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51.
Tomioka I Mizutani E Yoshida T Sugawara A Inai K Sasada H Sato E 《The Journal of reproduction and development》2007,53(4):835-842
The present study was conducted to demonstrate the spindle formation and behavior of chromosomes and microtubules during first division in reconstructed rat embryos produced by somatic cell nuclear transfer (SCNT) with cumulus cell nuclei. To demonstrate the effect of oocyte aging after ovulation on the cleavage of SCNT embryos, micromanipulation was carried out 11, 15 and 18 h after injection of hCG. SCNT oocytes were activated by incubation in culture medium supplemented with 5 microM ionomycin for 5 min followed by treatment with 2 mM 6-dimethylaminopurine (6-DMAP) in mR1ECM for 2-3 h. For immunocytochemical observation, the SCNT embryos were incubated with monoclonal anti-alpha-tubulin antibody and then fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG. Cleavage rates were significantly higher for oocytes collected after 15 and 18 h rather than for those collected 11 h after injection of hCG (56 and 53%, respectively vs. 28%; P<0.05). Premature chromosome condensation occurred before activation of the SCNT oocytes, but adequate spindle formation was only rarely observed. The distribution of microtubules in SCNT embryos after activation was different from those of fertilized and parthenogenic oocytes, i.e., a dense microtubule organization shaped like a ring was observed. Eighteen to 20 h post-activation, most SCNT embryos were in the 2-cell stage, but no nucleoli were clearly visible, which was quite different from the fertilized oocytes. In addition, first division with and without small cellular bodies containing DNA was observed in the rat SCNT embryos in some cases. The present study suggests that reorganization of transferred nuclei in rat SCNT embryos may be inadequate in terms of formation of the mitotic assembly and nucleolar reorganization. 相似文献
52.
A novel culture method of canine peripheral blood lymphocytes with concanavalin a and recombinant human interleukin-2 for adoptive immunotherapy 总被引:1,自引:0,他引:1
Kato M Watarai S Nishikawa S Iwasaki T Kodama H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2007,69(5):481-486
This study was designed to develop a novel culture method for the efficient proliferation of canine peripheral blood lymphocytes (cPBL) for adoptive immunotherapy. When cPBL were cultured in the presence of concanavalin A (Con A), proliferation of cPBL was induced and expression of interleukin-2 receptor (IL-2R) which enables to respond to exogenously added IL-2 was upregulated. And then, when cPBL were cultured with recombinant human interleukin-2 (rhIL-2) in addition to Con A, proliferation was accelerated and increased to about 10-fold after 1 week. The phenotypic analysis showed that the main population of the cultured cPBL was consisted of CD8+ positive lymphocytes. Among them, CD4+CD8+ double positive (DP) lymphocytes had significantly increased, and the ratio of CD4+ single positive (SP) lymphocytes to CD8+ SP lymphocytes (CD4+SP/CD8+SP) was decreased as compared to before culturing. To evaluate the cytotoxic activity of cPBL cultured with Con A and rhIL-2, furthermore, cytotoxic assay was carried out against xenogeneic melanoma cell line (MeWo), which resulted in MHC-unrestricted cytokilling. These results suggest that the culture method of cPBL by the use of Con A and rhIL-2 may be useful for generating lymphokine activated killer cells, and also this may be beneficial for adoptive immunotherapy of tumor-bearing dogs. 相似文献
53.
Onishi S Miyata H Inamoto T Qi WM Yamamoto K Yokoyama T Warita K Hoshi N Kitagawa H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2007,69(11):1123-1129
It is well known that some caspases in apoptosis is involved in determinant of terminal differentiation and maturation of various cells. Our previous study ultrastructurally clarified the differentiation into M cells from immature microvillous epithelial cells and the redifferentiation from M cells to microvillous epithelial cells in the follicle-associated epithelium (FAE) of rat Peyer's patch. In this study, the difference of epithelial apoptosis between the FAE of Peyer's patch and intestinal villi was immunohistochemically investigated in rat jejunoileum. As a result, cleaved caspase-3 was limited to several epithelial cells at the tip of FAE, whereas almost all of the epithelial cells were cleaved caspase-3 positive in intestinal villi. Cleaved caspase-9 was detected only in a few exfoliating or exfoliated epithelial cells of both FAE and intestinal villi. Nuclear DNA-fragmentation was detected only in several epithelial cells of the tip of FAE, while it was expressed from the middle regions in the intestinal villi. The DNase I expression of the epithelial cytoplasm was much weaker in FAE than in intestinal villi. Bcl-x expression was restricted in the apical cytoplasms of epithelial cells in the FAE, whereas it was restricted in whole cytoplasms in villous epithelial cells. These findings suggest that the progression of the apoptotic process in the epithelial cells of FAE is later than in the intestinal villi, so that the possibility of epithelial differentiation might be remained in the FAE, unlike in the intestinal villi. 相似文献
54.
55.
