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Host Cholesterol Synthesis Contributes to Growth of Intracellular Toxoplasma gondii in Macrophages OAK
NISHIKAWA, Yoshifumi; IBRAHIM, Hany M.; KAMEYAMA, Kyohko; SHIGA, Ikumi; HIASA, Jun; XUAN, Xuenan; 西川, 義文; 玄, 学南.
The intracellular protozoan Toxoplasma gondii lacks the ability to synthesize sterol and scavenges cholesterol from the lowdensity lipoprotein receptor (LDLR) pathway of its host to facilitate replication. Sterol biosynthesis inhibitors, however, have a demonstrated anti-Toxoplasma effect. In this study, we examined the host mevalonate pathway as a novel source of cholesterol for T. gondii and its effects on parasite growth in macrophages. Parasite growth did not significantly change in the absence of LDLR or when LDL was exogenously supplemented. Lovastatin and compactin, both inhibitors of hydroxymethylglutaryl-CoA (HMG-CoA) reductase in the mevalonate pathway, significantly inhibited T. gondii growth in both wild-type and LDLR-knockout macrophages....
Palavras-chave: Cholesterol; Lipid metabolism; Macrophage; Parasitology; Toxoplasma gondii.
Ano: 2011 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3583
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Identification, Cloning and Characterization of BmP41, a Common Antigenic Protein of Babesia microti OAK
MASATANI, Tatsunori; OOKA, Hideo; TERKAWI, Mohamad A.; CAO, Shinuo; LUO, Yuzi; ASADA, Masahito; HAYASHI, Kei; NISHIKAWA, Yoshifumi; XUAN, Xuenan; 西川, 義文; 玄, 学南.
Babesia microti is a rodent tick-borne blood parasite and the major causative agent of emerging human babesiosis. Here, we identified a candidate of common antigenic protein BmP41 of B. microti by serological screening of cDNA library of human-pathogenic Gray strain with antisera against rodent Munich strain. Immunofluorescent antibody test using mouse anti-recombinant BmP41 (rBmP41) serum revealed that native BmP41 was expressed in each of the developmental stages of B. microti merozoites. An enzyme-linked immunosorbent assay (ELISA) using rBmP41 detected specific antibodies in sera from hamsters infected with B. microti Gray strain and mice infected with B. microti Munich strain. Taken together, BmP41 could be a promising universal serological marker for...
Palavras-chave: Babesia microti; BmP41; Common antigen; ELISA; Immunoscreening.
Ano: 2013 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3917
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Characterization of Toxoplasma gondii 5' UTR with Encyclopedic TSS Information OAK
Yamagishi, J.; Watanabe, J.; Goo, Y. K.; Masatani, T.; Suzuki, Y.; Xuan, X.; 玄, 学南.
The 5′ UTR is widely involved in gene expression via post-transcriptional regulation. However, a detailed profile of the 5′ UTR for Toxoplasma gondii has not yet been demonstrated. To investigate the issue, we compared the predicted open reading frames (ORFs) and transcription start sites (TSSs) of T. gondii obtained by TSS-seq, a method that enables analysis of encyclopedic TSSs with next-generation sequencers. As a result, it was demonstrated that the mode length of the 5′ UTR is between 120 and 140 nucleotides (nts) when a subset of genes with predicted signal peptides was examined. However, when genes without the signal peptide were examined, the length was extended to approximately 600 nts. Because additional information on the predicted signal...
Ano: 2012 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3822
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Molecular Prevalence of Different Genotypes of Theileria orientalis Detected from Cattle and Water Buffaloes In Thailand OAK
Altangerel, Khukhuu; Sivakumar, Thillaiampalam; Inpankaew, Tawin; Jittapalapong, Sathaporn; Terkawi, Mohamad Alaa; Ueno, Akio; Xuan, Xuenan; Igarashi, Ikuo; Yokoyama, Naoaki; 玄, 学南; 五十嵐, 郁男; 横山, 直明.
Here we report on an epidemiological study regarding the molecular prevalence of different genotypes of Theileria orientalis present among domestic cattle and water buffalo populations bred in Thailand. A phylogenetic analysis based on the parasitic gene encoding a major piroplasm surface protein revealed the presence of 5 genotypes (Types 1, 3, 5, 7, and N-3) in cattle and 7 genotypes (Types 1, 3, 4, 5, 7, N-2, and N-3) in water buffaloes. Types 4, 7, and N-3 of T. orientalis were reported for the first time in water buffaloes. The previously reported C and Thai types from Thailand clustered as types 7 and 6, respectively, in the present analysis. Great similarities were observed among nucleotide sequences of isolates of the same genotype from cattle and...
