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Akira Yamamoto Akira Iwata Toshiki Saitoh Kotaro Tuchiya Tomoko Kanai Hajime Tsujimoto Atsuhiko Hasegawa Akira Ishihama Susumu Ueda 《Veterinary immunology and immunopathology》2002,90(3-4)
Feline granulocyte colony-stimulating factor (G-CSF) with an N-terminal histidine hexamer tag was expressed as inclusion bodies in E. coli. The G-CSF solubilized in 6 M guanidine solution was absorbed onto a Ni-NTA column and, after washing with decreasing concentrations of guanidine, eluted with imidazole in a soluble and apparently pure form. The activity of the recombinant feline G-CSF was 3×106 U/mg protein, as assayed by its stimulatory effect on NFS-60 cell proliferation. When a low level of purified feline G-CSF was administered once a day for two successive days to cats, the number of neutrophil increased 4-fold while the levels of other blood cell types remained virtually unchanged. Daily administration of G-CSF for a total of 11 days led to a more than 10-fold increase in neutrophils, an 8-fold increase in the number of monocytes and 2-fold increase in lymphocytes. No severe side effects or antibody production was observed in cats after administration of G-CSF. 相似文献
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Masahiro Hatamoto Ryo Aizawa Kogomi Koda Toshiki Fukuchi 《Journal of Pesticide Science》2021,46(2):198
Aminopyrifen is a novel 2-aminonicotinate fungicide with unique chemistry and a novel mode of action. The fungicide showed high antifungal activity mainly against Ascomycetes and its related anamorphic fungi under in vitro and pot conditions (EC50 values: 0.0039–0.23 mg/L and 1.2–12 mg/L, respectively). The active ingredient strongly inhibited germ-tube elongation of Botrytis cinerea below 0.1 mg/L and invasion into a plant. The compound exhibited no cross-resistance to commercial fungicides in B. cinerea. The antifungal agent showed high preventive efficacy and translaminar action. In the field, aminopyrifen controlled gray mold and powdery mildew at 150 mg/L. Our findings suggest that aminopyrifen is useful for protecting crops from various plant pathogens. 相似文献
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Zhihua Tao Minoru Sato Kegang Wu Hiromasa Kiyota Toshiyasu Yamaguchi Toshiki Nakano 《Fisheries Science》2012,78(4):923-926
We established a simple, rapid method for gizzerosine analysis in fish meal. Gizzerosine was extracted from fish meal with 0.1?N HCl solution. Samples and standard gizzerosine solutions were absorbed onto a paper disc, which was then set on electrophoresis paper for 18?min, and the paper was dried. Gizzerosine was visualized with Pauly??s reagent, and the intensity of the colored spots was digitized and calculated by image processing method software. We achieved successful separation of gizzerosine from other Pauly??s reagent-positive components in fish meal extracts. The linearity of gizzerosine estimation using this method was within the range 30?C1000?ng (R 2?=?0.99). Gizzerosine was satisfactorily detected and completely separated from histamine and other Pauly??s reagent-positive compounds. This method does not require expensive instruments or tedious pretreatment to eliminate interfering compounds, such as histamine or histidine. It also uses less reagent compared with high-performance liquid chromatography. Moreover, it is a simple, rapid, sensitive, and reproducible method. It is suitable for monitoring gizzerosine in fish meal products that contain as little as 10?ppm gizzerosine. 相似文献
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Enzymatic saccharifications of the hydrolysate collected as the water-soluble (WS) portion after hot-compressed water (HCW)
treatment of Japanese beech with xylanase and β-xylosidase were performed to investigate the hydrolytic properties of these enzymes. The WS portions with different molecular
weight distributions and initial concentrations of xylooligosaccharides were prepared as substrates, one of which contained
mainly smaller xylooligosaccharides [degree of polymerization (DP) = 2 or 3], while the other contained larger xylooligosaccharides
(DP ≥ 4). The highest xylose recovery was obtained from both of the WS portions treated with β-xylosidase rather than with xylanase. This suggests that β-xylosidase could have a higher activity than xylanase toward larger xylooligosaccharides (DP ≥ 4) as well as smaller xylooligosaccharides
(DP = 2 or 3) recovered from HCW treatment. As a result, the use of β-xylosidase was found to be effective for saccharifi cation of the WS portion, even with a short reaction time for the HCW
treatment. 相似文献
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Effects of voltage strength during electroporation on the development and quality of in vitro‐produced porcine embryos 
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下载免费PDF全文 K Nishio F Tanihara T‐V Nguyen T Kunihara M Nii M Hirata T Takemoto T Otoi 《Reproduction in domestic animals》2018,53(2):313-318
