bluuebunny/crossref_metadata_2025_split huggingface dataset:
Prion Protein Gene‐Deficient Cell Lines: Powerful Tools for Prion Biology
Authors: Sakudo Akikazu , Onodera Takashi , Ikuta Kazuyoshi 
Published: November 2013

AbstractPrion diseases are zoonotic infectious diseases commonly transmissible among animals via prion infections with an accompanying deficiency of cellular prion protein (PrPC) and accumulation of an abnormal isoform of prion protein (PrPSc), which are observed in neurons in the event of injury and disease. To understand the role of PrPC in the neuron in health and diseases, we have established an immortalized neuronal cell line HpL3–4 from primary hippocampal cells of prion protein (PrP) gene‐deficient mice by using a retroviral vector encoding Simian Virus 40 Large T antigen (SV40 LTag). The HpL3–4 cells exhibit cell‐type‐specific proteins for the neuronal precursor lineage. Recently, this group and other groups have established PrP‐deficient cell lines from many kinds of cell types including glia, fibroblasts and neuronal cells, which will have a broad range of applications in prion biology. In this review, we focus on recently obtained information about PrP functions and possible studies on prion infections using the PrP‐deficient cell lines.
Sporicidal Activity of the KMT Reagent in Its Vapor Phase against Geobacillus stearothermophilus Spores
Authors: Kida Nori , Mochizuki Yasushi , Taguchi Fumiaki 
Published: November 2013

AbstractIn an investigation of the sporicidal activity of the KMT reagent, a vapor phase study was performed using five kinds of carriers contaminated with Geobacillus stearothermophilus spores. When 25 ml of the KMT reagent was vaporized in a chamber (capacity; approximately 95 liters), the 2‐step heating method (vaporization by a combination of low temperature and high temperature) showed the most effective sporicidal activity in comparison with the 1‐step heating method (rapid vaporization). The 2‐step heating method appeared to be related to the sporicidal activity of vaporized KMT reagent, i.e., ethanol and iodine, which vaporized mainly when heated at a low temperature such as 55 C, and acidic water, which vaporized mainly when heated at a high temperature such as 300 C. We proposed that the KMT reagent can be used as a new disinfectant not only in the liquid phase but also in the vapor phase in the same way as peracetic acid and hydrogen peroxide.
Suppression of the Bacterial Antigen‐Specific T Cell Response and the Dendritic Cell Migration to the Lymph Nodes by Osteopontin
Authors: Begum Mst. Dilara , Umemura Masayuki , Kon Shigeyuki , Yahagi Ayano , Hamada Satoru , Oshiro Kiyotetsu , Gotoh Kazuyo , Nishizono Akira , Uede Toshimitsu , Matsuzaki Goro 
Published: November 2013

AbstractOsteopontin (OPN) has been reported to enhance the interferon (IFN)‐γ‐producing Th1‐type T cell response through the induction of interleukin (IL)‐12 and the suppression of IL‐10. We therefore investigated whether OPN could enhance Th1 induction by vaccination against bacterial antigen in vivo. Unexpectedly, the co‐inoculation of OPN suppressed the induction of IFN‐γ‐producing CD4+ T cells and T cell proliferative response after the subcutaneous heat‐killed Listeria monocytogenes (HKLM) immunization. These results suggest that OPN down‐regulates T cell priming. Since dendritic cells (DC) play a pivotal role in T cell priming, we next analyzed the effects of OPN on DC. The addition of OPN into the culture of either bone marrow‐derived immature DC or an immature DC line JAWSII showed no effects on the expression of MHC class II, CD80, and CD86 molecules before and after HKLM stimulation. Consistently, in vitro OPN‐treated DC showed a normal antigen‐presenting function to an established Listeria‐specific Th1‐type T cells. However, when the DC were transferred into the footpad with HKLM and OPN, the migration of the transferred DC into the regional LN was suppressed in comparison to the DC transferred with HKLM alone. Furthermore, the addition of OPN into the culture of the DC line and HKLM severely suppressed the HKLM‐induced expression of CCR7 chemokine receptor which is an important factor in the migration of DC into LN. All the results suggest the existence of an OPN‐mediated negative feedback mechanism in the T cell immune response through the regulation of DC migration.
Virological and Immunological Characteristics of Fatal Virus‐Associated Haemophagocytic Syndrome (VAHS)
Authors: Kashiwagi Yasuyo , Kawashima Hisashi , Sato Satoshi , Ioi Hiroaki , Amaha Masahiro , Takekuma Kouji , Hoshika Akinori , Oshiro Hisashi , Matsubayashi Jun , Mukai Kiyoshi 
Published: November 2013

