bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cholera Toxin Modulates the Systemic Immune Responses against Vibrio cholerae Surface Antigens after Repeated Inoculations
Authors: Fernandez‐Miyakawa Mariano E. , Brero Maria Luisa , Mateo Nancy A. 
Published: November 2013

AbstractThe immunomodulating properties of a low cholera toxin (CT) dose over the systemic antibody response against Vibrio cholerae antigens after a comparatively extensive period of time were evaluated. Groups of 10 mice were injected intraperitoneally three times at 0, 30 and 86 days with 500 μl of buffer or 108 viable recombinant V. cholerae bacteria (lacking cholera toxin A subunit) with or without 100 ng of CT. Sera were obtained from inoculated mice at 0, 14, 28, 37, 58, 80, 93, 114, 236 and 356 days after the first injection. Vibriocidal activity and IgM and IgG anti‐lipopolysaccharide (LPS) or outer membrane protein (OMP) antibodies levels were estimated by ELISA in sera of inoculated mice. Anti‐LPS IgG subclasses were measured 2 weeks after each immunization by ELISA. Treatment of mice with CT markedly influenced the immune response to LPS but not against OMP of V. cholerae. Simultaneous intraperitoneal administration of CT with V. cholerae resulted in marked enhancement of both IgM anti‐LPS and vibriocidal titers which subsisted for a relatively extensive period of time after repeated antigen administration. No differences were observed in IgM and IgG anti‐OMP titers after extended periods of time between CT and control treatments. A similar pattern of IgG anti‐LPS subclasses was observed in the serum samples analyzed. These results suggest that long term CT administration modulates the IgM anti‐V. cholerae LPS response and the serum vibriocidal activity.
Antimicrobial Activities of Cinnamyl Rifamycin Derivatives, T‐9 and T‐11, against Mycobacterium tuberculosis and Mycobacterium avium Complex (MAC) with Special Reference to the Activities against Intracellular MAC
Authors: Sato Katsumasa , Shimizu Toshiaki , Dimova Velitchka , Tomioka Haruaki 
Published: November 2013

AbstractThe mycobacterial activities of cinamyl rifamycin derivatives, T‐9 and T‐11, especially against extracellular and intracellular Mycobacterium avium complex residing within macrophages and type II pneumocytes were compared with those of other rifamycins. The activities of test rifamycins were found to be in the order rifalazil, rifabutin, T‐9, T‐11, and rifampicin.
Time‐Kill Assay of Itraconazole against Malassezia Species
Authors: Sugita Takashi , Tsubuku Hisae , Tajima Mami , Tsuboi Ryoji , Nishikawa Akemi 
Published: November 2013

AbstractThe in vitro pharmacodynamic activity of itraconazole against Malassezia was determined by time‐kill methods. Itraconazole showed fungistatic activity at concentrations greater than 2× minimum inhibitory concentration (MIC) for each of the isolates of all nine Malassezia species associated with humans. A concentration exceeding 4× MIC decreased the growth to less than the inoculum amount (1×105 cells/ml).
Human Immunodeficiency Virus‐Type 1 Specific Cellular Immunity in Chronic Infected Patients on Prolonged Highly Active Antiretroviral Treatment and on Structured Treatment Interruption
Authors: Matsuda Reikei , Boström Ann‐Charlotte , Fredriksson Malin , Fredriksson Eva‐Lena , Bratt Göran , Hejdeman Bo , Sandström Eric , Okuda Kenji , Wahren Britta 
Published: November 2013

