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H Sakurai Y Fujii Y Maki I Igarashi Y Omata A Saito K Ono N Suzuki 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》1991,53(5):823-831
Novel synthetic Obiopeptide (Obi-1), Glycil-penta-Glutamate, as an immunoregulator, injected intramuscularly to methylcholanthrene (MC) induced tumour bearing mice at 30-100 micrograms per mouse weekly, significantly inhibited tumour growth, compared to that of the untreated control. Numbers of mononuclear cells (MNC) in the spleen obtained from Obi-1 treated and untreated mice were reduced to 64% and 10% of the normal levels, respectively. In histopathological findings, variations of MNC in the spleen was observed in colonies of large MNC and lymphocytes in Obi-1 treated red marrow, while untreated one showed only scattered number of small lymphocytes. A significant O2- production was noted in monolayers of normal macrophages incubated with 0.5 mg Obi-1/ml for 48 hr. Cytotoxicity of spleen MNC cultured with Obi-1 against MC-tumour, Meth A, P-815 and YAC-1 target cells showed less than 10% cytotoxic activity in almost all experiments. 相似文献
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Fuminori Tanihara Maki Hirata Nhien T. Nguyen Quynh A. Le Takayuki Hirano Tatsuya Takemoto Michiko Nakai Dai‐ichiro Fuchimoto Takeshige Otoi 《Animal Science Journal》2019,90(1):55-61
Recently, we established the GEEP (“gene editing by electroporation of Cas9 protein”) method, in which the CRISPR/Cas9 system, consisting of a Cas9 protein and single guide RNA (sgRNA), is introduced into pig zygotes by electroporation and thus induces highly efficient targeted gene disruption. In this study, we examined the effects of sgRNA on the blastocyst formation of porcine embryos and evaluated their genome‐editing efficiency. To produce an animal model for diabetes, we targeted PDX‐1 (pancreas duodenum homeobox 1), a gene that is crucial for pancreas development during the fetal period and whose monoallelic disruption impairs insulin secretion. First, Cas9 protein with different sgRNAs that targeted distinct sites in the PDX‐1 exon 1 was introduced into in vitro‐fertilized zygotes by the GEEP method. Of the six sgRNAs tested, three sgRNAs (sgRNA1, 2, and 3) successfully modified PDX‐1 gene. The blastocyst formation rate of zygotes edited with sgRNA3 was significantly (p < 0.05) lower than that of control zygotes without the electroporation treatment. Our study indicates that the GEEP method can be successfully used to generate PDX‐1 mutant blastocysts, but the development and the efficiency of editing the genome of zygotes may be affected by the sgRNA used for CRISPR/Cas9 system. 相似文献
45.
Fuminori Tanihara Maki Hirata Satoru Iizuka Shinya Sairiki Masahiro Nii Nhien Thi Nguyen Quynh Anh Le Takayuki Hirano Takeshige Otoi 《Animal Science Journal》2019,90(6):712-718
The aim of this study was to investigate the ovarian follicular development, developmental competence of oocytes, and plasma anti‐Müllerian hormone (AMH) levels of Japanese wild boar crossbred (wild hybrid) gilts, whose litter size is inferior to that of European breeds. Ovary and plasma samples were collected from two different breeds of gilts (wild hybrid and Large White breeds). The ovaries from the wild hybrid gilts had a lower average numbers of secondary follicles and vesicular follicles in ovarian cross‐sections and of good quality oocytes collected from ovarian follicles as compared with those from Large White gilts (p < 0.05). The development rate to the blastocyst stage of good quality oocytes after in vitro maturation, fertilization and culture was also lower (p < 0.05) in wild hybrid gilts than in Large White gilts. Plasma AMH levels with >0.16 ng/ml were detected in 8.3% of the examined wild hybrid gilts and 33% of the Large White gilts. These results indicate that the low reproductive performance of wild hybrid breed may result in part from low numbers of vesicular follicles and good quality oocytes, and low developmental competence of oocytes. Moreover, plasma AMH levels may support low number of vesicular follicles in ovaries of wild hybrid gilts. 相似文献
46.