AbstractWe report three infants and one teenager with fatal virus‐associated haemophagocytic syndrome (VAHS). Two infants were admitted to our hospital because of cardio‐pulmonary arrest (CPA). The third infant was admitted to our department because of fever and liver dysfunction, and he was diagnosed as combined immunodeficiency with predominant T cell defect. The teenager was diagnosed as systemic lupus erythema (SLE) when she was 10 years old and admitted to our department because of fever and thrombocytopenia. The histological findings for the four patients' organs revealed many haemophagocytic cells. The patients were infected by Parainfluenza virus type 2 (PIV2), Enterovirus (EV), Cytomegalovirus (CMV), and Epstein‐Barr virus (EBV), respectively. Their laboratory data revealed elevated levels of ferritin and IL‐6, which also suggested virus‐associated haemophagocytic syndrome (VAHS). Although aggressive therapies were performed in all cases, the outcome was fatal. Further investigation would be needed to clarify the mechanism of VAHS and an effective therapeutic regimen is needed.
Autonomous Proliferation of HTLV−CD4+ T Cell Clones Derived from Human T Cell Leukemia Virus Type I (HTLV‐I)‐Associated Myelopathy Patients
Authors: Miyano‐Kurosaki Naoko , Kira Jun‐ichi , Barnor Jacob Samson , Maeda Naoyoshi , Misawa Naoko , Kawano Yuji , Tanaka Yuetsu , Yamamoto Naoki , Koyanagi Yoshio 
Published: November 2013

AbstractThat HTLV‐I infects CD4+ T cells and enhances their cell growth has been shown as successful long‐term in vitro proliferation in the presence of IL‐2. It is known that T cells isolated from HAM patients possess strong ability for cell proliferation in vitro and mRNA of various cytokines are abundantly expressed in CNS tissues of HAM patients. Hence, the cytokine‐induced proliferation could have an important role in pathogenesis and immune responses of HAM. In this study, we examined the relationship between cell proliferation and ability of in vitro cytokine production of CD4+ T cell clones isolated from HAM patients. We started a culture from a single cell to isolate cell clones immediately after drawing blood from the patients using limiting dilution method, which could allow the cell to avoid in vitro HTLV‐I infection after initiation of culture. Many cell clones were obtained and the rate of proliferation efficiency from a single cell was as high as 80%, especially in the 4 weeks' culture cells from HAM patients. These cells were classified as mainly Th0 phenotype that produce both IFN‐γ and IL‐4 after CD3‐stimulation. However, the frequency of proviral DNA in these cloned cells was significantly low. Our results indicate that the ability of cell proliferation in HAM patients is not restricted in HTLV‐I‐infected T cells. HTLV‐I‐uninfected CD4+ T cells, mainly Th0 cells, also have a strong ability to respond to IL‐2‐stimulation, showing that unusual immune activation on T cells has been observed in HAM patients.
Bacterial DNA Containing Methylated CpG Motifs Retains Immunostimulatory Activity in Synergy with Modified Lipopolysaccharides
Authors: Cho Yang Je , Ahn Bo Young , Song Eun Sun , Park Shin Ae , Lee Dong Hyeon , Kim Doo‐Sik , Lee Na Gyong 
Published: November 2013

