bluuebunny/crossref_metadata_2025_split huggingface dataset:
Changes in the Cell Structure of Flavobacterium psychrophilum with Length of Culture
Authors: Kondo Motoki , Kawai Kenji , Yagyu Ken‐ichi , Nakayama Kouichi , Kurohara Kenrou , Oshima Syun‐ichirou 
Published: November 2013

AbstractFlavobacterium psychrophilum, the pathogen of bacterial cold‐water disease, causes serious problems in ayu Plecoglossus altivelis culture. This study investigated the effect of the culture period of F. psychrophilum and on the structure of its cells. From the SDS‐PAGE of total proteins of cellular components, much difference was found between the 36 hr culture and the 48 and 72 hr cultures. A SEM observation of the cells showed many fragments, especially on the cell surface of the 36 hr culture. These fragments consisted of an outer membrane, seen by TEM observation, and may contain substances causing the virulence. Specific proteins observed by the SDS‐PAGE and fragments in the 36 hr culture may be related to the virulence of F. psychrophilum.
Bacterial Population in Russian Space Station “Mir”
Authors: Kawamura Yoshiaki , Li Ying , Liu Hongsheng , Huang Xinxiang , Li Zhiyu , Ezaki Takayuki 
Published: November 2013

AbstractWe had the opportunity to investigate the bacterial population in air samples, condensation water, and inner wall swabs from the Russian space station Mir. From the first and second air samples during the mission, 29 and 7 bacterial colonies were collected, respectively. The values were equivalent to 16.8 and 4.0 cfu/100 liter air, respectively. Condensation water was collected from three different sites. The total viable bacterial counts were 2.1 × 106, 5.2 × 102, and 3.0 × 101 cfu/ml. The phylogenetic position of each isolate was determined by total 16S rDNA sequencing. Bacteria from air samples were mainly Gram‐positive (35/36 colonies), and staphylococci occupied dominant specifically (23/36 colonies). On the other hand, Gram‐negative bacteria were mainly isolated from condensation water samples. Most strains were thought to be opportunistic pathogens or environmental bacteria (such as those that inhabit soil, water, or air) found on earth. However, 6 of 23 isolates were suspected to be new species according to phylogenetic analysis and quantitative DNA‐DNA hybridization data. The isolation of the other levels 3 and 2 bacteria, using specific selective media, was unsuccessful because all samples were heavily contaminated with fungi. To overcome this situation, PCR methods were applied to survey most levels 3 and 2 pathogenic bacteria in the condensation water samples. Up to 380 different primers for bacterial pathogens were used in this study. Only Mycobacterium avium 16S DNA sequences, however, could be amplified from the three water samples. The average bacteria count was estimated to be about 104 organisms/ml water.
Correlation between Human Herpesvirus 6 and 7 Infections after Living Related Liver Transplantation
Authors: Ihira Masaru , Yoshikawa Tetsushi , Suzuki Kyoko , Ohashi Masahiro , Suga Sadao , Asonuma Katsuhiro , Tanaka Koichi , Asano Yoshizo 
Published: November 2013