AbstractWe have followed 15 HIV‐1 chronically infected patients during prolonged highly active antiretroviral treatment (HAART) and subsequent long term structured treatment interruption (STI). We analyzed Nef, Tat, and p24 specific cellular immunity using IFN‐γ enzyme‐linked immunospot assays and T cell proliferation assays. Eight HAART patients showed IFN‐γ responses to at least one antigen, but no positive responses were seen during STI. We observed retained or increased p24 specific IFN‐γ responses in most patients during HAART with viral suppression. These results showed persisting HIV‐1 specific cellular immunity during HAART; however, in prolonged STI with viral rebound this immunity declined.
Genetic Diversity and Epizootiology ofChlamydophila psittaciPrevalent among the Captive and Feral Avian Species Based on VD2 Region ofompAGene
Authors: Chahota Rajesh , Ogawa Hirohito , Mitsuhashi Yoko , Ohya Kenji , Yamaguchi Tsuyoshi , Fukushi Hideto 
Published: November 2013

AbstractTo study genetic diversity and occurrence ofChlamydophila psittaci, a total of 1,147 samples from 11 avian orders including 53 genera and 113 species of feral and captive birds were examined usingompAgene based nested PCR. Three types of chlamydiae:C. psittaci(94.12%),C. abortus(4.41%) and unknownChlamydophilasp. (1.47%) were identified among 68 (5.93%) positive samples (Psittaciformes‐59,Ciconiiformes‐8 andPasseriformes‐1). Based on nucleotide sequence variations in the VD2 region ofompAgene, all 64 detectedC. psittacistrains were grouped into 4 genetic clusters. Clusters I, II, III and IV were detected from 57.35%, 19.12%, 10.29% and 7.35% samples respectively. A single strain of unknownChlamydophilasp. was found phylogenetically intermediate betweenChlamydophilaspecies infecting avian and mammalian hosts. AmongPsittaciformes, 28 out of 81 tested species including 10 species previously unreported were found to be chlamydiae positive. Chlamydiosis was detected among 8.97% sick and 48.39% dead birds as well 4.43% clinically normal birds. Therefore, it was observed that though various genetically diverse chlamydiae may cause avian chlamydiosis, only a fewC. psittacistrains are highly prevalent and frequently associated with clinical/subclinical infections.
The Postantifungal Effect (PAFE) of Itraconazole, in Comparison with Those of Miconazole and Fluconazole, on Candida Species
Authors: Uchida Katsuhisa , Abe Shigeru , Yamaguchi Hideyo 
Published: November 2013

AbstractTo study the postantifungal effect (PAFE) of itraconazole (ITCZ) against Candida species, we quantitated it using a newly introduced parameter T/C (T=time required for the drug‐treated culture to reach 5‐fold increase in turbidity; C=time required for the drug‐free control culture to reach 5‐fold increase in turbidity) referred as PAFE index and compared the results with those obtained for two other azole antifungal drugs fluconazole (FLCZ) and miconazole (MCZ). The mean values of PAFE index for ITCZ against three C. albicans strains were 1.28, 1.45, and 1.60 when exposed to the drug for 1 hr at concentrations of 1×MIC, 2×MIC and 4×MIC, respectively. These values are similar to those for MCZ and appreciably higher than those for FLCZ. PAFE index values for ITCZ against each single strain of three non‐albicans Candida species exposed to 2×MIC of the drug for 1 hr were: 1.22 with C. glabrata, 1.63 with C. parapsilosis, and 3.90 with C. tropicalis. Again, comparable values were obtained for MCZ and lower values for FLCZ. The drug concentration for exposure and the duration of exposure only slightly or scarcely affected the PAFE of ITCZ, although the magnitude of the PAFE considerably varied among different strains or species tested. In general, ITCZ, as well as MCZ, produced greater extents of PAFE than those for FLCZ against C. albicans and several non‐albicans Candida species.
Mechanism of Enhanced Hematopoietic Response by Soluble β‐Glucan SCG in Cyclophosphamide‐Treated Mice
Authors: Harada Toshie , Kawaminami Hiromi , Miura Noriko N. , Adachi Yoshiyuki , Nakajima Mitsuhiro , Yadomae Toshiro , Ohno Naohito 
Published: November 2013

