Serodiagnosis of Babesia gibsoni infection in dogs by an improved enzyme-linked immunosorbent assay with recombinant truncated P50     

Verdida RA  Hara OA  Xuan X  Fukumoto S  Igarashi I  Zhang S  Dong J  Inokuma H  Kabeya H  Sato Y  Moritomo T  Maruyama S  Claveria F  Nagasawa H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2004,66(12):1517-1521

The surface antigen P50 of Babesia gibsoni is an important candidate for the development of a diagnostic reagent for canine piroplasmosis. In order to establish an effective diagnostic method for practical use, the gene encoding truncated P50 (P50t) lacking a signal peptide and C-terminal hydrophobic regions were cloned and expressed in Escherichia coli as a fusion protein with glutathione S-transferase (GST). More than 90% portion of the GST-P50t was expressed as a soluble form, in contrast with GST-P50f (full-length), which was completely expressed as an insoluble form. This result indicates that removal of the hydrophobic signal peptide and C-terminus had dramatically improved its hydrophilicity. The purified GST-P50t was tested in an enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to B. gibsoni in dogs. The ELISA with GST-P50t clearly differentiated between B. gibsoni-infected dog sera and uninfected dog sera. In addition, the ELISA detected no cross-reactivity with sera from dogs experimentally infected with the closely related parasites, B. canis canis, B. canis vogeli, and B. canis rossi. Field serum samples collected from dogs in Japan and China were examined for the diagnosis of B. gibsoni infection by using the ELISA. 14.5% (9/62), 5.8% (7/120), and 5.4% (2/37) of tested samples were positive for dogs from Okinawa, Yamaguchi, and Osaka prefectures, Japan, respectively. On the other hand, 4.8% (2/41) of tested samples were positive for dogs from Nanjing, China. These results suggest that the GST-P50t could be a reliable reagent for practical use in ELISA for the serodiagnosis of canine piroplasmosis caused by B. gibsoni.  相似文献   

New method to detect oxolinic acid-resistant <Emphasis Type="Italic">Burkholderia glumae</Emphasis> infesting rice seeds using a mismatch amplification mutation assay polymerase chain reaction     

Yukiko?Maeda  Akinori?Kiba  Kouhei?Ohnishi  Yasufumi?HikichiEmail author 《Journal of General Plant Pathology》2004,70(4):215-217

Bacterial seedling rot and grain rot of rice caused by Burkholderia glumae are seed-borne diseases, traditionally controlled with oxolinic acid (OA). Ser83Arg and Ser83Ile substitutions in GyrA protein are commonly responsible for moderate and high resistance to OA, respectively, in field isolates of B. glumae. To detect OA-resistant B. glumae infesting rice seeds, a mismatch amplification mutation assay polymerase chain reaction protocol was developed using DNA from bacteria incubated in modified S-PG medium and primers based on the amino acid substitutions at position 83 in GyrA.  相似文献   

Decreased expression of matrix metalloproteinases and tissue inhibitors of metalloproteinase in the kidneys of hereditary nephrotic (ICGN) mice   总被引：3，自引：0，他引：3  

Uchio-Yamada K  Manabe N  Goto Y  Anann S  Yamamoto Y  Takano K  Ogura A  Matsuda J 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2005,67(1):35-41

Matrix metalloporoteinases (MMPs), which are dominantly regulated by tissue inhibitors of metalloproteinase (TIMPs), play important roles in extracellular matrix (ECM) degradation and are involved in the progression of kidney diseases. In glomeruli and tubulointerstitum of hereditary nephrotic (ICR-derived glomerulonephritis: ICGN) mouse kidneys, hyper-accumulation of ECM components occurred, and MMP activity decreased. In the present study, because lower levels of MMP activity may contribute to the progression of renal fibrosis in ICGN mice, Western blotting analysis and immunohistochemical staining for MMPs and TIMPs were performed to verify the expression levels of these proteins. Levels of MMP-2, MMP-9, MT1-MMP, TIMP-1 and TIMP-2 in the kidneys were decreased in ICGN mice in comparison with normal ICR mice. These results indicate that small amounts and low levels of activity of MMPs cause the progression of renal fibrosis in ICGN mice.  相似文献   

Analysis of sources of oxolinic acid-resistant field strains of <Emphasis Type="Italic">Burkholderia glumae</Emphasis> based on rep-PCR analysis and nucleotide sequences of <Emphasis Type="Italic">gyrB</Emphasis> and <Emphasis Type="Italic">rpoD</Emphasis>     

Yukiko Maeda  Mitsuo Horita  Hirosuke Shinohara  Akinori Kiba  Kouhei Ohnishi  Seiya Tsushima  Yasufumi Hikichi 《Journal of General Plant Pathology》2007,73(1):46-52

Repetitive sequence-based polymerase chain reaction (rep-PCR) analysis using BOX and ERIC as primers showed a highly divergent phylogeny among field strains of Burkholderia glumae. To elucidate the sources of oxolinic acid (OA) resistance in field strains of B. glumae isolated from rice seedlings cultivated in Mie, Toyama, and Iwate prefectures, Japan, the amino acid at position 83 of GyrA (GyrA83), which is involved in OA resistance, and the DNA patterns from the rep-PCR and the partial nucleotide sequences of gyrB and rpoD from various strains were analyzed. The ten Mie strains, in which GyrA83 was isoleucine (Ile), were divided into two groups based on the band patterns in rep-PCR analysis, although the nucleotide sequences of gyrB and rpoD were identical among the strains. Based on the band patterns in the rep-PCR analysis and the gyrB and rpoD sequences, two highly OA-resistant Toyama strains, Pg-13 and Pg-14, for which GyrA83 was serine (Ser) and Ile, respectively, were in the same lineage. This suggests that the bacteria might acquire OA resistance faster than phylogenic diversity as determined with the repetitive sequences BOX and ERIC and with gyrB and rpoD. Furthermore, three Iwate strains (H95, H101, and H104), isolated from seedlings of different cultivars grown in different years and having Ile at GyrA83, are probably in the same lineage, suggesting that OA-resistant bacteria might be transferred among different cultivars.  相似文献   

