes phosphatidylinositol‐specific phospholipase C (PI‐PLC), plcB gene encodes a broad‐range phospholipase C (PC‐PLC), and actA encodes a protein contributing to actin assembly in infected cells. The interaction of L. monocytogenes wild type (LO 28) strain and two derivative mutants, plcA− (BUG 206) and actA−/plcB− (LUT 12), with macrophages and T lymphocytes was investigated in a mouse model of listeriosis. Both mutants showed evidence of attenuation. The plcA− mutant, but not the plcB− mutant, expressed an increase in susceptibility to the anti‐listerial activity of macrophages. Both mutants showed a decreased ability to induce IL‐12 production by bone marrow macrophages when co‐stimulated with E. coli LPS or IFN‐γ. In vivo, L. monocytogenes plcA− mutant was found to be a more effective stimulator of T cells than the wild LO 28 strain.
A Structural Analysis of the Regularly Arranged Porin on the Outer Membrane of Campylobacter jejuni Based on Correlation Averaging
Authors: Amako Kazunobu , Baba Norio , Suzuki Nobuyuki , Wai Sun Nyunt , Umeda Akiko 
Published: November 2013

AbstractA negatively stained electron micrograph of regularly arranged porin proteins of Campylobacter jejuni on the isolated outer membrane of bacteria was analyzed in detail by the correlation averaging method using a computer‐assisted program. The results showed that the porin of C. jejuni had a trimeric structure separated by about 10.4±0.15 nm. In addition, the pores in the trimers were also separated by about 4.3 ±0.1 nm.
Immunological Studies of a 21 kDa Cellular Component Efficiently Incorporated into Rabies Virion Grown in a BHK‐21 Cell Culture
Authors: Sagara Junji , Tochikura Tadafumi S. , Yamamoto Toshiko , Tsukita Shoichiro , Tsukita Sachiko , Kawai Akihiko 
Published: November 2013

AbstractTo investigate cellular components incorporated into the rabies virion, monoclonal antibodies (MAbs) were screened based on their reactivity with additional virion components. Two of the MAbs we prepared recognized a virion‐associated 21 kDa polypeptide (referred to as VAP21) from a BHK‐21 cell. Since the MAbs precipitated the rabies virion and trypsin digestion eliminated the VAP21 antigen from the virion but alkaline treatment (pH 11) did not, VAP21 seems to be anchored into the viral envelope and exposed on the virion surface. Although quantitative immunoblot analyses indicated an apparently increased concentration of VAP21 in the virion, the ratio of the content of VAP21 to that of viral glycoprotein (G) was several times decreased as compared to the ratio of those in the cell. These data suggest that sorting of VAP21 occurs during the viral budding process on the cell but that it might be inefficient, probably due to a more intimate association of VAP21 with the viral envelope proteins. This assumption seems to be consistent with the results of immunofluorescence studies; that is, VAP21 displayed colocalized distribution with viral envelope antigens in the cell. From these results, it is suggested that VAP21 closely associates with the viral envelope proteins in the cell, and this association might cause passive but relatively efficient incorporation of VAP21 into the virion.
Metabolic Fate of [3H]1‐β‐d‐Arabinofuranosyl‐5‐[(E)‐2‐bromovinyl]uracil in Herpes Simplex Virus Type 1‐Infected Cells
Authors: Ashida Noriyuki , Sakata Shinji , Machida Haruhiko 
Published: November 2013