AbstractHuman herpesvirus 6 (HHV‐6) and human herpesvirus 7 (HHV‐7) are closely related to each other. Interaction between the two viruses at the time of primary HHV‐7 infection is suggested by in vivo and in vitro studies. However, interaction between the two viruses in organ transplant recipients has not been analyzed. We analyzed serially collected plasma samples obtained from 40 living related liver transplant recipients by serological assay (indirect immunofluorescence assay, IFA) and polymerase chain reaction (PCR). Significant increase or seroconversion of HHV‐6 IgG and HHV‐7 IgG antibody titers were observed in 45% and 58% of recipients respectively. Positive rate of IgM HHV‐6 antibody increased up to 35% at 4 weeks after transplantation. However, no remarkable peak in the positive rate of HHV‐7 IgM antibody was demonstrated. HHV‐6 and HHV‐7 DNA were detected in plasma in 15 (38%) and 16 (40%) of the 40 recipients respectively. HHV‐6 DNA was detected in 10 (26%) of the 38 recipients at 2 weeks after transplantation. The positive rate of the virus genome in plasma gradually decreased after that time. HHV‐7 DNA was detected in 5 (14%) of the 37 recipients at 2 weeks after transplantation; no obvious peak in the positive rate of HHV‐7 DNA was demonstrated. Antibody responses involving both HHV‐6 and HHV‐7, including either a significant increase in IgG antibody titers or positive identification of IgM antibody were observed in 17 (43%) of the 40 recipients. Thirteen out of the 17 recipients demonstrated concurrent antibody response against both viruses. HHV‐7 antibody response preceded the HHV‐6 antibody response in 2 of the remaining 4 recipients, whereas the opposite was true in the other 2 recipients. Both HHV‐6 and HHV‐7 DNA were detected in 7 (18%) of the 40 recipients. In 4 of those 7 recipients, DNA from both viruses was concurrently detected, 3 of whom had HHV‐7 DNA repeatedly detected after first detection of the virus DNA. The detection of HHV‐7 DNA preceded the detection of HHV‐6 DNA in 2 recipients, whereas HHV‐6 DNA appeared first in 1 recipient.
Investigation of Varicella‐Zoster Virus DNA in Lymphocyte Subpopulations by Quantitative PCR Assay
Authors: Ito Yoshinori , Kimura Hiroshi , Hara Shinya , Kido Shinji , Ozaki Takao , Nishiyama Yukihiro , Morishima Tsuneo 
Published: November 2013

AbstractTo investigate the nature of viremia during the acute phase of varicella, we studied the viral load in nine otherwise healthy children with varicella. Plasma and peripheral blood mononuclear cells (PBMC) were obtained, then PBMC were divided into CD4+T, CD8+T, and B lymphocytes and monocyte/macrophage fractions. The viral DNA in each component was quantified using a real‐time quantitative polymerase chain reaction assay. Varicella‐zoster virus (VZV) DNA was detected in plasma, PBMC and all subpopulations. The amount of viral DNA was similar in each PBMC subpopulation, suggesting that each lymphocyte fraction and monocytes carry similar amounts of VZV DNA during viremia.
Proteolytic Cleavage of Staphylococcal Exoproteins Analyzed by Two‐Dimensional Gel Electrophoresis
Authors: Kawano Yasushi , Kawagishi Mika , Nakano Miyo , Mase Kiyomi , Yamashino Takafumi , Hasegawa Tadao , Ohta Michio 
Published: November 2013

AbstractExtracellular proteases of Staphylococcus aureus are emerging as potential virulence factors that are relevant to the pathogenicity of staphylococcal infections. These proteases may also be involved in the proteolytic cleavage of other exoproteins released from this organism. To define the target exoproteins and their sites of cleavage by proteases, high‐resolution two‐dimensional polyacrylamide gel electrophoresis followed by N‐terminal amino acid sequencing of exoprotein spots was performed. Two to three hundred exoprotein spots were detected at the early‐stationary phase of cultures of S. aureus NCTC8325, and then at the late‐stationary stage most of these high molecular protein spots became invisible due to further proteolytic degradation. As the result of N‐terminal analysis, lipase, triacylglycerol lipase, orf619 protein and orf388 protein were detected as multiple spots at the early‐stationary phase. We found that these exoproteins were cleaved at 3, 7, 4 and 4 different sites, respectively, by proteases. According to the M.W. and pI of each peptide spot obtained from the gel and their matches with calculated values in addition to their N‐terminal sequences, we showed that the positions of putative peptides resulted from proteolytic cleavage of these proteins.
Burkholderia pseudomallei Interferes with Inducible Nitric Oxide Synthase (iNOS) Production: A Possible Mechanism of Evading Macrophage Killing
Authors: Utaisincharoen Pongsak , Tangthawornchaikul Nattaya , Kespichayawattana Wanapa , Chaisuriya Predawan , Sirisinha Stitaya 
Published: November 2013