AbstractSCG is a major 6‐branched 1,3‐β‐D‐glucan in Sparassis crispa Fr. SCG shows antitumor activity and also enhances the hematopoietic response in cyclophosphamide (CY)‐treated mice. In the present study, the molecular mechanism of the enhancement of the hematopoietic response was investigated. The levels of interferon‐ (IFN‐) γ, tumor necrosis factor‐ (TNF‐) α, granulocyte‐macrophage‐colony stimulating factor (GM‐CSF), interleukin‐ (IL‐) 6 and IL‐12p70 were significantly increased by SCG in CY‐treated mice. GM‐CSF production in the splenocytes from the CY‐treated mice was higher than that in normal mice regardless of SCG stimulation. Neutralizing GM‐CSF significantly inhibited the induction of IFN‐γ, TNF‐α and IL‐12p70 by SCG. The level of cytokine induction by SCG was regulated by the amount of endogenous GM‐CSF produced in response to CY treatment in a dose‐dependent manner. The expression of β‐glucan receptors, such as CR3 and dectin‐1, was up‐regulated by CY treatment. Blocking dectin‐1 significantly inhibited the induction of TNF‐α and IL‐12p70 production by SCG. Taken together, these results suggest that the key factors in the cytokine induction in CY‐treated mice were the enhanced levels of both endogenous GM‐CSF production and dectin‐1 expression.
Development of a Real‐Time PCR Assay for Detection of gyrA Mutations Associated with Reduced Susceptibility to Ciprofloxacin in Salmonella enterica Serovar Typhi and Paratyphi A
Authors: Morita Masatomo , Ito Kenitiro , Hirose Kenji , Takahashi Hideyuki , Shimuta Ken , Terajima Jun , Ohnishi Makoto , Harada Makoto , Matsuzaki Mitsuhiro , Watanabe Haruo , Izumiya Hidemasa 
Published: November 2013

AbstractA real‐time PCR assay with the cycling probe method was used to detect mutations at codons 83 and 87 in the DNA gyrase A subunit encoded by gyrA in Salmonella enterica serovar Typhi and Paratyphi A clinical isolates. The susceptibility estimated from the results of the gyrA mutation assay was consistent with that identified by the culture method using an E‐test. This assay allows rapid screening of S. enterica serovar Typhi and Paratyphi A with reduced susceptibility to ciprofloxacin.
Typing of bfpA Genes of Enteropathogenic Escherichia coli Isolated in Thailand and Japan by Heteroduplex Mobility Assay
Authors: Iida Mariko , Yamazaki Mitsugu , Yatsuyanagi Jun , Ratchtrachenchai Orn‐Anong , Subpasu Sarayoot , Okamura Noboru , Ito Kenitiro 
Published: November 2013

AbstractWe developed a rapid genetic approach for screening bfpA variants of enteropathogenic E. coli (EPEC) using a heteroduplex mobility assay (HMA). A total of 204 human EPEC strains were isolated in Thailand and Japan. Of 34 bfpA‐positive EPEC strains, bfpA variants were classified into 5 HMA‐types. Different HMA‐types were found in EPEC of the same serotypes. The results suggest that HMA is a simple and easy method to analyze polymorphism of bfpA gene, and can be used in laboratories without large apparatus such as sequencers.
CD8+ T Cells Play Disparate Roles in the Induction and the Effector Phases of Murine Experimental Allergic Conjunctivitis
Authors: Fukushima Atsuki , Yamaguchi Tomoko , Fukuda Ken , Sumi Tamaki , Kumagai Naoki , Nishida Teruo , Imai Shosuke , Ueno Hisayuki 
Published: November 2013