AbstractThe metabolic fate of 1‐β‐d‐arabinofuranosyl‐5‐[(E)‐2‐bromovinyl]uracil (BV‐araU) in herpes simplex virus type 1‐infected cells was studied using tritium‐labeled BV‐araU. [3H]BV‐araU was selectively taken‐up by infected cells. Approximately 10% of the total uptake of [3H]BV‐araU was recovered from the acid‐insoluble fraction at any time post‐infection. Both cellular uptake of [3H]BV‐araU and its incorporation into the acid‐insoluble fraction increased with increasing incubation time through 8 hr post‐infection. Uptake of [3H]BV‐araU and its incorporation into the acid‐insoluble fraction also increased proportionally to the duration of exposure to [3H]BV‐araU. An alkaline sucrose gradient sedimentation analysis revealed that the radioactive DNA obtained from cells pulse‐labeled with [3H]BV‐araU were small DNA fragments which remained at the top following a chasing period in isotope‐free medium, whereas that pulse‐labeled with [3H]thymidine was chased to a fraction of high molecular weight DNA. Nuclease P1 digestion reduced 99% of the [3H]BV‐araU‐labeled DNA extracted from infected cells to a low molecular weight. Following digestion of [3H]BV‐araU‐labeled DNA with micrococcal nuclease and spleen exonuclease, all of the radioactivity was recovered as [3H]BV‐araU 3′‐monophosphate. Thus, BV‐araU strongly inhibits the elongation of viral DNA strands as demonstrated by the alkaline sucrose gradient sedimentation analysis, whereas at least a portion of the [3H]BV‐araU is incorporated inside viral DNA strands in infected cells.
Salmonella typhi Ty2 OmpC Porin Induces Bactericidal Activity on U937 Monocytes
Authors: Blanco Luz P. , Toro Cecilia S. , Romero Jaime M. , Santiviago Carlos A. , Mora Guido C. 
Published: November 2013

AbstractThe immunogenic effect of Salmonella typhi OmpC porin during typhoid fever in humans was evaluated in vitro. Peripheral blood mononuclear cells from 17 patients were challenged with outer membrane preparations from Escherichia coli UH302 and UH302/pSTP2K2 strains, both lacking E. coli OmpF and OmpC porins, although UH302/pSTP2K2 expressed a plasmid‐encoded S. typhi Ty2 OmpC. The mononuclear cell supernatants, immunized in vitro with OmpC antigen, derived from 10 out of 17 patients activated U937 bactericidal capacity. In contrast, the supernatants from the immunization with outer membrane preparation lacking S. typhi Ty2 OmpC induced a significantly reduced bactericidal capacity of U937 cells. This procedure should prove useful for in vitro characterization of cellular immunogens from exclusive human pathogens.
Sequence Analysis of the actA Gene of Listeria monocytogenes Isolated from Human
Authors: Moriishi Kohji , Terao Michinori , Koura Minako , Inoue Satoshi 
Published: November 2013

AbstractThe region encoding proline‐rich units of actA genes was amplified from 24 strains of Listeria monocytogenes using polymerase chain reaction (PCR). PCR products of 13 strains showed the expected size of 623 bp, whereas those of 11 strains showed a short size of 518 bp. The shortening of these PCR products resulted from the deletion of one proline‐rich unit. These results indicate that ActA proteins are divided into at least two different types which are unrelated to bacterial serotypes.
Effect of Acidic Fibroblast Growth Factor (aFGF) on Phagocytosis in Mouse Peritoneal Macrophages
Authors: Ichinose Mitsuyuki , Sawada Masashi , Sasaki Kazuo , Oomura Yutaka 
Published: November 2013

AbstractThe effects of acidic fibroblast growth factor (aFGF) on phagocytosis in peritoneal macrophages from thioglycollate‐elicited mice were examined using flow cytometry. aFGF enhanced phagocytosis of fluorescein isothiocyanate‐labeled latex particles in a dose‐dependent manner. Basic fibroblast growth factor (bFGF) also enhanced phagocytosis. This study suggests that aFGF can modulate an important activity of macrophages.
Intervening Sequences in 16S rRNA Genes of Campylobacter sp.: Diversity of Nucleotide Sequences and Uniformity of Location
Authors: Etoh Yumiko , Yamamoto Ayako , Goto Nobuichi 
Published: November 2013

