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1.
Takaki Waritani Yoko Okuno Yoshinori Ashida Ryo Tsuchiya Kosaku Kobayashi Takatsugu Yamada 《Veterinary immunology and immunopathology》2001,80(3-4):333-338
Canine cationic trypsin was purified by salting-out, gel filtration and affinity chromatography. Purity was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight was ca. 28 kDa by SDS-PAGE.
Thirty hybridomas were obtained which produced mAb to canine cationic trypsin by the cell fusion technique. Twenty-two of these recognized cationic trypsin only, while eight hybridomas recognized both cationic and anionic trypsin. Several of the anti-canine cationic trypsin mAb were purified by salting-out and DEAE ion-change chromatography using ascites fluid of immunized BALB/c mice. The mAb proved to have very high specificity to canine cationic trypsin as shown by immunoblotting and it may be possible to use them to develop clinical assays. 相似文献
2.
Jaroenporn S Nagaoka K Ohta R Watanabe G Taya K 《The Journal of reproduction and development》2007,53(4):887-893
Prolactin (PRL) has been proposed to directly stimulate corticosterone release. However, the role of PRL on adrenocortical function in male HAA rats has not been fully clarified. The aim of this study was to investigate the effect of PRL on the secretion of corticosterone and progesterone using an in vitro cell culture system in male rats. Administration of PRL (10(-7) and 10(-6) M) resulted in dose-dependent increases in corticosterone and progesterone release. Cotreatment with PRL produced an increase in the stimulatory effects of ACTH-induced corticosterone and progesterone secretion. However, the PRL-induced corticosterone and progesterone releases were significantly reduced by treatment with AG490, a specific Janus kinase 2 (Jak2) inhibitor. In addition, administration of AG490 blunted the significant inhibition of ACTH-induced corticosterone and progesterone secretion by PRL. These results demonstrated that PRL could act directly on the adrenal gland to drive corticosterone and progesterone secretion in male rats. Additionally, the results emphasize that PRL stimulation of adrenal steroid release may be mediated through Jak2 activity. 相似文献
3.
鱼头和鱼内脏为鱼类在加工过程中废弃的主要组织,其中含有丰富的蛋白质和脂肪等有益成分,有必要进行回收利用。双鳍舵鲣鱼的头部和内脏约占其鱼体重量的30%,富含蛋白质(14.4%-22.6%)。为了将这些废弃物中的蛋白质进行回收,分别采用双鳍舵鲣鱼内脏本身含有的内源性蛋白酶(自溶)和四种外源性的商业蛋白酶Papain W-40(E1),Pancidase NP-2(E2),Sumichiimu FP(E3)和Protease M(E4)对废弃物中的蛋白质进行水解(50℃,1~4h),考察自溶和不同酶解方法对蛋白质的回收率以及对回收液氨基酸组成的影响。结果显示,自溶法对废弃物中蛋白质的回收效果最好。当水解温度为50℃,水解时间为2h时,自溶法的蛋白质回收率最高,达到76%;其次为E2酶解法,回收率达到70%;E3酶解法效果最差,回收率为63%。进一步研究表明,自溶回收液(AE)的氨基酸总量也最高,达到11756 mg/100g,而采用E1、E2、E3和E4水解方法得到的酶解回收液(EE1、EE2、EE3和EE4)中氨基酸含量分别仅为10294、10642、8750、10121 mg/100g。感观评定结果表明,自溶回... 相似文献
4.