AbstractAlthough CD4+ Th2 cells clearly play an essential role in the development of experimental allergic diseases, the functions CD8+ T cells may have in these diseases have been investigated less extensively and remain controversial. Here, we investigated the roles of CD8+ T cells in the development of experimental allergic conjunctivitis (EC). EC was induced in CD8α‐deficient (CD8KO) mice and wild‐type (WT) mice by active immunization with short ragweed pollen (RW) followed by challenge with RW‐containing eye drops. Alternatively, EC was induced by transferring RW‐primed splenocytes followed by RW challenge. With regard to actively immunized mice, CD8KO mice showed significantly less severe eosinophil infiltration of the conjunctiva and lower total IgE levels, although the levels of the other Igs were equivalent between the two strains. Cytokine production by cultured splenocytes also did not differ, but the WT conjunctivas showed upregulated IL‐5 and IL‐6 expression and greater upregulation of IL‐4 expression than the conjunctivas of CD8KO mice. Thus, CD8+ T cells may play a significant role during the induction phase by aiding IgE production and the generation of Th2 cytokines in the conjunctiva, thus promoting the development of EC. In contrast, splenocytes from CD8KO mice induced significantly more severe EC in WT mice than cells from WT mice. In addition, transfer of RW‐primed splenocytes induced significantly more severe eosinophil infiltration in CD8KO recipient mice. Thus, CD8+ T cells promote the development of EC during the induction phase, but suppress it during the effector phase.
Phylogenetic Analysis of Bacillus cereus Isolates from Severe Systemic Infections Using Multilocus Sequence Typing Scheme
Authors: Vassileva Maria , Torii Keizo , Oshimoto Megumi , Okamoto Akira , Agata Norio , Yamada Keiko , Hasegawa Tadao , Ohta Michio 
Published: November 2013

AbstractBacillus cereus strains from cases of severe or lethal systemic infections, including respiratory symptoms cases, were analyzed using multilocus sequence typing scheme of B. cereus MLST database. The isolates were evenly distributed between the two main clades, and 60% of them had allele profiles new to the database. Half of the collection's strains clustered in a lineage neighboring Bacillus anthracis phylogenetic origin. Strains from lethal cases with respiratory symptoms were allocated in both main clades. This is the first report of strains causing respiratory symptoms to be identified as genetically distant from B. anthracis. The phylogenetic location of the presented here strains was compared with all previously submitted to the database isolates from systemic infections, and were found to appear in the same clusters where clinical isolates from other studies had been assigned. It seems that the pathogenic strains are forming clusters on the phylogenetic tree.
Structural Requirements of Cholesterol for Binding to Vibrio cholerae Hemolysin
Authors: Ikigai Hajime , Otsuru Hiroshi , Yamamoto Koichiro , Shimamura Tadakatsu 
Published: November 2013

AbstractCholesterol is necessary for the conversion of Vibrio cholerae hemolysin (VCH) monomers into oligomers in liposome membranes. Using different sterols, we determined the stereochemical structures of the VCH‐binding active groups present in cholesterol. The VCH monomers are bound to cholesterol, diosgenin, campesterol, and ergosterol, which have a hydroxyl group at position C‐3 (3βOH) in the A ring and a C–C double bond between positions C‐5 and C‐6 (C–C Δ5) in the B ring. They are not bound to epicholesterol and dihydrocholesterol, which form a covalent link with a 3αOH group and a C–C single bond between positions C‐5 and C‐6, respectively. This result suggests that the 3βOH group and the C–C Δ5 bond in cholesterol are required for VCH monomer binding. We further examined VCH oligomer binding to cholesterol. However, this oligomer did not bind to cholesterol, suggesting that the disappearance of the cholesterol‐binding potential of the VCH oligomer might be a result of the conformational change caused by the conversion of the monomer into the oligomer. VCH oligomer formation was observed in liposomes containing sterols with the 3βOH group and the C–C Δ5 bond, and it correlated with the binding affinity of the monomer to each sterol. Therefore, it seems likely that monomer binding to membrane sterol leads to the assembly of the monomer. However, since oligomer formation was induced by liposomes containing either epicholesterol or dihydrocholesterol, the 3βOH group and the C–C Δ5 bond were not essential for conversion into the oligomer.
Genetic Variability of Human Cytomegalovirus UL132 Gene in Strains from Infected Infants
Authors: Sun Zheng Rong , Ji Yao Hua , Ruan Qiang , He Rong , Ma Yan Ping , Qi Ying , Mao Zhi Qin , Huang Yu Jing , Wang Yue Ping 
Published: November 2013