This study was conducted to determine suitable conditions for an experimental method in which the CRISPR/Cas9 system is introduced into in vitro‐produced porcine zygotes by electroporation. In the first experiment, when putative zygotes derived from in vitro fertilization (IVF) were electroporated by either unipolar or bipolar pulses, keeping the voltage, pulse duration and pulse number fixed at 30 V/mm, 1 msec and five repeats, respectively, the rate of blastocyst formation from zygotes electroporated by bipolar pulses decreased compared to zygotes electroporated by unipolar pulses. In the second experiment, the putative zygotes were electroporated by electroporation voltages ranging from 20 V/mm–40 V/mm with five 1‐msec unipolar pulses. The rate of cleavage and blastocyst formation of zygotes electroporated at 40 V/mm was significantly lower (p < .05) than that of zygotes electroporated at less than 30 V/mm. Moreover, the apoptotic nuclei indices of blastocysts derived from zygotes electroporated by voltages greater than 30 V/mm significantly increased compared with those from zygotes electroporated by voltages less than 25 V/mm (p < .05). When zygotes were electroporated with Cas9 mRNA and single‐guide RNA (sgRNA) targeting site in the FGF10 exon 3, the proportions of blastocysts with targeted genomic sequences were 7.7% (2/26) and 3.6% (1/28) in the embryos derived from zygotes electroporated at 25 V/mm and 30 V/mm, respectively. Our results indicate that electroporation at 25 V/mm may be an acceptable condition for introducing Cas9 mRNA and sgRNA into pig IVF zygotes under which the viability of the embryos is not significantly affected. 相似文献
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Akira Yamamoto Akira Iwata Toshiki Saito Fumiko Watanabe Susumu Ueda 《Veterinary immunology and immunopathology》2009,130(3-4):221-225
Canine granulocyte colony-stimulating factor (cG-CSF) with modification of cysteine at position 17 to serine was expressed in Brevibacillus choshinensis HPD31. cG-CSF secreted into the culture medium was purified by ammonium sulfate precipitation and consecutive column chromatography, using butyl sepharose and DEAE sepharose. Biological activity of the recombinant cG-CSF was 8.0 × 106 U/mg protein, as determined by its stimulatory effect on NFS-60 cell proliferation. Purified cG-CSF was subcutaneously administered once a day for two successive days to dogs (1, 5, 25, or 125 μg). Neutrophil count increased the following day in all dogs except those administered the lowest dose (1 μg). No severe side effects were observed in dogs after administration of cG-CSF. 相似文献
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Noritake K Matsuoka T Ohsawa T Shimomura K Sanbuissho A Uenoyama Y Maeda K Tsukamura H 《The Journal of reproduction and development》2011,57(3):409-415
It has recently been shown that neurokinin B, a tachykinin, is associated with GnRH pulse generation in sheep and goats. The aim of the present study was to clarify the role of tachykinin receptors in the control of LH secretion in rats. To this end, we evaluated the effect of CS-003, an antagonist for all three neurokinin receptors (NK1, NK2 and NK3 receptors), on pulsatile LH secretion in both sexes of rats with different routes of administration. Both oral and third ventricular administration of CS-003 suppressed LH secretion in both sexes of gonadectomized animals. Furthermore, intact male rats with oral administration of CS-003 showed decreased serum testosterone levels, which might be due to suppressed LH secretion. None of the three subtype-specific neurokinin receptor antagonists showed a significant effect on LH secretion in ovariectomized rats when each antagonist was singly administered. The present results suggest that neurokinins play a role in the control of pulsatile GnRH/LH secretion via multiple neurokinin receptors in both male and female rats. 相似文献
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Reconstruction of stratum corneum in organotypically cultured canine keratinocyte-derived CPEK cells
Yagihara H Okumura T Shiomi E Shinozaki N Kuroki S Sasaki Y Ito K Ono K Washizu T Bonkobara M 《Veterinary research communications》2011,35(7):433-437
The stratum corneum of epidermis is an essential barrier against the external environment and water loss. This study aimed
to develop an organotypic culture model that targets the reconstruction of the stratum corneum using canine keratinocyte-derived
CPEK cells. The CPEK cells cultured at the air-liquid interface became stratified and formed a stratum corneum-like layer
on stratum spinosum- and stratum granulosum-like layers. The CPEK cells in the stratum granulosum-like layer expressed the
cornified cell envelope (CCE)-related proteins loricrin and keratinocyte differentiation-associated protein. Organotypically
cultured CPEK cells were considered to form a CCE at the stratum granulosum-like layer, allowing the formation of a stratum
corneum-like layer. The organotypic culture of CPEK cells could be useful for studying the barrier function of canine stratum
corneum. 相似文献
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Takamitsu TSUKAHARA Yoko YOSHIDA Toshiki TSUSHIMA Takumi WATANABE Noritaka MATSUBARA Ryo INOUE Kazunari USHIDA 《Animal Science Journal》2011,82(2):302-306
Early weaning induces villous atrophy in the small intestine (SI) of piglets. Oral administration of live lactic acid bacteria (LAB) can improve villous shortening. In this study, we evaluated the oral administration of a heat‐killed and dried cell preparation of Enterococcus faecalis (a LAB) strain EC‐12 against villous atrophy in early‐weaned mice (Experiment 1) and pigs (Experiments 2 and 3). Twelve 16‐days‐old mice were divided into two groups in Experiment 1: gavage of EC‐12 (10 mg/kg body weight (BW)/day), or control. On day 21, SI was collected. Eighteen 21‐day‐old pigs were divided into two groups in Experiment 2: gavage of EC‐12 (10 mg/kg BW/day), or control. After 10 days, the villous height of jejunum was measured. Six 21‐day‐old pigs were divided into two groups in Experiment 3: the basal diet supplemented with EC‐12 at 0.05%‐fed group, or the basal diet‐fed group. After 10 days, the villous height of jejunum was measured. The villous heights in SI were significantly higher by EC‐12 administration in all experiments. EC‐12 successfully improved the villous atrophy in the early‐weaned mice and pigs when EC‐12 was administered orally. 相似文献