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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
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Sugisawa H Itou T Saito M Moritomo T Miura Y Sakai T 《Veterinary research communications》2003,27(6):453-461
Bovine colostrum and milk contain many immunomodulatory components. The low-molecular-weight fraction (<10 kDa) was separated from colostrum and milk by gel filtration chromatography, and its effect on the oxidative burst of bovine polymorphonuclear leukocytes (PMNL) was investigated in vitro. The oxidative burst activity induced by Staphylococcus aureus was considerably enhanced when PMNLs were incubated with this low-molecular-weight fraction. However, phorbol 12-myristate 13-acetate did not trigger a burst after priming with this fraction. The oxidative burst activity enhanced by this fraction was reduced after heating. These results confirmed that a low-molecular-weight substance(s) of less than 10 kDa, present in bovine milk and colostrum, enhances the oxidative burst activity of PMNL. 相似文献
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A novel biological activity of human recombinant interleukin-12 (rhIL-12) on canine peripheral blood mononuclear cells (PBMC) was investigated in vitro. Canine PBMC were cultured in the presence or absence of rhIL-12 for 3 days. The reactive oxygen species (ROS) production induced by opsonized-zymosan (OZ) was then measured by a luminol-dependent chemiluminescense assay and demonstrated that the ROS production was enhanced after culture with rhIL-12. A nitro blue tetrazolium test and flowcytometry analysis revealed that canine lymphocytes, eosinophils, and monocytes were capable of ROS production, but that monocytes had the highest capacity. These results suggest that rhIL-12 enhances ROS production from canine monocytes. 相似文献
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Tadahiko Kuriyama Sayaka Fusazaki Tadaaki Satou Tamotsu Nikaido 《Pesticide biochemistry and physiology》2005,81(3):176-187
The present study was undertaken to identify noncompetitive γ-aminobutyric acid receptor (GABAR) antagonists that are effective in nematodes, as well as to examine the hypothesis that the noncompetitive antagonism of the GABAR underlies the nematocidal activity of quassinoids against free-living nematodes of the Diplogastridae family. First, 14 known GABAR antagonists were screened for the effectiveness of their nematocidal activity in Diplogastridae. As a result, 3-isopropyl-1-phospha-2,6,7-trioxabicyclo[2.2.2]octane 1-sulfides (3-isopropyl-BPTs) were found to have high nematocidal activities, and 4-cyclohexyl-3-isopropyl-BPT (23) and 3-isopropyl-4-(2-propenyl)-BPT (27) were the two most potent analogues; these compounds are equipotent to samaderine B and more potent than the anthelmintic abamectin. 23-resistant nematodes, selected by challenge with 23, showed cross-resistance to samaderine B. 23 (10 μM) reduced [3H]23 binding to nematode membranes by 30.4%. Samaderine B (10 μM) resulted in a similar level of the inhibition, but had neither additive nor synergistic effects on the 23 inhibition of [3H]23 binding. These findings suggest that samaderine B shares a common binding site with the GABAR antagonist 23 in Diplogastridae. The results of comparative molecular field analysis, a method of three-dimensional quantitative structure-activity relationship analysis, supported this conclusion. 相似文献
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Nobutaka Someya Shinichi Numata Masami Nakajima Akira Hasebe Tadaaki Hibi Katsumi Akutsu 《Journal of General Plant Pathology》2003,69(4):276-282
The gene chiA, encoding for the endochitinase ChiA, was cloned from Serratia marcescens strain B2, a tomato epiphytic bacterium, and introduced into the epiphytic bacterium Erwinia ananas NR-1, isolated from rice phylloplane. The gene chiA was introduced under the control of two types of promoter into a broad-host-range plasmid vector. The vector contained various fragments with promoter activity isolated from E. ananas chromosomal DNA. The constructed vectors were designated pchiA-V1pcf9 and pchiA-V1pcf53 for their respective promoters. E. ananas NR-1 transformed with either of these vectors produced and secreted ChiA. The antifungal activity of ChiA produced by transformed E. ananas NR-1 was demonstrated in vitro by the inhibition of Pyricularia oryzae germ tube elongation such as bursting of the hyphal tip. Transformed E. ananas NR-1 suppressed the incidence of rice blast caused by P. oryzae under greenhouse conditions; however, the magnitude of the suppressive effect depended on which promoter was used. Both transformants and the nontransformant E. ananas NR-1 survived on rice phylloplane. It is expected that the rice epiphytic bacterium E. ananas NR-1 carrying a chitinolytic enzyme gene is an efficient biological control agent against rice blast. 相似文献