Nezu Y Sakaue Y Hara Y Tsuchida S Yokota F Takahashi K Tagawa M 《American journal of veterinary research》2002,63(6):804-810
OBJECTIVES: To determine whether small intestinal ischemia and reperfusion affects intestinal intramucosal pH (pHi), arterial and portal venous blood gas values, and intestinal blood flow (IBF) and to investigate relationships between regional intestinal tissue oxygenation and systemic variables in dogs. ANIMALS: 15 healthy adult Beagles. PROCEDURE: Occlusion of superior mesenteric artery (SMA) for 0, 30, or 60 minutes, followed by reperfusion for 180 minutes, was performed; IBF, pHi, arterial and portal venous blood gas values, arterial pressure, and heart rate were measured at various time points; and intestinal mucosal injury was histologically graded. RESULTS: Occlusion of the SMA induced significant decreases in pHi and IBF. After the release of the occlusion, IBF returned rapidly to baseline values, but improvement in pHi was slow. Arterial and portal venous blood gas analyses were less sensitive than tonometric measurements of pHi, and there was no correlation between results of blood gas analyses and tonometric measurements. Histologic score for intestinal mucosal injury increased significantly, depending on duration of ischemia, and there was a correlation between tonometric results and the histologic score. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that it is difficult to accurately evaluate local oxygenation disorders by monitoring at the systemic level, whereas clinically pHi is the only reliable indicator of inadequate regional intestinal tissue oxygenation in dogs. 相似文献
56.
Koichiro HEMMI Ikuo KOBAYASHI Makoto KAJISA Go KITAHARA Kiichi FUKUYAMA Hiroshi HARADA Shunichi KAMIMURA 《Animal Science Journal》2013,84(1):23-27
Two different artificial insemination (AI) protocols were investigated to determine the effect of the time interval from the onset of estrus (as detected by a pedometer) to the AI on the conception rate in Japanese Black cows. Seventy‐three cows were divided into two groups that received AI either after the induction of synchronized ovulation (synchronized AI group; n = 26) or at the time of observed estrus (control AI group; n = 47). Each group was further divided into two subgroups, which were artificially inseminated either 0–12 h (early AI group; n = 21) or 12–24 h (late AI group; n = 52) after the onset of estrus. There was no significant difference in the conception rate between the synchronized AI and control AI groups. The AI protocol and the detection of estrus using a threshold of counted steps (as measured by a pedometer) in this study could be effective for planned reproduction management without the observation of standing estrus in Japanese Black cows. 相似文献
57.
Effect of propylene glycol and undegradable protein source on rumen fermentation, blood metabolism and milk production in lactating dairy cows 总被引:6,自引:0,他引:6
Turigen BAIYILA Jifu HAN Shuhei KANDA Masahiro ITOH Yuichiro WASHIO Tamotu SUZUKI Hiroshi HORIKAWA Toshihiko KAMADA Hisao ITABASHI 《Animal Science Journal》2002,73(3):207-213
Eight lactating Holstein cows were divided into two groups ( n = 4) and used in a double reversal trial with three periods of 14 days each, to evaluate diets containing propylene glycol (PG) and ruminally undegradable protein (RUP) blend on milk yield and composition, ruminal fermentation and blood metabolism. The control diet contained 20% chopped Sudangrass hay, 20% cubed alfalfa hay, 12% corn silage, and 48% of the respective concentrate mixtures (dry matter basis). The experimental diet (PG + RUP) partially replaced the concentrate mixture from the control diet with 1.4% PG and 2.1% RUP. Both diets contained about 16% crude protein and 71% total digestible nutrients. Dry matter intake was similar between the two diets. Daily production of milk, milk lactose and milk solids-not-fat increased by 9.0%, 11.3% and 9.3%, respectively ( P < 0.1), for cows fed diets with PG + RUP; milk composition was unchanged. Although the concentration of ruminal total volatile fatty acids was unchanged, the proportion of propionic acid increased, and the proportion of acetic acid decreased with PG + RUP. The concentration of ruminal ammonia nitrogen and number of ciliate protozoa was not significantly affected by PG + RUP. The concentration of glucose in blood plasma increased, the concentration of urea nitrogen was unaffected, and the concentration of some essential free amino acids decreased with PG + RUP. It is suggested that these changes might be caused predominantly by PG, and the addition of PG may exert a favorable effect on milk production through increased metabolism. 相似文献
58.
Fujiko MINAMI-FUKUDA Makoto NAGAI Hikaru TAKAI Toshiaki MURAKAMI Tadashi OZAWA Shinobu TSUCHIAKA Sachiko OKAZAKI Yukie KATAYAMA Mami OBA Naomi NISHIURA Yukiko SASSA Tsutomu OMATSU Tetsuya FURUYA Satoshi KOYAMA Junsuke SHIRAI Hiroshi TSUNEMITSU Yoshiki FUJII Kazuhiko KATAYAMA Tetsuya MIZUTANI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2013,75(12):1651-1655
59.
60.
Munekazu Nakaichi Toshie Iseri Hiro Horikirizono Harumichi Itoh Hiroshi Sunahara Yuki Nemoto Kazuhito Itamoto Kenji Tani 《The Canadian veterinary journal. La revue veterinaire canadienne》2022,63(6):633
The pedigrees of 3 Dalmatian dogs afflicted with copper-associated hepatitis were investigated to discover the mode of inheritance. A composite family pedigree showed that the 3 affected Dalmatians were related. None of the parents of the affected dogs showed clinical symptoms of liver disease, and the disease had no sex predisposition. The estimated segregation ratio was approximately 3:1 based on surviving littermates. These findings suggested that the copper-associated hepatitis in these Dalmatians was an autosomal recessive mode of inheritance. In addition, some male Dalmatians imported from abroad might have been involved in the occurrence of this disease in Japan. 相似文献