AbstractBurkholderia pseudomallei is a causative agent of melioidosis, a life threatening disease which affects humans and animals in tropical and subtropical areas. This bacterium is known to survive and multiply inside cells such as macrophages. The mechanism of host defense against this bacterium is still unknown. In this study, we demonstrated that B. pseudomallei exhibited unique macrophage activation activity compared with Escherichia coli and Salmonella typhi. The mouse macrophage cell line (RAW 264.7) infected with B. pseudomallei at MOI of 0.1:1, 1:1 and 10:1 did not express a detectable level of inducible nitric oxide synthase (iNOS). Moreover, the B. pseudomallei infected cells released TNF‐α only when they were infected with high MOI (10:1). Unlike the cells infected with B. pseudomallei, the cells infected with E. coli, and S. typhi expressed iNOS even at MOI of 0.1:1. These infected cells also released a significantly higher level of TNF‐α at the low MOI ratio. The cells that were preactivated with IFN‐γ prior to being infected with B. pseudomallei exhibited an enhanced production of iNOS and TNF‐α release. The increased macrophage activation activity in the presence of IFN‐γ also correlated with the restriction of the intracellular bacteria survival. Moreover, IFN‐γ also prevented cell fusion and multinucleated cell formation induced by B. pseudomallei, a phenomenon recently described by our group. Altogether, these results indicate that internalization of B. pseudomallei failed to trigger substantial macrophage activation, a phenomenon which could prolong their survival inside the phagocytic cells and facilitate a direct cell to cell spreading of B. pseudomallei to neighboring cells.
Identification of Shiga Toxin‐Producing Escherichia coli Possessing Insertionally Inactivated Shiga Toxin Gene
Authors: Okitsu Tadayuki , Kusumoto Masahiro , Suzuki Rieko , Sata Shin , Nishiya Yoshiaki , Kawamura Yoshihisa , Yamai Shiro 
Published: November 2013

AbstractWe have investigated the Shiga toxin genes of Shiga toxin‐producing Escherichia coli (STEC) strains, using polymerase chain reaction (PCR) amplifying the full lengths of these genes. As a result, we found the Shiga toxin 2 gene which was insertionally inactivated by an insertion sequence (IS). This IS element was identical to IS1203v which has been also found in inactivated Shiga toxin 2 genes, and was inserted at the same site as in the previous paper. On the other hand, both Shiga toxin 2 genes were different (98.3% identity). These suggested that IS1203v independently inserted into each Shiga toxin 2 genes, and STEC strains possessing the insertionally inactivated Shiga toxin genes are most likely to have a wide distribution. Amplification of the full length of the Shiga toxin gene is one of the effective methods to detect the gene no matter where the IS element is included, i.e., the insertion can be reflected in the size of amplicon.
Region of Heat‐Stable Enterotoxin II of Escherichia coli Involved in Translocation across the Outer Membrane
Authors: Okamoto Keinosuke , Yamanaka Hiroyasu , Takeji Miho , Fujii Yoshio 
Published: November 2013