AbstractWe found and sequenced intervening sequences (IVSs) in the PCR‐amplicons of 16S rRNA genes of 3 strains of Campylobacter rectus, 2 strains of C. curvus and 2 strains of C. sputorum. The lengths of the IVSs were 140 to 233 bp. The IVSs of C. rectus were identical and had a sequence homology of 55 to 79% against those of C. curvus and C. helveticus. The IVSs of C. sputorum were 97.9‐100% homologous but poorly homologous to the other IVSs. In spite of the diversities of the lengths and the nucleotide sequences, all of the IVSs were located at the same position in the 16S rRNA genes.
Requirement of Cauliflower Mosaic Virus Open Reading Frame VI Product for Viral Gene Expression and Multiplication in Turnip Protoplasts
Authors: Kobayashi Kappei , Tsuge Seiji , Nakayashiki Hitoshi , Mise Kazuyuki , Furusawa Iwao 
Published: November 2013

AbstractCauliflower mosaic virus (CaMV) open reading frame (ORF) VI product (P6) has been shown to be the major constituent of viral inclusion body, to function as a post‐transcriptional transactivator, and to be essential for infectivity on whole plants. Although these findings suggest that P6 has an important role in viral multiplication, it is unknown whether P6 is required for viral multiplication in a single cell. To address this question, we transfected turnip protoplasts with an ORF VI frame‐shift (4 bp deletion) mutant (pCaFS6) of an infectious CaMV DNA clone (pCa122). The mutant was uninfectious. Co‐transfection of plasmids expressing P6 complemented the mutant. Overexpression of P6 elevated the infection rate in co‐transfection experiments with either pCa122 or pCaFS6. This would have been achieved by elevating the level of pregenomic 35S RNA, a putative polycistronic mRNA for ORFs I, II, III, IV and V, and by enhancing the accumulation of these five viral gene products. When CaMV ORFs I, II, III, IV and V were expressed from monocistronic constructs in which each of the ORFs was placed just downstream of the 35S promoter, the accumulation of ORF III, IV and V products depended on the co‐expression of P6. The accumulation of ORF I and II products was not detected, even in the presence of P6. These results suggest that P6 is involved in the stabilization of other viral gene products as well as in the activation of viral gene expression, and thus, is a prerequisite for CaMV multiplication.
The First Isolation of Arthroderma benhamiae in Japan
Authors: Kano Rui , Nakamura Yuka , Yasuda Kazuo , Watari Toshihiro , Watanabe Shinichi , Takahashi Hisashi , Tsujimoto Hajime , Hasegawa Atsuhiko 
Published: November 2013

AbstractA clinical isolate of Trichophyton mentagrophytes from rabbit was examined by polymerase chain reaction (PCR) analysis and a mating experiment. The species‐specific primers designed from the nucleotide sequences of the chitin synthase 1 (CHS1) gene in the teleomorph of Arthroderma benhamiae amplified a fragment from genomic DNA samples of A. benhamiae and the clinical isolate but not from those of A. simii and A. vanbreuseghemii. On the other hand, the species‐specific primers of A. simii and A. vanbreuseghemii did not amplify any fragment from the genomic DNA of the clinical isolates. When the isolate was respectively crossed with (+) or (‐) tester strains of A. benhamiae, A. simii and A. vanbreuseghemii, ascospores were produced in the crossing with the A. benhamiae (+) strain. Therefore, the isolate was identified to be A. benhamiae (‐), confirming the result of molecular analysis. This is the first report on the isolation of A. benhamiae in Japan.
Role of Macrophages in Acute Murine Cytomegalovirus Infection
Authors: Hamano Shinjiro , Yoshida Hiroki , Takimoto Hiroaki , Sonoda Koh‐hei , Osada Kazuhiro , He Xiangdong , Minamishima Yoichi , Kimura Genki , Nomoto Kikuo 
Published: November 2013