A new genus of the intestinal nematode, Lagostrongylus (Heligmonellidae) from lagomorphs: a review of related genera of subfamilies Heligmonellinae and Brevistriatinae     

S Fukumoto  M Kamiya  M Ohbayashi 《The Japanese journal of veterinary research》1986,34(3-4):241-250

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Epidemiology of abomasal nematodes of dairy cattle in Hokkaido, northern Japan     

S Fukumoto  K Etani  K Toi  M Hanadate  M Hidaka  K Yokoya  T Hiramatsu  T Iguchi  S Kudo  K Miyamoto 《Nippon juigaku zasshi. The Japanese journal of veterinary science》1990,52(2):379-385

The prevalence and intensity of infection with abomasal nematodiasis was studied in dairy cattle of Hokkaido, northern Japan, for successive two years. During the period of March in 1985 to September in 1987, a total number of 393 abomasa of Holstein-Friesian cows was examined for nematode parasites. Nematodes were detected from 75% of the cows. The prevalence of nematode species detected was Ostertagia ostertagi 250 (63.6%), Mecistocirrus digitatus 181 (46.1%), Trichostrongylus axei 85 (21.6%) and Haemonchus sp. 1 (0.3%). The prevalence and population composition of each growth stage varied seasonally in O. ostertagi and M. digitatus. The large percentage of arrested larvae, early L4 O. ostertagi and immature L5 M. digitatus, detected during the mid-winter and the increasing percentage of matured adult populations of both species in early spring revealed the occurrence of the autumn associated arrested development (hypobiosis) phenomenon in bovine abomasum nematodes of Japan.  相似文献   

Development of Increased Oxolinic Acid-resistance in Burkholderia glumae     

Yasufumi HIKICHI  Kyoko TSUJIGUCHI  Yukiko MAEDA  Tetsuro OKUNO 《Journal of General Plant Pathology》2001,67(1):58-62

Genomic DNA, partially digested with Sau3AI, of Burkholderia glumae Pg-13, resistant to oxolinic acid (OA), was ligated into pUC118, and Escherichia coli DH5α resistant to 2.5 μg/ml OA was transformed with the plasmid. After incubation on a medium supplemented with 20 μg/ml OA, a clone harboring pB′46 was selected. The nucleotide sequence of the 724-bp insert of pB′46 had no homology to any characterized gene. B. glumae Pg-5, resistant to 10 μg/ ml OA, was transformed with plasmid pUCD3101B′46 containing the insert. An obtained transformant, B25, was resistant to 50 μg/ml OA and had greater resistance to other quinolones than did Pg-5. Transformants of OA-sensitive B. glumae with pUCD3101B′46 had OA sensitivity similar to that of the parental isolates. Received 18 September 2000/ Accepted in revised form 18 October 2000  相似文献   

The role of interleukin-6 in the regulation of granulosa cell apoptosis during follicular atresia in pig ovaries     

Maeda A  Inoue N  Matsuda-Minehata F  Goto Y  Cheng Y  Manabe N 《The Journal of reproduction and development》2007,53(3):481-490

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Induction of apoptosis on K562 cell line and double strand breaks on colon cancer cell line expressing high affinity receptor for granulocyte macrophage-colony stimulating factor (GM-CSF)     

Roudkenar MH  Bouzari S  Kuwahara Y  Roushandeh AM  Baba T  Oloomi M  Fukumoto M 《Iranian Biomedical Journal》2008,12(1):1-6

BACKGROUND: Immunotoxins are comprised of both the cell targeting and the cell killing moieties. We previously established a new immunotoxin, i.e. Shiga toxin granulocyte macrophage-colony stimulating factor (StxA1-GM-CSF), comprises of catalytic domain of Stx, as a killing moiety and GM-CSF, as a cell targeting moiety. In this study, the ability of the immunotoxin to induce apoptosis and double strand breaks (DSB) on different cell lines was investigated. METHODS: The recombinant hybrid protein was expressed in bacterial expression system and purified with nickel-nitrilotriacetate acid resin. The K562 (erythroid leukemia) cell line and LS174 (colon carcinoma) were used in this study. The neutral comet assay was carried out for the detection of DSB and Hoechst staining was performed for apoptosis. RESULTS: StxA1-GM-CSF effectively induced apoptosis on K562 cell line and DNA Double Strand Break (DSB) were observed on colon cancer cell line treated with StxA1-GM-CSF. CONCLUSION: This novel action i.e. DNA damage might be a relevant mechanism of action for StxA1-GM-CSF that is designed to act as immunotoxin, although further investigation is required.  相似文献   

Molecular cloning and characterization of a thaumatin-like protein-encoding cDNA from rough lemon     

Bong-Gyu Kim  Takeshi Fukumoto  Satoshi Tatano  Kenji Gomi  Kouhei Ohtani  Yasuomi Tada  Kazuya Akimitsu 《Physiological and Molecular Plant Pathology》2009,74(1):3-10

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