AbstractHeat‐stable enterotoxin II of Escherichia coli (STII) is synthesized as a precursor form consisting of pre‐ and mature regions. The pre‐region is cleaved off from the mature region during translocation across the inner membrane, and the mature region emerges in the periplasm. The mature region, composed of 48 amino acid residues, is processed in the periplasm by DsbA to form an intramolecular disulfide bond between Cys‐10 and Cys‐48 and between Cys‐21 and Cys‐36. STII formed with these disulfide bonds is efficiently secreted out of the cell through the secretory system, including TolC. However, it remains unknown which regions of STII are involved in interaction with TolC. In this study, we mutated the STII gene and examined the secretion of these STIIs into the culture supernatant. A deletion of the part covering from amino acid residue 37 to the carboxy terminal end did not markedly reduce the efficiency of secretion of STII into the culture supernatant. On the other hand, the efficiency of secretion of the peptide covering from the amino terminal end to position 18 to the culture supernatant was significantly low. These observations indicated that the central region of STII from amino acid residue 19 to that at position 36 is involved in the secretion of STII into the milieu. The experiment using a dsbA‐deficient strain of E. coli showed that the disulfide bond between Cys‐21 and Cys‐36 by DsbA is necessary for STII to adapt to the structure that can cross the outer membrane.
Effector Mechanism and Clinical Response of BAK (BRM‐Activated Killer) Immuno‐Cell Therapy for Maintaining Satisfactory QOL of Advanced Cancer Patients Utilizing CD56‐Positive NIE (Neuro‐Immune‐Endocrine) Cells
Authors: Ebina Takusaburo , Ogama Naoko , Shimanuki Hiroko , Kubota Tomoka , Isono Noriko 
Published: November 2013

AbstractA new type of immuno‐cell therapy called BRM‐activated killer (BAK) therapy using non‐MHC‐restricted lymphocytes, CD56‐positive cells, was devised. Peripheral blood lymphocytes were selected by immobilization with anti‐CD3 monoclonal antibody and cultured for 2 weeks in the presence of IL‐2. Thereafter, they were reactivated by 1,000 U/ml of IFN‐α for 15 min. Twenty‐six outpatients with cancer whose performance status were over 80% on Karnofsky scale were selected for this study. About 6 × 109 BAK cells were returned by intravenous drip infusion, at one month intervals at an outpatient clinic to each of 20 advanced cancer patients in whom many metastatic lesions were found postoperatively, and to 6 patients with no postoperatively detectable metastases. The proportion of CD56‐positive cells increased from 20% to 50% with culture. CD56‐positive cells have strong cytotoxic activity and produced 20 ng/109 cells of β‐endorphin, an intracerebral hormone. During the course of BAK therapy, we adopted the Face scale as a QOL indicator. The QOL of all patients remained satisfactory or improved. β‐Endorphin is thought to make patients fell well and maintains good QOL because of its potent analgesic, sedative activity. From that facts that CD56 is a neural cell adhesion molecule and a member of the Ig superfamily, and that the CD56‐positive cell produces β‐endorphin, we concluded that the CD56‐positive cell is a multifunctional, integrated NIE (neuro‐immune‐endocrine) cell. Administration of BAK cells allowed all 20 advanced cancer patients with metastases to survive for over one year. All 6 patients receiving the same therapy for prevention of postoperative metastasis have been recurrence‐free for one to five years.
Gene Analysis of Vibrio cholerae NAGV14 Pilus and Its Distribution
Authors: Kuroki Hiromi , Toma Claudia , Nakasone Noboru , Yamashiro Tetsu , Iwanaga Masaaki 
Published: November 2013

AbstractAdhesive pilus of Vibrio cholerae O34, strain NAGV14, was genetically analyzed. The deduced amino acid (aa) sequence of the major pilin structural gene (VcfA) was 67% homologous to the MshA pilin in the N‐terminal region, but no homology was found in the C‐terminal region which contained the antigenic epitopes. Upstream and downstream flanking regions examined were highly homologous to mshB and mshC of the MSHA (mannose‐sensitive hemagglutinin) gene locus. A short leader sequence and a pair of cysteines near the C‐terminus which are the characteristics of type 4a pilus family were found. The major pilin structural gene of NAGV14 was compared to that of a strain V10 producing non‐adhesive pili. The deduced aa sequences showed 60% homology, and the distance between two cysteines in the C‐terminal region was different. A total of 177 V. cholerae strains were investigated for the presence of a type 4 pilus gene locus by PCR, and 95% were positive. The major pilin gene of NAGV14 was detected in 4 of 93 V. cholerae non‐O1, non‐O139 strains tested, but none of the V. cholerae O1 and O139 (72 and 12 strains, respectively). Our result suggested that a type 4 pilus gene locus similar to the MSHA gene locus is widely distributed among V. cholerae strains. We proposed naming this type 4 pilus gene locus the VCF (for V. cholerae flexible pili) gene locus.
3′‐Fluorine‐Substitution in 3‐Fluorocarbocyclic Oxetanocin A Augments the Drug's Inhibition of HHV‐6B Propagation in Chronically Infected Cultures
Authors: Wang Ping , Abe Kenji , Ojima Tomoko , Ohyashiki Junko H. , Satoh Hitoshi , Maruyama Tokumi , Nagata Hiroshi , Tanaka Hiroshi , Yamamoto Kohtaro 
Published: November 2013