AbstractWe previously described the immunostimulatory activity of CIA07, a combination of bacterial DNA fragments and modified LPS, and demonstrated that CIA07 has antitumor activity in a mouse bladder cancer model. In this study, we investigated whether methylation of the CpG motifs on the bacterial DNA fragments affects the immunostimulatory potential of CIA07. E. coli DNA fragments were methylated with CpG methylase, and then combined with modified LPS for experiments. Our results revealed that methylated CIA07 (mCIA07) and unmethylated CIA07 were equally active in inducing cytokine secretion from human whole blood cells. In addition, both methylated DNA fragments and mCIA07 retained the ability to activate expression and nuclear translocation of NF‐κB in RAW 264.7 cells. Finally, methylated DNA fragments and mCIA07 exhibited an antitumor activity comparable to those of their unmethylated counterparts in our mouse bladder cancer model. These data demonstrate that CpG methylation of E. coli DNA does not abrogate the immunostimulatory activity of DNA fragments or CIA07, suggesting that the synergistic activity by bacterial DNA in combination with LPS may be independent of the methylation status of CpG motifs.
Colonization and Turnover of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis in Otitis‐Prone Children
Authors: Yokota Shin‐ichi , Harimaya Atsushi , Sato Kiyoshi , Somekawa Yukihiro , Himi Tetsuo , Fujii Nobuhiro 
Published: November 2013

AbstractRecurrent otitis media are frequently intractable during childhood. It is unclear whether recurrent otitis media is caused by etiological bacteria colonization or by new infections. Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis were isolated from the nasopharynx of 7 otitis‐prone and 2 non‐prone children with recurrent otitis media. Plural bacterial species and strains were found in all children while affected by otitis media. The same strain was repeatedly isolated from all otitis‐prone children even after administration of antibiotics but was not from the non‐prone children. Antibiotic susceptibility did not differ significantly among the same repeatedly isolated strains. This pilot study suggests that the etiological bacteria tend to colonize and is hard to eliminate in otitis‐prone children.
Occurrence of RD9 Region and 500 bp Fragment among Clinical Isolates of Mycobacterium tuberculosis and Mycobacterium bovis
Authors: Das Samir , Das Suresh Chandra , Verma Rishendra 
Published: November 2013

AbstractThe present investigation dealt with the identification of Mycobacterium tuberculosis and M. bovis by RD9 region and 500 bp fragment PCR assays. Eight M. tuberculosis and 5 M. bovis characterized and identified from 40 human sputum and 41 bovine lung specimens and 20 M. tuberculosis and 9 M. bovis strains maintained at Mycobacteria Laboratory, Indian Veterinary Research Institute were included in this study. In this way, 28 M. tuberculosis and 14 M. bovis strains and, for comparison and control purpose, M. tuberculosis H37Rv, M. bovis BCG, M. canetti, M. smegmatis, M. phlei, M. chelonae, M. kansasii, M. xenopi and M. avium were subjected to RD9 and 500 bp amplification by PCR. All M. tuberculosis strains, M. tuberculosis H37 Rv and M. canetti yielded a product of 333 bp which showed presence of RD9 region in these strains, whereas all M. bovis yielded a product of 206 bp with RD9 PCR assay. There was no amplification product found in M. bovis BCG, M. xenopi, M. smegmatis, M. phlei, M. chelonae, M. kansasii, and M. avium. PCR based on 500 bp fragment showed a product of 500 bp in all M. bovis strains and M. bovis BCG. There was no amplification product of 500 bp found in M. canetti, M. smegmatis, M. phlei, M. chelonae, M. avium, M. kansasii, M. xenopi and was absent in all M. tuberculosis strains. The PCR assay results correlated 100% with the culture and biochemical results of the isolates. Our study suggested that PCR based on RD9 and 500 bp may effectively identify two closely related species of M. tuberculosis and M. bovis.
Identification of DDX1 as a JC Virus Transcriptional Control Region‐Binding Protein
Authors: Sunden Yuji , Semba Shingo , Suzuki Tadaki , Okada Yuki , Orba Yasuko , Nagashima Kazuo , Umemura Takashi , Sawa Hirofumi 
Published: November 2013