AbstractIt has been recognized that macrophages play an important role in controlling virus infection in experimental animal models. To evaluate the role of macrophages in acute murine cytomegalovirus infection, macrophages in the spleen and the liver were eliminated by an intravenous injection of liposomes containing a cytolytic agent, dichloromethylene diphosphonate. The depletion of macrophages led to a significant increase of virus titer in the spleen and lungs in both susceptible BALB/c and resistant C57BL/6 mice during the first three days after intravenous infection. In the spleen, the increase of virus titer in macrophage‐depleted BALB/c mice was much greater than that in NK cell‐depleted mice. These results suggest that macrophages contribute to protection mainly by the mechanisms which are independent of NK cells during the first three days after infection. The increase of virus titer in macrophage‐depleted C57BL/6 mice was as great as that in NK cell‐depleted mice because of the high contribution of NK cells to protection in C57BL/6 mice. In the liver in both strains of mice, the effects of macrophage depletion on virus titer were not as much as those in the spleen and lungs. Furthermore, the local depletion of peritoneal macrophages resulted in a great increase of virus titer in the spleen at three days after intraperitoneal infection. We conclude that macrophages greatly contribute to decreasing the virus load in some organs possibly through either or both intrinsic and extrinsic mechanisms in the early phase of primary infection with murine cytomegalovirus.
Antigenic Differences between Clinical and Environmental Isolates of Burkholderia pseudomallei
Authors: Sirisinha Stitaya , Anuntagool Narisara , Intachote Pakamas , Wuthiekanun Vanaporn , Puthucheary Savithri D. , Vadivelu Jammuna , White Nicholas J. 
Published: November 2013

AbstractBurkholderia pseudomallei is a free‐living organism that causes the potentially lethal tropical infection melioidosis. The disease is endemic in many parts of eastern Asia and northern Australia. The presence of two distinct biotypes in soil can be reliably distinguished by their ability to assimilate l‐arabinose. Whereas some soil isolates could utilize this substrate (Ara+), the remaining soil isolates and all clinical isolates tested so far could not (Ara−). Only the Ara− isolates were virulent in animal models. We have raised a murine monoclonal antibody (MAb) that can readily distinguish Ara− from Ara+ biotypes. The MAb reacted with a high molecular weight component present only on the Ara− biotype. With this MAb, clinical and soil Ara−isolates gave identical positive reactions in agglutination, immunofluorescence, ELISA and immunoblot assays. Using these same assay systems, the soil Ara+ biotype did not react with the MAb. Similar but distinct immunoblot patterns were also noted when these two Ara biotypes were probed with sera from patients with melioidosis or with polyclonal immune rabbit sera. These data showed that the Ara− biotype from both clinical and environmental isolates is antigenically different from its Ara+ environmental counterpart. The SDS‐PAGE protein and lectin‐binding profiles of both groups of Ara− isolates were also found to be different from those of the Ara+ biotype.
Leishmania donovani Infection of a Susceptible Host Results in Apoptosis of Th1‐Like Cells: Rescue of Anti‐Leishmanial CMI by Providing Th1‐Specific Bystander Costimulation
Authors: Das Gobardhan , Vohra Harpreet , Saha Bhaskar , Agrewala Javed N. , Mishra Gyan C. 
Published: November 2013

AbstractA protective immune response against Leishmania donovani infection is mediated by T‐helper type 1 (Th1) cells. Th1 induced cell‐mediated immunity (CMI), as assessed by anti‐leishmanial DTH response, is lost in a susceptible host such as BALB/c mice. Although the impaired Th1 function eventuates in unhindered parasite growth and in manifestation of the susceptible phenotype, the mechanism of down‐regulation of the Th1 function is yet to be elucidated. Here, we provide evidence that the parasite downregulates the expression of a Th1‐specific costimulatory molecule, M150, on the surface of infected BALB/c mice‐derived macrophages. Th cells are rendered unresponsive to anti‐CD3 Ab‐mediated stimulation after interaction with infected macrophages. The anergized T cells produce much less IL‐2, IL‐4 and IFN‐γ compared to those T cells which were costimulated using normal macrophages. The defect in proliferation, anti‐CD3 Ab induced unresponsiveness and IFN‐γ but not IL‐4 production can be restored by providing bystander costimulation through M150. These results not only unfold a novel immune evasion strategy used by the parasite but also clarify the mechanism of Th1 cell debilitation during the disease. Recovery of Th1 cytokine production by bystander costimulation through M150 may help in formulating a new strategy for the elimination of intracellular parasites.