AbstractAn infection of TaY cells, which originated from an adult T‐cell leukemia, with an HHV‐6B OK isolate resulted in a chronically infected culture, termed TaY(OK). Cell cloning analysis revealed that the TaY(OK) culture consisted of a mixture of cells permissive and refractory to the infection, and that the permissive cells were continuously produced from the refractory cell population. Since the chronically infected culture has been maintained for over 2 years without the addition of uninfected TaY cells, we used it for an evaluation of the antiviral potency of nucleoside analogs, especially carbocyclic oxetanocins (COXTs). MTT (3‐[4,5‐dimethylthiazol‐2‐yl]‐2,5‐diphenyltetrazolium bromide) assays showed a lack of toxicity of ganciclovir (GCV), COXTs, and their derivatives, to TaY(OK) cells at 1 μm. Therefore we compared the antiviral potencies of these drugs at 1 μm by monitoring the viral loads produced during a 1‐day period during the course of the drug treatment. Among the drugs tested, 3′‐fluorocarbocyclic oxetanocin A (3′‐C.OXT‐A) was the most effective for inhibiting the virus production, and at concentrations ranging from 0.5 μm to 10 μm, the inhibition of the viral production was dose‐dependent. A comparison of the chemical structures of the derivatives with that of C.OXT‐A, which is the parental molecule, suggested that the 3′‐fluorine‐modification might account for the higher anti‐HHV‐6 activity and lower cytotoxicity.
A Sandwich ELISA for Detection of Soluble GPI‐80, a Glycosylphosphatidyl‐Inositol (GPI)‐Anchored Protein on Human Leukocytes Involved in Regulation of Neutrophil Adherence and Migration—Its Release from Activated Neutrophils and Presence in Synovial Fluid of Rheumatoid Arthritis Patients—
Authors: Huang Jibiao , Takeda Yuji , Watanabe Tadashi , Sendo Fujiro 
Published: November 2013

AbstractGPI‐80, mainly distributed on human neutrophils and monocytes, is a novel glycosylphosphatidylinositol (GPI)‐anchored protein regulating neutrophil adherence and migration. We prepared a polyclonal antibody specific to GPI‐80 by immunizing rabbits with the purified GPI‐80 and established a sandwich ELISA for detecting soluble GPI‐80. We found that soluble GPI‐80 was released from fMLP activated neutrophils and was present at high concentrations in synovial fluids but not sera of rheumatoid arthritis patients, suggesting that GPI‐80 may play a role in inflammatory diseases. The detection of soluble GPI‐80 may help us to understand its physiological and pathological functions.
Regulation of Fas‐Mediated Apoptosis by NF‐κB Activity in Human Hepatocyte Derived Cell Lines
Authors: Marusawa Hiroyuki , Hijikata Makoto , Watashi Koichi , Chiba Tsutomu , Shimotohno Kunitada 
Published: November 2013