Ano: 2011 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3592
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Identification of a novel B. gibsoni 27-kDa protein as a serodiagnostic antigen OAK
Terkawi, M. A.; Aboge, G.; Jia, H.; Goo, Y-K.; Ooka, H.; Yamagishi, Junya; Nishikawa, Yoshifumi; Kawazu, Shin-ichiro; Fujisaki, K.; Xuan, Xuenan; 山岸, 潤也; 西川, 義文; 河津, 信一郎; 玄, 学南.
A novel gene encoding 27-kDa protein was identified by the screening of Babesia gibsoni cDNA library with acutely infected dog serum. The BgP27 is a single copy gene with a predicted open reading frame of 762 bp and 254 amino acids. The phylogenic analysis of the deduced amino acid of BgP27 demonstrated considerable identities with members of Plasmodium berghei circumsporozoite protein family that ranged between 18.4% and 22.8%. The BgP27 was expressed as a glutathione S-transferase fusion protein in Escherichia coli. The serum raised in mice against the recombinant protein specifically reacted with a 27-kDa protein in the extracts of B. gibsoni parasites. Confocal laser scanning microscopic observation showed high fluorescent reactivity with both...
Palavras-chave: Babesia gibsoni; Enzyme-linked immunosorbent assay; Deiagnostic performance.
Ano: 2008 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/2232
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Cloning and Characterization of a 2-Cys Peroxiredoxin from Babesia gibsoni OAK
MASATANI, Tatsunori; ASADA, Masahito; ICHIKAWA-SEKI, Madoka; USUI, Miho; TERKAWI, Mohamad A.; HAYASHI, Kei; KAWAZU, Shin-ichiro; XUAN, Xuenan; 河津, 信一郎; 玄, 学南.
Peroxiredoxins (Prxs) are a family of antioxidant enzymes. Here, we cloned a 2-Cys Prx, BgTPx-1, from the canine Babesia parasite B. gibsoni. Sequence identity between BgTPx-1 and 2-Cys Prx of B. bovis was 81% at the amino acid level. Enzyme activity assay by using recombinant BgTPx-1 (rBgTPx-1) indicated that BgTPx-1 has antioxidant activity. Antiserum from a mouse immunized with rBgTPx-1 reacted with parasite lysates and detect a protein with a monomeric size of 22 kDa and also a 44 kDa protein, which might be an inefficiently reduced dimer. BgTPx-1 was expressed in the cytoplasm of B. gibsoni merozoites. These results suggest that the BgTPx-1 may play a role to control redox balance in the cytoplasm of B. gibsoni.
Palavras-chave: Antioxidant activity; Babesia gibsoni; Canine; Peroxiredoxin.
Ano: 2014 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3925
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Development and evaluation of an enzyme-linked immunosorbent assay using recombinant p23 for the detection of antibodies to Cryptosporidium parvum in cattle OAK
Bannai, Hiroshi; Nishikawa, Yoshifumi; Seo, Jin-yong; Nakamura, Chinatsu; Zhang, Sofa; Kimata, Isao; Takashima, Yasuhiro; Li, Junyou; Igarashi, Ikuo; Xuan, Xuenan; 西川, 義文; 五十嵐, 郁男; 玄, 学南.
An enzyme-linked immunosorbent assay (ELISA) based on the recombinant p23 of Cryptosporidium parvum was established for the detection of antibodies against C. parvum in cattle. The sensitivity and specificity of the ELISA were evaluated with the standard indirect fluorescent antibody test (IFAT) using sporozoites as antigens. Of 77 bovine sera collected from China, 20 (26.0%) were identified as positive by the IFAT. The same samples were tested with the ELISA. The optical densities at 415 nm were compared to the IFAT results and statistically analyzed to designate a provisional cut-off point. As a result, the cut-off point was concluded to be 0.08, which was considered to be the best condition in the light of its sensitivity (80%) and specificity (73.7%)....
Palavras-chave: Cryptosporidium parvum; P23; ELISA; IFAT.