AbstractTo investigate the mechanism behind JC virus (JCV) cell specificity we performed electrophoretic mobility shift assays (EMSA) using probes derived from the JCV transcriptional control region (JCV‐TCR). Using nuclear extracts from the JCV‐susceptible neuroblastoma cell line IMR‐32, EMSA revealed a 670 kDa JCV‐TCR‐binding protein complex designated as #3‐bp. This complex could not be detected in nuclear extracts from non‐susceptible cell lines. Using column chromatographic purification and microsequencing, we identified cleavage stimulation factor (CstF) as a component of #3‐bp. However, as CstF is present in many cell types, we speculated that the IMR‐32‐specific component(s) of #3‐bp bind CstF. We performed a yeast two‐hybrid assay using CstF‐77 as the bait against a HeLa cDNA‐subtracted IMR‐32 cDNA library. This analysis detected binding between CstF‐77 and the RNA helicase DDX1. Subsequently, biotinylated DNA affinity precipitation and chromatin immunoprecipitation assays also confirmed that DDX1 binds specifically to JCV‐TCR. Our findings indicate that an association between DDX1 and the JCV‐TCR may play a significant role in JCV infection in IMR‐32 cells.
Cytolethal Distending Toxin (Cdt)‐Producing Escherichia coli Isolated from a Child with Bloody Diarrhea in Japan
Authors: Hinenoya Atsushi , Nagita Akira , Asakura Masahiro , Tsukamoto Teizo , Ramamurthy Thandavarayan , Nair Gopinath Balakrish , Takeda Yoshifumi , Yamasaki Shinji 
Published: November 2013

AbstractIn a retrospective analysis by PCR, the cdtI gene encoding the cytolethal distending toxin (Cdt) was detected in Escherichia coli O2:H12 strain isolated from the bloody diarrheal stool specimen of a child. To our knowledge, this is the first report showing the possible association of Cdt‐producing E. coli in Japan, particularly in a child with bloody diarrhea.
Prediction of Efficient Virological Response to Pegylated Interferon/Ribavirin Combination Therapy by NS5A Sequences of Hepatitis C Virus and Anti‐NS5A Antibodies in Pre‐Treatment Sera
Authors: El‐Shamy Ahmed , Sasayama Mikiko , Nagano‐Fujii Motoko , Sasase Noriko , Imoto Susumu , Kim Soo Ryang , Hotta Hak 
Published: November 2013

AbstractA considerable number of patients infected with Hepatitis C virus subtype 1b (HCV‐1b) do not respond to pegylated interferon/ribavirin combination therapy. In this study we explored a useful factor(s) to predict treatment outcome. A total of 47 HCV‐1b‐infected patients were treated with pegylated interferon/ribavirin for 48 weeks. Sera of the patients were examined for the entire NS5A sequence of the HCV genome, HCV RNA titers and anti‐NS5A antibodies. According to their responses, the patients were divided into two groups, early viral responders who cleared the virus by week 16 (EVR[16w]) and those who did not (Non‐EVR[16w]). The mean number of mutations in the V3 region (aa 2356 to 2379) or that in the V3 region plus its N‐terminally flanking region, which we refer to as interferon/ribavirin resistance‐determining region (IRRDR; aa 2334 to 2379), of NS5A obtained from the pretreatment sera was significantly larger for EVR(16w) compared with Non‐EVR(16w). Moreover, HCV‐1b isolates with ≥5 mutations in V3 or those with ≥6 mutations in IRRDR were almost exclusively found in EVR(16w). Also, the presence of detectable levels of anti‐NS5A antibodies in the pretreatment sera was closely associated with EVR(16w). In conclusion, a high degree of sequence variation in V3 (≥5) or IRRDR (≥6) and the presence of detectable levels of anti‐NS5A antibodies in the pretreatment sera would be useful factors to predict EVR(16w). On the other hand, a less diverse sequence in V3 (≤4) or IRRDR (≤5) together with the absence of detectable anti‐NS5A antibodies could be a predictive factor for Non‐EVR(16w).
Isolation and Characterization of Virulent Yellowtail Ascites Virus
Authors: Hirayama Takeshi , Nagano Ichiro , Shinmoto Hajime , Yagyu Ken‐ichi , Oshima Syun‐ichirou 
Published: November 2013