AbstractNuclear Factor‐κB (NF‐κB) is known to be one of the key regulators of genes involved in survival signaling. The purpose of this study is to investigate the role of NF‐κB activity in signaling events related to cell survival in hepatocytes, which has been supposed to be one of the most sensitive organs against Fas‐induced cytotoxicity. The functions of NF‐κB activity on Fas‐mediated apoptosis in different human cell lines were investigated by a magnetic concentration system for cells with exogenous protein production. We demonstrated that the activation of NF‐κB was triggered by anti‐Fas treatment in hepatocyte derived cell lines, HepG2 and Huh‐7 cells. Overexpression of kinase‐inactive NF‐κB‐inducing kinase (NIK) and IκB kinase (IKK) inhibited the activation of NF‐κB introduced by anti‐Fas treatment in these cells. Notably, inactivation of NF‐κB by the production of IκB‐α protein made these cells more susceptible to apoptosis induced by Fas stimulation. In contrast, transient expression of NIK showed a suppressive effect on Fas‐mediated apoptosis in the same cell lines. These findings suggest the involvement of NF‐κB in suppression of Fas‐mediated apoptosis in human hepatocyte derived cell lines, in which concomitant activation of this transcriptional factor was observed through the stimulation of Fas signaling.
Engraftment of Tonsillar Mononuclear Cells in Human Skin/SCID Mouse Chimera —Validation of a Novel Xenogeneic Transplantation Model for Autoimmune Diseases—
Authors: Yamanaka Noboru , Yamamoto Yoshikazu , Kuki Kiyonori 
Published: November 2013

AbstractPustulosis palmaris et plantaris (PPP) has been considered as one of the typical tonsillar focal infections, based on the marked clinical improvement of the skin lesions after tonsillectomy. Despite the accumulation of data showing the clinical efficacy of tonsillectomy for this skin lesion, fundamental etiological and pathophysiological issues have yet to be addressed. One primary obstacle hindering investigators has been the lack of an appropriate animal model for this human skin disorder. In the early stage of PPP, it has been reported that lymphocytes, predominantly CD4+ T lymphocytes, infiltrate the palmar and plantar skins. However, the origin and mechanism of infiltration by these lymphocytes is not clear and there are very few reports on whether tonsillar mononuclear cells react directly with the skin. We have been intrigued by the ability to engraft human cells onto severe combined immunodeficiency (SCID) mice, together with the opportunity for long‐term graft survival and ability to adoptively transfer various human immunocompetent cells. In this review, we addressed the existing deficiencies in our understanding of the relationship between tonsils and PPP by using emerging transplantation technology involving SCID mice.
Mechanism of Selective Inhibition of Respiratory Syncytial Virus by a Benzodithiin Compound (RD3–0028)
Authors: Sudo Kenji , Konno Kenji , Watanabe Wataru , Shigeta Shiro , Yokota Tomoyuki 
Published: November 2013

AbstractRD3–0028, a compound with a benzodithiin structure, was found to be a potent inhibitor of respiratory syncytial virus (RSV) replication. Its action is specific; no activity is seen against influenza A virus, measles virus, herpes simplex virus type 1 or 2, or human cytomegalovirus. A time‐dependent drug addition experiment indicated that the antiviral activity occurs in the late stage of the RSV replication cycle, since this compound completely inhibited syncytium formation even when added up to 16 hr after the infection of cell monolayers at an MOI of 3. RD3–0028 had no direct virucidal effect on RSV. Western blotting analysis showed that RD3–0028 significantly decreased the amount of RSV proteins released into the cell culture medium. Moreover, five independent isolates of the RSV long strain were selected for growth in RD3–0028 (5–20 μg/ml). These resistant viruses were more than 80‐fold less sensitive to RD3–0028 than the long strain. The F gene segment of each of these viruses was sequenced and in each case the mutant RNA segment contained at least one sequence alteration, converting asparagine 276 to tyrosine (F1 protein). These results suggest that RD3–0028 inhibits RSV replication by interfering with intracellular processing of the RSV fusion protein, or a step immediately thereafter, leading to loss of infectivity.