Ano: 2006 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/142
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Molecular Detection of Theileria Species in Sheep from Northern China OAK
CAO, Shinuo; ZHANG, Shoufa; JIA, Lijun; XUE, Shujiang; YU, Longzheng; KAMYINGKIRD, Ketsarin; MOUMOUNI, Paul Franck Adjou; MOUSSA, Ahmed Abd El Moniem; ZHOU, Mo; ZHANG, Yuanming; TERKAWI, Mohamad Alaa; MASATANI1), Tatsunori; NISHIKAWA, Yoshifumi; XUAN, Xuenan; 西川, 義文; 玄, 学南.
Ovine theileriosis is a tick-borne disease that restricts the development of small ruminant husbandry in northern China. In this study, we report on a molecular epidemiological survey of ovine Theileria spp. in 198 blood samples taken from sheep in northern China. The DNA samples were screened by a nested polymerase chain reaction (PCR) targeting the 18S rRNA gene of ovine Theileria spp. The prevalence of ovine Theileria spp. in Yanji, Nongan, Longjing, Toudao and Jinchang was 80%, 40%, 37%, 24% and 32%, respectively. The sequencing analyses approved the present of the T. orientalis and/or T. luwenshuni in these regions. Taken together, we have demonstrated a high incidence of Theileria spp. in northern China that calls for the need to design effective...
Palavras-chave: China; Ovine Theileria spp; Sheep; Theileria luwenshuni.
Ano: 2013 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3924
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Molecular Epidemiological Survey of Theileria orientalis in Thua Thien Hue Province, Vietnam OAK
KHUKHUU, Altangerel; LAN, Dinh Thi Bich; LONG, Phung Thang; UENO, Akio; LI, Yan; LUO, Yuzi; MACEDO, Alan Caine Costa de; MATSUMOTO, Kotaro; INOKUMA, Hisashi; KAWAZU, Shin-Ichiro; IGARASHI, Ikuo; XUAN, Xuenan; YOKOYAMA, Naoaki; 松本, 高太郎; 猪熊, 壽; 河津, 信一郎; 五十嵐, 郁男; 玄, 学南; 横山, 直明.
Theileria orientalis is a benign bovine protozoan parasite that occasionally causes serious economic loss in the livestock industry. We report the findings of a molecular epidemiological survey of T. orientalis in 94 Vietnamese yellow cattle, 43 water buffaloes, 21 sheep, 21 goats and 85 blood-sucking ticks of cattle in the Thua Thien Hue province of Vietnam. The major piroplasm surface protein (MPSP) gene of T. orientalis was detected using polymerase chain reaction from 13 cattle (13.8%), 11 water buffaloes (25.6%), 1 sheep (4.8%) and 9 ticks (10.6%). Phylogenetic analysis using MPSP gene sequences showed the presence of seven genotypes, four previously categorized genotypes (Types 1, 3, 5 and 7) and three new genotypes (Types N-1, N-2 and N-3).
Palavras-chave: Cattle; Genotype; MPSP; Theileria orientalis; Vietnam.
Ano: 2011 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3585
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Construction of the recombinant pseudorabies viruses expressing Cryptosporidium parvum an immunodominant surface protein, p23 OAK
Takashima, Yasuhiro; Xuan, Xuenan; Nagasawa, Hideyuki; Matsumoto, Yasunobu; Igarashi, Ikuo; Fujisaki, Kozo; Mikami, Takeshi; Otsuka, Haruki; 玄, 学南; 五十嵐, 郁男.
To develop a vaccine against cryptosporidiosis in animals, we constructed recombinant pseudorabies virus (PrV), a member of the Herpesviridae Alphaherpesvirus subfamily, expressing an immunodominant surface protein p23 of Cryptosporidium parvum sporozoites. Because of the wide host range of PrV, it has the possibility as the vector to delivery the foreign genes to several species of animals containing experiment animal. In the recombinant constructed in this study, the p23 gene under the control of CAG promoter was integrated into the thymidine kinase (TK) gene of PRV. Antibody against p23 recognized p23 expressed in CPK cells infected with the recombinant, as the approximate 23 kDa specific band in Western blotting analysis. This study showed the...
Palavras-chave: Cryptosporidium parvum; P23; Herpes; Pseudorabies virus; Subunit vaccine.
Ano: 2000 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/129
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Expression Patterns of the Implantation-associated Genes in the Uterus during the Estrous Cycle in Mice OAK
LEE, Dong-Soo; YANAGIMOTO UETA, Yoshiko; XUAN, Xuenan; IGARASHI, Ikuo; FUJISAKI, Kozo; SUGIMOTO, Chihiro; TOYODA, Yutaka; SUZUKI, Hiroshi; 玄, 学南; 五十嵐, 郁男; 鈴木, 宏志.