AbstractYellowtail ascites virus (YTAV) is the causative agent of ascites and deformity in fish and causes serious losses to the fish‐farming industry of yellowtail fry and fingerling Seriola quinqueradiata in Japan. In 2006, cultured yellowtail died from ascites in Kochi, Japan. We isolated and characterized a virus from the diseased fish. Based on the pathogenicity, culture characteristics, morphological features, RT‐PCR results targeting VP2/NS region, phylogeny based on the VP1 amino acid sequence, and immunochemical reactivity of structural proteins, the virus isolate was identified as YTAV (designated as YTAV‐06). YTAV‐06 was a more virulent isolate than YTAV Y‐6, isolated originally from yellowtail with ascites. To our knowledge, this is the first report describing that YTAV isolates may vary in their virulence.
Trafficking of QD‐Conjugated MPO‐ANCA in Murine Systemic Vasculitis and Glomerulonephritis Model Mice
Authors: Hoshino Akiyoshi , Nagao Tomokazu , Ito‐Ihara Toshiko , Ishida‐Okawara Akiko , Uno Kazuko , Muso Eri , Nagi‐Miura Noriko , Ohno Naohito , Tokunaka Kazuhiro , Naoe Shiro , Hashimoto Hiroshi , Yasuhara Masato , Yamamoto Kenji , Suzuki Kazuo 
Published: November 2013

AbstractIn systemic vasculitis, the serum level of myeloperoxidase (MPO)‐specific anti‐neutrophil cytoplasmic autoantibodies (MPO‐ANCA) is significantly elevated with the progression of disease. We have established a model of murine systemic vasculitis by administration of MPO‐ANCA and fungal mannoprotein to C57BL/6 mice. We examined the role of MPO and MPO‐ANCA in the pathogenesis of glomerulonephritis and systemic vasculitis in this model using quantum dots (QDs). We demonstrated that QD‐conjugated MPO‐ANCA (ANCA‐QD) visualized the translocation of MPO on the neutrophil membrane surface after stimulation with proinflammatory cytokines. We also observed that MPO translocation on neutrophils in both patients with rapid progressive glomerulonephritis and these model mice without any stimulation, suggesting that MPO translocation is certain to contribute to the development of glomerular lesion. In addition, blood flow on the kidney surface vessel was significantly decelerated in both SCG/Kj mice and this model, suggesting that ANCA induces the damage of blood vessel. These results indicate that MPO‐ANCA and surface‐translocated MPO on the activated neutrophils coordinately plays essential roles in the initial steps of the glomerulonephritis.
Contribution of Multi‐Antimicrobial Resistance to the Population of Antimicrobial ResistantEscherichia coliIsolated from Apparently Healthy Pigs in Japan
Authors: Harada Kazuki , Asai Tetsuo , Kojima Akemi , Sameshima Toshiya , Takahashi Toshio 
Published: November 2013

AbstractThe aim of this study was to clarify the involvement of tetracycline usage in resistance rates against other antimicrobials. Antimicrobial susceptibility testing was carried out on 545 porcineEscherichia coliisolates throughout Japan. As the result of analyzing by regions, resistance rates against kanamycin, oxytetracycline and trimethoprim in the Kanto/Koshinetu district were higher than those in some other districts. High resistance rates against kanamycin or trimethoprim in oxytetracycline‐resistant isolates were also observed in the Kanto/Koshinetu district The prevalence of multi‐antimicrobial resistance through co‐selection of resistances against kanamycin or trimethoprim by tetracycline usage could be the cause of regional differences in these resistances in porcineE. coli. By a communicative surveillance, kanamycin‐ and trimethoprim‐resistance rates were likely to be elevated with tetracycline usage. Thus, usage of specific antimicrobial(s) is a remarkable viewpoint to control antimicrobial resistant bacteria.
WT1 (Wilms' Tumor 1) Peptide Immunotherapy for Renal Cell Carcinoma
Authors: Iiyama Tatsuo , Udaka Keiko , Takeda Sachihiko , Takeuchi Tamotsu , Adachi Yoshihiro C. , Ohtsuki Yuji , Tsuboi Akihiro , Nakatsuka Shin‐ichi , Elisseeva Olga A. , Oji Yusuke , Kawakami Manabu , Nakajima Hiroko , Nishida Sumiyuki , Shirakata Toshiaki , Oka Yoshihiro , Shuin Taro , Sugiyama Haruo 
Published: November 2013

