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Registros recuperados: 37
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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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Characterization of P15 Antigen of Cryptosporidium parvum Expressed by a Recombinant Vaccinia Virus OAK
Xuan, Xuenan; Zhang, Sofa; Kamio, Tsugihiko; Tsushima, Y.; Kamada, Takenori; Nishikawa, Yoshifumi; Otsuka, Haruki; Karanis, Panagiotis; Igarashi, Ikuo; Nagasawa, Hideyuki; Fujisaki, Kozo; Mikami, Takeshi; 玄, 学南; 西川, 義文; 五十嵐, 郁男.
Palavras-chave: C. parvum; P15; Vaccinia virus; Subunit vaccine.
Ano: 1999 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/314
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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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Clinical usefulness of antibodies against Babesia gibsoni detected by ELISA with recombinant P50 OAK
Miyama, Takako; Inokuma, Hisashi; Itamoto, Kazuhito; Okuda, Masaru; Verdida, Rodolfo A.; Xuan, Xuenan; 猪熊, 壽; 玄, 学南.
The clinical usefulness of antibodies against Babesia gibsoni detected by ELISA with recombinant P50 was examined in dogs in an area where B. gibsoni infection was endemic. Only 8 among 14 dogs with acute type B. gibsoni infection without a previous history of infection were positive. This high percentage of false-negative results is thought to be a weak point of ELISA as a diagnostic tool. However, 14 other anemic dogs with a confirmed history of B. gibsoni infection were all positive, thus confirming the higher sensitivity of ELISA in detecting a history of infection.
Palavras-chave: Babesia gibsoni; Diagnosis; ELISA.
Ano: 2006 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/925
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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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CLONING AND EXPRESSION OF AN ANTIGEN OF BABESIA GIBSONI IN ESCHERICHIA COLI AND ITS USE FOR THE IMMUNODIAGNOSIS OF CANINE BABESIOSIS OAK
Fukumoto, Shinya; Sekine, Yukiko; Kimbita, Elikira; Huang, Xiaohong; Xuan, Xuenan; Inoue, Noboru; Yokoyama, Naoaki; Igarashi, Ikuo; Fujisaki, Kozo; Sugimoto, Chihiro; Nagasawa, Hideyuki; Mikami, Takeshi; Suzuki, Hiroshi; 福本, 晋也; 玄, 学南; 井上, 昇; 横山, 直明; 五十嵐, 郁男; 鈴木, 宏志.
Palavras-chave: Babesia gibsoni; Babesia canis; CDNA library; ELISA; P30 gene.
Ano: 2002 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/624
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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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CURRENT SERODIAGNOSTIC MEASURES FOR EQUINE BABESIOSIS OAK
Hirata, Haruyuki; Yokoyama, Naoaki; Xuan, Xuenan; Igarashi, Ikuo; 横山, 直明; 玄, 学南; 五十嵐, 郁男.
Palavras-chave: Babesia equi; Babesia caballi; Diagnosis; ELISA; ICT.
Ano: 2004 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/638
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Detection of Antibodies to Babesia equi by The Latex Agglutination Test with Recombinant Merozoite Antigen-2 OAK
Tanaka, Tetsuya; Xuan, Xuenan; Igarashi, Ikuo; Nagasawa, Hideyuki; Fujisaki, Kozo; Suzuki, Naoyoshi; Mikami, Takeshi; 玄, 学南; 五十嵐, 郁男.
Palavras-chave: Babesia equi; EMA-2; Baculovirus; Latex agglutination test.
Ano: 1999 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/329
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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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Diagnosis of Babesia caballi Infection in Horses by Polymerase Chain Reaction OAK
Xuan, Xuenan; Igarashi, Ikuo; Avarzed, A.; Ikadai, Hiromi; Inoue, Noboru; Nagasawa, Hideyuki; Fujisaki, Kozo; Toyoda, Yutaka; Suzuki, Naoyoshi; Mikami, Takeshi; 玄, 学南; 五十嵐, 郁男; 井上, 昇.
A set of primers were designed according to the published sequence of the gene encoding a rhoptry protein of Babesia caballi, and used to amplify parasite DNA from the blood samples obtained from carrier horses by polymerase chain reaction(PCR)method.The PCR method was sensitive enough to detect parasite DNA from 2.5 μl blood sample with a parasitemia of 0.000001%. The PCR method was compared with fluorescent antibody test(IFAT) in order to evaluate the diagnosis effciency for B. caball infection in horses. Of 142 field samples from Mongolia, 28(20%) and 96(69%)samples were identified positively by PCR and IFAT, respectively. Although the sensitivity of PCR was lower than IFAT, it was noted that the 5 IFAT-negative samples were PCR-positive, suggesting...
Palavras-chave: Babesia caballi; PCR; IFAT.
Ano: 1998 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/281
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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 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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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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Identification of a novel gene encoding a secreted antigen 1 of Babesia gibsoni and evaluation of its use in serodiagnosis OAK
Jia, Honglin; Zhou, Jinlin; Ikadai, Hiromi; Matsuu, Aya; Suzuki, Hiroshi; Fujisaki, Kozo; Xuan, Xuenan; 鈴木, 宏志; 五十嵐, 郁男; 玄, 学南.
Serum from a dog immunized with blood plasma from a B. gibsoni-infected dog, putatively containing secreted antigens, was used to screen a cDNA expression library. A novel gene encoding BgSA1 was identified from the isolated clones. The serum raised in mice immunized with the recombinant BgSA1 expressed in Escherichia coli could recognize a native parasite protein with a molecular mass of 59 kDa. Comparing with the previously established ELISA with recombinant P50 as antigen, the ELISA with recombinant BgSA1 as the antigen was more sensitive when they were used to detect field samples. Moreover, a sandwich ELISA with anti-BgSA1 antibodies could detect the circulating BgSA1 in a serial blood plasma from a dog experimentally infected with B. gibsoni. These...
Palavras-chave: Babesia gibsoni; Secreted Antigen 1; Identification; Serodiagnosis.
Ano: 2006 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/1655
Registros recuperados: 37
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