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Kanae SHIOKAWA Chandika D. GAMAGE Nobuo KOIZUMI Yoshihiro SAKODA Kenta SHIMIZU Yoshimi TSUDA Kumiko YOSHIMATSU Jiro ARIKAWA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2016,78(2):221-230
The applicability of the recombinant LipL32 for serodiagnosis of leptospiral infection in field rodents was
assessed in this study. An immunodominant region of LipL32 was determined by monoclonal antibodies, and then,
truncated LipL32 (tLipL32) was designed to contain the region (87–188th amino acid). The tLipL32 was compared
between two recombinant expression hosts Escherichia coli and Pichia
pastoris in ELISA. With field rat sera, tLipL32 expressed by P. pastoris
(tLipL32p) had high antigenicity without background reactions, while tLipL32 expressed by E.
coli (tLipL32e) showed high background reactions, which were reduced by pre-adsorption of sera with
E. coli. To evaluate tLipL32-ELISA, field rat sera were tentatively divided into a
Leptospira infection positive (12 sera) and a negative group (12 sera) based on the results
from flaB gene PCR of kidney samples and WB with whole Leptospira cell.
Consequently, the sensitivity of tLipL32p-ELISA for field rat sera was 83% . A similar result was obtained
from tLipL32e-ELISA with adsorbed sera, (92%). However, sensitivity of tLipL32e-ELISA using sera without an
adsorption treatment was 50%. Regardless of the expression host, tLipL32-ELISA had 100% specificity and
sensitivity in experimentally infected laboratory rats. These results suggest that recombinant LipL32
expressed by P. pastoris is more applicable for serodiagnosis in field rats due to a lack of
background reaction. 相似文献
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Nurun Nahar Khanam Junichi Kihara Yuichi Honda Toshihide Tsukamoto Sakae Arase 《Journal of General Plant Pathology》2005,71(4):285-288
When detached broad bean leaves were preinoculated with virulent strain B304 of Botrytis cinerea 24 h before a challenge inoculation with strain B304, lesion formation by B304 was significantly inhibited in red light but not in the dark. In leaves that were preinoculated with avirulent strain 021 and then challenged by B304, however, lesion formation was not inhibited even under red light. Such differences in lesion formation after the challenge inoculation with an avirulent strain were also observed with lesions caused by Alternaria alternata, a nonpathogen of broad bean and by avirulent strain 021R in the presence of germination fluid from spores of strains B304 and 021R. These results suggest the possibility that virulent B. cinerea produced a suppressor involved in induced susceptibility and an elicitor involved in resistance induced by red light during spore germination. 相似文献
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Teruo NONOMURA Yoshinori MATSUDA Motoaki TSUDA Kazuto URANAKA Hideyoshi TOYODA 《Journal of General Plant Pathology》2001,67(3):224-227
Commercially available tomato cultivars were hydroponically cultured for inoculation, with Ralstonia solanacearum (K-101), which causes bacterial wilt, by pouring an inoculum suspension into the nutrient solution. Cultivar susceptibility
to the bacteria was evaluated, based on the highest percentage of wilting. Because the length of time for wilt appearance
varied among cultivars, some cultivars appeared to be suppressive to the translocation and/or multiplication of the invading
pathogen. Thus, this hydroponic inoculation system is effective for examining levels of susceptibility in tomato cultivars
to bacterial wilt.