AbstractTumor‐specific immunotherapy with a Wilms' tumor 1 (WT1) peptide has been on clinical trial for leukemia, myelodysplastic syndrome, breast and lung cancers and is producing promising results. In this study, we treated three patients with renal cell carcinoma with an anchor modified, HLA‐A*2402 binding WT1 peptide which was emulsified in Freund's incomplete adjuvant. In two patients tumor growth was suppressed and clinical response was evaluated as stable disease by the RECIST criteria after 3 months of weekly immunizations. Notably, development of new metastases has stopped in these patients for a prolonged period. No deleterious side effects were observed. Peptide‐specific T cells were expanded in PBMCs of the patients and a substantial fraction of them bore the surface phenotype consistent with a CD8+ cytotoxic effector population. Although established tumors did not regress further, considering the component of the vaccine, i.e. peptide alone, the stabilization effect suggested the potential of WT1 peptide to develop into a more effective vaccine. To our knowledge, this is the first report of WT1 immunotherapy for renal cell carcinoma. Hopefully, the results will stimulate more extensive clinical studies.
Serological Evidence of Leptospiral Infection in Pig Populations in Different Districts in Japan
Authors: Naito Michiko , Sakoda Yoshihiro , Kamikawa Takayuki , Nitta Yoshiki , Hirose Kazuhiko , Sakashita Mitsuaki , Kurokawa Satoru , Kida Hiroshi 
Published: November 2013

AbstractSerum samples were collected from 938 pigs of 24 farms in Hokkaido, Kagoshima, and Okinawa prefectures in Japan in 2001–2005. Enzyme‐linked immunosorbent assay (ELISA) was used for the detection of antibodies to LipL32 antigen which is common to Leptospira interrogans. Samples positive in ELISA were then investigated by microscopic agglutination test for the identification of causal leptospires. Antibodies specific to leptospires of serovars Copenhageni, Bratislava, Australis and Javanica were detected in serum samples of pigs from each of the three districts. In addition, antibodies to leptospires of serovars Autumnalis and Tarassovi were predominantly detected in those from Kagoshima. The present study, thus, revealed that leptospires belonging to different serovars prevail in the pig population in Japan. In addition, it is the first detection of antibodies to leptospires belonging to serovars Javanica and Tarassovi in pigs in Japan.
Characteristics of a Pink‐Pigmented Bacterium Isolated from Biofilm in a Cooling Tower in Tokyo, Japan
Authors: Furuhata Katsunori , Goto Keiichi , Kato Yuko , Saitou Keiko , Sugiyama Jun‐ichi , Hara Motonobu , Yoshida Shin‐ichi , Fukuyama Masafumi 
Published: November 2013

AbstractStrain K‐20, a Gram‐negative, non‐motile, non‐spore‐forming and strictly aerobic rod, which produces a pale pink pigment, was isolated from biofilm in a cooling tower in Tokyo, Japan. The taxonomic feature of the strain was studied using phenotypic tests and phylogenetic analysis. Phylogenetic analysis of 16S rRNA gene sequences showed that the strain was related to Roseomonas gilardii subsp. rosea, Roseomonas gilardii subsp. gilardii, Roseomonas cervicalis and Roseomonas mucosa at 94.3–94.6 sequence similarities. Growth occurred at 25–40 C and pH 5.0–10.0, optimal at 35 C and pH 7.0. Growth did not occur in the presence of ≧2% NaCl. The API 20NE identification system gave a positive result for urease, L‐arabinose, potassium gluconate, adipic acid, malic acid and trisodium citrate (API code number 0201465). The predominant fatty acids of strain K‐20 were C18:1Δ11 (50.8%) and C16:1 (17.2%). Cells contained ubiquinone 10 (Q‐10) as the major quinone and the G+C content was 72.0 mol%. Based on phenotypic, chemotaxonomic and phylogenetic data, it was assumed that strain K‐20 (=JCM 14634) is a novel species of the genus Roseomonas.