AbstractHuman cytomegalovirus (HCMV) displays genetic polymorphisms. HCMV infects a number of organs and cell types, leading to the hypothesis that HCMV disease and tissue tropism may be related to specific sequence variability. A gene in UL/b′ of HCMV, UL132 open reading frame (ORF), encodes glycoprotein (gpUL132) which is identified as a low‐abundance structural component of HCMV. In this study, the sequence variability of the UL132 gene was studied in 30 clinical strains. The results showed that a large number of nucleotide non‐synonymous substitutions occurred in the UL132 ORF, particularly in the 5′ half, in comparison to the UL132 of reference strain, Toledo. The UL132 variants of the clinical strains were clustered clearly into three major groups in the phylogenetic tree: G1(10/30), G2(9/30), and G3(11/30). The precise definition of UL132 genotypes and their putative functions would be helpful in a better understanding of the HCMV.
New Epitope Peptides Derived from Hepatitis C Virus (HCV) 2a Which Have the Capacity to Induce Cytotoxic T Lymphocytes in HLA‐A2+ HCV‐Infected Patients
Authors: Wang Yi , Takao Yukari , Harada Mamoru , Yutani Shigeru , Ide Tatsuya , Sata Michio , Itoh Kyogo , Yamada Akira 
Published: November 2013

AbstractBecause cytotoxic T lymphocytes (CTLs) play an important role in the specific immunotherapy of hepatitis C virus (HCV) infection, a series of CTL epitopes has been defined from HCV genotype 1a or 1b protein. Here, we attempted to identify HCV2a‐derived epitopes that are capable of inducing HLA‐A2‐restricted and peptide‐specific CTLs. Peripheral blood mononuclear cells (PBMCs) of HLA‐A2+ HCV2a‐infected patients or healthy donors were stimulated in vitro with each of the HCV2a‐derived peptides, which were prepared based on the HLA‐A2‐binding motif, and their peptide‐specific and HLA‐A2‐restricted cytotoxicities were examined. The HCV2a 432–441, HCV2a 716–724, and HCV2a 2251–2260 peptides were found to efficiently induce peptide‐specific CTLs from the PBMCs of HLA‐A2+ HCV2a‐infected patients. Cytotoxicity was mainly mediated by CD8+ T cells in a HLA class I‐restricted manner. These results indicate that the HCV2a 432–441, HCV2a 716–724, and HCV2a 2251–2260 peptides might be applicable for peptide‐based immunotherapy of HLA‐A2+ HCV2a‐infected patients.
Recombinant Vaccinia DIs Expressing Simian Immunodeficiency Virus Gag and Pol in Mammalian Cells Induces Efficient Cellular Immunity as a Safe Immunodeficiency Virus Vaccine Candidate
Authors: Okamura Tomotaka , Someya Kenji , Matsuo Kazuhiro , Hasegawa Atsuhiko , Yamamoto Naoki , Honda Mitsuo 
Published: November 2013