Ryo Ezaki Kennosuke Ichikawa Mei Matsuzaki Hiroyuki Horiuchi 《The Journal of Poultry Science》2022,59(2):182
In chickens, primordial germ cells (PGCs) are effective targets for advanced genome editing, including gene knock-in. Although a long-term culture system has been established for chicken PGCs, it is necessary to select a gene-editing tool that is efficient and precise for editing the PGC genome while maintaining its ability to contribute to the reproductive system. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) and CRISPR-mediated precise integration into the target chromosome (CRIS-PITCh) methods are superior as the donor vector is easier to construct, has high genome editing efficiency, and does not select target cells, compared to the homologous recombination method, which has been conventionally used to generate knock-in chickens. In this study, we engineered knock-in chicken PGCs by integrating a fluorescent protein gene cassette as a fusion protein into the chicken vasa homolog (CVH) locus of chicken PGCs using the CRIS-PITCh method. The knock-in PGCs expressed the fluorescent protein in vitro and in vivo, facilitating the tracking of PGCs. Furthermore, we characterized the efficiency of engineering double knock-in cell lines. Knock-in cell clones were obtained by limiting dilution, and the efficiency of engineering double knock-in cell lines was confirmed by genotyping. We found that 82% of the analyzed clones were successfully knocked-in into both alleles. We suggest that the production of model chicken from the knock-in PGCs can contribute to various studies, such as the elucidation of the fate of germ cells and sex determination in chicken. 相似文献
5.
Isolation and culture of rabbit primordial germ cells 总被引:2,自引:0,他引:2
Kakegawa R Teramura T Takehara T Anzai M Mitani T Matsumoto K Saeki K Sagawa N Fukuda K Hosoi Y 《The Journal of reproduction and development》2008,54(5):352-357
Primordial germ cells (PGCs) are embryonic precursors of the gametes of adult animals and are considered stem cells of the germline. Since their proliferation in vitro correlates well with the schedule of developmental changes in vivo, they might be interesting research tools for genomic imprinting, germ-cell tumors and fertility. Furthermore, once primordial germ cells are separated and placed on a feeder layer with cytokines, they become cultured pluripotent cell lines called embryonic germ (EG) cells. EG cells share several important characteristics with embryonic stem (ES) cells as they can also contribute to the germ line of chimeras. To investigate the characteristics of PGCs and establish rabbit EG (rEG) cells, we cultured rabbit PGCs (rPGCs) in vitro with various combinations of leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF) and forskolin on inactivated mouse embryonic fibroblast (MEF) feeder layers. The present study found PGC proliferation in early cultures and induction of rEG-like colonies. These cells expressed pluripotent markers, such as alkaline phosphatase activity, OCT-4, Sox-2 and SSEA-1, in the undifferentiated state; however, the cells did not develop into a teratoma when injected into the kidney capsules of SCID mice, although the restricted differentiation potentials to neural cells were determined via embryoid body formation. From these characteristics and further characterization of the germ stem cell markers Vasa, SCP-1 and SCP-3, we suggested that these were hybrid cells with characteristics somewhere between PGC and EG cells. 相似文献
6.
Watanabe M Hisasue M Souma T Ohshiro S Yamada T Tsuchiya R 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2008,70(10):1095-1099
Detection of hemotropic Mycoplasma spp. infection was attempted in cats by PCR using whole blood without DNA extraction. A total 46 of 54 (85%) cats with suspected Mycoplasma spp. infection showed a positive reaction, corresponding completely with the results of standard PCR testing. The direct PCR assay was sensitive enough to detect more than 0.0061% parasitemia for ;C. M. haemominutum' and 0.0075% parasitemia for M. haemofelis. These data indicate that the direct PCR assay might be sufficient for use as a tool in clinical examinations. 相似文献
7.
Hiroto Miura Takuya Hashimoto Yukiko Kawanishi Hiroki Kawauchi Ryo Inoue Noriaki Shoji Kunihiko Saito Mario Sekiya Yosuke Saito Jumpei Yasuda Chiemi Yonezawa Tetsushiro Endo Hirotaka Kasuya Yutaka Suzuki Yasuo Kobayashi Satoshi Koike 《Animal Science Journal》2021,92(1):e13601
The rumen microbiota comprises a vast range of bacterial taxa, which may affect the production of high-quality meat in Japanese Black cattle. The aim of this study was to identify core rumen microbiota in rumen fluid samples collected from 74 Japanese Black cattle raised under different dietary conditions using 16S rRNA gene amplicon sequencing. In the rumen of fattening Japanese Black cattle, 10 bacterial taxa, showing >1% average relative abundance and >95% prevalence, irrespective of the dietary conditions and the fattening periods, were identified as the core rumen bacterial taxa, which accounted for approximately 80% of the rumen microbiota in Japanese Black cattle. Additionally, population dynamics of the core rumen bacterial taxa revealed two distinct patterns: Prevotella spp. and unclassified Bacteroidales decreased in the mid-fattening period, whereas unclassified Clostridiales, unclassified Ruminococcaceae, Ruminococcus spp., and unclassified Christensenellaceae increased during the same period. Therefore, the present study reports the wide distribution of the core rumen bacterial taxa in Japanese Black cattle, and the complementary nature of the population dynamics of these core taxa, which may ensure stable rumen fermentation during the fattening period. 相似文献
8.