Received 13 December 2000/ Accepted in revised form 27 March 2001 相似文献
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The effect of heating on the hygroscopicity of Japanese cedar wood was investigated as a simple evaluation of thermal degradation
in large-dimension timber being kiln-dried at high temperatures (>100°C). Small wood pieces were heated at 120°C in the absence
of moisture (dry heating) and steamed at 60°, 90°, and 120°C with saturated water vapor over 2 weeks, and their equilibrium
moisture contents (M) at 20°C and 60% relative humidity (RH) were compared with those of unheated samples. No significant change was induced by
steaming at 60°C, while heating above 90°C caused loss in weight (WL) and reduction in M of wood. The effects of steaming were greater than those of dry heating at the same heating temperature. After extraction
in water, the steamed wood showed additional WL and slight increase in M because of the loss of water-soluble decomposition residue. The M of heated wood decreased with increasing WL, and such a correlation became clearer after the extraction in water. On the basis of experimental correlation, the WL of local parts in large-dimension kiln-dried timber was evaluated from their M values. The results indicated that the thermal degradation of inner parts was greater than that of outer parts. 相似文献
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Atsushi IDETA Yoshito AOYAGI Kanami TSUCHIYA Yuuki NAKAMURA Kou HAYAMA Atsushi SHIRASAWA Kenichiro SAKAGUCHI Naomi TOMINAGA Yoshiyuki NISHIMIYA Sakae TSUDA 《The Journal of reproduction and development》2015,61(1):1-6
Embryos obtained via superovulation are necessary for mammalian artificial reproduction, and viability is a key determinant of success. Nonfreezing storage at 4 C is possible, but currently used storage solutions can maintain embryo viability for only 24–48 h. Here we found that 10 mg/ml antifreeze protein (AFP) dissolved in culture medium 199 with 20% (v/v) fetal bovine serum and 25 mM HEPES could keep bovine embryos alive for 10 days at 4 C. We used a recombinant AFP isolated from the notched-fin eelpout (Zoarces elongatus Kner). Photomicroscopy indicated that the AFP–embryo interaction was enhanced at 37 C. Embryos pre-warmed with the AFP solution at 37 C for 60 min maintained high viability, whereas those that were not pre-warmed could live no longer than 7 days. Thus, short-term storage of bovine embryos was achieved by a combination of AFP-containing medium and controlled pre-warming. 相似文献
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Much evidence has suggested that sex steroid hormone-induced growth of uterine cells is mediated by polypeptide growth factors synthesized in uterine tissues. The present study aimed to clarify the effect of insulin-like growth factor-I (IGF-I) on the proliferation of mouse endometrial stromal cells obtained from immature mice. IGF-I and IGF-I receptor (type I) mRNAs were detected in the endometrial stromal cells. IGF-I increased bromodeoxyuridine (BrdU) uptake in the endometrial stromal cells, indicating an increase in DNA replication. E2 increased IGF-I mRNA levels in the endometrial stromal cells. IGF-I receptor is a tyrosine kinase receptor, and treatment with genistein, a tyrosine kinase inhibitor, reduced IGF-I-induced BrdU-uptake in the endometrial stromal cells. IGF-I signaling pathways involve mitogen-activated protein (MAP) kinase and phosphatidylinositol-3 kinase (PI-3 kinase). Treatment with 10(-7) M of the MAP kinase inhibitor PD098059 and 10(-5) M of the PI-3 kinase inhibitor LY294002 decreased IGF-I-induced BrdU-uptake in the endometrial stromal cells. However, LY294002 (10(-5) M) also decreased the BrdU-uptake in the absence of IGF-I treatment. These results suggest that endometrial IGF-I is involved in the proliferation of endometrial stromal cells in a paracrine or autocrine manner, and that the MAP kinase pathway is involved in DNA replication of endometrial stromal cells. 相似文献
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A method for identification and typing of wood rotting fungi using the melting temperature [T(m)] of DNA fragments coding rRNA (rDNA) was examined. The T(m)s of four DNA fragments, inter transcribed spacer (ITS) I, ITS II, and two partial fragments of 28S rDNA from each of 20 species of wood rotting fungi, were measured by melting curve analysis. The T(m) variation was large enough between species to enable identification based on the T(m) values. A pair of T(m)s of the ITS I region (between the primers ITS1 and ITS2) and the ITS II region (between the primers ITS3 and ITS4) had the highest resolution for identifying wood rotting fungi. To assess about the diversity of the T(m), intraspecific diversity of these DNA fragment sequences was evaluated using test strain sequences and data from the GenBank/EMBL/DDBJ biological database. The intraspecific diversity of T(m) was considered to be small because the nucleotide diversity of each fragment was small within the species. 相似文献