AbstractA highly attenuated vaccinia virus substrain of Dairen‐I (DIs) shows promise as a candidate vector for eliciting positive immunity against immune deficiency virus. DIs was randomly obtained by serial 1‐day egg passages of a chorioarantoic membrane‐adapted Dairen strain (DIE), resulting in substantial genomic deletion, including various genes regulating the virus‐host‐range. To investigate the impact of that deletion and of the subsequent insertion of a foreign gene into that region of DIs on the ability of the DIs recombinant to induce antigen‐specific immunity, we generated a recombinant vaccinia DIs expressing full‐length gag and pol genes of simian immunodeficiency virus (SIV) (rDIsSIV gag/pol) and studied the biological and immunological characteristics of the recombinant natural mutant. The rDIsSIV gag/pol developed a tiny plaque on the chick embryo fibroblast (CEF). Viral particles of rDIsSIV gag/pol as well as SIV Gag‐like particles were electromicroscopically detected in the cytoplasm. Interestingly, the recombinant DIs strain grows well in CEF cells but not in mammalian cells. While rDIsSIV gag/pol produces SIV proteins in mammalian HeLa and CV‐1 cells, recombinant modified vaccinia Ankara strain (MVA) expressing SIV gag and pol genes (MVA/SIV239gag/pol) clearly replicates in HeLa and CV‐1 cell lines under synchronized growth conditions and produces the SIV protein in all cell lines. Moreover, intradermal administration of rDIsSIV gag/pol or of MVA/SIV239gag/pol elicited similar levels of IFN‐γ spot‐forming cells specific for SIV Gag. If the non‐productive infection characteristically induced by recombinant DIs is sufficient to trigger immune induction, as we believe it is, then a human immunodeficiency virus vaccine employing the DIs recombinant would have the twin advantages of being both effective and safe.
Analysis of Bacterial Community Structure in the Natural Circulation System Wastewater Bioreactor by Using a 16S rRNA Gene Clone Library
Authors: Niu Shi‐Quan , Fukushima Jun , Jiang Ying , Ishikawa Yuichi , Ueda Tohru , Matsumoto Satoshi 
Published: November 2013

AbstractA variety of physical and chemical parameters are routinely monitored during operation of the Natural Circulation System, a wastewater purification bioreactor in which only natural materials and no synthetic chemicals are used. However, the microbial community structures existing in the Natural Circulation System have not been well characterized. Thus, bacterial community structure and composition in this system were studied using clone library analysis of 16S ribosomal RNA genes amplified using PCR with universal bacterial primer sets. The PCR products were then subcloned into the pGEM‐T vector. Each unique restriction fragment length polymorphism pattern, created by using two pairs of restriction endonucleases, was designated as an operational taxonomic unit (OTU). The Natural Circulation System comprises five tanks, the second and third of which play a major role in the bioreactor. Clone library profiles and principal component analysis revealed differences in the bacterial community structures of the second (anaerobic chamber) and the third (aerobic chamber) tanks. However, the β‐proteobacteria, Bacteroidetes/Chlorobi and γ‐proteobacteria groups were dominant in both tanks. Bacterial composition was more complex in the second tank (107 OTUs) than in the third tank (68 OTUs). Of a total of 154 OTUs in the clone libraries, only 21 were common to the two tanks. The results obtained in this study should provide important information for future research into and management of the Natural Circulation System wastewater bioreactor.
Screening of Upregulated Genes in Suckling Mouse Central Nervous System during the Disease Stage of Rabies Virus Infection
Authors: Ubol Sukathida , Kasisith Jitra , Mitmoonpitak Channarong , Pitidhamabhorn Dhanesh 
Published: November 2013

AbstractThe pathogenesis of hydrophobia remains unclear. The aim of this study was to identify the differentially upregulated genes that correlated with disease development in an experimental mouse model to provide better understanding of pathological mechanisms in rabies. The present work employed Clontech mouse array 1.2 II containing 1,176 gene transcripts. Suckling mice were intracerebrally infected with canine rabies virus. The gene expression profiles on day 2, 4 and 6 post inoculation were followed. The results show genes whose expression increased at least twofold above the control, mock‐infected brain. The numbers of genes showing altered expression level were 29, 109 and 98 genes on day 2, 4 and 6, respectively. The genes with altered expression were classified into eight major groups, namely immune response, metabolism, receptor and transporter, growth factors, death mediated factors, transcription and translation factors, proteases, and kinases. The numbers of upregulated genes during the disease stage was much higher than during the asymptomatic stage. This suggested that direct interaction between RABV and target cells induced massive destruction of a cellular homeostasis which may lead to functional termination of the CNS.