Tsuyoshi Ito Yoshitaka Katoh Yuko Shimada Aya Ohnuma-Koyama Naofumi Takahashi Maki Kuwahara Takanori Harada 《Journal of toxicologic pathology》2016,29(1):39-43
Extraskeletal osteosarcoma is extremely rare in mice. This case report demonstrates a spontaneous murine extraskeletal osteosarcoma that exhibited various histological growth patterns in an ICR mouse. At necropsy, the tumor mass was located in the abdominal wall and was 45 × 30 × 25 mm in size. Histopathologically, the tumor showed the following four growth patterns: a solid pattern of polygonal cells embedded in an osteoid eosinophilic matrix with calcification, an irregular sheet pattern of short spindle cells accompanying some eosinophilic multinucleated cells, a fascicular pattern of spindle cells and a cystic pattern lined by short spindle cells. Immunohistochemically, most of the tumor cells were positive for vimentin, proliferating cell nuclear antigen and osterix. The multinucleated cells mentioned above were desmin positive and were regarded as regenerative striated muscles but not tumor cells. Since no clear continuity with normal bone tissues was observed, the tumor was diagnosed as an “extraskeletal osteosarcoma.” 相似文献
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Current-year shoots are mostly made of primary tissues, whereas first-order branches comprise mainly secondary tissues. Differences in tissue composition of these units reflect differences in functional design. We compared the allometry of current-year shoots and first-order branches in eight deciduous broad-leaved tree species and examined the functional differences underlying the design of current-year shoots and first-order branches. Allometric relationships of first-order branches tended to be compatible with predictions of the pipe model and elastic similarity model. That is, allometric constants of the relationships between leaf mass and stem diameter at the branch base and between stem diameter and stem mass were 2.0 and 0.33-0.38, respectively, indicating that the functional regulation of stem form of first-order branches can be predicted by the two models. However, allometric relationships of current-year shoots were not compatible with the predictions of the pipe and elastic similarity models. Thus, the allometric constant of the relationship between leaf mass and stem diameter at the base of current-year shoots was larger than 2.0, and the allometric constant of the relationship between stem length and stem diameter of shoots was larger than 1.0 in all species examined. However, current-year shoots had an allometric constant of leaf mass against stem length that was less than 1.0, suggesting a functional demand on shoot design to reduce self-shading. Also, allometric constants of stem length against stem diameter at the shoot base were larger in monopodial species than in sympodial species, whereas allometric constants of leaf mass per shoot against stem length were smaller in monopodial species than in sympodial species. We propose that the allometries of current-year shoots reflect their function as disposable units for temporary leaf arrangement. 相似文献
49.
Ishii M Tay TW Matsui T Kidokoro T Mizukami T Kanai Y Hayashi Y Kurohmaru M 《The Journal of reproduction and development》2006,52(3):461-468
The alphavbeta3 and alphavbeta5 integrins are known as transmembrane receptors capable of binding to the RGD amino acid peptide sequence. In mouse early gonadogenesis, some proteins containing the RGD sequence are deposited into extracellular space and participate in morphogenesis. We analyzed the expression patterns of the alphavbeta3 and alphavbeta5 integrins in mouse developing gonads (10.5-13.5 days post coitum) using whole-mount in situ hybridization. The alphav integrin mRNA was homogenously expressed in developing gonadal regions. On the other hand, the beta3 integrin mRNA was found only in large and round cells (presumptive germ cells), whereas beta5 integrin was localized in gonadal somatic cells, with the exception of coelomic epithelial cells. The beta3 integrin-expressed cells were determined to be primordial germ cells because the number of these cells was drastically reduced in busulfan-treated gonads. In this study, we demonstrated that the alphavbeta3 and alphavbeta5 integrins are widely localized in the mouse developing gonads and discussed their presumptive functions on mouse gonadogenesis. 相似文献
50.
Yoshizawa K Sasaki T Uehara N Kuro M Kimura A Kinoshita Y Miki H Yuri T Tsubura A 《Journal of toxicologic pathology》2012,25(1):27-35
Seven-week-old male Lewis rats received a single intraperitoneal injection of N-ethyl-N-nitrosourea (ENU) (100, 200, 400 or 600 mg/kg), and retinal damage was evaluated 7 days after the treatment. Sequential morphological features of the retina and retinal DNA damage, as determined by a TUNEL assay and phospho-histone H2A.X (γ-H2AX), were analyzed 3, 6, 12, 24 and 72 hr, 7 days, and/or 30 days after 400 mg/kg ENU treatment. Activation of the nuclear enzyme poly (ADP-ribose) polymerase (PARP) was analyzed immunohistochemically by poly (ADP-ribose) (PAR) expression in response to DNA damage of the retina. All rats that received ≥ 400 mg/kg of ENU developed retinal degeneration characterized by the loss of photoreceptor cells in both the central and peripheral retina within 7 days. In the 400 mg/kg ENU-treated rats, TUNEL-positive signals were only located in the photoreceptor cells and peaked 24 hr after ENU treatment. The γ-H2AX signals in inner retinal cells appeared at 24 hr and peaked at 72 hr after ENU treatment, and the PAR signals selectively located in the photoreceptor cell nuclei appeared at 12 hr and peaked at 24 hr after ENU treatment. However, degeneration was restricted to photoreceptor cells, and no degenerative changes in inner retinal cells were seen at any time points. Retinal thickness and the photoreceptor cell ratio in the central and peripheral retina were significantly decreased, and the retinal damage ratio was significantly increased 7 days after ENU treatment. In conclusion, ENU induced retinal degeneration in adult rats that was characterized by photoreceptor cell apoptosis through PARP activity. 相似文献