The mRNA expression patterns of EGF, HB-EGF, Amphiregulin, EGF receptor, IGF-1, CSF-1, IL-1 alpha, IL-1 beta, IL-1 receptor type 1, IL-1 receptor antagonist, LIF, COX-1, COX-2, Mucin-1, calcitonin, and rat USAG-1 mouse homologue, all of which are involved in the process of conceptus implantation to the endometrium, were examined during the estrous cycle by means of real-time quantitative PCR. COX-2, HB-EGF, LIF, Mucin-1, CSF-1, IL-1 alpha, IL-1 beta, and IL-1 receptor antagonist were temporally regulated during the estrous cycle and highly expressed during the estrous stage. In the case of COX-1, EGF, IGF-1, and EGF receptor, the highest mRNA expression was during the diestrous stage. In contrast, the rat USAG-1 mouse homologue mRNA expression did not...
Palavras-chave: Estrous cycle; Implantation; Mice; Ovarian hormone.
Ano: 2005 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3022
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PCR Detection and Genetic Diversity of Bovine Hemoprotozoan Parasites in Vietnam OAK
SIVAKUMAR, Thillaiampalam; LAN, Dinh Thi Bich; LONG, Phung Thang; YOSHINARI, Takeshi; TATTIYAPONG, Muncharee; GUSWANTO, Azirwan; OKUBO, Kazuhiro; IGARASHI, Ikuo; INOUE, Noboru; XUAN, Xuenan; YOKOYAMA, Naoaki; 井上, 昇; 玄, 学南; 横山, 直明.
Hemoprotozoan infections often cause serious production losses in livestock. In the present study, we conducted a PCR-based survey of Babesia bovis, Babesia bigemina, Theileria annulata, Theileria orientalis, Trypanosoma evansi and Trypanosoma theileri, using 423 DNA samples extracted from blood samples of cattle (n=202), water buffaloes (n=43), sheep (n=51) and goats (n=127) bred in the Hue and Hanoi provinces of Vietnam. With the exception of T. annulata and T. evansi, all other parasite species (B. bovis, B. bigemina, T. orientalis and T. theileri) were detected in the cattle populations with B. bovis being the most common among them. Additionally, four water buffaloes and a single goat were infected with B. bovis and B. bigemina, respectively. The Hue...
Palavras-chave: Epidemiology; Hemoprotozoa; Livestock; PCR; Vietnam.
Ano: 2013 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4015
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Babesia gibsoni internal transcribed spacer 1 region is highly conserved amongst isolates from dogs across Japan OAK
LIU, Mingming; CAO, Shinuo; VUDRIKO, Patrick; SUZUKI, Hiroshi; SOMA, Takehisa; XUAN, Xuenan; 鈴木, 宏志; 玄, 学南.
Babesia gibsoni is a tick-borne apicomplexan parasite of dogs that often causes fever and hemolytic anemia with highly variable clinical outcome. In this study, we sequenced the 254bp Internal Transcribed Spacer 1 region (ITS1) of 54 B. gibsoni isolates from 14 different geographical regions of Japan. The 54 isolates shared high sequence identity with each other and with B. gibsoni isolates reported in GenBank database (97.2–100%). Consistent with previous reports, phylogenetic analysis showed that B. gibsoni isolates from Japan formed the same clade with those from U.S.A., Australia, India and Taiwan. Our finding indicates that B. gibsoni ITS1 region is highly conserved among isolates from dogs in Japan, making it a useful genetic marker for molecular...
Palavras-chave: Babesia gibsoni; Conserved gene; Internal transcribed spacer 1 (ITS1); Japan.
Ano: 2016 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4302
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Exposure of C57BL/6J Male Mice to an Electric Field Improves Copulation Rates with Superovulated Females OAK
HORI, Takuya; YAMSAARD, Thicomeporn; UETA, Yoshiko Yanagimoto; HARAKAWA, Shinji; KANEKO, Etsushi; MIYAMOTO, Akio; XUAN, Xuenan; TOYODA, Yutaka; SUZUKI, Hiroshi; 宮本, 明夫; 玄, 学南; 鈴木, 宏志.
