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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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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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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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Epidemiological Survey of Babesia bovis and Babesia bigemina Infections of Cattle in Philippines OAK
YU, Longzheng; TERKAWI, Mohmad Alaa; CRUZ-FLORES, Mary Jane; G. CLAVERIA, Florencia; ABOGE, Gabriel Oluga; YAMAGISHI, Junya; GOO, Youn-Kyoung; CAO, Shinuo; MASATANI, Tatsunori; NISHIKAWA, Yoshifumi; XUAN, Xuenan; 西川, 義文; 玄, 学南.
A total of 250 blood samples were collected from clinically healthy cattle in five provinces of Philippines. DNA was extracted from the samples and analyzed by nested PCR assays for an epidemiological survey of Babesia bovis and Babesia bigemina infections. Out of the 250 samples, 27 (10.8%) and 16 (6.4%) were positive for B. bovis infection and B. bigemina infection, respectively. Mixed infections were detected in a total of 4 samples (1.6%). Our data provide baseline information regarding the epidemiology of B. bovis and B. bigemina infections in cattle in Philippines, which can be utilized in developing proper strategies for disease control and management.
Palavras-chave: Babesia bigemina; Babesia bovis; NPCR; Philippines.
Ano: 2013 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3919
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Epidemiological Survey of Theileria Parasite Infection of Cattle in Northeast China by Allele-Specific PCR OAK
YU, Longzheng; ZHANG, Shoufa; LIANG, Wanfeng; JIN, Chunmei; JIA, Lijun; LUO, Yuzi; LI, Yan; CAO, Shinuo; YAMAGISHI, Junya; NISHIKAWA, Yoshifumi; KAWANO, Suguru; FUJISAKI, Kozo; XUAN, Xuenan; 西川, 義文; 玄, 学南.
An epidemiological survey on a Theileria parasite infection of cattle in Northeast China was carried out using allele-specific PCR and DNA sequence analysis of the major piroplasm surface protein (MPSP) gene. The results showed that 14 of 104 blood samples were positive for Theileria by PCR. Among the positive cases, co-infection with various combinations of C- and I-type parasites was detected in 12 samples; no B- and Thai-type parasites were detected by allele-specific PCR. Phylogenetic analysis based on the MPSP gene sequences revealed that Theileria parasites with the MPSP types 1, 2, and 4 were distributed in Northeast China.
Palavras-chave: Major piroplasm surface protein; PCR; Theileria.
Ano: 2011 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/3586
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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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Expression of a Gene Encoding Trypanosoma congolense Putative Abc1 Family Protein is Developmentally Regulated(Parasitology) OAK
BATICADOS, Waren N.; WITOLA, William H.; INOUE, Noboru; KIM, Jung-Yeon; KUBOKI, Noritaka; XUAN, Xuenan; YOKOYAMA, Naoaki; SUGIMOTO, Chihiro.
During the attempt to seek T. congolense species-specific diagnostic antigens, we discovered one cDNA clone (P74) encoding 74 kDa putative abc1 protein (p74) from T. congolense PCF cDNA library. It has been suggested that members of the abc1 family are novel chaperonins and essential for both the mitochondrial electron transfer in the bc 1 complex and the coenzyme Q biosynthesis. Although abc1 protein in yeast has a nuclear or mitochondrial subcellular location, neither nuclear localization signal nor mitochondrial targeting signal was found within p74. Northern blot analysis revealed that the transcription level of P74 mRNA in bloodstream form (BSF) cells were 4 times higher than that in procyclic form cells. Western blot analysis also indicated that p74...
Palavras-chave: Trypanosoma congolense; Abc1 family protein; Coenzyme Q.
Ano: 2005 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4165
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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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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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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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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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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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種特異的ELISAによる日本の犬におけるNeospora caninumの血清疫学調査(寄生虫病学) OAK
久保田, 直樹; 坂田, 義美; 宮崎, 直美; 板本, 和仁; 坂内, 天; 西川, 義文; 玄, 学南; 猪熊, 壽; NISHIKAWA, Yoshifumi; XUAN, Xuenan; INOKUMA, Hisashi.
組換抗原を用いた種特異的ELISAにより,犬のNeospora caninum感染について全国的血清疫学調査を実施した.1,206頭中全国30県から126頭(10.4%)が陽性を示した.陽性率は,雄よりも雌で有意に高く,また他品種と比較して有意に高い陽性率を示す品種がみられた.子宮蓄膿症および糖尿病の病歴を有する犬の陽性率は,他の疾病を有する犬または疾病のない犬の陽性率に比べて高値であった. A seroepidemiological survey of Neospora caninum infection among dogs in Japan was conducted using species-specific enzyme-linked immunosorbent assay with recombinant surface antigen (Nc-SAG1t). Among 1,206 dogs examined, 126 dogs (10.4%) from 30 prefectures from Hokkaido to Okinawa were positive to N. caninum infections, which were more frequently detected in females than males. Siberian Huskies showed the highest positive rate compared with the other breeds. Dogs with pyometra and diabetes mellitus showed the higher positive rates than dogs with other...
Palavras-chave: Canine; Nbospora caninum; Species-specific ELISA.
Ano: 2008 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4003
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