Shohei KOBAYASHI Natsuko KAWANO Kenji MIYADO Ryo OHTA Takahiro AKIMOTO Taichi HATAKEYAMA Maiko KAWAGUCHI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2022,84(1):153
Tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) is widely used as a flame retardant and is known to exhibit anti-androgenic effects in vitro and in vivo. To assess the reproductive toxicity potency of TDCIPP, we investigated the effects of 7 days of TDCIPP oral administration on epididymal sperm motion and concentration in adult male Wistar–Imamichi rats. Thirty-five days after the final administration, sperm parameters were evaluated by computer-assisted sperm analysis. Results showed that sperm swimming progression and vigor and sperm concentration in TDCIPP-treated rats were unexpectedly higher than those in control rats. TDCIPP did not significantly affect the percentage of motile sperms or sperm swimming pattern. These results contribute to the understanding of the biological effects of TDCIPP. 相似文献
9.
Yuichi Takai Tomoya Sano Takeshi Watanabe Ryo Fukuda 《Journal of toxicologic pathology》2013,26(3):319-323
This report describes the morphological and immunohistochemical characteristics of an
adrenocortical carcinoma with distant metastasis in a Sprague-Dawley rat. Macroscopically,
a single large mass was observed in the adrenal gland, and multiple nodules were noted in
the lung, liver and thyroid. Histologically, the adrenal tumor consisted of a solid growth
of eosinophilic round cells with nuclear atypia. Vascular invasion was present, and
multiple metastatic lesions were also observed in the lungs, liver, and mediastinal lymph
nodes. Immunohistochemically, the nuclei of these tumor cells were positive for
Steroidogenic Factor-1 (SF-1). In the thyroid, tumor cells histologically resembling
adrenal cells were immunohistochemically negative for SF-1 but positive for calcitonin;
thus the lesion was diagnosed as thyroid C-cell carcinoma. From these results, the present
case was diagnosed as adrenocortical carcinoma with distant metastases. SF-1 could be a
valuable marker for the differential diagnosis of adrenocortical tumors versus other
endocrine tumors such as C-cell carcinoma. 相似文献
10.
Ryo Kubota Mark A. Schell Gabriel D. Peckham Joanne Rue Anne M. Alvarez Caitilyn Allen Daniel M. Jenkins 《Journal of General Plant Pathology》2011,77(3):182-193
New rapid diagnostic methods are urgently needed to discriminate the quarantine pathogen Ralstonia solanacearum (Rs) race 3 biovar 2 (R3B2) from other populations of Rs that lack the adaptation to cause bacterial wilt disease in temperate
regions. We used an in silico bioinformatic approach to identify several genome sequences potentially specific to R3B2 strains. Primer sets were designed
to PCR-amplify sequences in these regions, and four sets were ultimately shown to be >99% accurate for detection of R3B2 strains.
On the basis of these results, several primers were designed to enable development of a loop-mediated isothermal amplification
assay that was rapid, technologically simple, and essentially 100% accurate for identification of R3B2 when applied to a comprehensive
collection of geographically diverse Rs strains. We fortuitously found that a sequence in one of the “R3B2-specific” regions
has ~90% identity to a sequence present in strains of the blood disease bacterium (BDB), a member of the Rs species complex
that infects banana. Alignments of these sequences allowed design of a second PCR primer set that proved 100% accurate for
identification of BDB strains when tested on the 22 BDB strains available to us. These results demonstrate the power of in silico genomic subtraction for rapid identification of population-specific DNA sequences and for the development of simple, reliable
detection methods for Rs subpopulations. 相似文献