It is well-known that there are considerable strain differences in the relative copulation rates between male and superovulated female mice. In particular, the C57BL/6J strain of mice has a lower rate of successful copulation. We examined the effect of exposure to an electric field on sexual behavior in C57BL/6J male mice. When C57BL/6J males were exposed to a 50 Hz, 45 kV/m electric field for 30 min per day for 11 days and placed in a cage with a superovulated female of the same strain, the successful copulation rates of males was significantly improved compared with unexposed males (P<0.05). These results suggest that the exposure of C57BL/6J male mice to an electric field improves their sub-fertility activity in mating with superovulated females.
Palavras-chave: Mice; Male; Electric field; Copulation; Superovulation; Sexual behavior.
Ano: 2005 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3021
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Starfish, Asterias amurensis and Asterina pectinifera, as Potential Sources of Th1 Immunity-Stimulating Adjuvants OAK
KAWASE, Osamu; GOO, Youn-Kyoung; JUJO, Hayato; NISHIKAWA, Yoshifumi; XUAN, Xuenan; 西川, 義文; 玄, 学南.
Saponin is the generic name of steroid or triterpene glycosides, and the capacities of some saponins to stimulate both Th1 immune response and production of cytotoxic T cells are useful as vaccine components against intracellular pathogens. Because saponins have been found commonly in starfish, we assessed the potential of starfish, Asterias amurensis and Asterina pectinifera, as adjuvant sources. Crude starfish saponins had hemolytic activities (EC50=10 to 100 μg/ml) and thin layer chromatography indicated heterogeneity of their constituents. When starfish saponis were subcutaneously injected into mice with ovalbumin (OVA), OVA-specific IgG, especially IgG2a instead of IgG1 was produced in mouse blood, suggesting starfish saponins stimulated Th1 type...
Palavras-chave: Adjuvant; Saponin; Starfish; Th1 immunity; Vaccine.
Ano: 2011 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3587
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Babesia bovis Dihydroorotate Dehydrogenase (BboDHODH) is a Novel Molecular Target of Drug for Bovine Babesiosis OAK
KAMYINGKIRD, Ketsarin; CAO, Shinuo; MASATANI, Tatsunori; MOUMOUNI, Paul Franck Adjou; VUDRIKO, Patrick; MOUSA, Ahmed Abd El Moniem; TERKAWI, Mohamad Alaa; NISHIKAWA, Yoshifumi; IGARASHI, Ikuo; XUAN, Xuenan; 西川, 義文; 五十嵐, 郁男; 玄, 学南.
The emergence of drug resistance and adverse side effects of current bovine babesiosis treatment suggest that the search of new drug targets and development of safer and effective compounds are required. This study focuses on dihydroorotate dehydrogenase (DHODH), the fourth enzyme of pyrimidine biosynthesis pathway as a potential drug target for bovine babesiosis. Recombinant Babesia bovis DHODH protein (rBboDHODH) was produced in Escherichia coli and used for characterization and measurement of enzymatic activity. Furthermore, the effects of DHODH inhibitors were evaluated in vitro. The recombinant B. bovis DHODH histidine fusion protein (rBboDHODH) had 42.4-kDa molecular weight and exhibited a specific activity of 475.7 ± 245 Unit/mg, a Km = 276.2 µM for...
Palavras-chave: Atovaquone; Babesia bovis; Chemotherapeutic target agent; Dihydroorotate dehydrogenase; Leflunomide.
Ano: 2014 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3918
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The Influence of the regulation of Toxoplasma gondii TgMIC2 transgene on host cell infection OAK
Buates, S.; Xuan, Xuenan; Igarashi, Makoto; Sugimoto, Chihiro; Inoue, Noboru; 玄, 学南; 五十嵐, 慎; 井上, 昇.
Toxoplasma gondii microneme protein 2 (TgMIC2), an apically stored adhesin, was shown to be a key participant involved in the initial attachment and invasion to a host cell. In this study, we had established the clonal line of T. gondii tachyzoites which over-expressed TgMIC2 using the tetracycline repressor (TetR)-based inducible gene expression system. The TgMIC2 over-expression significantly reduced tachyzoite propagation in vitro (p < 0.002) and significantly reduced the parasite virulence in mice (p = 0.002). The over-expression of exogenous TgMIC2 caused the increase of mRNA expression level of endogenous TgMIC2. Wild type parasites mainly expressed TgMIC2115 protein, the microneme store and the surface form of TgMIC2. Over-expression of TgMIC2...
Palavras-chave: Toxoplasma gondii; TgMIC2; Over-expression.
Ano: 2008 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/2236
Registros recuperados: 37
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