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Yoshiyuki MAKIZUMI Shin-ichi TAKEDA Yuichi MATSUZAKI Ryoji NAKAUNE Hiroshi HAMAMOTO Katsumi AKUTSU Tadaaki HIBI 《Journal of General Plant Pathology》2002,68(4):338-341
Two genes, BMR1 and BMR3, encoding new members of the ABC superfamily in Botrytis cinerea, were cloned and characterized. The topologies of the encoded proteins, BMR1 and BMR3, were both (NBF-TMD6)2, similar to most of the toxicant-efflux ABC transporters. These genes had different induced expression patterns after treatment
with various toxicants, suggesting they may have different roles in toxicant-resistance.
Received 24 September 2002/ Accepted in revised form 2 December 2002 相似文献
48.
Nobutaka SOMEYA Masami NAKAJIMA Tadaaki HIBI Isamu YAMAGUCHI Katsumi AKUTSU 《Journal of General Plant Pathology》2002,68(2):177-182
An antagonistic bacterium, Serratia marcescens strain B2, controlled rice blast after being sprayed onto rice phylloplane, as did the bacterial suspension when poured into
rhizosphere soil of rice plants. Three days after root treatment, rice blast conidia were sprayed onto rice foliage. A week
after pathogen inoculation, rice blast was suppressed and lesions caused by the pathogen decreased in size. Brown deposits
were observed around sites of pathogen infection after root treatment. Induced resistance was not associated with an increase
in the activitiy of peroxidase, phenylalanine ammonia lyase, tyrosine ammonia lyase, β-1,3-glucanase, β-1,4-glycosidase, N-acetylhexosaminidase or chitinase. However, lipoxygenase levels were elevated after the root treatment with strain B2 following
inoculation with the pathogen. Strain B2 was not detected in rice foliage after root treatment. These data suggest that strain
B2 induced resistance against rice blast caused by Pyricularia oryzae.
Received 1 November 2001/ Accepted in revised form 25 January 2002 相似文献
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Moritomo Y Shimojo K Miyadera K Khalaj M Asano Y Kunieda T Ogawa H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2008,70(3):293-296
A coagulopathy with subcutaneous bleeding and muscular or peritracheal/periesophageal bleeding occurred in two male Japanese Brown calves of the same dam. One of the affected calves died three days after the onset of bleeding and the other survived normally until being slaughtered despite once suffering from subcutaneous hematoma. Hemostatic tests of the latter case showed prolonged activated partial thromboplastin time (APTT), and severely reduced factor VIII activity. In addition, von Willebrand factor activity, determined by the human platelet aggregation test, was within the normal range; therefore, the calf was diagnosed with hemophilia A. These are the first bovine cases of hemophilia A definitely diagnosed clinicopathologically. 相似文献
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Nakajima Masami Goto Masao Akutsu Katsumi Hibi Tadaaki 《European journal of plant pathology / European Foundation for Plant Pathology》2004,110(2):223-226
Copper-containing bactericides have been used to control bacterial canker of kiwifruit, caused by Pseudomonas syringae pv. actinidiae. However, the efficacy of copper has been reduced by the occurrence of copper-resistant strains. Analysis of the DNA sequence of a cluster region containing the copper-resistance genes from P. syringae pv. actinidiae suggested the presence of three possible different systems for copper resistance: copper-trapping, copper-efflux and copper-transport systems. Transposon insertional inactivation analysis indicated that the copper-trapping system was essential for copper resistance. 相似文献